"Forens-OMICS": a cross disciplinary implementation of omics sciences to in vivo and post-mortem ageing investigations for forensic applications

该研究项目的目的是应用现代生物学和生物化学实验室中的一些最尖端技术来解决法医科学家可能被要求回答的两个最重要的问题，以解决犯罪，即受害者的死后间隔（PMI）（从他/她死亡的时间）和受害者的年龄（AAD，死亡年龄）。尽管有几种不同的方法可用于进行这些估计，但它们通常依赖于进行分析的法医科学家的专业知识，因此个体间的可重复性和客观性较差。此外，到目前为止开发的用于估计PMI的大多数技术依赖于必须对软组织进行的评估（例如，在死亡后的短时间内），并且旨在解决AAD的技术通常依赖于骨骼的完整性来寻找特定的形态骨骼特征（例如，颅缝，耻骨联合），可以提供有关受害者的实际年龄的信息。从这些假设出发，很明显，实际方法的局限性是什么，即缺乏从分离的遗骸或不完整的尸体中获得的老化信息（如尸体碎片），这是一个非常常见的情况下，冷案件的受害者，自然灾害，战争受害者，恐怖袭击，人口贩运的受害者，也为考古发掘。如今，科技进步使研究人员可以从极少量的材料，例如极细小的骨碎片，提取大量宝贵的生物资料。这包括遗传信息，以及蛋白质和代谢物信息，这些信息通常总结在“组学”学科的共同术语下。有趣的是，这里引用的所有生物分子都可以根据包含它们的生物组织的体内和死后老化现象带来特定的“签名”，并且可以以非靶向方式在小生物样品中研究这些签名，以评估它们与老化现象（PMI和AAD）的联系及其预测能力。“Forens-OMICS”一词在此表明，几种“组学”技术首次在全球范围内得到应用，其目的是解决衰老现象，主要用于法医学应用，但也用于考古学应用。与美利坚合众国的几个人类学机构新近建立的几个合作项目将允许收集和取样大量年龄较大的人骨范围（从非常年轻到老年人）和以广泛的PMI为特征的个体（从几个月到四十多年）。收集的骨骼样本将用于提取DNA，蛋白质和代谢物，并对这些特定的生物分子进行高通量分析，以推断与PMI和AAD相关的可量化特征。然后，所有恢复的信息将与先进的生物信息学工具结合在一起，以开发一个数学模型，该模型将估计PMI和AAD，并预测该方法的预测能力，并澄清哪些生物分子（或其组合）将提供最准确的估计。总之，新发现的生物标志物将用于开发一种商业试剂盒，可供法医，警察或研究人员使用，以简单，快速，无偏见和可靠的方式进行这些估计。该工具包还将使“非组学”专家能够获得基本的调查线索，这将有助于改善未解决的犯罪和未来涉及存在未解决的或高度碎片化的遗骸的犯罪的结果。

The aim of this research project is to apply some of the most cutting-edge technologies available in modern biology and biochemistry laboratories to address two of the most important questions that a forensic scientist could be asked to answer in order to solve a crime, namely the post-mortem interval (PMI) of the victim (the time elapsed from his/her death) and the age of the victim (AAD, age-at-death). Despite the availability of several different approaches that can be used to perform these estimations, they usually rely on the expertise of the forensic scientist that performs the analysis, thus suffering from poor inter-individual reproducibility and objectivity. Moreover, most of the techniques developed so far to estimate PMI rely on evaluations that has to be performed on soft tissues (e.g. after short periods from death), and techniques aimed at addressing AAD often rely on the completeness of the skeleton to look for specific morphological skeletal traits (e.g., cranial sutures, pubic symphysis) that can give information about the chronological age of the victim. Starting from these assumptions, it is clear and evident what the limits of the actual methodologies are, that is the lack of ageing information obtainable from skeletonised remains or from incomplete bodies (such as body fragments), which is a very common situation for cold cases victims, natural disasters, war victims, terroristic attacks, human trafficking victims, and also for archaeological excavations.Nowadays, technological progress allows researchers to extract an invaluable amount of biological information starting from very small amount of materials such as very small fragment of bone. This includes genetic information, as well as protein and metabolite information, and these are generally summarised under the common term of "omics" disciplines. Interestingly, all the biomolecules cited here can bring a specific "signature" depending on in-vivo and post-mortem ageing phenomena of the biological tissue that contains them, and these signatures could be investigated within a small biological sample in a non-targeted way, in order to evaluate their linkage with ageing phenomena (both PMI and AAD) and their predictive power. The term "Forens-OMICS" indicates here the first global application of several "omics" technologies aimed to address ageing phenomena primarily for forensic applications, but also for archaeological ones.To achieve these aims, several newly established collaborations with several anthropological facilities in the United States of America will allow the collection and the sampling of a significant number of human bones of either individuals with a wide age range (from very young to elderly individuals) and of individuals characterised by a wide range of PMIs (from a couple of months to over forty years). The bone samples collected will be then used to extract DNA, proteins and metabolites, and high-throughput analyses will be performed on each of these specific biomolecules to extrapolate quantifiable features associated with both PMI and AAD. All the recovered information will then be combined together with advanced bioinformatics tools, in order to develop a mathematical model that will estimate PMI and AAD, and that will predict the predictive power of the methodology, as well as clarifying which of the biomolecules (or combination thereof) will provide the most accurate estimations. To conclude, the newly found biomarkers will be used to develop a commercial kit that could be used by forensic examiners, police officers or researchers to make these estimations in an easy, quick, un-biased and reliable way. The kit will also allow "non-omics" experts to get fundamental investigative clues that will help to improve the outcome of unsolved crimes and of future crimes involving the presence of skeletonised or highly fragmented remains.