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

这个项目建立在以前的研究基础上，这些研究利用生物学和生化实验室的先进技术来解决法医学中的两个关键问题：确定死亡间隔(PMI)和估计受害者的死亡年龄(AAD)。现有的这些估计方法往往依赖于科学家的专业知识，导致结果不一致和主观性。此外，PMI估计技术需要在死亡后不久进行软组织评估，而AAD的确定依赖于完整的骨骼特征。在处理骨骼残骸或残缺不全的遗体时，这些限制是显而易见的，在悬案、自然灾害和其他情况下很常见。现在，技术进步使研究人员能够使用“组学”学科(基因组学、蛋白质组学、新陈代谢组学)从微小的样本中提取有价值的生物信息，如微小的骨骼碎片。这些骨生物分子在小的生物样本中表现出与体内和死后老化过程相关的明显特征。在本课题组此前开展的研究中，将应用《组学》技术研究人类遗骸的老化问题命名为《Forens-OMICS》。最初的工作包括以非针对性的方式研究多个分子特征(DNA、蛋白质、脂类、代谢物)，以确定推测的衰老预测标记。作为先前在“FORENS-OMICS”中进行的工作的继续，该项目现在的目标是使用提供定量精确度和重现性的有针对性的方法来验证以前确定的标记，这是法医情景中的一项关键要求。经过验证的标记物将被合并到用户友好的小组中，使用法医实验室中广泛存在的仪器进行无偏见、可靠和快速的评估。除了验证先前确定的用于PMI和AAD估计的生物标记物外，该项目还通过将“Forens-OMICS”方法应用于通常在犯罪现场发现的多种生物液来扩大其最初从骨骼遗骸中估计PMI/AAD的范围，旨在估计现场污渍沉积以来的时间(TSD)。通过建立特定标记物(RNA、微生物、蛋白质、代谢物、脂类)与时间之间的关系，通过多组学方法进行准确的TSD估计成为可能。为了实现这些目标，该项目将与美国的人类学机构合作，提供更多的人骨材料，用于在一组独立的样本上验证识别的标志物。18世纪和19世纪的历史人类骨骼也将被包括在内，以评估“Forens-OMICS”方法对较老标本的适用性，以便将来也在考古背景下实施。获得的信息将使用先进的生物信息学工具进行整合，通过对经过验证的生物标记物进行有针对性的分析，为PMI和AAD估计开发改进的数学模型。此外，将“Forens-OMICS”方法应用于生物体液将为多组学策略在新的法医学背景下的应用铺平道路，并将为从业者提供一种新的工具，以一种新颖和准确的方式评估TSD。

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.