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Dental Metapeptidomics in Forensic and Archaeological Science: Applications to Biological Profiling, Microbiome and Diet
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Os esqueletos humanos constituem uma fonte fundamental de informação em investigações forenses e arqueológicas, permitindo obter dados relevantes tanto para a identificação de indivíduos como para a compreensão de modos de vida passados. Os dentes assumem especial importância devido à sua resistência à degradação pós-morte, preservando péptidos que funcionam como cápsulas do tempo. Dada esta relevância, as amostras dentárias exigem abordagens minimamente destrutivas, capazes de salvaguardar a sua integridade e, ao mesmo tempo, possibilitar a análise molecular. Este projeto centrou-se no desenvolvimento de um protocolo rápido, eficaz e minimamente destrutivo para a caracterização abrangente de peptidomas dentários arqueológicos, com recurso à espectrometria de massa de alta resolução combinada com metapeptidómica. O protocolo permitiu a reconstrução de perfis biológicos humanos, bem como a deteção de péptidos de origem bacteriana, animal e vegetal, refletindo interações com multiespécies relacionadas com o ambiente e a dieta.
A determinação do sexo biológico foi alcançada com sucesso em 25 amostras arqueológicas através da deteção dos péptidos AMELX-1 (SIRPPYPSY) e AMELY-2 (SM(ox)IRPPY). Foram ainda identificados novos péptidos de amelogenina por sequenciação de novo, incluindo um péptido derivado de AMELY presente de forma consistente em 88% das amostras de indivíduos do sexo masculino, demonstrando elevado potencial como um novo biomarcador para a determinação sexual de indivíduos, sujeito a validação futura. Em contraste, as tentativas de estimar a idade aquando da morte com base na deamidação de asparagina e glutamina não apresentaram correlações fiáveis, salientando a necessidade de investigação adicional acerca dos fatores que influenciam estas modificações.
O protocolo desenvolvido permitiu ainda a deteção de péptidos bacterianos provenientes de espécies patogénicas, como M. tuberculosis, M. bovis, S. aureus, K. pneumoniae e P. aeruginosa, demonstrando o potencial da metapeptidómica dentária para investigar contextos epidemiológicos passados. Além disso, a identificação de péptidos associados à dieta e ao ambiente, provenientes de espécies como G. gallus, S. scrofa, B. taurus, C. hircus, O. aries, T. aestivum, Z. mays, N. tabacum, O. sativa, A. thaliana e C. reinhardtii, proporcionou informações importantes sobre dietas passadas, práticas culturais e exposições ambientais.
Em suma, este estudo reforça a aplicabilidade da metapeptidómica dentária e de protocolos minimamente destrutivos como ferramentas altamente informativas em investigações forenses e arqueológicas, contribuindo para a elaboração de perfis biológicos, exploração de exposições a agentes bacterianos e ambientais e estudo de práticas culturais e alimentares em populações antigas.
Skeletonized human remains represent a fundamental source of information in forensic and archaeological investigations, providing insights into individual identification and past lifeways. Dental structures are particularly valuable due to their resistance to post-mortem degradation, preserving peptides that act as molecular time capsules. Given this relevance, dental specimens demand minimally destructive approaches that safeguard integrity while enabling molecular analysis. This project focused on developing a rapid, effective, and minimally destructive workflow for the comprehensive characterization of ancient dental peptidomes using high-resolution mass spectrometry combined with metapeptidomics. The protocol enabled the reconstruction of human biological profiles, while also detecting bacterial, animal, and plant-derived peptides, thereby reflecting multispecies interactions related to environment and diet. Biological sex determination was successfully achieved in 25 archaeological samples through the detection of both AMELX-1 (SIRPPYPSY) and AMELY-2 (SM(ox)IRPPY). Novel amelogenin peptides were identified by de novo sequencing, including an AMELY-derived peptide consistently present in 88% of the biological male samples, suggesting strong potential as a new biomarker for biological sex determination, pending future validation. In contrast, attempts to estimate age-at-death based on asparagine and glutamine deamidation did not yield reliable correlations, emphasizing the need for further research into the factors influencing these modifications. The workflow also enabled the detection of bacterial peptides from pathogenic species such as M. tuberculosis, M. bovis, S. aureus, K. pneumoniae, and P. aeruginosa, highlighting the potential of dental metapeptidomics to investigate past epidemiological contexts. Moreover, dietary- and environmental-related peptides from species including G. gallus, S. scrofa, B. taurus, C. hircus, O. aries, T. aestivum, Z. mays, N. tabacum, O. sativa, A. thaliana and C. reinhardtii provided insights into ancient diets, cultural practices, and environmental exposures. Overall, this study supports the applicability of minimally destructive dental metapeptidomics as a highly informative tool in forensic and archaeological research, contributing to biological profiling, exploration of bacterial and environmental exposures, and cultural and dietary practices in ancient populations.
Skeletonized human remains represent a fundamental source of information in forensic and archaeological investigations, providing insights into individual identification and past lifeways. Dental structures are particularly valuable due to their resistance to post-mortem degradation, preserving peptides that act as molecular time capsules. Given this relevance, dental specimens demand minimally destructive approaches that safeguard integrity while enabling molecular analysis. This project focused on developing a rapid, effective, and minimally destructive workflow for the comprehensive characterization of ancient dental peptidomes using high-resolution mass spectrometry combined with metapeptidomics. The protocol enabled the reconstruction of human biological profiles, while also detecting bacterial, animal, and plant-derived peptides, thereby reflecting multispecies interactions related to environment and diet. Biological sex determination was successfully achieved in 25 archaeological samples through the detection of both AMELX-1 (SIRPPYPSY) and AMELY-2 (SM(ox)IRPPY). Novel amelogenin peptides were identified by de novo sequencing, including an AMELY-derived peptide consistently present in 88% of the biological male samples, suggesting strong potential as a new biomarker for biological sex determination, pending future validation. In contrast, attempts to estimate age-at-death based on asparagine and glutamine deamidation did not yield reliable correlations, emphasizing the need for further research into the factors influencing these modifications. The workflow also enabled the detection of bacterial peptides from pathogenic species such as M. tuberculosis, M. bovis, S. aureus, K. pneumoniae, and P. aeruginosa, highlighting the potential of dental metapeptidomics to investigate past epidemiological contexts. Moreover, dietary- and environmental-related peptides from species including G. gallus, S. scrofa, B. taurus, C. hircus, O. aries, T. aestivum, Z. mays, N. tabacum, O. sativa, A. thaliana and C. reinhardtii provided insights into ancient diets, cultural practices, and environmental exposures. Overall, this study supports the applicability of minimally destructive dental metapeptidomics as a highly informative tool in forensic and archaeological research, contributing to biological profiling, exploration of bacterial and environmental exposures, and cultural and dietary practices in ancient populations.
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Palavras-chave
Dental Metapeptidomics Biological Sex determination Dietary reconstruction Archaeology Forensics Microbiome