Zhai Ting, Mazzucato Patrizia, Ricciardi Catherine, Christiani David C, Liang Liming, Samson Leona D, Chaim Isaac A, Nagel Zachary D
Department of Environmental Health, Harvard T.H. Chan School of Public Health, Boston, MA, 02115, USA.
Department of Biological Engineering, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA.
medRxiv. 2025 Jun 16:2025.06.13.25329369. doi: 10.1101/2025.06.13.25329369.
Rare genetic DNA repair deficiency syndromes can lead to immunodeficiency, neurological disorders, and cancer. In the general population, inter-individual variation in DNA repair capacity (DRC) influences susceptibility to cancer and several age-related diseases. Genome wide association studies and functional analyses show that defects in multiple DNA repair pathways jointly increase disease risk, but previous technologies did not permit comprehensive analyses of DNA repair in populations. To overcome these limitations, we used fluorescence multiplex host cell reactivation (FM-HCR) assays that directly quantify DRC across six major DNA repair pathways. We assessed DRC in phytohemagglutinin-stimulated primary lymphocytes from 56 healthy individuals and validated assay reproducibility in 10 individuals with up to five independent blood draws. We furthermore developed generalized analytical pipelines for systematically adjusting for batch effects and both experimental and biological confounders. Our results reveal significant inter-individual variation in DRC for each of 10 reporter assays that measure the efficiency of distinct repair processes. Our data also demonstrate that correlations between the activities of different DNA repair pathways are relatively weak. This finding suggests that each pathway may independently influence susceptibility to the health effects of DNA damage. We furthermore developed a pipeline for analyzing comet repair kinetics and related our new functional data to previously reported comet assay data for the same individuals. Our pioneering analysis underscores the sensitivity of FM-HCR assays for detecting subtle biological differences between individuals and establishes standardized methodologies for population studies. Our findings and open source analytical tools advance precision medicine by enabling comprehensive exploration of genetic, demographic, clinical, and lifestyle factors and supporting targeted interventions to enhance DNA repair and maintain genomic integrity, thereby promoting personalized healthcare and disease prevention.
罕见的遗传性DNA修复缺陷综合征可导致免疫缺陷、神经紊乱和癌症。在普通人群中,个体间DNA修复能力(DRC)的差异会影响患癌易感性以及几种与年龄相关的疾病。全基因组关联研究和功能分析表明,多种DNA修复途径的缺陷共同增加了疾病风险,但以往的技术无法对人群中的DNA修复进行全面分析。为了克服这些局限性,我们使用了荧光多重宿主细胞再激活(FM-HCR)检测方法,该方法可直接量化六种主要DNA修复途径的DRC。我们评估了56名健康个体经植物血凝素刺激的原代淋巴细胞中的DRC,并在10名个体中进行了多达5次独立采血,验证了检测的可重复性。此外,我们还开发了通用分析流程,以系统地调整批次效应以及实验和生物学混杂因素。我们的结果显示,在测量不同修复过程效率的10种报告基因检测中,每种检测的DRC在个体间都存在显著差异。我们的数据还表明,不同DNA修复途径的活性之间的相关性相对较弱。这一发现表明,每条途径可能独立影响对DNA损伤健康影响的易感性。此外,我们还开发了一个分析彗星修复动力学的流程,并将我们的新功能数据与之前报道的同一批个体的彗星试验数据相关联。我们的开创性分析强调了FM-HCR检测在检测个体间细微生物学差异方面的敏感性,并建立了人群研究的标准化方法。我们的研究结果和开源分析工具通过全面探索遗传、人口统计学、临床和生活方式因素,并支持针对性干预以增强DNA修复和维持基因组完整性,从而推动个性化医疗和疾病预防,推进了精准医学的发展。
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