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Development of a novel forensic age estimation strategy for aged blood samples by combining piRNA and miRNA markers

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Abstract

In forensic investigations, age estimation is vital for determining whether a suspect is under or over the legally defined adult age. With breakthroughs in RNA sequencing technology, small noncoding RNAs have provided new ways to solve problems related to the age estimation of trace or aged samples, owing to their small molecular weight and better stability. In our previous study, we had applied miRNAs for the age estimation of bloodstains; however, further improvement of the existing model is needed. PIWI-interacting RNAs (PiRNAs), which are 24–32 nt noncoding small RNA molecules involved in the PIWI-piRNA pathway, play an important role in the aging process. In this study, we explored the possibility of simultaneously analyzing piRNAs and miRNAs for better age estimation purpose. Through massively parallel sequencing, five age-related piRNAs were identified in blood samples that had been stored for eight years. Further real-time PCR analysis revealed that two piRNAs (piR-000753 and piR-020548) showed relatively higher efficiency in age estimation. Additionally, two age-related miRNAs (miR-324-3p and miR-330-5p) were used to build the estimation model. Among all algorithms tested, gradient boosting showed the lowest mean absolute error (MAE) and root mean square error (RMSE) values (3.171 and 4.403 years, respectively) for the validation dataset (n = 110). The errors of the model were less than 5 years and 10 years for 81.82% and 96.36% of the samples, respectively. The results suggest that the combined use of piRNA and miRNA markers may increase the accuracy of age estimation, and our new model has great potential for application in forensic casework.

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Acknowledgements

This work was supported by National Natural Science Foundation of China (No. 82002006 and 82030058), Organization Department of Beijing Talents Project (2018400685627G339), the Open project of Shanghai Key Laboratory of Forensic Medicine, Key lab of Forensic Science, Ministry of Justice, China (Academy of Forensic Science)(KF202206) and Qingchuang Talents Induction Program of Shandong Higher Education Institution (2022 Forensic medicine innovation team).

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  1. Chen Fang and Peng Zhou contributed equally to this work.

Authors and Affiliations

  1. Shandong Provincial Hospital Affiliated to Shandong First Medical University, Jinan, 250021, People’s Republic of China

    Chen Fang, Peng Zhou, Min Li, Chunjiang Yu, Deping Meng & Xiaoqun Xu

  2. Shanxi Medical University, 030001, Taiyuan, People’s Republic of China

    Jiangwei Yan

  3. Department of Pathology and Forensic Medicine, School of Clinical and Basic Medical Sciences, Shandong First Medical University& Shandong Academy of Medical Sciences, Jinan, 250117, People’s Republic of China

    Chen Fang, Peng Zhou, Ran Li, Jinghan Guo, Huixian Qiu, Jingjuan Zhang, Min Li, Chunjiang Yu, Deping Meng, Xiaoqun Xu & Jiangwei Yan

  4. Institute of Analysis and Testing, Beijing Academy of Science and Technology (Beijing Center for Physical and Chemical Analysis), Beijing, 100094, People’s Republic of China

    Xu Liu & Di Guan

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  1. Chen Fang
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Correspondence to Chen Fang or Jiangwei Yan.

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The study was Ethics Committee of Shandong First Medical University (approval number 2022–729).

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Fang, C., Zhou, P., Li, R. et al. Development of a novel forensic age estimation strategy for aged blood samples by combining piRNA and miRNA markers. Int J Legal Med 137, 1327–1335 (2023). https://doi.org/10.1007/s00414-023-03028-8

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