"Forens-OMICS" on Human Remains and Biological Fluids for Age-at-Death, Post-Mortem Interval and Time Since Deposition Estimation in Forensic Contexts

This project builds upon previous research that utilized advanced technologies in biology and biochemistry labs to address two crucial questions in forensic science: determining the post-mortem interval (PMI) and estimating the age-at-death (AAD) of a victim. Existing methods for these estimations often rely on the expertise of scientists, resulting in inconsistent results and subjectivity. Additionally, PMI estimation techniques require soft tissue evaluations soon after death, while AAD determination relies on intact skeletal features. These limitations are evident when dealing with skeletal remains or incomplete bodies, common in cold cases, natural disasters, and other scenarios.

Technological advancements now allow researchers to extract valuable biological information from minute samples, such as tiny bone fragments, using "omics" disciplines (genomics, proteomics, metabolomics). These bone biomolecules exhibit distinct signatures related to in-vivo and post-mortem aging processes within small biological samples. In the previous research conducted by our group, the application of "omics" technologies to study the aging of human remains was named "Forens-OMICS". The initial work involved studying multiple molecular signatures (DNA, proteins, lipids, metabolites) in a non-targeted manner to identify putative predictive markers for aging.

As a continuation of the previously conducted work in "Forens-OMICS", this project now aims to validate the previously identified markers using targeted approaches that provide quantitative precision and reproducibility, a crucial requirement in forensic scenarios. The validated markers will then be combined into user-friendly panels for unbiased, reliable, and quick estimations using the instrumentation widely available in forensic laboratories.

In addition to validating the previously identified biomarkers for PMI and AAD estimation, this project also expands its original scope of PMI/AAD estimation from skeletal remains by applying the "Forens-OMICS" approach to multiple biological fluids usually recovered at crime scenes, aiming to estimate the time since deposition (TsD) of a stain at the scene. By establishing a relationship between specific markers (RNA, microbes, proteins, metabolites, lipids) and time, accurate TsD estimation via multi-omics approaches becomes possible. This is crucial for linking DNA-identified stains to crimes or estimating PMI from cadaveric fluids.

To achieve these goals, the project will be conducted in collaboration with anthropological facilities in the United States, which will provide additional human bone material for the validation of the identified markers on a set of independent samples. Historical human bones from the 18th and 19th centuries will also be included to assess the "Forens-OMICS" method applicability to older specimens, for its future implementation also on archaeological contexts. The obtained information will be integrated using advanced bioinformatics tools to develop improved mathematical models for PMI and AAD estimation via the targeted analysis of the validated biomarkers. Additionally, the application of the "Forens-OMICS" method to biological fluids will pave the way for the application of multi-omics strategies in new forensic contexts and will provide practitioners with a new tool for assessing TsD in a novel and precise way.