Daida Kensuke, Yoshino Hiroyo, Malik Laksh, Baker Breeana, Ishiguro Mayu, Genner Rylee, Paquette Kimberly, Li Yuanzhe, Nishioka Kenya, Masuzugawa Satoshi, Hirano Makito, Takahashi Kenta, Kolmogorov Mikhail, Billingsley Kimberley J, Funayama Manabu, Blauwendraat Cornelis, Hattori Nobutaka
Integrative Neurogenomics Unit, Laboratory of Neurogenetics, National Institute on Aging, National Institutes of Health, Bethesda, MD, USA.
Center for Alzheimer's and Related Dementias (CARD), National Institute on Aging and National Institute of Neurological Disorders and Stroke, National Institutes of Health, Bethesda, MD, USA.
Ann Neurol. 2025 Apr;97(4):753-765. doi: 10.1002/ana.27155. Epub 2024 Dec 19.
Variants in PRKN and PINK1 are the leading cause of early-onset autosomal recessive Parkinson's disease, yet many cases remain genetically unresolved. We previously identified a 7 megabases complex structural variant in a pair of monozygotic twins using Oxford Nanopore Technologies (ONT) long-read sequencing. This study aims to determine if ONT long-read sequencing can detect a second variant in other unresolved early-onset Parkinson's disease (EOPD) cases with 1 heterozygous PRKN or PINK1 variant.
ONT long-read sequencing was performed on EOPD patients with 1 reported PRKN/PINK1 pathogenic variant, with onset age under 50. Positive controls included EOPD patients with 2 known PRKN pathogenic variants. Initial testing involved short-read targeted panel sequencing for single nucleotide variants and multiplex ligation-dependent probe amplification for copy number variants.
A total of 47 patients were studied (PRKN "one-variant," n = 23; PINK1 "one-variant," n = 12; PRKN "two-variants," n = 12). ONT long-read sequencing identified a second pathogenic variant in 26% of PRKN "one-variant" patients (6/23), but none in PINK1 "one-variant" patients (0/12). Detected variants included 1 complex inversion, 2 structural variant overlaps, and 3 duplications. In the PRKN "two-variants" group, both variants were identified in all patients (100%, 12/12).
ONT long-read sequencing effectively identifies pathogenic structural variants in the PRKN locus missed by conventional methods. It should be considered for unresolved EOPD cases when a second variant is not detected through conventional approaches. ANN NEUROL 2025;97:753-765.
PRKN和PINK1基因的变异是早发性常染色体隐性帕金森病的主要病因，但许多病例在基因层面仍未得到解决。我们之前使用牛津纳米孔技术（ONT）长读长测序在一对同卵双胞胎中鉴定出一个7兆碱基的复杂结构变异。本研究旨在确定ONT长读长测序能否在其他未解决的早发性帕金森病（EOPD）病例中检测到第二个变异，这些病例已有1个杂合的PRKN或PINK1变异。
对发病年龄在50岁以下、已报告有1个PRKN/PINK1致病变异的EOPD患者进行ONT长读长测序。阳性对照包括有2个已知PRKN致病变异的EOPD患者。初始检测包括针对单核苷酸变异的短读长靶向测序和针对拷贝数变异的多重连接依赖探针扩增。
共研究了47例患者（PRKN“单变异”组，n = 23；PINK1“单变异”组，n = 12；PRKN“双变异”组，n = 12）。ONT长读长测序在26%的PRKN“单变异”患者（6/23）中鉴定出第二个致病变异，但在PINK1“单变异”患者中未发现（0/12）。检测到的变异包括1个复杂倒位、2个结构变异重叠和3个重复。在PRKN“双变异”组中，所有患者均鉴定出两个变异（100%，12/12）。
ONT长读长测序有效地鉴定出了传统方法遗漏的PRKN基因座中的致病变异。当通过传统方法未检测到第二个变异时，对于未解决的EOPD病例应考虑使用该方法。《神经病学纪事》2025年；97：753 - 765。
