.2025 Oct 17;11(42):eadr1524.

doi: 10.1126/sciadv.adr1524. Epub 2025 Oct 15.

Impact of intermittent lead exposure on hominid brain evolution

Renaud Joannes-Boyau^{1 2 3}, Janaina Sena de Souza⁴, Manish Arora⁵, Christine Austin⁵, Kira Westaway⁶, Ian Moffat^{7 8}, Wei Wang⁹, Wei Liao⁹, Yingqi Zhang³, Justin W Adams^{2 10 11}, Luca Fiorenza¹⁰, Flora Dérognat¹, Marie-Helene Moncel¹², Gary T Schwartz¹³, Marian Bailey¹, Filipe F Dos Santos^{14 15}, Gabriela D A Guardia¹⁴, Rafael L V Mercuri^{14 16}, Pedro A F Galante¹⁴, Aline M A Martins^{17 18}, Blake L Tsu¹⁹, Christopher A Barnes¹⁹, John Yates 3rd¹⁸, Luiz Pedro Petroski²⁰, Sandra M Sanchez-Sanchez⁴, Jose Oviedo²¹, Roberto H Herai²⁰, Bernardo Lemos^{21 22}, Matthew Tonge¹, Alysson R Muotri^{4 17 23}

Affiliations

¹ Geoarchaeology and Archaeometry Research Group, Southern Cross University, Military Road, Lismore, NSW 2480, Australia.
² Palaeo-Research Institute, University of Johannesburg, Auckland Park, Johannesburg 2006, South Africa.
³ Institute of Vertebrate Paleontology and Paleoanthropology, Chinese Academy of Sciences, Beijing 100044, China.
⁴ Department of Pediatrics and Cellular & Molecular Medicine, School of Medicine, University of California San Diego, La Jolla, CA 92093, USA.
⁵ Department of Environmental Medicine, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
⁶ School of Natural Sciences, Faculty of Science and Engineering, Macquarie University, Sydney, NSW 2109, Australia.
⁷ Archaeology, College of Humanities, Arts and Social Sciences, Flinders University, Adelaide, SA, Australia.
⁸ McDonald Institute for Archaeological Research, University of Cambridge, Cambridge, UK.
⁹ Institute of Cultural Heritage, Shandong University, Qingdao 266237 China.
¹⁰ Department of Anatomy and Developmental Biology, Biomedicine Discovery Institute, Monash University, Melbourne, VIC 3800 Australia.
¹¹ Department of Geosciences, Museums Victoria, 11 Nicholson St., Carlton, VIC 3053, Australia.
¹² Département Hommes et Environnement, Museum National d'Histoire Naturelle, CNRS 7194, Institut de Paléontologie Humaine, 1 rue René Panhard, 75013 Paris, France.
¹³ Institute of Human Origins, School of Human Evolution and Social Change, Arizona State University, Tempe, AZ 85287, USA.
¹⁴ Hospital Sirio-Libanes, Sao Paulo 01308-060, Brazil.
¹⁵ Departamento de Bioquimica, Instituto de Quimica, Universidade de Sao Paulo, Sao Paulo, Brazil.
¹⁶ Interunidades em Bioinformática, Universidade de Sao Paulo, Sao Paulo, Brazil.
¹⁷ ISSCOR - Integrated Space Stem Cell Orbital Research, Sanford Consortium of Regenerative Medicine, UCSD, La Jolla, CA 92093, USA.
¹⁸ Department of Molecular Medicine, The Scripps Research Institute, La Jolla, CA 92037, USA.
¹⁹ NGeneBioAI Inc., 9810 Summers Ridge Rd. #100, San Diego, CA 92121, USA.
²⁰ Pontifícia Universidade Católica do Paraná, School of Medicine and Life Sciences, Graduate Program in Health Sciences (PUCPR/PPGCS), Laboratory of Bioinformatics and Neurogenetics (LaBiN), Curitiba, Paraná 80215-901, Brazil.
²¹ Department of Pharmacology and Toxicology, R Ken Coit College of Pharmacy, University of Arizona, Tucson, AZ 85721, USA.
²² Program in Molecular and Integrative Physiological Science and Department of Environmental Health, Harvard T. H. Chan School of Public Health, Boston, MA 02115, USA.
²³ University of California San Diego, Kavli Institute for Brain and Mind, Center for Academic Research and Training in Anthropogeny (CARTA), Archealization Center (ArchC), La Jolla, CA 92093, USA.

PMID:41091888
PMCID: PMC12527068
DOI: 10.1126/sciadv.adr1524

Impact of intermittent lead exposure on hominid brain evolution

Renaud Joannes-Boyau et al. Sci Adv.2025.

.2025 Oct 17;11(42):eadr1524.

doi: 10.1126/sciadv.adr1524. Epub 2025 Oct 15.

Authors

Affiliations

¹ Geoarchaeology and Archaeometry Research Group, Southern Cross University, Military Road, Lismore, NSW 2480, Australia.
² Palaeo-Research Institute, University of Johannesburg, Auckland Park, Johannesburg 2006, South Africa.
³ Institute of Vertebrate Paleontology and Paleoanthropology, Chinese Academy of Sciences, Beijing 100044, China.
⁴ Department of Pediatrics and Cellular & Molecular Medicine, School of Medicine, University of California San Diego, La Jolla, CA 92093, USA.
⁵ Department of Environmental Medicine, Icahn School of Medicine at Mount Sinai, New York, NY 10029, USA.
⁶ School of Natural Sciences, Faculty of Science and Engineering, Macquarie University, Sydney, NSW 2109, Australia.
⁷ Archaeology, College of Humanities, Arts and Social Sciences, Flinders University, Adelaide, SA, Australia.
⁸ McDonald Institute for Archaeological Research, University of Cambridge, Cambridge, UK.
⁹ Institute of Cultural Heritage, Shandong University, Qingdao 266237 China.
¹⁰ Department of Anatomy and Developmental Biology, Biomedicine Discovery Institute, Monash University, Melbourne, VIC 3800 Australia.
¹¹ Department of Geosciences, Museums Victoria, 11 Nicholson St., Carlton, VIC 3053, Australia.
¹² Département Hommes et Environnement, Museum National d'Histoire Naturelle, CNRS 7194, Institut de Paléontologie Humaine, 1 rue René Panhard, 75013 Paris, France.
¹³ Institute of Human Origins, School of Human Evolution and Social Change, Arizona State University, Tempe, AZ 85287, USA.
¹⁴ Hospital Sirio-Libanes, Sao Paulo 01308-060, Brazil.
¹⁵ Departamento de Bioquimica, Instituto de Quimica, Universidade de Sao Paulo, Sao Paulo, Brazil.
¹⁶ Interunidades em Bioinformática, Universidade de Sao Paulo, Sao Paulo, Brazil.
¹⁷ ISSCOR - Integrated Space Stem Cell Orbital Research, Sanford Consortium of Regenerative Medicine, UCSD, La Jolla, CA 92093, USA.
¹⁸ Department of Molecular Medicine, The Scripps Research Institute, La Jolla, CA 92037, USA.
¹⁹ NGeneBioAI Inc., 9810 Summers Ridge Rd. #100, San Diego, CA 92121, USA.
²⁰ Pontifícia Universidade Católica do Paraná, School of Medicine and Life Sciences, Graduate Program in Health Sciences (PUCPR/PPGCS), Laboratory of Bioinformatics and Neurogenetics (LaBiN), Curitiba, Paraná 80215-901, Brazil.
²¹ Department of Pharmacology and Toxicology, R Ken Coit College of Pharmacy, University of Arizona, Tucson, AZ 85721, USA.
²² Program in Molecular and Integrative Physiological Science and Department of Environmental Health, Harvard T. H. Chan School of Public Health, Boston, MA 02115, USA.
²³ University of California San Diego, Kavli Institute for Brain and Mind, Center for Academic Research and Training in Anthropogeny (CARTA), Archealization Center (ArchC), La Jolla, CA 92093, USA.

PMID:41091888
PMCID: PMC12527068
DOI: 10.1126/sciadv.adr1524

Abstract

Gene-environmental interactions shape the evolution of brain architecture and function. Neuro-oncological ventral antigen 1 (NOVA1) is one gene that distinguishes modern humans from extinct hominids. However, the evolutionary pressures that selected the modernNOVA1 allele remain elusive. Here, we show using fossil teeth that several hominids (Australopithecus africanus,Paranthropus robustus, earlyHomo sp.,Gigantopithecus blacki,Pongo sp.,Homo neanderthalensis, andHomo sapiens) were consistently exposed to lead over 2 million years, contradicting the idea that lead exposure is solely a modern phenomenon. Moreover, lead exposure on human brain organoids carrying the archaicNOVA1 variant disruptsFOXP2 expression in cortical and thalamic organoids, a gene crucial for the development of human speech and language abilities. Overall, the fossil, cellular, and molecular data support that lead exposure may have contributed to the impact of social and behavioral functioning during evolution, likely affording modern humans a survival advantage.

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Figures

**Fig. 1.. Reconstructing primate’s exposure to environmental lead.**
(A) World map with indication of the country from which the archaeological and modern samples were obtained. (B) Sketch of a sectioned fossil tooth analyzed by laser ablation combined with inductively coupled plasma mass spectrometry (LA-ICP-MS). The laser is rastered across the sectioned surface, and ejected material is transported to the mass spectrometer. Green and dashed lines depict the biogenic pattern of metal uptake within growth rings [adapted from (65)]. (C) Photo (left) of a sectionedG. blacki third molar from the site of Queque in China. The²⁰⁸Pb/⁴³Ca distribution from the marked area is shown on the right. The geochemical map reveals a large number of biogenic banding relating to repeated acute exposures to lead.

**Fig. 2.. Lead banding in twoG. blacki fossil teeth.**
²⁰⁸Pb/⁴³Ca distribution in second molar (A) and third molar (B) ofG. blacki teeth from the site of Queque in Southern China dated to the middle Pleistocene. The Pb banding follows the architecture of the dental tissue growth pattern, which is evidence of its biogenic origin and incorporation during tooth growth. Both teeth show an impressive number of repeated acute exposures to lead.

**Fig. 3.. Lead banding in fossil hominid teeth across the Pleistocene.**
²⁰⁸Pb/⁴³Ca distribution in (A)*G. blacki* third molar from the site of Chuifeng in China. (B)*A. africanus* canine from the site of Sterkfontein in South Africa. (C)*G. blacki* second molar from the site of Queque in China. (D)*Pongo* sp. canine from the site of Shanzu in China. (E)*H. sapiens* first molar from the site of Mohui in China. (F)*H. neanderthalensis* molar from the site of Payre in France.

**Fig. 4.. Characterization of cortical brain organoids ofNOVA1^hu/hu andNOVA1^ar/ar.**
(A) Schematic representation of the brain cortical organoid (COs) differentiation protocol. (B) Representative bright-field images of human COs captured at day 16, proliferative stage of neuroprogenitor cells (*NOVA1*^hu/hu andNOVA1^ar/ar). (C) Representative immunohistochemistry images of 60-day-old brain organoids (*NOVA1*^hu/hu). COs contain proliferative regions (SOX2⁺), expressing the cortical marker (FOXG1⁺) surrounded by mature neurons (MAP2⁺). Scale bars, 50 μm. (D) Cell cycle quantification of brain organoids carrying theNOVA1^ar/ar genetic variant. CTL, control group.n ≥ 2. One-way analysis of variance (ANOVA) Tukey test and Kruskal-Wallis test, followed by Dunn’s test. n.s., not significant. *P < 0.05. (E) Volcano plot showing the 616 up-regulated (blue) and 429 down-regulated (red) genes under theNOVA1^ar/ar condition compared to theNOVA1^hu/hu control after applying the false discovery rate (FDR) < 0.05 and log₂ fold change (|L2FC|) > 1 filters. (F) Bubble plot showing the top enriched Gene Ontology Biological Process (GO:BP) terms (FDR < 0.05; bigger bubbles mean more enrichment) for both the up-regulated (blue) and down-regulated (red) genes shown in (E). Terms are further organized into major color-coded functional “groups.” (G) Protein-protein interactions (PPIs) showing the differentially expressed genes (DEGs) that represent the major color-coded functional groups shown in (F). Higher STRINGdb scores (minimum cutoff: 0.7, “high confidence”) mean less transparency and thicker edges. Higher |L2FC| values mean bigger nodes, while lower FDR values mean darker (blue: up-regulated; red: down-regulated) nodes.

**Fig. 5.. GO enrichment and gene network analysis of lead-induced changes inNOVA1 variants.**
Bubble plots showing the top 20 enriched GO:BP terms (FDR < 0.05; bigger bubbles mean more enrichment) for both the up-regulated (blue) and down-regulated (red) genes in (A)*NOVA1*^hu/hu treated with 10 μM lead. (B)*NOVA1*^hu/hu treated with 30 μM lead. (C)*NOVA1*^ar/ar treated with 10 μM lead. (D)*NOVA1*^ar/ar treated with 30 μM lead. In all figures, the terms are further organized into major color-coded functional groups; PPIs showing the DEGs representing the major color-coded functional groups are shown in the respective bubble plots. Higher STRINGdb scores (minimum cutoff: 0.4, “medium default confidence”) mean less transparency and thicker edges. Higher |L2FC| values mean more prominent nodes, while lower FDR values mean darker (blue, up-regulated; red, down-regulated) nodes in (E)*NOVA1*^ar/ar treated with 10 μM lead and (F)*NOVA1*^ar/ar treated with 30 μM lead. Figures S12 and S13 are related to this figure.

**Fig. 6.. Lead exposure changes the single-cell transcriptome inNOVA1^ar/ar COs.**
(A andB) Uniform Manifold Approximation and Projection (UMAP) of 99,623 cells from integrated datasets of 60-day-old COs. The integrated dataset is colored by six cellular groups (*NOVA1*^hu/hu CTL,n = 16,315 cells;*NOVA1*^hu/hu 10 μM,n = 16,890 cells;*NOVA1*^hu/hu 30 μM,n = 16,268;*NOVA1*^ar/ar CTL,n = 15,411 cells;*NOVA1*^ar/ar 10 μM,n = 17,937 cells;*NOVA1*^ar/ar 30 μM,n = 16,802 cells). (C andD) UMAP ofNOVA1^hu/hu andNOVA1^ar/ar COs treated with different lead concentrations showing eight different clusters. (E) Overlap of protein-coding genes up- and down-regulated inNOVA1^ar/ar versusNOVA1^hu/hu. (F) Bubble plot presenting pathways altered inNOVA1^ar/ar. (G) Overlap of protein-coding genes up- and down-regulated inNOVA1^ar/ar 10 μM group versusNOVA1^ar/ar. (H) Bubble plot presenting pathways altered inNOVA1^ar/ar 10 μM group. (I) Overlap of protein-coding genes up- and down-regulated inNOVA1^ar/ar 30 μM group versusNOVA1^ar/ar. (J) Bubble plot presenting pathways altered inNOVA1^ar/ar 30 μM group. (K) Bubble plot presentingFOXP2 expression under all conditions. Figure S14 is related to this figure.

**Fig. 7.. Lead affects neural pathways in COs at the single-cell proteomics level.**
(A) Schematic representation of the single-cell proteomics protocol. (B) Volcano plot ofNOVA1^hu/hu versusNOVA1^ar/ar. A total of 840 peptides were recovered in 427 proteins after peptide filtering (Q threshold = 0.01, |L2FC| threshold = 0.445). (C) Volcano plot ofNOVA1^ar/ar versusNOVA1^ar/ar 10 μM group. A total of 710 peptides were recovered in 366 proteins after peptide filter (Q threshold = 0.01, |L2FC| threshold = 0.424). (D) Volcano plot ofNOVA1^ar/ar versusNOVA1^ar/ar 30 μM group. A total of 684 peptides were recovered in 352 proteins after peptide filter (Q threshold = 0.01, |L2FC| threshold = 0.447). (E) Network showing the most important proteins on the pathway. CFL1, Cofilin-1 (required for neural tube morphogenesis and neural crest cell migration); CRMP1, dihydropyrimidinase-related protein 1 (plays a role in axon guidance and, during the axon guidance process, acts downstream of SEMA3A to promote FLNA dissociation from F-actin, which results in the rearrangement of the actin cytoskeleton and the collapse of the growth cone); DPYSL3, dihydropyrimidinase-related protein 3 (plays a role in axon guidance, neuronal growth cone collapse, and cell migration); TUBB2B, tubulin-β 2B chain (plays a critical role in proper axon guidance in central and peripheral axon tracts and is implicated in neuronal migration); MBP, myelin basic protein (plays an important role in the early developing brain long before myelination). Figures S16 to S18 are related to this figure. [(B) to (D)] Full blue arrowheads, down-regulated; empty blue arrowheads, down-regulated and outside plot limits; full orange arrowheads, up-regulated; empty orange arrowheads, up-regulated and outside plots limit; full black dots, not significant; empty black dots, not significant and outside plot limits.

**Fig. 8.. Thalamic brain organoid single-cell transcriptome cellular characterization and clusters.**
(A) Schematic representation of the TO differentiation protocol. (B) Bright-field image of TO at 20 and 50 days. (C andD) UMAP of 71,942 cells from integrated datasets of 60-day-old TO. The integrated dataset is colored by four cellular groups (*NOVA1*^hu/hu CTL,n = 18,932 cells;*NOVA1*^hu/hu 10 μM,n = 16,657 cells;*NOVA1*^ar/ar CTL,n = 18,507 cells;*NOVA1*^ar/ar 10 μM,n = 17,846 cells). (E andF) UMAP ofNOVA1^hu/hu andNOVA1^ar/ar TOs was treated with different lead concentrations, showing nine different clusters. (G) Immunohistochemistry image of 60-day-oldNOVA1^hu/hu andNOVA1^ar/ar TOs. They are TCF7L2⁺ and PAX6⁺ organoids. Scale bar, 50 μm. (H andI) The expression ofTCF7L2 andLHX1 are thalamic markers visualized on the UMAP. Figure S19 is related to this figure.

**Fig. 9.. Exposure to lead increasedFOXP2 gene expression and altered the alternative splicing inNOVA1^ar/ar in TOs.**
(A) Immunohistochemistry image of 60-day-oldNOVA1^hu/hu andNOVA1^ar/ar TOs. TOs are TCF7L2⁺, FOXP2^+, and MAP2⁺. Scale bars, 50 μm. (B) Bubble plot showing the gene expression ofFOXP2 in COs and TO. (C) Relative expression [reverse transcription quantitative polymerase chain reaction (PCR)] ofFOXP2 in TO ofNOVA1^hu/hu,*NOVA1*^ar/ar, andNOVA1^ar/ar 10 μM group.n = 6 replicates. **P < 0.01. (D andE) UMAP ofNOVA1^hu/hu,*NOVA1*^hu/hu 10 μM,*NOVA1*^ar/ar, andNOVA1^ar/ar 10 μM group showing the cellular expression ofFOXP2. (F)*NOVA1*^hu/hu genes that altered their splicing after lead exposure. (G) Predicted expression of alternative splicing isoforms based on splice junctions after exposure to 10 and 30 μM in theNOVA1^ar/ar TO. Reference Sequence accession codes represent gene isoforms, and it is shown, for each condition, a sashimi plot (curvy lines indicate splice junction reads and the number above the spliced reads), indicating the predicted expressed transcript isoform (expression values are indicated by reads per kilobase per million mapped reads). Exons without supporting splice junction reads were not considered expressed and were depleted from the figure to improve visualization. Alternative spliced exons are highlighted in yellow. (H) Interaction network of genes that had an alteration in their splicing andNOVA1,*FOXP2*,*POU3F2*, andGRID2.

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References

1. Trumble B. C., Finch C. E., The exposome in human evolution: From dust to diesel. Q. Rev. Biol. 94, 333–394 (2019). - PMC - PubMed
1. Hong S., Candelone J.-P., Patterson C. C., Boutron C. F., Greenland ice evidence of hemispheric lead pollution two millennia ago by greek and roman civilizations. Science 265, 1841–1843 (1994). - PubMed
1. Smith D. R., Flegal A. R., Lead in the biosphere: Recent trends. Ambio 24, 21–23 (1995).
1. Bushnell P. J., Bowman R. E., Effects of chronic lead ingestion on social development in infant rhesus monkeys. Neurobehav. Toxicol. 1, 207–219 (1979). - PubMed
1. Needleman H. L., Gatsonis C. A., Low-level lead exposure and the IQ of children: A meta-analysis of modern studies. JAMA 263, 673–678 (1990). - PubMed

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Impact of intermittent lead exposure on hominid brain evolution

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Impact of intermittent lead exposure on hominid brain evolution

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