[0044] In some embodiments of any of the aspects, the cell membrane coating comprises cell membrane components from a CAR-T cell, a cancer cell, and a macrophage. In some embodiments of any of the aspects, the source cells comprise a CAR-T cell, a cancer cell, and a macrophage. In some embodiments of any of the aspects, the cell membrane coating comprises cell membrane components from a CAR-T cell, a cancer cell, and a THP-1 macrophage. In some embodiments of any of the aspects, the source cells comprise an CAR-T cell, a cancer cell, and a THP-1 macrophage. In some embodiments of any of the aspects, the cell membrane coating comprises cell membrane components from a CAR-T cell, a leukemia cell, and a THP-1 macrophage. In some embodiments of any of the aspects, the source cells comprise an CAR-T cell, a leukemia cell, and a THP- 1 macrophage. In some embodiments of any of the aspects, the cell membrane coating comprises cell membrane components from an anti-CD 123 CAR-T cell, a AML cell, and a THP-1 macrophage. In some embodiments of any of the aspects, the source cells comprise an anti-CD 123 CAR-T cell, a AML cell, and a THP-1 macrophage.

[0045] In some embodiments of any of the aspects, the cell membrane coating does not comprise a source cell. In some embodiments of any of the aspects, the cell membrane coating does not comprise a source cell nucleus. In some embodiments of any of the aspects, the cell membrane coating does not comprise source cell DNA.

[0046] In some embodiments of any of the aspects, when the cell membrane coating comprises cell membrane components from multiple source cells, the cell membrane coating is a mixture, fusion, or hybridization of cell membrane components from two or more source cells. In some embodiments of any of the aspects, the the mixture, fusion, or hybridization of cell membrane components has an equal amount of cell membrane components from each source cell. In some embodiments of any of the aspects, the the mixture, fusion, or hybridization of cell membrane components has an unequal amount of cell membrane components from each source cell.

[0047] In some embodiments of any of the aspects, when the cell membrane coating comprises cell membrane components from multiple source cells, the cell membrane coating is a mixture, fusion, or hybridization of cell membranes from two or more source cells. In some embodiments of any of the aspects, the the mixture, fusion, or hybridization of cell membranes has an equal amount of cell membrane components from each source cell.

[0048] The amount of cell membrane components can be the amount as measured by protein concentrations or lipid concentrations. The amount of cell membrane components can be the amount as measured by protein concentrations. The amount of cell membrane components can be the amount as measured by lipid concentrations. [0049] In some embodiments of any of the aspects, the cell membrane components further comprise a detectable label. In some embodiments of any of the aspects, the cell membrane components obtained from each source cell further comprise a detectable label and/or comprise the ability to generate a detectable signal (e.g. by catalyzing reaction converting a compound to a detectable product). In some embodiments of any of the aspects, the cell membrane components obtained from each source cell each further comprises a detectable label and/or comprise the ability to generate a detectable signal (e.g. by catalyzing reaction converting a compound to a detectable product), e.g., each detectable label is distinguishable from the detectable labels comprised by the cell membrane components obtained from the other source cells. In some embodiments of any of the aspects, the engineered cell comprises or expresses a detectable label and/or comprise the ability to generate a detectable signal (e.g. by catalyzing reaction converting a compound to a detectable product) which is not present in the cell membrane coating.

[0050] Detectable labels can comprise, for example, a light-absorbing dye, a fluorescent dye, or a radioactive label. Detectable labels, methods of detecting them, and methods of incorporating them into reagents are well known in the art.

[0051] In some embodiments of any of the aspects, detectable labels can include labels that can be detected by spectroscopic, photochemical, biochemical, immunochemical, electromagnetic, radiochemical, or chemical means, such as fluorescence, chemifluoresence, or chemiluminescence, or any other appropriate means. The detectable labels used in the methods described herein can be primary labels (where the label comprises a moiety that is directly detectable or that produces a directly detectable moiety) or secondary labels (where the detectable label binds to another moiety to produce a detectable signal, e.g., as is common in immunological labeling using secondary and tertiary antibodies). The detectable label can be linked by covalent or non-covalent means to the reagent. Alternatively, a detectable label can be linked such as by directly labeling a molecule that achieves binding to the reagent via a ligand-receptor binding pair arrangement or other such specific recognition molecules. Detectable labels can include, but are not limited to radioisotopes, biolumine scent compounds, chromophores, antibodies, chemiluminescent compounds, fluorescent compounds, metal chelates, and enzymes.

[0052] In other embodiments, the detectable label is a fluorescent compound. When the fluorescently label is exposed to light of the proper wavelength, its presence can then be detected due to fluorescence. In some embodiments of any of the aspects, a detectable label can be a fluorescent dye molecule, or fluorophore including, but not limited to fluorescein, phycoerythrin, phycocyanin, o- phthaldehyde, fluorescamine, Cy3™, Cy5™, allophy cocyanine, Texas Red, peridenin chlorophyll, cyanine, tandem conjugates such as phycoerythrin-Cy5™, green fluorescent protein, rhodamine, fluorescein isothiocyanate (FITC) and Oregon Green™, rhodamine and derivatives (e.g., Texas red and tetrarhodimine isothiocynate (TRITC)), biotin, phycoerythrin, AMCA, CyDyes™, 6- carboxyfhiorescein (commonly known by the abbreviations FAM and F), 6-carboxy-2',4',7',4,7- hexachlorofmorescein (HEX), 6-carboxy-4',5'-dichloro-2',7'-dimethoxyfmorescein (JOE or J), N,N,N',N'-tetramethyl-6carboxyrhodamine (TAMRA or T), 6-carboxy-X-rhodamine (ROX or R), 5- carboxyrhodamine-6G (R6G5 or G5), 6-carboxyrhodamine-6G (R6G6 or G6), and rhodamine 110; cyanine dyes, e.g. Cy3, Cy5 and Cy7 dyes; coumarins, e.g umbelliferone; benzimide dyes, e.g. Hoechst 33258; phenanthridine dyes, e.g. Texas Red; ethidium dyes; acridine dyes; carbazole dyes; phenoxazine dyes; porphyrin dyes; polymethine dyes, e.g. cyanine dyes such as Cy3, Cy5, etc; BODIPY dyes and quinoline dyes. In some embodiments of any of the aspects, a detectable label can be a radiolabel including, but not limited to³H,¹²⁵1,³⁵S,¹⁴C,³²P, and³³P. In some embodiments of any of the aspects, a detectable label can be an enzyme including, but not limited to horseradish peroxidase and alkaline phosphatase. An enzymatic label can produce, for example, a chemiluminescent signal, a color signal, or a fluorescent signal. Enzymes contemplated for use to detectably label an antibody reagent include, but are not limited to, malate dehydrogenase, staphylococcal nuclease, delta-V-steroid isomerase, yeast alcohol dehydrogenase, alpha- glycerophosphate dehydrogenase, triose phosphate isomerase, horseradish peroxidase, alkaline phosphatase, asparaginase, glucose oxidase, beta-galactosidase, ribonuclease, urease, catalase, glucose-VI-phosphate dehydrogenase, glucoamylase and acetylcholinesterase. In some embodiments of any of the aspects, a detectable label is a chemiluminescent label, including, but not limited to lucigenin, luminol, luciferin, isoluminol, theromatic acridinium ester, imidazole, acridinium salt and oxalate ester. In some embodiments of any of the aspects, a detectable label can be a spectral colorimetric label including, but not limited to colloidal gold or colored glass or plastic (e.g., polystyrene, polypropylene, and latex) beads.

[0053] In some embodiments of any of the aspects, a detectable label can be a detectable tag, such as c-Myc, HA, VSV-G, HSV, FLAG, V5, HIS, or biotin. Other detection systems can also be used, for example, a biotin-streptavidin system. In this system, a first molecule is biotinylated. The quantity of biotin present is determined using a streptavidin-peroxidase conjugate and a chromagenic substrate. Such streptavidin peroxidase detection kits are commercially available, e.g. from DAKO; Carpinteria, CA.

[0054] A reagent can also be detectably labeled using fluorescence emitting metals such as¹⁵²Eu, or others of the lanthanide series. These metals can be attached to a target using such metal chelating groups as diethylenetriaminepentaacetic acid (DTP A) or ethylenediaminetetraacetic acid (EDTA). [0055] The one or more cell types that may be included in the methods or compositions described herein can comprise any mammalian cell type selected from cells that make up the mammalian body, including germ cells, somatic cells, and stem cells. The term “germ cells” refers to any line of cells that give rise to gametes (eggs and sperm). The term “somatic cells” refers to any biological cells forming the body of a multicellular organism; any cell other than a gamete, germ cell, gametocyte or undifferentiated stem cell. Examples of somatic cells include fibroblasts, chondrocytes, osteoblasts, tendon cells, mast cells, wandering cells, immune cells, pericytes, inflammatory cells, endothelial cells, myocytes (cardiac, skeletal and smooth muscle cells), adipocytes (i.e., lipocytes or fat cells), parenchyma cells (neurons and glial cells, nephron cells, hepatocytes, pancreatic cells, lung parenchyma cells) and non-parenchymal cells (e.g., sinusoidal hepatic endothelial cells, Kupffer cells and hepatic stellate cells). The term “stem cells” refers to cells that have the ability to divide for indefinite periods and to give rise to virtually all of the tissues of the mammalian body, including specialized cells. The stem cells include pluripotent cells, which upon undergoing further specialization become multipotent progenitor cells that can give rise to functional or somatic cells. Examples of stem and progenitor cells include hematopoietic stem cells (adult stem cells; i.e., hemocytoblasts) from the bone marrow that give rise to red blood cells, white blood cells, and platelets; mesenchymal stem cells (adult stem cells) from the bone marrow that give rise to stromal cells, fat cells, and types of bone cells; epithelial stem cells (progenitor cells) that give rise to the various types of skin cells; neural stem cells and neural progenitor cells that give rise to neuronal and glial cells; and muscle satellite cells (progenitor cells) that contribute to differentiated muscle tissue. [0056] In some embodiments, the engineered cell and/or the recipient cell (e.g., the cell which is coated with the cell membrane coating to provide the engineered cell) are mammalian cells. In some embodiments, the engineered cell and/or the recipient cell (e.g., the cell which is coated with the cell membrane coating to provide the engineered cell) are murine cells. In some embodiments, the engineered cell and/or the recipient cell (e.g., the cell which is coated with the cell membrane coating to provide the engineered cell) are primate cells. In some embodiments, the engineered cell and/or the recipient cell (e.g., the cell which is coated with the cell membrane coating to provide the engineered cell) are human cells. In some embodiments, the engineered cell and/or the recipient cell (e.g., the cell which is coated with the cell membrane coating to provide the engineered cell) are obtained from or derived from the same species of organism. In some embodiments, the engineered cell and/or the recipient cell (e.g., the cell which is coated with the cell membrane coating to provide the engineered cell) are obtained from or derived from different species.

[0057] In some embodiments, the engineered cell and/or the recipient cell (e.g., the cell which is coated with the cell membrane coating to provide the engineered cell) are primary cells. In some embodiments, the engineered cell and/or the recipient cell (e.g., the cell which is coated with the cell membrane coating to provide the engineered cell) are cell lines. In some embodiments, the engineered cell and/or the recipient cell (e.g., the cell which is coated with the cell membrane coating to provide the engineered cell) are cells differentiated from stem cells, progenitor cells, iPSCs, or the like in vitro.

[0058] In some embodiments, the engineered cell and/or the recipient cell (e.g., the cell which is coated with the cell membrane coating to provide the engineered cell) are obtained from or derived from diseased primary cells or a subject having a disease affecting the engineered and/or receipient cell type.

[0059] In some embodiments, the source cell(s) and/or reference cell(s) are mammalian cells. In some embodiments, the source cell(s) and/or reference cell(s) are murine cells. In some embodiments, the source cell(s) and/or reference cell(s) are primate cells. In some embodiments, the source cell(s) and/or reference cell(s) are human cells. In some embodiments, the source cell(s) and/or reference cell(s) are obtained from or derived from the same species of organism. In some embodiments, the source cell(s) and/or reference cell(s) are obtained from or derived from different species.

[0060] In some embodiments, the source cell(s) and/or reference cell(s) are primary cells. In some embodiments, the source cell(s) and/or reference cell(s) are cell lines. In some embodiments, the source cell(s) and/or reference cell(s) are cells differentiated from stem cells, progenitor cells, iPSCs, or the like in vitro.

[0061] In some embodiments, the source cell(s) and/or reference cell(s) are obtained from or derived from diseased primary cells or a subject having a disease affecting the engineered and/or receipient cell type.

[0062] In some embodiments of any of the aspects, the one or more source cells, the engineered cell comprising a cell membrane coating, and/or the recipient cell (e.g., the cell which is coated with the cell membrane coating to provide the engineered cell) are obtained from a subject. In some embodiments of any of the aspects, the one or more source cells, the engineered cell comprising a cell membrane coating, and/or the recipient cell (e.g., the cell which is coated with the cell membrane coating to provide the engineered cell) are obtained from a subject and the engineered cell is administered to that same subject. In some embodiments of any of the aspects, the one or more source cells are obtained from a subject and the engineered cell is administered to that same subject.

In some embodiments of any of the aspects, the one or more source cells, the engineered cell comprising a cell membrane coating, and/or the recipient cell (e.g., the cell which is coated with the cell membrane coating to provide the engineered cell) are obtained from a first subject and the engineered cell is administered to a second subject.

[0063] In some embodiments of any of the aspects, the engineered cell and/or the recipient cell is not autologous to a subject. In some embodiments of any of the aspects, the engineered cell and/or the receipient cell is autologous to a subject.

[0064] In some embodiments of any of the aspects, the source cell is autologous to a subject.

[0065] In some embodiments of any of the aspects, the engineered cell, recipient cell, and/or source cell are cells from a sample. The term “sample” as used herein denotes a sample taken or isolated from a biological organism, e.g., a blood or plasma sample from a subject. In some embodiments of any of the aspects, the present invention encompasses several examples of a biological sample. In some embodiments of any of the aspects, the biological sample is cells, or tissue, or peripheral blood, or bodily fluid. Exemplary biological samples include, but are not limited to, a biopsy, a tumor sample, biofluid sample; blood; serum; plasma; urine; sperm; mucus; tissue biopsy; organ biopsy; synovial fluid; bile fluid; cerebrospinal fluid; mucosal secretion; effusion; sweat; saliva; and/or tissue sample etc. The term also includes a mixture of the above-mentioned samples. The term “sample” also includes untreated or pretreated (or pre-processed) biological samples. In some embodiments of any of the aspects, a sample can comprise cells from a subject. [0066] The sample can be obtained by removing a sample from a subject, but can also be accomplished by using a previously isolated sample (e.g. isolated at a prior timepoint and isolated by the same or another person). In some embodiments of any of the aspects, the sample can be an untreated sample. As used herein, the phrase “untreated sample” refers to a sample that has not had any prior sample pre-treatment except for dilution and/or suspension in a solution. Exemplary methods for treating a sample include, but are not limited to, centrifugation, fdtration, sonication, homogenization, heating, freezing and thawing, and combinations thereof. In some embodiments of any of the aspects, the sample can be a frozen test sample, e.g., a frozen tissue. The frozen sample can be thawed before employing methods, assays and systems described herein. After thawing, a frozen sample can be centrifuged before being subjected to methods, assays and systems described herein. In some embodiments of any of the aspects, the sample is a clarified sample, for example, by centrifugation and collection of a supernatant comprising the clarified sample. In some embodiments of any of the aspects, a sample can be a pre-processed sample, for example, supernatant or filtrate resulting from a treatment selected from the group consisting of centrifugation, filtration, thawing, purification, and any combinations thereof. In some embodiments of any of the aspects, the sample can be treated with a chemical and/or biological reagent. Chemical and/or biological reagents can be employed to protect and/or maintain the stability of the sample, including biomolecules (e.g., nucleic acid and protein) therein, during processing. One exemplary reagent is a protease inhibitor, which is generally used to protect or maintain the stability of protein during processing. The skilled artisan is well aware of methods and processes appropriate for pre-processing of biological samples.

[0067] In some embodiments of any of the aspects, the methods, assays, and systems described herein can further comprise a step of obtaining or having obtained a sample from a subject. In some embodiments of any of the aspects, the subject can be a human subject.

[0068] In some embodiments of any of the aspects, the engineered T cell or recipient T cell is a CD8+ T cell.

[0001] In some embodiments of any of the aspects, the engineered T cell or recipient T cell is a CAR-T cell, e.g., a cell comprising or expressing a chimeric antigen receptor (CAR). CAR-T-cell and related therapies relate to adoptive cell transfer of immune cells (e.g., T-cells) expressing a CAR that binds specifically to a target (e.g., CD123) to treat a subject. The cells are engineered and/or genetically modified to express the CAR. The structure of CARs and methods for making them and CAR-T cells are well known in the art. Further discussion of CAR-T therapies can be found, e.g., in Wei et al. Journal of Hematology and Oncology 2019 12:62; Maus et al. Blood 2014 123:2624-35; Reardon et al. Neuro-Oncology 2014 16:1441-1458; Hoyos et al. Haematologica 2012 97:1622; Byrd et al. J Clin Oncol 2014 32:3039-47; Maher et al. Cancer Res 2009 69:4559-4562; and Tamada et al. Clin Cancer Res 2012 18:6436-6445; each of which is incorporated by reference herein in its entirety. [0069] In some embodiments of any of the aspects, the CAR can be specific for or active against a diseased cell. In some embodiments of any of the aspects, the CAR can be specific for or active against a diseased cell of the same type as from which the cell membrane components were obtained or derived from. Exemplary targets for CARs include but are not limited to CD123, CD19, CD20, BCMA, CLL-1, CD22, HER-2, CEA, EpCAM, TAG-72, EGFRvIII, IL13Ra2, EphA2, CD44v6, NKG2D, CD133, TNFRSF17, SDC1, MS4A1, TNFRSF8, CD33, CD38, CD5, NCAM1, CD70, ULBP1, ULBP2, IL1RAP, CEACAM5, MET, EGFR, ERBB2, GPC3, MSLN, Mucl, PDCD1, CD274, KDR, FOLH1, FAP, CA9, FOLR1, LI CAM, ROR1, CD23, CD44, CD174, SLAMF7, GD2, PSCA, GPNMB, CD276, CD133, CSPG4, and TEM1.

[0070] In some embodiments of any of the aspects, the CAR is an anti-CD 123 CAR, e.g., it can bind specifically to CD 123. CD 123, also known as Cell Differentiation 123 or Interleukin-3 receptor is a receptor for IL-3. The sequences of CD123 are known in the art for a variety of species, e.g., for human (see NCBI Gene ID 3563). Anti-CD 123 reagents, including CAR-T cells are known in the art and include MB-102 CAR-T cells from Mustang Bio Inc., RIVA-CAR T 123, 7G3 MAb, CSL362 (taclotuzumab) MAb, and G4723A MAb. Such reagents are further described in the art at, e.g, Testa et al. Cancers 2019 11:1358; which is incorporated by reference herein in its entirety.

[0071] In some embodiments of any of the aspects, the engineered cell present in a composition, or combination, of the disclosure exhibit an increased utility that is not exhibited when the corresponding recipient cell occurs alone or when said recipient cell is present at a naturally occurring concentration. In some embodiments of any of the aspects, the compositions of the disclosure- comprising engineered cells as taught herein— exhibit markedly different characteristics/properties compared to their closest naturally occurring counterpart, e.g., the corresponding recipient cell. That is, the compositions of the disclosure exhibit markedly different functional and/or structural characteristics/properties, as compared to their closest naturally occurring counterpart. For instance, the engineered cells of the disclosure are structurally different from a recipient cell as it naturally exists in a subject, for at least the following reasons: said engineered cell comprises a cell membranecoating, said engineered cell can be isolated and purified, such that it is not found in the milieu of the subject, said engineered cell can be present at concentrations that do not occur in the subject, and said engineered cell can be associated with acceptable carriers that do not occur in the subject. Further, the engineered cells of the disclosure are functionally different from a recipient cell as it naturally exists in a subject, for at least the following reasons: said engineered cell when applied in an isolated and purified form can lead to therapeutic effects and immune evasion, said engineered cell now has a new utility as a supplement capable of administration to a subject, wherein the recipient cell could not have such a utility in its natural state in a subject.

[0072] An engineered cell comprising a cell membrane coating can be prepared by a method comprising: a) obtaining cell membrane components from one or more source cells; b) optionally, mixing or fusing the cell membrane components of two or more source cells; and c) contacting a recipient cell with the cell membrane components to obtain an engineered cell comprising a cell membrane coating.

[0073] Cell membrane components can be obtained from one or more source cells by any method known in the art that disrupts cellular membrane integrity or lyses a cell, e.g., contacting with detergents, freezing and then thawing the source cell(s) (i.e., freeze-thaw), bead milling, use of a blender, liquid homogenization, sonication, and the like. In some embodiments of any of the aspects, the cell membrane components are obtained by freeze-thaw of the source cell(s). In some embodiments of any of the aspects, obtaining the cell membrane components can further comprise washing or purifying the cell membrane components, e.g., affinity purification, centriguation, size filtration, extraction, etc.

[0074] Cell membrane components, e.g., cell membrane components from two different source cell types can be mixed or fused by physical mixing, fluid flow, sonication, or any other method of mixing known in the art. In some embodiments of any of the aspects, the mixing or fusing of the cell membrane components comprises sonication. In some embodiments of any of the aspects, the mixing or fusing of the cell membrane components comprises fluid flow. In some embodiments of any of the aspects, the mixing or fusing of the cell membrane components comprises sonication and fluid flow. [0075] In some embodiments of any of the aspects, the method of preparing an engineered cell comprising a cell membrane coating can be conducted in whole or in part in one or more fluidic (e.g., microfluidic) devices. In some embodiments of any of the aspects, the method of preparing an engineered cell comprising a cell membrane coating can be conducted in a fluidic (e.g., microfluidic) device. In some embodiments of any of the aspects, the steps b) and c) of a method of preparing an engineered cell comprising a cell membrane coating can be conducted in whole or in part in one or more fluidic (e.g., microfluidic) devices. In some embodiments of any of the aspects, the steps b) and c) of a method of preparing an engineered cell comprising a cell membrane coating can be conducted in a fluidic (e.g., microfluidic) device.

[0076] In one aspect of any of the embodiments, described herein is a fluidic (e.g., microfluidic) device 100 comprising, in the following order along a flow path(s): one or more initial input ports 101; a flow channel for mixing materials provided to the one or more initial input ports 102; one or more recipient cell input ports 103; a flow channel 104 comprising sonication input 105, or capable of receiving sonication input; and one or more output ports 106. An exemplary embodiment is depicted in Fig. 2.

[0077] As used herein “fluidic device” refers to a device of any size or orientation which comprises one or more fluid channels, e,g., flow channels, and is suitable for the culture of living cells. A fluidic device can be capable of moving any amount of fluid within the fluid flow ranges described herein below, e.g. a fluidic device can be a microfluidic device or a device capable of moving larger volumes of fluid. In some embodiments of any of the aspects, the fluidic device can be an microfluidic device. The term “microfluidic” as used herein relates to components where moving fluid is constrained in or directed through one or more channels wherein one or more dimensions are 1 mm or smaller (microscale). Microfluidic channels may be larger than microscale in one or more directions, though the channel(s) will be on the microscale in at least one direction. In some instances the geometry of a microfluidic channel may be configured to control the fluid flow rate through the channel (e.g. increase channel height to reduce shear). Microfluidic channels can be formed of various geometries to facilitate a wide range of flow rates through the channels.

[0078] “Channels" are pathways (whether straight, curved, single, multiple, in a network, etc.) through a medium (e.g., silicon) that allow for movement of liquids and gasses. Channels thus can connect other components, i.e., keep components "in communication" and more particularly, "in fluidic communication" and still more particularly, "in liquid communication." Such components include, but are not limited to, liquid-intake ports and gas vents. Microchannels are channels with dimensions less than 1 millimeter and greater than 1 micron. Channels can include capillaries, tubes, pathways, and/or grooves deposed within or upon a medium or substrate.

[0079] As used herein, the term “port” refers to a portion of the fluidic device described herein which provides a means for fluid and/or cells to enter and/or exit the device and/or to enter and/or exit portions of the device. The port can be of a size and shape to accept and/or secure a connection with tubes, connections, or adaptors of a fluidic or microfluidic device and allow passage of fluid and/or cells when attached to a fluidic or microfluidic device.

[0080] Fluids can be provided in a fluid source connected to an input port 101 and/or 103. A fluid source can be a reservoir or other container comprising a volume of fluid such that the fluid can be caused to move from the fluid source and through the one or more channels of the fluidic device. The fluid source can be coupled to the one or more channels of the fluidic device by any means of conducting a fluid, e.g. tubing, piping, channels, or the like. The fluidic device and/or the fluid source can comprise ports.

[0081] Either positive or negative fluid pressure, or both, can be used to cause the fluid to flow through the channels 102 and/or 104. In accordance with some embodiments of the invention, the fluid in fluid source can be pressurized and a valve can be provided between the fluid source and the channel 102 and/or 104 to control the flow of fluid into the channel. In accordance with some embodiments of the invention, a vacuum source can be connected to the outlet port of the channel to draw the fluid through the channel. In accordance with some embodiments of the invention, gravity can be used to cause the fluid to flow through the channel. For example, the fluid source can be elevated above the device and the fluid collection reservoir can be placed below the device to provide fluid pressure that causes fluid to flow through the channel. A valve at the fluid source or in the fluid flow path can be used to control the rate of fluid flow. In accordance with some embodiments of the invention, one or more pumps can be used cause the fluid to flow from the fluid source through the channel.

[0082] In some embodiments of any of the aspects, the method can comprise providing fluid to one or more channels by connecting the fluid channel(s) to a fluid source, e.g, a fluid source comprised by a fluidics system. In some embodiments of any of the aspects, a device described herein can comprise a connection of a fluid channel(s) to a fluid source, e.g, a fluid source comprised by a fluidics system. In some embodiments of any of the aspects, the connection can be via a port in the fluidic device.

[0083] In some embodiments of any of the aspects, control of the fluid flow from the fluid source through a fluid channel can be automated. In an embodiment in which control of the flow of solution from the fluid source is automated, a syringe pump or solenoid can be used. In other embodiments, one or more computing devices or systems may be used to control fluid flow. Alternatively or additionally, a computing device may be coupled to a fluid source or port in order to control the flow of fluid from the fluid source. Exemplary, but non-limiting, embodiments of automated devices are described in US Patent Publication US 2014/0038279; which is incorporated by reference herein its entirety.

[0084] In some embodiments of any of the aspects, the fluid flow rate through the one or more channels of the fluidic device can be from 1 to 100 mU/h. In some embodiments of any of the aspects, the fluid flow rate through the one or more channels of the fluidic device can be from 5 to 50 mU/h. In some embodiments of any of the aspects, the fluid flow rate through the one or more channels of the fluidic device can be from 2 to 75 mU/h.

[0085] The structures of the fluidic devices described herein can be formed, such as by etching, 3-D printing, machining, or micro-machining. In some embodiments of any of the aspects, the device described herein is etching -free.

[0086] The fluidic devices described herein can be made of a biocompatible flexible material or a biocompatible non-flexible material according to the design and application requirements. The fluidic devices and/or portions thereof can be made of a flexible material, including but not limited to, a biocompatible material such as polydimethyl siloxane (PDMS), polyurethane or polyimide. The fluidic devices and/or portions thereof can also be made of non-flexible materials like glass, silicon, polysulfone, hard plastic, and the like, as well as combinations of these materials. [0087] A biocompatible polymer refers to materials which do not have toxic or injurious effects on biological functions. Biocompatible polymers include natural or synthetic polymers. Examples of biocompatible polymers include, but are not limited to, collagen, poly(alpha esters) such as poly(lactate acid), poly(glycolic acid), polyorthoesters and polyanhydrides and their copolymers, polyglycolic acid and polyglactin, cellulose ether, cellulose, cellulosic ester, fluorinated polyethylene, phenolic, poly-4-methylpentene, polyacrylonitrile, polyamide, polyamideimide, polyacrylate, polybenzoxazole, polycarbonate, polycyanoarylether, polyester, polyestercarbonate, polyether, polyetheretherketone, polyetherimide, polyetherketone, polyethersulfone, polyethylene, polyfluoroolefm, polyimide, polyolefin, polyoxadiazole, polyphenylene oxide, polyphenylene sulfide, polypropylene, polystyrene, polysulfide, polysulfone, polytetrafluoroethylene, polythioether, polytriazole, polyurethane, polyvinyl, polyvinylidene fluoride, regenerated cellulose, silicone, urea- formaldehyde, polyglactin, or copolymers or physical blends of these materials.

[0088] A biocompatible material can also be, for example, ceramic coatings on a metallic substrate. But any type of coating material and the coating can be made of different types of materials: metals, ceramics, polymers, hydrogels or a combination of any of these materials. Biocompatible materials include, but are not limited to an oxide, a phosphate, a carbonate, a nitride or a carbonitride. Among the oxide the following ones are preferred: tantalum oxide, aluminum oxide, iridium oxide, zirconium oxide or titanium oxide. Substrates are made of materials such as metals, ceramics, polymers or a combination of any of these. Metals such as stainless steel, Nitinol, titanium, titanium alloys, or aluminum and ceramics such as zirconia, alumina, or calcium phosphate are of particular interest.

[0089] As described herein, engineered cells comprising a cell membrane coating as described herein provide improved safety and efficacy, e.g., by immune evasion. Accordingly, in one aspect of any of the embodiments, described herein is a method of treating a subject, the method comprising administering an engineered cell comprising a cell membrane coating as described herein to the subject.

[0090] In one aspect of any of the embodiments, described herein is a method of administering a cell to a subject, the method comprising administering an engineered cell comprising a cell membrane coating as described herein to the subject.

[0091] In one aspect of any of the embodiments, described herein is an engineered cell comprising a cell membrane coating for use in a method of administering a cell to a subject. In one aspect of any of the embodiments, described herein is an engineered cell comprising a cell membrane coating for use in a method of treating a subject.

[0092] In some embodiments of any of the aspects, the subject has or is in need of treatment for cancer and/or the method of treatment is a method of treating cancer. In some embodiments of any of the aspects, the cancer is a hematological cancer or hematopoietic cancer. In some embodiments of any of the aspects, the cancer is leukemia. In some embodiments of any of the aspects, the cancer is acute myeloid leukemia.

[0093] As used herein, the term “cancer” relates generally to a class of diseases or conditions in which abnormal cells divide without control and can invade nearby tissues. Cancer cells can also spread to other parts of the body through the blood and lymph systems. There are several main types of cancer. Carcinoma is a cancer that begins in the skin or in tissues that line or cover internal organs. Sarcoma is a cancer that begins in bone, cartilage, fat, muscle, blood vessels, or other connective or supportive tissue. Leukemia is a cancer that starts in blood-forming tissue such as the bone marrow, and causes large numbers of abnormal blood cells to be produced and enter the blood. Lymphoma and multiple myeloma are cancers that begin in the cells of the immune system. Central nervous system cancers are cancers that begin in the tissues of the brain and spinal cord.

[0094] In some embodiments of any of the aspects, the cancer is a primary cancer. In some embodiments of any of the aspects, the cancer is a malignant cancer. As used herein, the term “malignant” refers to a cancer in which a group of tumor cells display one or more of uncontrolled growth (i.e., division beyond normal limits), invasion (i.e.. intrusion on and destruction of adjacent tissues), and metastasis (i.e. , spread to other locations in the body via lymph or blood). As used herein, the term “metastasize” refers to the spread of cancer from one part of the body to another. A tumor formed by cells that have spread is called a “metastatic tumor” or a “metastasis.” The metastatic tumor contains cells that are like those in the original (primary) tumor. As used herein, the term “benign” or “non-malignant” refers to tumors that may grow larger but do not spread to other parts of the body. Benign tumors are self-limited and typically do not invade or metastasize.

[0095] A “cancer cell” or “tumor cell” refers to an individual cell of a cancerous growth or tissue. A tumor refers generally to a swelling or lesion formed by an abnormal growth of cells, which may be benign, pre -malignant, or malignant. Most cancer cells form tumors, but some, e.g., leukemia, do not necessarily form tumors. For those cancer cells that form tumors, the terms cancer (cell) and tumor (cell) are used interchangeably.

[0096] As used herein the term "neoplasm" refers to any new and abnormal growth of tissue, e.g., an abnormal mass of tissue, the growth of which exceeds and is uncoordinated with that of the normal tissues. Thus, a neoplasm can be a benign neoplasm, premalignant neoplasm, or a malignant neoplasm.

[0097] A subject that has a cancer or a tumor is a subject having objectively measurable cancer cells present in the subject’s body. Included in this definition are malignant, actively proliferative cancers, as well as potentially dormant tumors or micrometastatses. Cancers which migrate from their original location and seed other vital organs can eventually lead to the death of the subject through the functional deterioration of the affected organs. [0098] Examples of cancer include but are not limited to, carcinoma, lymphoma, blastoma, sarcoma, leukemia, basal cell carcinoma, biliary tract cancer; bladder cancer; bone cancer; brain and CNS cancer; breast cancer; cancer of the peritoneum; cervical cancer; choriocarcinoma; colon and rectum cancer; connective tissue cancer; cancer of the digestive system; endometrial cancer; esophageal cancer; eye cancer; cancer of the head and neck; gastric cancer (including gastrointestinal cancer); glioblastoma (GBM); hepatic carcinoma; hepatoma; intra-epithelial neoplasm.; kidney or renal cancer; larynx cancer; leukemia; liver cancer; lung cancer (e.g., small-cell lung cancer, non small cell lung cancer, adenocarcinoma of the lung, and squamous carcinoma of the lung); lymphoma including Hodgkin’s and non-Hodgkin’s lymphoma; melanoma; myeloma; neuroblastoma; oral cavity cancer (e.g., lip, tongue, mouth, and pharynx); ovarian cancer; pancreatic cancer; prostate cancer; retinoblastoma; rhabdomyosarcoma; rectal cancer; cancer of the respiratory system; salivary gland carcinoma; sarcoma; skin cancer; squamous cell cancer; stomach cancer; testicular cancer; thyroid cancer; uterine or endometrial cancer; cancer of the urinary system; vulval cancer; as well as other carcinomas and sarcomas; as well as B-cell lymphoma (including low grade/follicular non-Hodgkin’s lymphoma (NHL); small lymphocytic (SL) NHL; intermediate grade/follicular NHL; intermediate grade diffuse NHL; high grade immunoblastic NHL; high grade lymphoblastic NHL; high grade small non-cleaved cell NHL; bulky disease NHL; mantle cell lymphoma; AIDS-related lymphoma; and Waldenstrom’s Macroglobulinemia); chronic lymphocytic leukemia (CLL); acute lymphoblastic leukemia (ALL); Hairy cell leukemia; chronic myeloblastic leukemia; and post-transplant lymphoproliferative disorder (PTLD), as well as abnormal vascular proliferation associated with phakomatoses, edema (such as that associated with brain tumors), and Meigs’ syndrome [0099] A “cancer cell” is a cancerous, pre-cancerous, or transformed cell, either in vivo, ex vivo, or in tissue culture, that has spontaneous or induced phenotypic changes that do not necessarily involve the uptake of new genetic material. Although transformation can arise from infection with a transforming virus and incorporation of new genomic nucleic acid, or uptake of exogenous nucleic acid, it can also arise spontaneously or following exposure to a carcinogen, thereby mutating an endogenous gene. Transformation/cancer is associated with, e.g., morphological changes, immortalization of cells, aberrant growth control, foci formation, anchorage independence, malignancy, loss of contact inhibition and density limitation of growth, growth factor or serum independence, tumor specific markers, invasiveness or metastasis, and tumor growth in suitable animal hosts such as nude mice.

[00100] In some embodiments of any of the aspects, the subject is a subject at risk of cytokine release syndrome (CRS). Cytokine release syndrome is a condition that may occur after treatment with some types of immunotherapy, such as monoclonal antibodies and CAR-T cells. Cytokine release syndrome is caused by a large, rapid release of cytokines into the blood from immune cells affected by the immunotherapy. Signs and symptoms of cytokine release syndrome include fever, nausea, headache, rash, rapid heartbeat, low blood pressure, and trouble breathing. Most patients have a mild reaction, but sometimes, the reaction may be severe or life threatening. Further details on CRS and risk factors for CRS can be found in the art, e.g., in Yan et al. Front. Immunol., 23 February 2021 I doi.org/10.3389/fimmu.2021.611366; Frey Best Pract Res Clin Haematol. 2017 Dec;30(4):336-340; and Hong et al. Bone Marrow Transplantation volume 56, pages 570-580 (2021); each of which is incorporated by reference herein in its entirety.

[00101] In some embodiments of any of the aspects, the methods described herein relate to treating a subject having or diagnosed as having cancer with a composition (e.g., an engineered cell comprising a cell membrane coating) described herein. Subjects having cancer can be identified by a physician using current methods of diagnosing cancer. Symptoms and/or complications of cancer which characterize these conditions and aid in diagnosis are well known in the art and include but are not limited to, for example, a lump/mass/tumor, swelling, fatique, swollen lymph nodes, frequent infections, or pain. Tests that may aid in a diagnosis of, e.g. cancer include, but are not limited to, x- rays, MRI, ultrasound, a biopsy, or tests for the function/activity of affected organs or systems. A family history of cancer or exposure to risk factors for cancer (e.g. smoke, radiation, pollutants, mutation, etc.) can increase the risk of a subject having cancer.

[00102] The compositions and methods described herein can be administered to a subject having or diagnosed as having cancer. In some embodiments of any of the aspects, the methods described herein comprise administering an effective amount of compositions described herein to a subject in order to alleviate a symptom of a cancer. As used herein, "alleviating a symptom” of a cancer is ameliorating any condition or symptom associated with the cancer, e.g, a reduction in cancer cell growth or numbers. As compared with an equivalent untreated control, such reduction is by at least 5%, 10%, 20%, 40%, 50%, 60%, 80%, 90%, 95%, 99% or more as measured by any standard technique. A variety of means for administering the compositions described herein to subjects are known to those of skill in the art. Such methods can include, but are not limited to oral, parenteral, intravenous, intramuscular, subcutaneous, transdermal, airway (aerosol), pulmonary, cutaneous, topical, injection, or intratumoral administration. Administration can be local or systemic. In some embodiments of any of the apsects, the administration is subcutaneous.

[00103] The term “effective amount" as used herein refers to the amount of a composition needed to alleviate at least one or more symptom of the disease or disorder, and relates to a sufficient amount of pharmacological composition to provide the desired effect. The term "therapeutically effective amount" therefore refers to an amount of a composition that is sufficient to provide a particular therapeutic effect when administered to a typical subject. An effective amount as used herein, in various contexts, would also include an amount sufficient to delay the development of a symptom of the disease, alter the course of a symptom disease (for example but not limited to, slowing the progression of a symptom of the disease), or reverse a symptom of the disease. Thus, it is not generally practicable to specify an exact “effective amount". However, for any given case, an appropriate “effective amount" can be determined by one of ordinary skill in the art using only routine experimentation.

[00104] Effective amounts, toxicity, and therapeutic efficacy can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the LD50 (the dose lethal to 50% of the population) and the ED50 (the dose therapeutically effective in 50% of the population). The dosage can vary depending upon the dosage form employed and the route of administration utilized. The dose ratio between toxic and therapeutic effects is the therapeutic index and can be expressed as the ratio LD50/ED50. Compositions and methods that exhibit large therapeutic indices are preferred. A therapeutically effective dose can be estimated initially from cell culture assays. Also, a dose can be formulated in animal models to achieve a circulating plasma concentration range that includes the IC50 (i.e.. the concentration of a cell, system, and/or drug, which achieves a half-maximal inhibition of symptoms) as determined in cell culture, or in an appropriate animal model. Levels in plasma can be measured, for example, by high performance liquid chromatography. The effects of any particular dosage can be monitored by a suitable bioassay, e.g., assay for cancer cell growth, among others. The dosage can be determined by a physician and adjusted, as necessary, to suit observed effects of the treatment.

[00105] Effective amounts, toxicity, and therapeutic efficacy can be determined by standard pharmaceutical procedures in cell cultures or experimental animals, e.g., for determining the minimal effective dose and/or maximal tolerated dose. The dosage can vary depending upon the dosage form employed and the route of administration utilized. A therapeutically effective dose can be estimated initially from cell culture assays. Also, a dose can be formulated in animal models to achieve a dosage range between the minimal effective dose and the maximal tolerated dose. The effects of any particular dosage can be monitored by a suitable bioassay, e.g., assay for cancer growth and/or size among others. The dosage can be determined by a physician and adjusted, as necessary, to suit observed effects of the treatment.

[00106] In some embodiments, the technology described herein relates to a pharmaceutical composition comprising an engineered cell comprising a cell membrane coating as described herein, and optionally a pharmaceutically acceptable carrier. In some embodiments, the active ingredients of the pharmaceutical composition comprise an engineered cell comprising a cell membrane coating as described herein. In some embodiments, the active ingredients of the pharmaceutical composition consist essentially of an engineered cell comprising a cell membrane coating as described herein. In some embodiments, the active ingredients of the pharmaceutical composition consist of an engineered cell comprising a cell membrane coating as described herein.

[00107] Pharmaceutically acceptable carriers and diluents include saline, aqueous buffer solutions, solvents and/or dispersion media. The use of such carriers and diluents is well known in the art. Some non-limiting examples of materials which can serve as pharmaceutically-acceptable carriers include: (1) sugars, such as lactose, glucose and sucrose; (2) starches, such as com starch and potato starch; (3) cellulose, and its derivatives, such as sodium carboxymethyl cellulose, methylcellulose, ethyl cellulose, microcrystalline cellulose and cellulose acetate; (4) powdered tragacanth; (5) malt; (6) gelatin; (7) lubricating agents, such as magnesium stearate, sodium lauryl sulfate and talc; (8) excipients, such as cocoa butter and suppository waxes; (9) oils, such as peanut oil, cottonseed oil, safflower oil, sesame oil, olive oil, com oil and soybean oil; (10) glycols, such as propylene glycol;

(11) polyols, such as glycerin, sorbitol, mannitol and polyethylene glycol (PEG); (12) esters, such as ethyl oleate and ethyl laurate; (13) agar; (14) buffering agents, such as magnesium hydroxide and aluminum hydroxide; (15) alginic acid; (16) pyrogen-free water; (17) isotonic saline; (18) Ringer's solution; (19) ethyl alcohol; (20) pH buffered solutions; (21) polyesters, polycarbonates and/or polyanhydrides; (22) bulking agents, such as polypeptides and amino acids (23) semm component, such as semm albumin, HDL and LDL; (22) C2-C12 alcohols, such as ethanol; and (23) other non toxic compatible substances employed in pharmaceutical formulations. Wetting agents, coloring agents, release agents, coating agents, sweetening agents, flavoring agents, perfuming agents, preservative and antioxidants can also be present in the formulation. The terms such as "excipient", "carrier", "pharmaceutically acceptable carrier" or the like are used interchangeably herein. In some embodiments, the carrier inhibits the degradation of the active agent, e.g. an engineered cell comprising a cell membrane coating as described herein.

[00108] In some embodiments, the pharmaceutical composition as described herein can be a parenteral dose form. Since administration of parenteral dosage forms typically bypasses the patient's natural defenses against contaminants, parenteral dosage forms are preferably sterile or capable of being sterilized prior to administration to a patient. Examples of parenteral dosage forms include, but are not limited to, solutions ready for injection, dry products ready to be dissolved or suspended in a pharmaceutically acceptable vehicle for injection, suspensions ready for injection, and emulsions. In addition, controlled-release parenteral dosage forms can be prepared for administration of a patient, including, but not limited to, DUROS^®-type dosage forms and dose-dumping.

[00109] Suitable vehicles that can be used to provide parenteral dosage forms of compositions as disclosed within are well known to those skilled in the art. Examples include, without limitation: sterile water; water for injection USP; saline solution; glucose solution; aqueous vehicles such as but not limited to, sodium chloride injection, Ringer's injection, dextrose Injection, dextrose and sodium chloride injection, and lactated Ringer's injection; water-miscible vehicles such as, but not limited to, ethyl alcohol, polyethylene glycol, and propylene glycol; and non-aqueous vehicles such as, but not limited to, com oil, cottonseed oil, peanut oil, sesame oil, ethyl oleate, isopropyl myristate, and benzyl benzoate. Compounds that alter or modify the solubility of a pharmaceutically acceptable salt of a molecule as disclosed herein can also be incorporated into the parenteral dosage forms of the disclosure, including conventional and controlled-re lease parenteral dosage forms.

[00110] Pharmaceutical compositions comprising an engineered cell comprising a cell membrane coating can also be formulated to be suitable for oral administration, for example as discrete dosage forms, such as, but not limited to, tablets (including without limitation scored or coated tablets), pills, caplets, capsules, chewable tablets, powder packets, cachets, troches, wafers, aerosol sprays, or liquids, such as but not limited to, syrups, elixirs, solutions or suspensions in an aqueous liquid, a non- aqueous liquid, an oil-in-water emulsion, or a water-in-oil emulsion. Such compositions contain a predetermined amount of the pharmaceutically acceptable salt of the disclosed compounds, and may be prepared by methods of pharmacy well known to those skilled in the art. See generally,

Remington: The Science and Practice of Pharmacy, 21st Ed., Lippincott, Williams, and Wilkins, Philadelphia PA. (2005).

[00111] Conventional dosage forms generally provide rapid or immediate drug release from the formulation. Depending on the pharmacology and pharmacokinetics of the drug, use of conventional dosage forms can lead to wide fluctuations in the concentrations of the drug in a patient's blood and other tissues. These fluctuations can impact a number of parameters, such as dose frequency, onset of action, duration of efficacy, maintenance of therapeutic blood levels, toxicity, side effects, and the like. Advantageously, controlled-release formulations can be used to control a drug's onset of action, duration of action, plasma levels within the therapeutic window, and peak blood levels. In particular, controlled- or extended-release dosage forms or formulations can be used to ensure that the maximum effectiveness of a drug is achieved while minimizing potential adverse effects and safety concerns, which can occur both from under-dosing a drug (i.e., going below the minimum therapeutic levels) as well as exceeding the toxicity level for the drug. In some embodiments, the a system, or cell comprising or encoding a system can be administered in a sustained release formulation.

[00112] Controlled-release pharmaceutical products have a common goal of improving therapy over that achieved by their non-controlled release counterparts. Ideally, the use of an optimally designed controlled-release preparation in medical treatment is characterized by a minimum of substance being employed to cure or control the condition in a minimum amount of time. Advantages of controlled- release formulations include: 1) extended activity of the therapeutic; 2) reduced dosage frequency; 3) increased patient compliance; 4) usage of less total therapeutic; 5) reduction in local or systemic side effects; 6) minimization of therapeutic accumulation; 7) reduction in blood level fluctuations; 8) improvement in efficacy of treatment; 9) reduction of potentiation or loss of therapeutic activity; and 10) improvement in speed of control of diseases or conditions. Kim, Chemg-ju, Controlled Release Dosage Form Design, 2 (Technomic Publishing, Lancaster, Pa.: 2000).

[00113] Most controlled-release formulations are designed to initially release an amount of a therapeutic (active ingredient) that promptly produces the desired therapeutic effect, and gradually and continually release other amounts of therapeutic to maintain this level of therapeutic or prophylactic effect over an extended period of time. In order to maintain this constant level of therapeutic in the body, the therapeutic must be released from the dosage form at a rate that will replace the amount of drug being metabolized and excreted from the body. Controlled-release of an active ingredient can be stimulated by various conditions including, but not limited to, pH, ionic strength, osmotic pressure, temperature, enzymes, water, and other physiological conditions or compounds.

[00114] A variety of known controlled- or extended-release dosage forms, formulations, and devices can be adapted for use with the salts and compositions of the disclosure. Examples include, but are not limited to, those described in U.S. Pat. Nos.: 3,845,770; 3,916,899; 3,536,809; 3,598,123; 4,008,719; 5674,533; 5,059,595; 5,591 ,767; 5,120,548; 5,073,543; 5,639,476; 5,354,556; 5,733,566; and 6,365,185 B1 ; each of which is incorporated herein by reference. These dosage forms can be used to provide slow or controlled-release of one or more active ingredients using, for example, hydroxypropylmethyl cellulose, other polymer matrices, gels, permeable membranes, osmotic systems (such as OROS^® (Alza Corporation, Mountain View, Calif. USA)), or a combination thereof to provide the desired release profde in varying proportions.

[00115] Im some embodiments of any of the aspects, the engineered cell comprising a cell membrane coating described herein is administered as a monotherapy, e.g., another treatment for the condition (e.g., cancer) is not administered to the subject.

[00116] In some emboidments of any of the aspects, the methods described herein can further comprise administering a second agent and/or treatment to the subject, e.g. as part of a combinatorial therapy. Non-limiting examples of a second agent and/or treatment can include radiation therapy, surgery, gemcitabine, cisplastin, paclitaxel, carboplatin, bortezomib, AMG479, vorinostat, rituximab, temozolomide, rapamycin, ABT-737, PI-103; alkylating agents such as thiotepa and CYTOXAN® cyclosphosphamide; alkyl sulfonates such as busulfan, improsulfan and piposulfan; aziridines such as benzodopa, carboquone, meturedopa, and uredopa; ethylenimines and methylamelamines including altretamine, triethylenemelamine, trietylenephosphoramide, triethiylenethiophosphoramide and trimethylolomelamine; acetogenins (especially bullatacin and bullatacinone); a camptothecin (including the synthetic analogue topotecan); bryostatin; callystatin; CC-1065 (including its adozelesin, carzelesin and bizelesin synthetic analogues); cryptophycins (particularly cryptophycin 1 and cryptophycin 8); dolastatin; duocarmycin (including the synthetic analogues, KW-2189 and CB1- TM1); eleutherobin; pancratistatin; a sarcodictyin; spongistatin; nitrogen mustards such as chlorambucil, chlomaphazine, cholophosphamide, estramustine, ifosfamide, mechlorethamine, mechlorethamine oxide hydrochloride, melphalan, novembichin, phenesterine, prednimustine, trofosfamide, uracil mustard; nitrosureas such as carmustine, chlorozotocin, fotemustine, lomustine, nimustine, and ranimnustine; antibiotics such as the enediyne antibiotics (e.g., calicheamicin, especially calicheamicin gammall and calicheamicin omegall (see, e.g., Agnew, Chem. Inti. Ed. Engl., 33: 183-186 (1994)); dynemicin, including dynemicin A; bisphosphonates, such as clodronate; an esperamicin; as well as neocarzinostatin chromophore and related chromoprotein enediyne antiobiotic chromophores), aclacinomysins, actinomycin, authramycin, azaserine, bleomycins, cactinomycin, carabicin, caminomycin, carzinophilin, chromomycinis, dactinomycin, daunorubicin, detorubicin, 6-diazo-5-oxo-L-norleucine, ADRIAMYCIN® doxorubicin (including morpholino- doxorubicin, cyanomorpholino-doxorubicin, 2-pyrrolino-doxorubicin and deoxydoxorubicin), epirubicin, esorubicin, idarubicin, marcellomycin, mitomycins such as mitomycin C, mycophenolic acid, nogalamycin, olivomycins, peplomycin, potfiromycin, puromycin, quelamycin, rodorubicin, streptonigrin, streptozocin, tubercidin, ubenimex, zinostatin, zorubicin; anti-metabolites such as methotrexate and 5-fluorouracil (5-FU); folic acid analogues such as denopterin, methotrexate, pteropterin, trimetrexate; purine analogs such as fludarabine, 6-mercaptopurine, thiamiprine, thioguanine; pyrimidine analogs such as ancitabine, azacitidine, 6-azauridine, carmofur, cytarabine, dideoxyuridine, doxifluridine, enocitabine, floxuridine; androgens such as calusterone, dromostanolone propionate, epitiostanol, mepitiostane, testolactone; anti-adrenals such as aminoglutethimide, mitotane, trilostane; folic acid replenisher such as frolinic acid; aceglatone; aldophosphamide glycoside; aminolevulinic acid; eniluracil; amsacrine; bestrabucil; bisantrene; edatraxate; defofamine; demecolcine; diaziquone; elformithine; elliptinium acetate; an epothilone; etoglucid; gallium nitrate; hydroxyurea; lentinan; lonidainine; maytansinoids such as maytansine and ansamitocins; mitoguazone; mitoxantrone; mopidanmol; nitraerine; pentostatin; phenamet; pirarubicin; losoxantrone; podophyllinic acid; 2-ethylhydrazide; procarbazine; PSK® polysaccharide complex (JHS Natural Products, Eugene, Oreg.); razoxane; rhizoxin; sizof iran; spirogermanium; tenuazonic acid; triaziquone; 2,2',2"-trichlorotriethylamine; trichothecenes (especially T-2 toxin, verracurin A, roridin A and anguidine); urethan; vindesine; dacarbazine; mannomustine; mitobronitol; mitolactol; pipobroman; gacytosine; arabinoside ("Ara-C"); cyclophosphamide; thiotepa; taxoids, e.g., TAXOL® paclitaxel (Bristol-Myers Squibb Oncology, Princeton, N.J.), ABRAXANE® Cremophor- firee, albumin-engineered nanoparticle formulation of paclitaxel (American Pharmaceutical Partners, Schaumberg, Ill.), and TAXOTERE® doxetaxel (Rhone -Poulenc Rorer, Antony, France); chloranbucil; GEMZAR® gemcitabine; 6-thioguanine; mercaptopurine; methotrexate; platinum analogs such as cisplatin, oxaliplatin and carboplatin; vinblastine; platinum; etoposide (VP-16); ifosfamide; mitoxantrone; vincristine; NAVELBINE.RTM. vinorelbine; novantrone; teniposide; edatrexate; daunomycin; aminopterin; xeloda; ibandronate; irinotecan (Camptosar, CPT-11)

(including the treatment regimen of irinotecan with 5-FU and leucovorin); topoisomerase inhibitor RFS 2000; difluoromethylomithine (DMFO); retinoids such as retinoic acid; capecitabine; combretastatin; leucovorin (LV); oxaliplatin, including the oxaliplatin treatment regimen (FOLFOX); lapatinib (Tykerb.RTM.); inhibitors of PKC-alpha, Raf, H-Ras, EGFR (e.g., erlotinib (Tarceva®)) and VEGF-A that reduce cell proliferation and pharmaceutically acceptable salts, acids or derivatives of any of the above.

[00117] In addition, the methods of treatment can further include the use of radiation or radiation therapy. Further, the methods of treatment can further include the use of surgical treatments.

[00118] In certain embodiments, an effective dose of an engineered cell comprising a cell membrane coating as described herein can be administered to a patient once. In certain embodiments, an effective dose of a composition comprising an engineered cell comprising a cell membrane coating can be administered to a patient repeatedly. For systemic administration, subjects can be administered a therapeutic amount of a composition comprising a system, or cell comprising or encoding a system, such as, e.g. 0.1 mg/kg, 0.5 mg/kg, 1.0 mg/kg, 2.0 mg/kg, 2.5 mg/kg, 5 mg/kg, 10 mg/kg, 15 mg/kg, 20 mg/kg, 25 mg/kg, 30 mg/kg, 40 mg/kg, 50 mg/kg, or more.

[00119] In some embodiments, after an initial treatment regimen, the treatments can be administered on a less frequent basis. For example, after treatment biweekly for three months, treatment can be repeated once per month, for six months or a year or longer. Treatment according to the methods described herein can reduce levels of a marker or symptom of a condition, e.g. cancer cell growth by at least 10%, at least 15%, at least 20%, at least 25%, at least 30%, at least 40%, at least 50%, at least 60%, at least 70%, at least 80 % or at least 90% or more.

[00120] The dosage of a composition as described herein can be determined by a physician and adjusted, as necessary, to suit observed effects of the treatment. With respect to duration and frequency of treatment, it is typical for skilled clinicians to monitor subjects in order to determine when the treatment is providing therapeutic benefit, and to determine whether to increase or decrease dosage, increase or decrease administration frequency, discontinue treatment, resume treatment, or make other alterations to the treatment regimen. The dosing schedule can vary from once a week to daily depending on a number of clinical factors, such as the subject's sensitivity to the active ingredient. The desired dose or amount of activation can be administered at one time or divided into subdoses, e.g., 2-4 subdoses and administered over a period of time, e.g., at appropriate intervals through the day or other appropriate schedule. In some embodiments, administration can be chronic, e.g., one or more doses and/or treatments daily over a period of weeks or months. Examples of dosing and/or treatment schedules are administration daily, twice daily, three times daily or four or more times daily over a period of 1 week, 2 weeks, 3 weeks, 4 weeks, 1 month, 2 months, 3 months, 4 months, 5 months, or 6 months, or more. A composition comprising a system, or cell comprising or encoding a system can be administered over a period of time, such as over a 5 minute, 10 minute, 15 minute, 20 minute, or 25 minute period.

[00121] The dosage ranges for the administration an engineered cell comprising a cell membrane coating according to the methods described herein depend upon, for example, the form of an engineered cell comprising a cell membrane coating, its potency, and the extent to which symptoms, markers, or indicators of a condition described herein are desired to be reduced, for example the percentage reduction desired for cancer cell growth. The dosage should not be so large as to cause adverse side effects. Generally, the dosage will vary with the age, condition, and sex of the patient and can be determined by one of skill in the art. The dosage can also be adjusted by the individual physician in the event of any complication.

[00122] The efficacy of an engineered cell comprising a cell membrane coating in, e.g. the treatment of a condition described herein, or to induce a response as described herein (e.g. cancer) can be determined by the skilled clinician. However, a treatment is considered “effective treatment," as the term is used herein, if one or more of the signs or symptoms of a condition described herein are altered in a beneficial manner, other clinically accepted symptoms are improved, or even ameliorated, or a desired response is induced e.g., by at least 10% following treatment according to the methods described herein. Efficacy can be assessed, for example, by measuring a marker, indicator, symptom, and/or the incidence of a condition treated according to the methods described herein or any other measurable parameter appropriate, e.g. cancer cell growth. Efficacy can also be measured by a failure of an individual to worsen as assessed by hospitalization, or need for medical interventions (i.e., progression of the disease is halted). Methods of measuring these indicators are known to those of skill in the art and/or are described herein. Treatment includes any treatment of a disease in an individual or an animal (some non-limiting examples include a human or an animal) and includes: (1) inhibiting the disease, e.g., preventing a worsening of symptoms (e.g. pain or inflammation); or (2) relieving the severity of the disease, e.g., causing regression of symptoms. An effective amount for the treatment of a disease means that amount which, when administered to a subject in need thereof, is sufficient to result in effective treatment as that term is defined herein, for that disease. Efficacy of an agent can be determined by assessing physical indicators of a condition or desired response, (e.g. a reduction in cancer cell growth). It is well within the ability of one skilled in the art to monitor efficacy of administration and/or treatment by measuring any one of such parameters, or any combination of parameters. Efficacy can be assessed in animal models of a condition described herein, for example treatment of cancer. When using an experimental animal model, efficacy of treatment is evidenced when a statistically significant change in a marker is observed, e.g. cancer cell growth.

[00123] Another aspect of the technology described herein relates to kits for preparing an engineered cell comprising a cell membrane coating and/or treating a subject (e.g., a subject in need of cell therapy), among others. Described herein are kit components that can be included in one or more of the kits described herein.

[00124] In some embodiments, the kit comprises an engineered cell comprising a cell membrane coating. As will be appreciated by one of skill in the art, the cell can be supplied in a lyophilized form or a concentrated form that can diluted or suspended in liquid prior to use, e.g., with cells. Preferred formulations include those that are non-toxic to the cells and/or does not affect growth rate or viability and can be supplied in aliquots or in unit doses.

[00125] In some embodiments, the kit comprises microfluidic device as described herein.

[00126] In some embodiments, the components described herein can be provided singularly or in any combination as a kit. Such a kit includes the components described herein, e.g., an engineered cell, a microfluidic device, a source cell, cell membrane components, and/or a recipient cell.

[00127] In some embodiments, the compositions in the kit can be provided in a watertight or gas tight container which in some embodiments is substantially free of other components of the kit. For example, a composition can be supplied in more than one container, e.g., it can be supplied in a container having sufficient reagent for a predetermined number of cell culture events, e.g., 1, 2, 3 or greater. One or more components as described herein can be provided in any form, e.g., liquid, dried or lyophilized form. It is preferred that the components described herein are substantially pure and/or sterile. When the components described herein are provided in a liquid solution, the liquid solution preferably is an aqueous solution, with a sterile aqueous solution being preferred.

[00128] In addition, the kit optionally comprises informational material. The informational material can be descriptive, instructional, marketing or other material that relates to the methods described herein. The informational material of the kits is not limited in its form. In one embodiment, the informational material can include information about production of the polypeptide or system, concentration, date of expiration, batch or production site information, and so forth. In one embodiment, the informational material relates to methods for using or administering the components of the kit.

[00129] The kit can be provided with its various elements included in one package, e.g., a fiber- based, e.g., a cardboard, or polymeric, e.g., a Styrofoam box. The enclosure can be configured so as to maintain a temperature differential between the interior and the exterior, e.g., it can provide insulating properties to keep the reagents at a preselected temperature for a preselected time.

[00130] In one respect, the present invention relates to the herein described compositions, methods, and respective componcnt(s) thereof, as essential to the technology, yet open to the inclusion of unspecified elements, essential or not ("comprising). In some embodiments of any of the aspects, other elements to be included in the description of the composition, method or respective component thereof are limited to those that do not materially affect the basic and novel characteristic(s) of the technology (e.g., the composition, method, or respective component thereof “consists essentially of’ the elements described herein). This applies equally to steps within a described method as well as compositions and components therein. In other embodiments of any of the aspects, the compositions, methods, and respective components thereof, described herein are intended to be exclusive of any element not deemed an essential element to the component, composition or method (e.g., the composition, method, or respective component thereof “consists of’ the elements described herein). This applies equally to steps within a described method as well as compositions and components therein.

[00131] For convenience, the meaning of some terms and phrases used in the specification, examples, and appended claims, are provided below. Unless stated otherwise, or implicit from context, the following terms and phrases include the meanings provided below. The definitions are provided to aid in describing particular embodiments, and are not intended to limit the claimed invention, because the scope of the invention is limited only by the claims. Unless otherwise defined, all technical and scientific terms used herein have the same meaning as commonly understood by one of ordinary skill in the art to which this invention belongs. If there is an apparent discrepancy between the usage of a term in the art and its definition provided herein, the definition provided within the specification shall prevail.

[00132] For convenience, certain terms employed herein, in the specification, examples and appended claims are collected here.

[00133] The terms “decrease”, “reduced”, “reduction”, or “inhibit” are all used herein to mean a decrease by a statistically significant amount. In some embodiments, “reduce,” “reduction" or “decrease" or “inhibit” typically means a decrease by at least 10% as compared to a reference level (e.g. the absence of a given treatment or agent) and can include, for example, a decrease by at least about 10%, at least about 20%, at least about 25%, at least about 30%, at least about 35%, at least about 40%, at least about 45%, at least about 50%, at least about 55%, at least about 60%, at least about 65%, at least about 70%, at least about 75%, at least about 80%, at least about 85%, at least about 90%, at least about 95%, at least about 98%, at least about 99% , or more. As used herein,

“reduction” or “inhibition” does not encompass a complete inhibition or reduction as compared to a reference level. “Complete inhibition” is a 100% inhibition as compared to a reference level. A decrease can be preferably down to a level accepted as within the range of normal for an individual without a given disorder.

[00134] The terms “increased”, “increase”, “enhance”, or “activate” are all used herein to mean an increase by a statically significant amount. In some embodiments, the terms “increased”, “increase”, “enhance”, or “activate” can mean an increase of at least 10% as compared to a reference level, for example an increase of at least about 20%, or at least about 30%, or at least about 40%, or at least about 50%, or at least about 60%, or at least about 70%, or at least about 80%, or at least about 90% or up to and including a 100% increase or any increase between 10-100% as compared to a reference level, or at least about a 2-fold, or at least about a 3 -fold, or at least about a 4-fold, or at least about a 5-fold or at least about a 10-fold increase, or any increase between 2-fold and 10-fold or greater as compared to a reference level. In the context of a marker or symptom, an “increase” is a statistically significant increase in such level. [00135] In some embodiments of any of the aspects, a cell or biomolecule(s) “obtained from” a specified organism refers to a cell or biomolecule(s) isolated by physical or chemical means from that organism, and the progeny of that originally isolated cell which retain the characteristics of that cell. In some embodiments of any of the aspects, a cell or biomolecule(s) “derived from” a specified organism is descended from a cell or biomolecule(s) obtained from the specified organism but which as undergone changes ex vivo, e.g, genetic engineering, differentiation, chemical modification, or dedifferentiation. Accordingly, the cell or biomolecule(s) “derived” from a specified organism can be identified as having key genetic or phenotype characteristics of the source species but many have alterations or additions not found in the source species naturally.

[00136] As used herein, a "subject" means a human or animal. Usually the animal is a vertebrate such as a primate, rodent, domestic animal or game animal. Primates include chimpanzees, cynomologus monkeys, spider monkeys, and macaques, e.g., Rhesus. Rodents include mice, rats, woodchucks, ferrets, rabbits and hamsters. Domestic and game animals include cows, horses, pigs, deer, bison, buffalo, feline species, e.g., domestic cat, canine species, e.g., dog, fox, wolf, avian species, e.g., chicken, emu, ostrich, and fish, e.g., trout, catfish and salmon. In some embodiments, the subject is a mammal, e.g., a primate, e.g., a human. The terms, “individual,” “patient” and “subject” are used interchangeably herein.

[00137] Preferably, the subject is a mammal. The mammal can be a human, non-human primate, mouse, rat, dog, cat, horse, or cow, but is not limited to these examples. Mammals other than humans can be advantageously used as subjects that represent animal models of cancer. A subject can be male or female.

[00138] A subject can be one who has been previously diagnosed with or identified as suffering from or having a condition in need of treatment (e.g. cancer) or one or more complications related to such a condition, and optionally, have already undergone treatment for the condition or the one or more complications related to the condition Alternatively, a subject can also be one who has not been previously diagnosed as having a condition or one or more complications related to the condition.

For example, a subject can be one who exhibits one or more risk factors for the condition or one or more complications related to the condition or a subject who does not exhibit risk factors.

[00139] A “subject in need” of treatment for a particular condition can be a subject having that condition, diagnosed as having that condition, or at risk of developing that condition.

[00140] As used herein, the terms “protein" and “polypeptide" are used interchangeably herein to designate a series of amino acid residues, connected to each other by peptide bonds between the alpha-amino and carboxy groups of adjacent residues. The terms "protein", and "polypeptide" refer to a polymer of amino acids, including modified amino acids (e.g., phosphorylated, glycated, glycosylated, etc.) and amino acid analogs, regardless of its size or function. "Protein" and “polypeptide” are often used in reference to relatively large polypeptides, whereas the term "peptide" is often used in reference to small polypeptides, but usage of these terms in the art overlaps. The terms "protein" and "polypeptide" are used interchangeably herein when referring to a gene product and fragments thereof. Thus, exemplary polypeptides or proteins include gene products, naturally occurring proteins, homologs, orthologs, paralogs, fragments and other equivalents, variants, fragments, and analogs of the foregoing.

[00141] As used herein, the term “nucleic acid” or “nucleic acid sequence” refers to any molecule, preferably a polymeric molecule, incorporating units of ribonucleic acid, deoxyribonucleic acid or an analog thereof. The nucleic acid can be either single -stranded or double-stranded. A single -stranded nucleic acid can be one nucleic acid strand of a denatured double- stranded DNA. Alternatively, it can be a single-stranded nucleic acid not derived from any double -stranded DNA. In one aspect, the nucleic acid can be DNA. In another aspect, the nucleic acid can be RNA. Suitable DNA can include, e.g., genomic DNA or cDNA. Suitable RNA can include, e.g., mRNA.

[00142] In some embodiments of any of the aspects, a polypeptide, nucleic acid, or cell as described herein can be engineered. As used herein, “engineered" refers to the aspect of having been manipulated by the hand of man. For example, a polypeptide is considered to be “engineered" when at least one aspect of the polypeptide, e.g., its sequence, has been manipulated by the hand of man to differ from the aspect as it exists in nature. As is common practice and is understood by those in the art, progeny of an engineered cell are typically still referred to as “engineered" even though the actual manipulation was performed on a prior entity.

[00143] In some embodiments of any of the aspects, the engineered cell and/or the cell membrane coating described herein is exogenous. In some embodiments of any of the aspects, the engineered cell and/or the cell membrane coating described herein described herein is ectopic. In some embodiments of any of the aspects, the engineered cell and/or the cell membrane coating described herein described herein is not endogenous.

[00144] The term "exogenous" refers to a substance present in a cell other than its native source. The term "exogenous" when used herein can refer to a nucleic acid (e.g. a nucleic acid encoding a polypeptide) or a polypeptide that has been introduced by a process involving the hand of man into a biological system such as a cell or organism in which it is not normally found and one wishes to introduce the nucleic acid or polypeptide into such a cell or organism. Alternatively, “exogenous” can refer to a nucleic acid or a polypeptide that has been introduced by a process involving the hand of man into a biological system such as a cell or organism in which it is found in relatively low amounts and one wishes to increase the amount of the nucleic acid or polypeptide in the cell or organism, e.g., to create ectopic expression or levels. In contrast, the term "endogenous" refers to a substance that is native to the biological system or cell. As used herein, “ectopic” refers to a substance that is found in an unusual location and/or amount. An ectopic substance can be one that is normally found in a given cell, but at a much lower amount and/or at a different time. Ectopic also includes substance, such as a polypeptide or nucleic acid that is not naturally found or expressed in a given cell in its natural environment.

[00145] As used herein, the terms "treat,” "treatment," "treating,” or “amelioration” refer to therapeutic treatments, wherein the object is to reverse, alleviate, ameliorate, inhibit, slow down or stop the progression or severity of a condition associated with a disease or disorder, e.g. cancer. The term “treating" includes reducing or alleviating at least one adverse effect or symptom of a condition, disease or disorder associated with a cancer. Treatment is generally “effective" if one or more symptoms or clinical markers are reduced. Alternatively, treatment is “effective" if the progression of a disease is reduced or halted. That is, “treatment" includes not just the improvement of symptoms or markers, but also a cessation of, or at least slowing of, progress or worsening of symptoms compared to what would be expected in the absence of treatment. Beneficial or desired clinical results include, but are not limited to, alleviation of one or more symptom(s), diminishment of extent of disease, stabilized (/. e. , not worsening) state of disease, delay or slowing of disease progression, amelioration or palliation of the disease state, remission (whether partial or total), and/or decreased mortality, whether detectable or undetectable. The term "treatment" of a disease also includes providing relief from the symptoms or side-effects of the disease (including palliative treatment).

[00146] In some embodiments of any of the aspects, described herein is a prophylactic method of treatment. As used herein “prophylactic” refers to the timing and intent of a treatment relative to a disease or symptom, that is, the treatment is administered prior to clinical detection or diagnosis of that particular disease or symptom in order to protect the patient from the disease or symptom. Prophylactic treatment can encompass a reduction in the severity or speed of onset of the disease or symptom, or contribute to faster recovery from the disease or symptom. Accordingly, the methods described herein can be prophylactic relative to CRS. In some embodiments of any of the aspects, prophylactic treatment is not prevention of all symptoms or signs of a disease.

[00147] As used herein, the term “pharmaceutical composition” refers to the active agent in combination with a pharmaceutically acceptable carrier e.g. a carrier commonly used in the pharmaceutical industry. The phrase "pharmaceutically acceptable" is employed herein to refer to those compounds, materials, compositions, and/or dosage forms which are, within the scope of sound medical judgment, suitable for use in contact with the tissues of human beings and animals without excessive toxicity, irritation, allergic response, or other problem or complication, commensurate with a reasonable benefit/risk ratio. In some embodiments of any of the aspects, a pharmaceutically acceptable carrier can be a carrier other than water. In some embodiments of any of the aspects, a pharmaceutically acceptable carrier can be a cream, emulsion, gel, liposome, nanoparticle, and/or ointment. In some embodiments of any of the aspects, a pharmaceutically acceptable carrier can be an artificial or engineered carrier, e.g., a carrier that the active ingredient would not be found to occur in in nature. [00148] As used herein, the term "administering," refers to the placement of a compound as disclosed herein into a subject by a method or route which results in at least partial delivery of the agent at a desired site. Pharmaceutical compositions comprising the compounds disclosed herein can be administered by any appropriate route which results in an effective treatment in the subject. In some embodiments, administration comprises physical human activity, e.g., an injection, act of ingestion, an act of application, and/or manipulation of a delivery device or machine. Such activity can be performed, e.g., by a medical professional and/or the subject being treated.

[00149] As used herein, “contacting" refers to any suitable means for delivering, or exposing, an agent to at least one cell. Exemplary delivery methods include, but are not limited to, direct delivery to cell culture medium, perfusion, injection, or other delivery method well known to one skilled in the art. In some embodiments, contacting comprises physical human activity, e.g., an injection; an act of dispensing, mixing, and/or decanting; and/or manipulation of a delivery device or machine.

[00150] The term “statistically significant" or “significantly" refers to statistical significance and generally means a two standard deviation (2SD) or greater difference.

[00151] Other than in the operating examples, or where otherwise indicated, all numbers expressing quantities of ingredients or reaction conditions used herein should be understood as modified in all instances by the term “about.” The term “about” when used in connection with percentages can mean ±1%.

[00152] As used herein, the term “comprising” means that other elements can also be present in addition to the defined elements presented. The use of “comprising” indicates inclusion rather than limitation.

[00153] The term "consisting of' refers to compositions, methods, and respective components thereof as described herein, which are exclusive of any element not recited in that description of the embodiment.

[00154] As used herein the term "consisting essentially of' refers to those elements required for a given embodiment. The term permits the presence of additional elements that do not materially affect the basic and novel or functional characteristic(s) of that embodiment of the invention.

[00155] As used herein, the term “specific binding” refers to a chemical interaction between two molecules, compounds, cells and/or particles wherein the first entity binds to the second, target entity with greater specificity and affinity than it binds to a third entity which is a non-target. In some embodiments, specific binding can refer to an affinity of the first entity for the second target entity which is at least 10 times, at least 50 times, at least 100 times, at least 500 times, at least 1000 times or greater than the affinity for the third nontarget entity. A reagent specific for a given target is one that exhibits specific binding for that target under the conditions of the assay being utilized.

[00156] The singular terms "a," "an," and "the" include plural referents unless context clearly indicates otherwise. Similarly, the word "or" is intended to include "and" unless the context clearly indicates otherwise. Although methods and materials similar or equivalent to those described herein can be used in the practice or testing of this disclosure, suitable methods and materials are described below. The abbreviation, "e.g." is derived from the Latin exempli gratia, and is used herein to indicate a non-limiting example. Thus, the abbreviation "e.g." is synonymous with the term "for example." [00157] Groupings of alternative elements or embodiments of the invention disclosed herein are not to be construed as limitations. Each group member can be referred to and claimed individually or in any combination with other members of the group or other elements found herein. One or more members of a group can be included in, or deleted from, a group for reasons of convenience and/or patentability. When any such inclusion or deletion occurs, the specification is herein deemed to contain the group as modified thus fulfilling the written description of all Markush groups used in the appended claims.

[00158] Unless otherwise defined herein, scientific and technical terms used in connection with the present application shall have the meanings that are commonly understood by those of ordinary skill in the art to which this disclosure belongs. It should be understood that this invention is not limited to the particular methodology, protocols, and reagents, etc., described herein and as such can vary. The terminology used herein is for the purpose of describing particular embodiments only, and is not intended to limit the scope of the present invention, which is defined solely by the claims. Definitions of common terms in immunology and molecular biology can be found in The Merck Manual of Diagnosis and Therapy, 20th Edition, published by Merck Sharp & Dohme Corp., 2018 (ISBN 0911910190, 978-0911910421); Robert S. Porter et al. (eds.), The Encyclopedia of Molecular Cell Biology and Molecular Medicine, published by Blackwell Science Ltd., 1999-2012 (ISBN 9783527600908); and Robert A. Meyers (ed.), Molecular Biology and Biotechnology: a Comprehensive Desk Reference, published by VCH Publishers, Inc., 1995 (ISBN 1-56081-569-8); Immunology by Wemer Luttmann, published by Elsevier, 2006; Janeway's Immunobiology, Kenneth Murphy, Allan Mowat, Casey Weaver (eds.), W. W. Norton & Company, 2016 (ISBN 0815345054, 978-0815345053); Lewin's Genes XI, published by Jones & Bartlett Publishers, 2014 (ISBN- 1449659055); Michael Richard Green and Joseph Sambrook, Molecular Cloning: A Laboratory Manual, 4th ed., Cold Spring Harbor Laboratory Press, Cold Spring Harbor, N.Y., USA (2012) (ISBN 1936113414); Davis et al., Basic Methods in Molecular Biology, Elsevier Science Publishing, Inc., New York, USA (2012) (ISBN 044460149X); Laboratory Methods in Enzymology: DNA, Jon Lorsch (ed.) Elsevier, 2013 (ISBN 0124199542); Current Protocols in Molecular Biology (CPMB), Frederick M. Ausubel (ed.), John Wiley and Sons, 2014 (ISBN 047150338X, 9780471503385), Current Protocols in Protein Science (CPPS), John E. Coligan (ed.), John Wiley and Sons, Inc., 2005; and Current Protocols in Immunology (CPI) (John E. Coligan, ADA M Kruisbeek, David H Margulies, Ethan M Shevach, Warren Strobe, (eds.) John Wiley and Sons, Inc., 2003 (ISBN 0471142735, 9780471142737), the contents of which are all incorporated by reference herein in their entireties. [00159] One of skill in the art can readily identify a chemotherapeutic agent of use (e.g. see Physicians' Cancer Chemotherapy Drug Manual 2014, Edward Chu, Vincent T. DeVita Jr., Jones & Bartlett Learning; Principles of Cancer Therapy, Chapter 85 in Harrison's Principles of Internal Medicine, 18th edition; Therapeutic Targeting of Cancer Cells: Era of Molecularly Targeted Agents and Cancer Pharmacology, Chs. 28-29 in Abeloff s Clinical Oncology, 2013 Elsevier; and Fischer D S (ed): The Cancer Chemotherapy Handbook, 4th ed. St. Louis, Mosby-Year Book, 2003).

[00160] In some embodiments of any of the aspects, the disclosure described herein does not concern a process for cloning human beings, processes for modifying the germ line genetic identity of human beings, uses of human embryos for industrial or commercial purposes or processes for modifying the genetic identity of animals which are likely to cause them suffering without any substantial medical benefit to man or animal, and also animals resulting from such processes.

[00161] In all embodiments where a sample is obtained or has been obtained or provided, the sample can be sample taken, obtained, or provided via minimally invasive methods and/or involves only a minor intervention. In some embodiments of any of the aspects, a sample is taken, obtained, or provided by one or more of a blood draw or prick, an epidermal or mucus membrane swab, buccal sampling, saliva sample, a epidermal skin sampling technique, and/or collection of a secreted or expelled bodily fluid (e.g., mucus, urine, sweat, etc), fecal sampling, semen/seminal fluid sampling, or clippings (e.g., of hair or nails). In some emodiments of any of the aspects, the sample comprises, consists of, or consists essentially of blood (or any fraction or component thereof), serum, urine, mucus, epithelial cells, saliva, buccal cells, a secreted or expelled bodily fluid, and/or hair or nail clippings.

[00162] Other terms are defined herein within the description of the various aspects of the invention.

[00163] All patents and other publications; including literature references, issued patents, published patent applications, and co-pending patent applications; cited throughout this application are expressly incorporated herein by reference for the purpose of describing and disclosing, for example, the methodologies described in such publications that might be used in connection with the technology described herein. These publications are provided solely for their disclosure prior to the filing date of the present application. Nothing in this regard should be construed as an admission that the inventors are not entitled to antedate such disclosure by virtue of prior invention or for any other reason. All statements as to the date or representation as to the contents of these documents is based on the information available to the applicants and does not constitute any admission as to the correctness of the dates or contents of these documents.

[00164] The description of embodiments of the disclosure is not intended to be exhaustive or to limit the disclosure to the precise form disclosed. While specific embodiments of, and examples for, the disclosure are described herein for illustrative purposes, various equivalent modifications are possible within the scope of the disclosure, as those skilled in the relevant art will recognize. For example, while method steps or functions are presented in a given order, alternative embodiments may perform functions in a different order, or functions may be performed substantially concurrently. The teachings of the disclosure provided herein can be applied to other procedures or methods as appropriate. The various embodiments described herein can be combined to provide further embodiments. Aspects of the disclosure can be modified, if necessary, to employ the compositions, functions and concepts of the above references and application to provide yet further embodiments of the disclosure. Moreover, due to biological functional equivalency considerations, some changes can be made in protein structure without affecting the biological or chemical action in kind or amount. These and other changes can be made to the disclosure in light of the detailed description. All such modifications are intended to be included within the scope of the appended claims.

[00165] Specific elements of any of the foregoing embodiments can be combined or substituted for elements in other embodiments. Furthermore, while advantages associated with certain embodiments of the disclosure have been described in the context of these embodiments, other embodiments may also exhibit such advantages, and not all embodiments need necessarily exhibit such advantages to fall within the scope of the disclosure.

[00166] In some embodiments, the present technology may be defined in any of the following numbered paragraphs:

1. An engineered cell comprising a cell membrane coating on the surface of the cell, the cell membrane coating comprising cell membrane components of one or more source cells.

2. The engineered cell of any of the preceding paragraphs, wherein the cell membrane coating comprises cell membrane components of at least two source cells.

3. The engineered cell of any of the preceding paragraphs, wherein the cell membrane coating comprises cell membrane components of at least three source cells.

4. The engineered cell of any of the preceding paragraphs, wherein the at least two source cells are different types of cells.

5. The engineered cell of any of the preceding paragraphs, wherein the one or more source cells comprise a T cell (e.g., a CAR-T cell), a diseased cell, and/or a macrophage (e.g., a THP-1 macrophage).

6. The engineered cell of paragraph 5, wherein the diseased cell is a cancer cell, leukemia cell, or an acute myeloid leukemia (AML) cell.

7. The engineered cell of any of the preceding paragraphs, wherein the cell membrane components include one or more of: a lipid, a lipid bilayer, transmembrane proteins, membrane-displayed biomolecules, and cell surface proteins. 8. The engineered cell of any of the preceding paragraphs, wherein the cell membrane coating comprises a mixture, fusion, or hybridization of cell membrane components from two or more source cells.

9. The engineered cell of paragraph 8, wherein the mixture, fusion, or hybridization of cell membrane components has an equal amount of cell membrane components from each source cell, e.g., as measured by protein concentrations.

10. The engineered cell of any of the preceding paragraphs, wherein the cell membrane components obtained from each source cell further comprises a detectable label.

11. The engineered cell of any of the preceding paragraphs, wherein the cell membrane components obtained from each source cell each further comprise a detectable label, wherein each detectable label is distinguishable from the detectable labels comprised by the cell membrane components obtained from the other source cells.

12. The engineered cell of any of the preceding paragraphs, wherein the cell membrane coating further comprises a detectable label.

13. The engineered cell of any of the preceding paragraphs, wherein the engineered cell further comprises a detectable label.

14. The engineered cell of any of the preceding paragraphs, wherein the engineered cell is a T cell or CD8+ T cell.

15. The engineered cell of any of the preceding paragraphs, wherein the engineered cell is a CAR-T cell.

16. The engineered cell of any of the preceding paragraphs, wherein the CAR-T cell comprises a CAR specific for or active against the diseased cell.

17. The engineered cell of any of the preceding paragraphs, wherein the CAR is an anti-CD 123 CAR.

18. The engineered cell of any of the preceding paragraphs, wherein the one or more source cells and/or the engineered cell are obtained from a subject.

19. The engineered cell of any of the preceding paragraphs, wherein the engineered cell is not autologous to a subject.

20. The engineered cell of any of the preceding paragraphs, wherein the engineered cell is autologous to a subject.

21. A method of producing an engineered cell comprising a cell membrane coating, the method comprising: a. Obtaining cell membrane components from one or more source cells; b. Optionally, mixing or fusing the cell membrane components of two or more source cells; and c. Contacting a recipient cell with the cell membrane components to obtain and engineered cell comprising a cell membrane coating. 22. The method of paragraph 21, wherein the cell membrane components are obtained by freeze-thaw of the source cells.

23. The method of any of paragraphs 21-22, wherein the mixing or fusing of the cell membrane components comprises sonication and/or fluid flow.

24. The method of any of paragraphs 21-23, wherein the method, or at least steps b) and c), is conducted in a microfluidic device.

25. A microfluididic device comprising, in the following order along a flow path(s): a. One or more initial input ports; b. A flow channel for mixing materials provided to the one or more initial input ports; c. One or more recipient cell input ports; d. A flow channel comprising sonication input, or capable of receiving sonication input; and e. One or more output ports.

26. A method of treating a subject, the method comprising administering an engineered cell of any of paragraphs 1-20 to the subject.

27. The method of paragraph 26, wherein the subject has or is in need of treatment for cancer.

28. The method of paragraph 27, wherein the cancer is a hematological cancer or hematopoietic cancer.

29. The method of paragraph 27, wherein the cancer is leukemia.

30. The method of paragraph 27, wherein the cancer is acute myeloid leukemia.

31. The method of any of paragraphs 26-30, wherein the subject is at risk of cytokine release syndrome (CRS).

[00167] In some embodiments, the present technology may be defined in any of the following numbered paragraphs:

4. The engineered cell of any of paragraphs 2-3, wherein the at least two source cells are different types of cells.

5. The engineered cell of any of the preceding paragraphs, wherein the one or more source cells comprise a T cell, a diseased cell, and/or a macrophage.

6. The engineered cell of paragraph 5, wherein the T cell is a CAR-T cell.

7. The engineered cell of paragraph 5, wherein the diseased cell is a cancer cell. The engineered cell of paragraph 7, wherein the cancer cell is a leukemia cell or an acute myeloid leukemia (AML) cell. The engineered cell of paragraph 5, wherein the macrophage is a THP-1 macrophage. The engineered cell of any of the preceding paragraphs, wherein the cell membrane components include one or more of: a lipid; a lipid bilayer; transmembrane proteins; membrane-displayed biomolecules; and cell surface proteins. The engineered cell of any of the preceding paragraphs, wherein the cell membrane coating comprises a mixture, fusion, or hybridization of cell membrane components from two or more source cells. The engineered cell of paragraph 11, wherein the mixture, fusion, or hybridization of cell membrane components has an equal amount of cell membrane components from each source cell. The engineered cell of paragraph 12, wherein the amount of cell membrane components is the amount as measured by protein concentrations. The engineered cell of any of the preceding paragraphs, wherein the cell membrane components obtained from each source cell further comprises a detectable label. The engineered cell of any of the preceding paragraphs, wherein the cell membrane components obtained from each source cell each further comprise a detectable label, wherein each detectable label is distinguishable from the detectable labels comprised by the cell membrane components obtained from the other source cells. The engineered cell of any of the preceding paragraphs, wherein the cell membrane coating further comprises a detectable label. The engineered cell of any of the preceding paragraphs, wherein the engineered cell further comprises a detectable label. The engineered cell of any of the preceding paragraphs, wherein the engineered cell is a T cell. The engineered cell of paragraph 18, wherein the T cell is a CD8+ T cell. The engineered cell of paragraph 18, wherein the T cell is a CAR-T cell. The engineered cell of paragraph 20, wherein the CAR-T cell comprises a CAR specific for or active against the diseased cell. The engineered cell of paragraph 21, wherein the CAR is an anti-CD123 CAR. The engineered cell of any of the preceding paragraphs, wherein the one or more source cells and/or the engineered cell are obtained from a subject. The engineered cell of any of the preceding paragraphs, wherein the engineered cell is not autologous to a subject. 25. The engineered cell of any of the preceding paragraphs, wherein the engineered cell is autologous to a subject.

26. A method of producing an engineered cell comprising a cell membrane coating, the method comprising: a) obtaining cell membrane components from one or more source cells; b) optionally, mixing or fusing the cell membrane components of two or more source cells; and c) contacting a recipient cell with the cell membrane components to obtain an engineered cell comprising a cell membrane coating.

27. The method of paragraph 26, wherein the cell membrane components are obtained by freeze- thaw of the source cells.

28. The method of any of paragraphs 26-27, wherein the mixing or fusing of the cell membrane components comprises sonication and/or fluid flow.

29. The method of any of paragraphs 26-28, wherein the method, or at least steps b) and c), is conducted in a microfluidic device.

30. A microfluididic device comprising, in the following order along a flow path(s): a) one or more initial input ports; b) a flow channel for mixing materials provided to the one or more initial input ports; c) one or more recipient cell input ports; d) a flow channel comprising sonication input, or capable of receiving sonication input; and e) one or more output ports.

31. A method of treating a subject, the method comprising administering an engineered cell of any of paragraphs 1-25 to the subject.

32. The method of paragraph 31, wherein the subject has or is in need of treatment for cancer.

33. The method of paragraph 32, wherein the cancer is a hematological cancer or hematopoietic cancer.

34. The method of paragraph 32, wherein the cancer is leukemia.

35. The method of paragraph 32, wherein the cancer is acute myeloid leukemia.

36. The method of any of paragraphs 31-35, wherein the subject is at risk of cytokine release syndrome (CRS).

[00168] The technology described herein is further illustrated by the following examples which in no way should be construed as being further limiting.

EXAMPLES

Example 1: High-Throughput Tailorable Cell Membrane Hybridization and Camouflaging for

Personalized “One-for-All” Acute Myeloid Leukemia Theranostics [00169] Rationale and Hypothesis

[00170] Acute myeloid leukemia (AML) is one of the most common types of cancers of the bone marrow and blood in adults, accounting for about one-third of new leukemia cases every year.¹ As treatments for AML are largely unchanged for the last four decades, AML remains a disease with poor prognosis, with a five-year survival rate of around 25%.^{1 3} In recent years, many emerging AML therapeutic approaches have been actively explored, including the chimeric antigen receptor (CAR) T cell mmunotherapy.⁴’⁵ CAR T cell immunotherapy has shown some efficacy in treating hematological malignancies, particularly B cell malignancies, in which the malignant cells express specific targetable antigens. However, the same CAR T cell immunotherapy has not been as effective in combating AML, where most of the surface antigens targetable in AML are also expressed by the healthy hematopoietic stem and progenitor cells. As such, targeting these myeloid antigens on AML cells may also eradicate the normal cells, leading to intolerable myeloablation, infections, and bleeding complications. In addition, the use of CAR T cell immunotherapy in treating hematological malignancies may induce adverse side effects, most notably the potentially life-threatening cytokine release syndrome (CRS).^6,7 CRS is characterized by an abnormal elevation of serum cytokines, such as interferon-g (IFNy), interleukin-6 (IL-6), IL-8, and IL-10, and elevated inflammatory responses following CAR T cell infusion. To date, the collective issues of a lack of AML-specific target antigens and the occurrence of CRS remain unresolved and have considerably limited the application of CAR T cell immunotherapy in AML. Furthermore, these challenges have prevented effective real time monitoring of immunotherapeutic efficacy and tracking of residual leukemic cells, which are crucial to suppress AML relapse and improve patient outcome.

[00171] Recent years have seen increasing efforts in leveraging nanomedicine to augment cancer immunotherapy, especially in enhancing tumor targeting and reducing off-target toxicity.^8,9 Additionally, nanomedicine-mediated phototherapies, such as photothermal and photodynamic therapies, has been increasingly combined with immunotherapy to realize synergistic cancer treatments with improved outcome.^10,11 Conventional nanomedicine, however, suffers from various shortcomings which impede its clinical translation.^12,13 These include laborious and time-consuming formulation process, low product throughput, substantial batch-to-batch variations, inefficient tumor localization, non-specific organ distribution, and fast clearance from the body.

[00172] To overcome the fundamental limitations of CAR T cell immunotherapy for AML treatment and conventional nanomedicine, described herein is the use of two emerging nanomedicine formulation approaches: cell membrane coating and microfluidic technologies. Unlike existing formulation methods that rely primarily on synthetic materials and complicated chemical processing, the cell membrane coating technique, which so far has been used to modify nanoparticles^14,15 and bacteria,¹⁶ relies on natural materials and simple processing steps. Based on this strategy, theranostic agents are coated with natural cellular components, which endow these agents with the biological properties and functionalities similar to the source cells from which the membrane is derived, such as homotypic targeting, cytokine binding and isolation, immune evasion, and extended in vivo residence time. Moreover, cell membrane coating technology enables the use of cellular materials derived from patients, essentially personalizing patient treatments and minimizing excessive immune response. Meanwhile, microfluidic platforms provide a highly controlled approach to process cell membranes and construct membrane-coated theranostic agents, significantly simplifying the formulation process, shortening the formulation time, and increasing product volume, quality, and reproducibility. Therefore, it is contemplated herein that the efficacy and safety of AML therapy can be significantly improved by adopting cell membrane coating and microfluidic technologies to formulate personalized cellular theranostic agents capable of binding specifically to AML cells, destroying these malignant cells, and removing excessive serum cytokines from the body.

[00173] Objectives and Methodology

[00174] Described herein is the construction of a high-throughput microfluidic platform which permits the hybridization of, e.g., three different patient-derived cell membranes and the coating of the hybridized membrane onto cytotoxic T cells, to generate multifunctional personalized cellular agents for combinatorial AML theranostics. The “one-for-all” cellular agents have several novel features: 1) hybridized membrane from CAR T cell membrane (for homing to AML cells), AML cell membrane (for highly specific homotypic binding with AML cells, essentially eliminating the need for a specific target antigen and minimizing off-target effect), and macrophage membrane (for cytokine absorption, improved tumor accumulation, and prolonged in vivo circulation lifetime) to realize triple-targeting capability, and 2) T cells camouflaged within the hybridized cell membrane, which will be labeled with a probe for realizing combinatorial immuno- and phototherapies, visualizing treatment response, and locating residual and metastatic leukemic cells. The work described herein is illustrated in Figs. 1A-1C.

[00175] Fabrication of microfluidic devices and formulation of hybridized membrane-coated cellular theranostic agents.

[00176] Design and fabrication of microfluidic devices. Microfluidic devices for membrane hybridization and cellular agent synthesis will be fabricated using soft lithography. Flow rates within the microfluidic devices will be simulated and optimized in COMSOL to realize a uniform mixing and fusion of different membranes.

[00177] Synthesis of triple membrane-coated cellular agents. CD8+ T cells will be isolated from the peripheral blood mononuclear cells. CAR T cells targeting CD 123+ AML cells, which are currently being actively explored for phase I/II clinical trials, will be generated through lentiviral transduction and characterized for their surface expression, specificity, and cytotoxicity. The membrane of these CAR T cells, along with those of KGla AML cells and THP-1 macrophages, will be obtained through a freeze-thaw method. Each membrane will be labeled with a distinct fluorescent dye (i.e., DiD, Dil, and DiO) for downstream characterization. The different cell membranes at a ratio of 1 : 1 : 1 (based on membrane protein concentrations) will be mixed and fused within the microfluidic device at optimized flow rates under sonication to produce hybridized membrane, which will then be used to coat the CD8+ T cells within the microfluidic device.

[00178] Characterization of the physical properties of the cellular agents. Transmission electron microscopy will be used to confirm membrane coating on T cells and estimate the size of the cellular agents. The surface charge of the cellular agents will be characterized using a zeta potential analyzer. The hybridization of the three cell membranes will be verified through the colocalization of their respective fluorescent dyes and proteomic analysis. The protein expression of the hybridized membrane and membrane-coated T cells will be compared using SDS PAGE and western blot. The viability of the cellular agents over time will be assessed through cell proliferation assay. The agent stability in water, saline, and serum over time will be examined.

[00179] In vitro validation of the performance of cellular agents using cells.

[00180] Evaluation of the cellular targeting, biocompatibility, and therapeutic efficacy of the cellular agents. Subsequent to physical property characterization, the theranostic agents will be synthesized to incorporate indocyanine green (ICG) dye, in place of DiD, Dil, and DiO dyes, within the hybridized membrane. The FDA-approved ICG will serve as a near-infrared (NIR) fluorescence imaging probe, photothermal agent, and photosensitizer. The inclusion of ICG is expected to confer the cellular agents combinatorial theranostic modalities. The efficiency of photothermal conversion and reactive oxygen species generation of the cellular agents over time will be characterized under NIR laser excitation. The specific targeting of the cellular agents will be confirmed by using confocal fluorescence microscopy to image agent co-localization with different fluorescently-labeled co cultured cells, particularly KG la AML cells, LCL lymphoblastoid cells, BMEC-1 bone marrow endothelial cells, and THP-1 macrophages. The biocompatibility and efficacy of immuno-, photothermal, and photodynamic therapies of the cellular agents will be evaluated based on cellular morphological changes, cell proliferation assay, and apoptosis/necrosis assay, in the absence (as a control) and presence of the cellular agents and NIR laser excitation. The secretion of IFNy and TNFa after cell co-cultures will be characterized using flow cytometry to investigate the activation of effector pathways against AML cells.

[00181] Assessment of the cytokine binding of the cellular agents in vitro. Cytokine binding will be examined by incubating the cellular agents with various cytokines typically elevated in patient serum following CAR T cell administration (i.e., IFNy, IL-6, IL-8, and IL-10) over time. Each agent- cytokine mixture will then be centrifuged and the cytokine concentration in the supernatant will be quantified using ELISA. The bound cytokines will be estimated from the difference between the initial cytokine concentration and the supernatant concentration. Cytokine solutions in the absence of cellular agents will be used as a control.

[00182] In vivo validation of the performance of personalized cellular agents using AML mouse model.

[00183] Evaluation of the therapeutic efficacy, circulation lifetime, hiodistrihution, and toxicity of the cellular agents. A xenogeneic mouse model of systemic AML will first be established using a KG la leukemic cell line (KGla-ffLuc-eGFP cells) and confirmed through the detection of leukemia in the peripheral blood after several days. Cellular agents will be injected intravenously into AML mice (n = 3) and healthy mice (control, n = 3), in the absence and presence of NIR light excitation.

For control, the AML mice (n = 3) will be treated with saline. The tumor progression will be tracked using bioluminescence and fluorescence imaging and flow cytometry of peripheral blood at various time points. The circulation lifetime of the cellular agents will be estimated from the fluorescence measurement of the blood collected at different time points post agent injection. The biodistribution of the cellular agents will be evaluated in vivo through fluorescence imaging and ex vivo from the fluorescence of the tissues of the main organs (i.e., heart, lung, liver, spleen, and kidney) of the AML and control mice sacrificed 48 h post agent injection. The toxicity of the cellular agents will be assessed through histopathologic examination of the tissues stained with hematoxylin and eosin. [00184] Characterization of the cytokine binding of the cellular agents in vivo. Cellular agents will be administered intravenously into AML mice (n = 3) and healthy mice (control, n = 3). The AML mice (n = 3) will also be injected with saline as control. Blood samples will be collected from the mice at predetermined time points and the plasma will be isolated through centrifugation. Cytokines (i.e., IFNy, IL-6, IL-8, and IL-10) in the plasma will then be quantified by ELISA.

[00185] Synthesis and characterization of personalized cellular agents. Personalized cellular agents will first be prepared using AML cells, T cells, and macrophages extracted from the AML mice. Following physical property characterization, subsequent study using the personalized cellular agents will be performed similarly to what is described above. To investigate the potential of the personalized cellular agents in mitigating AML recurrence and metastasis, the treated AML mice will be re-challenged with AML cells, followed by the intravenous injection of cellular agents and evaluation of their antileukemic activity. All statistical analysis in this proposal will be performed using student’s t-test.

[00186] Discussion

[00187] The novelty of the methods and compositions described herein includes: 1) tailorable and high-throughput hybridization of three different types of cell membrane through microfluidics, 2) engineering of immune cells through membrane camouflaging, and 3) simultaneous enhanced AML targeting, minimized CRS toxicity, and AML combinatorial theranostics within a single platform. These features have not been previously demonstrated. It is expected that the establishment of the current platform, which relies entirely on patient-derived and FDA-approved materials, will catalyze a clinically and commercially translatable technology to address the huge unmet need in AML immunotherapy to improve patient treatment response and survival. Furthermore, due to its customizability and controllability, the present approach can be easily adopted for a wide range of cell types to improve the immunotherapy and combinatorial therapies of other liquid and solid malignancies.

[00188] References

1. Cancer Facts & Figures 2020. American Cancer Society.

2. Yang X, Wang J (2018). Precision Therapy for Acute Myeloid Leukemia. Journal of Hematology & Oncology 11:3.

3. Lai C, Doucette K, Norsworthy K (2019). Recent Drug Approvals for Acute Myeloid Leukemia. Journal of Hematology & Oncology 12:100.

4. Cummins KD, Gill S (2019). Chimeric Antigen Receptor T-Cell Therapy for Acute Myeloid Leukemia: How Close to Reality? Haematologica 104(7): 1302-1308.

5. Mardiana S, Gill S (2020). CAR T Cells for Acute Myeloid Leukemia: State of the Art and Future Directions. Frontiers in Oncology 10:697.

6. Bonifant CL, Jackson HJ, Brentjens RJ, Curran KJ (2016). Toxicity and Management in CAR T- Cell Therapy. Molecular Therapy Oncolytics 3:16011.

7. Wei J, Liu Y, Wang C, Zhang Y, Tong C, Dai G, Wang W, Rasko JEJ, Melenhorst J, Qian W, Liang A, Han W (2020). The Model of Cytokine Release Syndrome in CAR T-Cell Treatment for B- Cell Non-Hodgkin Lymphoma. Signal Transduction and Targeted Therapy 5:134.

8. Shi Y, Lammers T (2019). Combining Nanomedicine and Immunotherapy. Accounts of Chemical Research 52(6): 1543-1554.

9. Irvine DJ, Dane EL (2020). Enhancing Cancer Immunotherapy with Nanomedicine. Nature Reviews Immunology 20:321-334.

10. Ng CW, Li J, Pu K (2018). Recent Progresses in Phototherapy-Synergized Cancer Immunotherapy. Advanced Functional Materials 28(46): 1804688.

11. Zhou S, Li D, Lee C, Xie J (2020). Nanoparticle Phototherapy in the Era of Cancer Immunotherapy. Trends in Chemistry 2(12): 1082-1095.

12. Wilhelm S, Tavares AJ, Dai Q, Ohta S, Audet J, Dvorak HF, Chan WCW (2016). Analysis of Nanoparticle Delivery to Tumours. Nature Reviews Materials 1:16014.

13. Meng H, Leong W, Leong KW, Chen C, Zhao Y (2018). Walking the Line: The Fate of Nanomaterials at Biological Barriers. Biomaterials 174:41-53.

14. Fang RH, Kroll AV, Gao W, Zhang L (2018). Cell Membrane Coating Nanotechnology.

Advanced Materials 30(23): 1706759. 15. Zhen X, Cheng P, Pu K (2019). Recent Advances in Cell Membrane -Camouflaged Nanoparticles for Cancer Phototherapy. Small 15(1): 1804105.

16. Cao Z, Cheng S, Wang X, Pang Y, Liu J (2019). Camouflaging Bacteria by Wrapping with Cell Membranes. Nature Communications 10:3452.

EXAMPLE 2: Microfluidic-Mediated Formulation of Cell Membrane-Coated Nanostructures

[00189] Gold (Au) nanostructures (nanonuggets (NN) or nanostars (NS)) were coated with macrophage membrane vesiscles via microfluidic-mediated physical mixing (Figs. 4A-4D). The size, surface charge, protein expression, hydrodynamic size, and zeta potential of the coated nanostructures were analyzed (Figs. 5-9). The coated nanostructures were taken up by cancer cells (Fig. 10A-10C) and demonstrated increased immune evasion abilities (Figs. 1 lA-11C).

[00190] Gold (Au) nanostructures (nanonuggets (NN) or nanostars (NS)) were coated with cancer cell membrane vesiscles via microfluidic -mediated physical mixing (Figs. 12A-12D). The size, surface charge, protein expression, hydrodynamic size, and zeta potential of the coated nanostructures were analyzed (Figs. 13-17).

Claims

CLAIMS What is claimed herein is:

2. The engineered cell of any of the preceding claims, wherein the cell membrane coating comprises cell membrane components of at least two source cells.

3. The engineered cell of any of the preceding claims, wherein the cell membrane coating comprises cell membrane components of at least three source cells.

4. The engineered cell of any of claims 2-3, wherein the at least two source cells are different types of cells.

5. The engineered cell of any of the preceding claims, wherein the one or more source cells comprise a T cell, a diseased cell, and/or a macrophage.

6. The engineered cell of claim 5, wherein the T cell is a CAR-T cell.

7. The engineered cell of claim 5, wherein the diseased cell is a cancer cell.

8. The engineered cell of claim 7, wherein the cancer cell is a leukemia cell or an acute myeloid leukemia (AML) cell.

9. The engineered cell of claim 5, wherein the macrophage is a THP-1 macrophage.

10. The engineered cell of any of the preceding claims, wherein the cell membrane components include one or more of: a lipid; a lipid bilayer; transmembrane proteins; membrane-displayed biomolecules; and cell surface proteins.

11. The engineered cell of any of the preceding claims, wherein the cell membrane coating comprises a mixture, fusion, or hybridization of cell membrane components from two or more source cells.

12. The engineered cell of claim 11, wherein the mixture, fusion, or hybridization of cell membrane components has an equal amount of cell membrane components from each source cell.

13. The engineered cell of claim 12, wherein the amount of cell membrane components is the amount as measured by protein concentrations.

14. The engineered cell of any of the preceding claims, wherein the cell membrane components obtained from each source cell further comprises a detectable label.

15. The engineered cell of any of the preceding claims, wherein the cell membrane components obtained from each source cell each further comprise a detectable label, wherein each detectable label is distinguishable from the detectable labels comprised by the cell membrane components obtained from the other source cells.

16. The engineered cell of any of the preceding claims, wherein the cell membrane coating further comprises a detectable label.

17. The engineered cell of any of the preceding claims, wherein the engineered cell further comprises a detectable label.

18. The engineered cell of any of the preceding claims, wherein the engineered cell is a T cell.

19. The engineered cell of claim 18, wherein the T cell is a CD8+ T cell.

20. The engineered cell of claim 18, wherein the T cell is a CAR-T cell.

21. The engineered cell of claim 20, wherein the CAR-T cell comprises a CAR specific for or active against the diseased cell.

22. The engineered cell of claim 21, wherein the CAR is an anti-CD 123 CAR.

23. The engineered cell of any of the preceding claims, wherein the one or more source cells and/or the engineered cell are obtained from a subject.

24. The engineered cell of any of the preceding claims, wherein the engineered cell is not autologous to a subject.

25. The engineered cell of any of the preceding claims, wherein the engineered cell is autologous to a subject.

27. The method of claim 26, wherein the cell membrane components are obtained by freeze-thaw of the source cells.

28. The method of any of claims 26-27, wherein the mixing or fusing of the cell membrane components comprises sonication and/or fluid flow.

29. The method of any of claims 26-28, wherein the method, or at least steps b) and c), is conducted in a microfluidic device.

31. A method of treating a subject, the method comprising administering an engineered cell of any of claims 1-25 to the subject.

32. The method of claim 31, wherein the subject has or is in need of treatment for cancer.

33. The method of claim 32, wherein the cancer is a hematological cancer or hematopoietic cancer.

34. The method of claim 32, wherein the cancer is leukemia.

35. The method of claim 32, wherein the cancer is acute myeloid leukemia.

36. The method of any of claims 31-35, wherein the subject is at risk of cytokine release syndrome (CRS).