Shoag Jonathan E, Srinivasa Amoolya, Loh Caitlin A, Liu Mei Hong, Lassen Emilie, Melanaphy Shana, Costa Benjamin M, Grońska-Pęski Marta, Jabara Nisrine T, Picciotto Shany, Choi Una, Bohorquez Anyull D, Barbieri Christopher E, Callum Pamela, Skytte Anne-Bine, Evrony Gilad D
Department of Urology, University Hospitals Cleveland Medical Center, Case Western Reserve University School of Medicine, Cleveland, OH, USA.
Center for Human Genetics and Genomics, New York University Grossman School of Medicine, New York, NY, USA.
Nat Commun. 2025 Mar 15;16(1):2546. doi: 10.1038/s41467-025-57507-0.
Mutations that accumulate in the human male germline with age are a major driver of genetic diversity and contribute to genetic diseases. However, aging-related male germline mutation rates have not been measured directly in germline cells (sperm) at the level of individuals. We developed a study design in which we recalled 23 sperm donors with prior banked samples to provide new sperm samples. The old and new sequential sperm samples were separated by long timespans, ranging from 10 to 33 years. We profiled these samples by high-fidelity duplex sequencing and demonstrate that direct high-fidelity sequencing of sperm yields cohort-wide mutation rates and patterns consistent with prior family-based (trio) studies. In every individual, we detected an increase in sperm mutation burden between the two sequential samples, yielding individual-specific measurements of germline mutation rate. Deep whole-genome sequencing of sequential sperm samples from two individuals followed by targeted validation measured remarkably stable mosaicism of clonal mutations that likely arose during embryonic and germline development, suggesting that age did not substantially impact the diversity of spermatogonial stem cell pools in these individuals. Our application of high-fidelity and deep whole-genome sequencing to sequential sperm samples provides insight into aging-related mutation processes in the male germline.
随着年龄增长在人类男性生殖系中积累的突变是遗传多样性的主要驱动因素，并导致遗传疾病。然而，与衰老相关的男性生殖系突变率尚未在个体水平的生殖系细胞（精子）中直接测量。我们设计了一项研究，召回了23名之前储存过样本的精子捐献者，以提供新的精子样本。新旧连续精子样本之间间隔了很长时间，从10年到33年不等。我们通过高保真双链测序对这些样本进行了分析，并证明对精子进行直接高保真测序可得出与先前基于家系（三联体）研究一致的队列范围的突变率和模式。在每个个体中，我们检测到两个连续样本之间精子突变负担增加，得出了个体特异性的生殖系突变率测量结果。对两名个体的连续精子样本进行深度全基因组测序，随后进行靶向验证，测量到在胚胎和生殖系发育过程中可能出现的克隆突变具有显著稳定的嵌合性，这表明年龄对这些个体中生精干细胞库的多样性没有实质性影响。我们将高保真和深度全基因组测序应用于连续精子样本，为男性生殖系中与衰老相关的突变过程提供了见解。
