PDF(Pubmed)
Abstract：
UNASSIGNED: Mutations within the genes PRKN and PINK1 are the leading cause of early onset autosomal recessive Parkinson\'s disease (PD). However, the genetic cause of most early-onset PD (EOPD) cases still remains unresolved. Long-read sequencing has successfully identified many pathogenic structural variants that cause disease, but this technology has not been widely applied to PD. We recently identified the genetic cause of EOPD in a pair of monozygotic twins by uncovering a complex structural variant that spans over 7 Mb, utilizing Oxford Nanopore Technologies (ONT) long-read sequencing. In this study, we aimed to expand on this and assess whether a second variant could be detected with ONT long-read sequencing in other unresolved EOPD cases reported to carry one heterozygous variant in PRKN or PINK1.
UNASSIGNED: ONT long-read sequencing was performed on patients with one reported PRKN/PINK1 pathogenic variant. EOPD patients with an age at onset younger than 50 were included in this study. As a positive control, we also included EOPD patients who had already been identified to carry two known PRKN pathogenic variants. Initial genetic testing was performed using either short-read targeted panel sequencing for single nucleotide variants and multiplex ligation-dependent probe amplification (MLPA) for copy number variants.
UNASSIGNED: 48 patients were included in this study (PRKN \"one-variant\" n = 24, PINK1 \"one-variant\" n = 12, PRKN \"two-variants\" n = 12). Using ONT long-read sequencing, we detected a second pathogenic variant in six PRKN \"one-variant\" patients (26%, 6/23) but none in the PINK1 \"one-variant\" patients (0%, 0/12). Long-read sequencing identified one case with a complex inversion, two instances of structural variant overlap, and three cases of duplication. In addition, in the positive control PRKN \"two-variants\" group, we were able to identify both pathogenic variants in PRKN in all the patients (100%, 12/12).
UNASSIGNED: This data highlights that ONT long-read sequencing is a powerful tool to identify a pathogenic structural variant at the PRKN locus that is often missed by conventional methods. Therefore, for cases where conventional methods fail to detect a second variant for EOPD, long-read sequencing should be considered as an alternative and complementary approach.
UNASSIGNED: ONT long-read sequencing was performed on patients with one reported PRKN/PINK1 pathogenic variant. EOPD patients with an age at onset younger than 50 were included in this study. As a positive control, we also included EOPD patients who had already been identified to carry two known PRKN pathogenic variants. Initial genetic testing was performed using either short-read targeted panel sequencing for single nucleotide variants and multiplex ligation-dependent probe amplification (MLPA) for copy number variants.
UNASSIGNED: 48 patients were included in this study (PRKN \"one-variant\" n = 24, PINK1 \"one-variant\" n = 12, PRKN \"two-variants\" n = 12). Using ONT long-read sequencing, we detected a second pathogenic variant in six PRKN \"one-variant\" patients (26%, 6/23) but none in the PINK1 \"one-variant\" patients (0%, 0/12). Long-read sequencing identified one case with a complex inversion, two instances of structural variant overlap, and three cases of duplication. In addition, in the positive control PRKN \"two-variants\" group, we were able to identify both pathogenic variants in PRKN in all the patients (100%, 12/12).
UNASSIGNED: This data highlights that ONT long-read sequencing is a powerful tool to identify a pathogenic structural variant at the PRKN locus that is often missed by conventional methods. Therefore, for cases where conventional methods fail to detect a second variant for EOPD, long-read sequencing should be considered as an alternative and complementary approach.
摘要：
■基因PRKN和PINK1内的突变是早发性常染色体隐性遗传帕金森病（PD）的主要原因。然而,大多数早发性PD（EOPD）病例的遗传原因仍未解决。长读测序已成功鉴定出许多致病结构变异，但该技术尚未广泛应用于PD。我们最近通过发现跨越7Mb的复杂结构变体，在一对单卵双胞胎中确定了EOPD的遗传原因，利用牛津纳米孔技术(ONT)长读测序。在这项研究中,我们旨在对此进行扩展,并评估在其他未解决的EOPD病例中是否可以通过ONT长读数测序检测到第二个变异,这些病例报道在PRKN或PINK1中携带一个杂合变异.
■对有1例PRKN/PINK1致病性变异的患者进行ONT长读测序。本研究包括发病年龄小于50岁的EOPD患者。作为阳性对照,我们还纳入了已被鉴定携带两种已知PRKN致病变种的EOPD患者.使用针对单核苷酸变体的短读取靶向组测序和针对拷贝数变体的多重连接依赖性探针扩增(MLPA)进行初始遗传测试。
■48例患者纳入本研究(PRKN\"单变异\"n=24,PINK1\"单变异\"n=12,PRKN\"双变异\"n=12)。使用ONT长读取测序，我们在6名PRKN“单变异”患者中检测到第二个致病变异(26%，6/23），但在PINK1“单变异”患者中没有（0%，0/12）。长读数测序确定了一个具有复杂反转的病例，结构变体重叠的两个实例，和三个重复的情况。此外,在阳性对照PRKN“双变”组中，我们能够在所有患者中鉴定出PRKN的两种致病变异(100%，12/12).
■该数据突出表明，ONT长读测序是一种强大的工具，可用于识别PRKN基因座上的致病性结构变异，而常规方法通常会错过这种变异。因此,对于常规方法无法检测到EOPD的第二种变体的情况，长读数测序应被视为一种替代和补充方法.
■对有1例PRKN/PINK1致病性变异的患者进行ONT长读测序。本研究包括发病年龄小于50岁的EOPD患者。作为阳性对照,我们还纳入了已被鉴定携带两种已知PRKN致病变种的EOPD患者.使用针对单核苷酸变体的短读取靶向组测序和针对拷贝数变体的多重连接依赖性探针扩增(MLPA)进行初始遗传测试。
■48例患者纳入本研究(PRKN\"单变异\"n=24,PINK1\"单变异\"n=12,PRKN\"双变异\"n=12)。使用ONT长读取测序，我们在6名PRKN“单变异”患者中检测到第二个致病变异(26%，6/23），但在PINK1“单变异”患者中没有（0%，0/12）。长读数测序确定了一个具有复杂反转的病例，结构变体重叠的两个实例，和三个重复的情况。此外,在阳性对照PRKN“双变”组中，我们能够在所有患者中鉴定出PRKN的两种致病变异(100%，12/12).
■该数据突出表明，ONT长读测序是一种强大的工具，可用于识别PRKN基因座上的致病性结构变异，而常规方法通常会错过这种变异。因此,对于常规方法无法检测到EOPD的第二种变体的情况，长读数测序应被视为一种替代和补充方法.
