- Article
- Published:
A humanized system for pharmacologic control of gene expression
- Victor M. Rivera1,
- Tim Clackson1,
- Sridaran Natesan1,
- Roy Pollock1,
- Jane F. Amara1,
- Terence Keenan1,
- Shannon R. Magari1,
- Tom Phillips1,
- Nancy L. Courage1,
- Franklin Cerasoli Jr.1,
- Dennis A. Holt1 &
- …
- Michael Gilman1
Nature Medicinevolume 2, pages1028–1032 (1996)Cite this article
3998Accesses
518Citations
16Altmetric
Abstract
Gene therapy was originally conceived as a medical intervention to replace or correct defective genes in patients with inherited disorders. However, it may have much broader potential as an alternative delivery platform for protein therapeutics, such as cytokines, hormones, antibodies and novel engineered proteins. One key technical barrier to the widespread implementation of this form of therapy is the need for precise control over the level of protein production. A suitable system for pharmacologic control of therapeutic gene expression would permit precise titration of gene product dosage, intermittent or pulsatile treatment, and ready termination of therapy by withdrawal of the activating drug. We set out to design such a system with the following properties: (1) low baseline expression and high induction ratio; (2) positive control by an orally bioavailable small–molecule drug; (3) reduced potential for immune recognition through the exclusive use of human proteins; and (4) modularity to allow the independent optimization of each component using the tools of protein engineering. We report here the properties of this system and demonstrate its use to control circulating levels of human growth hormone in mice implanted with engineered human cells.
This is a preview of subscription content,access via your institution
Access options
Subscription info for Japanese customers
We have a dedicated website for our Japanese customers. Please go tonatureasia.com to subscribe to this journal.
Buy this article
- Purchase on SpringerLink
- Instant access to the full article PDF.
¥ 4,980
Prices may be subject to local taxes which are calculated during checkout
Similar content being viewed by others
References
Gossen, M., Bonin, A. & Bujard, H. Control of gene activity in higher eukaryotic cells by prokaryotic regulatory elements.Trends Biol. Sci.18, 471–475 (1993).
Spencer, D.M. Creating conditional mutants in mammals.Trends Genet.12, 181–187 (1996).
Ptashne, M. How eukaryotic transcriptional activators work.Nature335, 683–689 (1988).
Brent, R. & Ptashne, M. A eukaryotic transcriptional activator bearing the DNA specificity of a prokaryotic repressor.Cell43, 729–736 (1985).
Hope, I.A. & Struhl, K. Functional dissection of a eukaryotic transcriptional activator protein, GCN4 of yeast.Cell46, 885–894 (1986).
Fields, S. & Song, O.-K. A novel genetic system to detect protein-protein interactions.Nature340, 245–246 (1989).
Spencer, D.M., Wandless, T.J., Schreiber, S.L. & Crabtree, G.R. Controlling signal transduction with synthetic ligands.Science262, 1019–1024 (1993).
Austin, D.J., Crabtree, G.R. & Schreiber, S.L. Proximity versus allostery: The role of regulated protein dimerization in biology.Chem. Biol.1, 131–136 (1994).
Pruschy, M.N.et a Mechanistic studies of a signaling pathway activated by the organic dimerizer FK1012.Chem. Biol.1, 163–172 (1994).
Spencer, D.M., Graef, I., Austin, D.J., Schreiber, S.L. & Crabtree, G.R. A general strategy for producing conditional alleles of Src-like tyrosine kinases.Proc. Natl. Acad. Sci. USA92, 9805–9809 (1995).
Holsinger, L.J., Spencer, D.M., Austin, D.J., Schreiber, S.L. & Crabtree, G.R. Signal transduction in T lymphocytes using a conditional allele of Sos.Proc. Natl Acad. Sci. USA92, 9810–9814 (1995).
Belshaw, P.J., Ho, S.N., Crabtree, G.R. & Schreiber, S.L. Controlling protein association and subcellular localization with a synthetic ligand that induces het-erodimerization of proteins.Proc. Natl. Acad. Sci. USA93, 4604–4607 (1996).
Ho, S.N., Biggar, S.R., Spencer, D.M., Schreiber, S.L. & Crabtree, G.R. Controlling transcription with synthetic ligands.Nature (in the press).
Bierer, B.E.et al. Two distinct signal transmission pathways in T lymphocytes are inhibited by complexes formed between an immunophilin and either FK506 or rapamycin.Proc. Natl. Acad. Sci. USA87, 9231–9235 (1990).
Brown, E.J.et al. A mammalian protein targeted by G1-arresting rapamycin-receptor complex.Nature369, 756–758 (1994).
Chiu, M.L., Katz, H. & Berlin, V. RAPT1, a mammalian homolog of yeast Tor, interacts with the FKBP12/rapamycin complex.Proc. Natl. Acad. Sci. USA91, 12574–12578 (1994).
Sabatini, D.M., Erdjument-Bromage, H., Lui, M., Tempst, P. & Snyder, S.H. RAFT1: A mammalian protein that binds to FKBP12 in a rapamycin-dependent fashion and is homologous to yeast TORs.Cell78, 35–43 (1994).
Sabers, C.J.et al. Isolation of a protein target of the FKBP12-rapamycin complex in mammalian cells.J. Biol. Chem.270, 815–822 (1995).
Stan, R.et al. Interaction between FKBP12-rapamycin and TOR involves a conserved serine residue.J. Biol. Chem.269, 32037–32030 (1994).
Kahan, B., Chang, J. & Sehgal, S. Preclinical evaluation of a new potent im-munosuppressive agent, rapamycin.Transplantation52, 185–191 (1991).
Kay, J., Kromwel, L., Doe, S. & Denyer, M. Inhibition of T and B lymphocyte proliferation by rapamycin.Immunology72, 544–549 (1991).
Stepkowski, S., Chen, H., Daloze, P. & Kahan, B., mycin, apotent immuno-suppressive drug for vascularized heart, kidney, and small bowel transplantation ir- the rat.Transplantation51, 22–26 (1991).
Pomerantz, J.L., Sharp, P.A. & CO.Structure-based design of transcription factors.Science267, 93–96 (1995).
Schmitz, M.L. & Baeurle, P.A. The p65 subunit is responsible for the strong transcription activating potential of NF-KB.EMBO J.10, 3805–3817 (1991).
Chen, J., Zheng, X.F., Brown, E.J. & Schreiber, S.L. Identification of an 11-kDa FKBP12-rapamycin-binding domain within the 289-kDa FKBP12-rapamycin-associated protein and characterization of a critical serine residue.Proc. Natl. Acad. Sci. USA92, 4947–4951 (1995).
Heartlein, M.W.et al. Long-term production and delivery of human growth hormone.Proc. Natl. Acad. Sci. USA91, 10967–10971 (1994).
Riddell, S.R.et al. T-cell mediated rejection of gene-modified HIV-specific cyto-toxic T lymphocytes in HIV-infected patients.Nature Med.2, 216–223 (1996).
Stern, L.J. & Wiley, D.C. Antigenic peptide binding by class I and class II histo-compatibility proteins.Structure2, 245–251 (1994).
Luengo, J.I.et al. Structure-activity studies of rapamycin analogs: Evidence that the C-7 methoxy is part of the effector domain and positioned at the FKBP12/FRAP interface.Chem. Biol2, 471–481 (1995).
Belshaw, P.J., Schoepfer, J., Liu, K., Morrison, K. & Schreiber, S.L. Rational design of orthogonal receptor-ligand combinations.Angew. Chem. Int. Ed. Eng.34, 2129–2132 (1995).
Choi, J., Chen, J., Schreiber, S.L. & Clardy, J. Structure of the FKBP12-rapamycin complex interacting with the binding domain of human FRAP.Science273, 239–242 (1996).
Attar, R.M. & Gilman, M.Z. Expression cloning of a novel zinc-finger protein that binds to the c-fos serum response element.Mol. Cell. Biol.12, 2432–2443 (1992).
Standaert, R.F., Galat, A., Verdine, G.L. & Schreiber, S.L. Molecular cloning and overexpression of the human FK506-binding protein FKBP.Nature346, 671–674 (1990).
Boshart, M.et al A very strong enhancer is located upstream of an immediate early gene of human cytomegalovirus.Cell41, 521–530 (1985).
Siebenlist, U.et al Promoter region of interleukin-2 gene undergoes chro-matin structure changes and confers inducibility on chloramphenicol acetyl-transferase gene during activation of T cells.Mol. Cell Biol6, 3042–3049 (1986).
Selden, R.F., Burke-Howie, K., Rowe, M.E., Goodman, H.M. & Moore, D.D. Human growth hormone as a reporter gene in regulation studies employing transient gene expression.Mol Cell Biol.6, 3173–3179 (1986).
Morgenstern, J.P. & Land, H. Advanced mammalian gene transfer: High titre retroviral vectors with multiple drug selection markers and a complementary helper-free packaging cell line.Nucleic Acids Res.18, 3587–3596 (1990).
Muller, A.J.et al. BCR first exon sequences specifically activate BCR/ABL tyrosine kinase oncogene of Philadelphia chromosome-positive human leukemias.Mol Cell Biol11, 1785–1792 (1991).
Pear, W.S., Nolan, G.P., Scott, M.L. & Baltimore, D. Production of high-titer helper-free retroviruses by transient transfection.Proc. Natl. Acad. Sci. USA90, 8392–8396 (1993).
Author information
Authors and Affiliations
ARIAD Gene Therapeutics, 26 Landsdowne Street, Cambridge, Massachusetts, 02139, USA
Victor M. Rivera, Tim Clackson, Sridaran Natesan, Roy Pollock, Jane F. Amara, Terence Keenan, Shannon R. Magari, Tom Phillips, Nancy L. Courage, Franklin Cerasoli Jr., Dennis A. Holt & Michael Gilman
- Victor M. Rivera
Search author on:PubMed Google Scholar
- Tim Clackson
Search author on:PubMed Google Scholar
- Sridaran Natesan
Search author on:PubMed Google Scholar
- Roy Pollock
Search author on:PubMed Google Scholar
- Jane F. Amara
Search author on:PubMed Google Scholar
- Terence Keenan
Search author on:PubMed Google Scholar
- Shannon R. Magari
Search author on:PubMed Google Scholar
- Tom Phillips
Search author on:PubMed Google Scholar
- Nancy L. Courage
Search author on:PubMed Google Scholar
- Franklin Cerasoli Jr.
Search author on:PubMed Google Scholar
- Dennis A. Holt
Search author on:PubMed Google Scholar
- Michael Gilman
Search author on:PubMed Google Scholar
Rights and permissions
About this article
Cite this article
Rivera, V., Clackson, T., Natesan, S.et al. A humanized system for pharmacologic control of gene expression.Nat Med2, 1028–1032 (1996). https://doi.org/10.1038/nm0996-1028
Received:
Accepted:
Issue date:
Share this article
Anyone you share the following link with will be able to read this content:
Sorry, a shareable link is not currently available for this article.
Provided by the Springer Nature SharedIt content-sharing initiative
This article is cited by
A programmable protease-based protein secretion platform for therapeutic applications
- Xinyi Wang
- Liping Kang
- Haifeng Ye
Nature Chemical Biology (2024)
A simeprevir-inducible molecular switch for the control of cell and gene therapies
- Stacey E. Chin
- Christina Schindler
- Natalie J. Tigue
Nature Communications (2023)
A single-component, light-assisted uncaging switch for endoproteolytic release
- Mingguang Cui
- Seunghwan Lee
- Dongmin Lee
Nature Chemical Biology (2023)
Designer installation of a substrate recruitment domain to tailor enzyme specificity
- Rodney Park
- Chayanid Ongpipattanakul
- Brian Kuhlman
Nature Chemical Biology (2023)
Mandipropamid as a chemical inducer of proximity for in vivo applications
- Michael J. Ziegler
- Klaus Yserentant
- Richard Wombacher
Nature Chemical Biology (2022)


