应用 RT-QuIC 检测在帕金森病患者的胃肠道中验证α-突触核蛋白的种籽活性：体内和尸检研究。

In vivo and autopsy validation of alpha-synuclein seeding activity using RT-QuIC assay in the gastrointestinal tract of patients with Parkinson's disease.

机构信息

Department of Neurology, Chungnam National University Sejong Hospital, Chungnam National University, 20, Bodeum 7-ro, Sejong-si, Republic of Korea; Department of Neurology, Chungnam National University College of Medicine, 282, Munhwa-ro, Jung-gu, Daejeon, Republic of Korea.

Laboratory Animal Center, Division of Research Strategy, Korea Brain Research Institute, Daegu, Republic of Korea.

出版信息

Parkinsonism Relat Disord. 2022 Oct;103:23-28. doi: 10.1016/j.parkreldis.2022.08.012. Epub 2022 Aug 17.

DOI:10.1016/j.parkreldis.2022.08.012

PMID:36029607

Abstract

OBJECTIVE

In the present study, real-time quaking-induced conversion (RT-QuIC) assay was used to evaluate pathologic alpha-synuclein (AS) seeding activity in formalin-fixed paraffin-embedded (FFPE) tissue from the gastrointestinal (GI) tract of Parkinson's disease (PD) patients.

METHODS

This study was conducted in two parts: Part I. a preliminary autopsy study that included four autopsy-confirmed patients with synucleinopathy (2 PD, 1 dementia with Lewy bodies [DLB], and 1 multiple system atrophy [MSA]) and two normal autopsy controls. Frozen and FFPE tissues of the brain were obtained. Part II. a clinical case-control study that included 20 clinically diagnosed PD patients and matched controls. Surgically resected FFPE tissues from the upper and lower GI tracts were used. The RT-QuIC assay was performed to evaluate pathologic seed amplification using frozen or FFPE tissues. The presence or absence of AS aggregation was confirmed by conventional phosphorylated AS (pAS) immunohistochemistry (IHC).

RESULTS

In Part I, RT-QuIC assay showed pathologic AS amplification in frozen and FFPE brain tissues of PD and DLB patients, and FFPE stomach tissue of PD patients but not in the MSA patient and controls. In Part II, pathologic seeding activity was found in 10% (2/20) of the stomach tissues of clinical PD patients but in none of the matched controls. IHC showed pAS-positive staining in 55% of patients (11/20) and 15% of controls (3/20).

CONCLUSION

The present study results showed that the RT-QuIC assay using FFPE tissue of the GI tract was inadequate as a biomarker in PD.

摘要

目的

本研究采用实时液芯震动转换（RT-QuIC）分析方法，评估帕金森病（PD）患者胃肠道（GI）组织福尔马林固定石蜡包埋（FFPE）样本中病理性α-突触核蛋白（AS）的种子生成活性。

方法

本研究分两部分进行：第一部分，包括 4 例经尸检证实的神经退行性疾病患者（2 例 PD、1 例路易体痴呆[DLB]和 1 例多系统萎缩[MSA]）和 2 例正常尸检对照的初步尸检研究。获取了脑的冷冻和 FFPE 组织。第二部分，包括 20 例临床诊断为 PD 的患者和匹配对照的临床病例对照研究。使用手术切除的上、下 GI 组织的 FFPE 样本。采用 RT-QuIC 分析方法，使用冷冻或 FFPE 组织评估病理性种子扩增。采用常规磷酸化 AS（pAS）免疫组织化学（IHC）检测 AS 聚集的存在或不存在。

结果

在第一部分中，RT-QuIC 分析显示 PD 和 DLB 患者的冷冻和 FFPE 脑组织以及 PD 患者的 FFPE 胃组织中存在病理性 AS 扩增，但 MSA 患者和对照组则没有。在第二部分中，发现 10%（20 例中的 2 例）临床 PD 患者的胃组织存在病理性种子活性，但对照组中没有。IHC 显示 55%（20 例中的 11 例）患者和 15%（20 例中的 3 例）对照存在 pAS 阳性染色。