Ultrasensitive detection of aggregatedα-synuclein using quiescent seed amplification assay for the diagnosis of Parkinson’s disease  

作　　者：Hengxu Mao Yaoyun Kuang Du Feng Xiang Chen Lin Lu Wencheng Xia Tingting Gan Weimeng Huang Wenyuan Guo Hancun Yi Yirong Yang Zhuohua Wu Wei Dai Hui Sun Jieyuan Wu Rui Zhang Shenqing Zhang Xiuli Lin Yuxuan Yong Xinling Yang Hongyan Li Wenjun Wu Xiaoyun Huang Zhaoxiang Bian Hoi Leong Xavier Wong Xin-Lu Wang Michael Poppell Yi Ren Cong Liu Wen-Quan Zou Shengdi Chen Ping-Yi Xu 

机构地区：[1]Department of Neurology,the First Affiliated Hospital of Guangzhou Medi-cal University,Guangzhou 510120,China [2]School of Basic Medical Science,Guangzhou Medical University,Guangzhou 511436,China [3]Interdisciplinary Research Center on Biology and Chemistry,Shanghai Institute of Organic Chemistry,Chinese Academy of Sciences,Shanghai 201210,China [4]Deptartment of Neurology,The First Affiliated Hospital of Gannan Medical University,Ganzhou 341000,China [5]Institute of Neurology,Jiangxi Academy of Medical Clinical Sciences,The First Affiliated Hospital,Jiangxi Medical College,Nan-chang University,Nanchang 330006,China [6]Bio-X Institutes,Key Laboratory for the Genetics of Developmental and Neuropsychiatric Disorders(Ministry of Education),Shanghai Jiao Tong University,Shanghai 200030,China [7]The Second Affiliated Hospital of Xinjiang Medical University,Urumqi 830054,China [8]Department of Neurology,Xinjiang Uygur Autonomous Region People’s Hospital,Urumqi 830054,China [9]Department of Neurology,Zhongshan City People’s Hospital,Zhongshan 528400,China [10]Dongguan Songshan Lake Central Hospital,Dongguan 523000,China [11]Jockey Club School of Chinese Medicine,Baptist University Road,Hong Kong 999077,China [12]Department of Nuclear Medicine,the First Affiliated Hospital of Guangzhou Medical University,Guangzhou 510120,China [13]Department of Biomedical Sciences,College of Medicine,Florida State University,Tallahassee 32306,USA [14]State Key Laboratory of Chemical Biology,Shanghai Institute of Organic Chemistry,Chinese Academy of Sciences,Shanghai 200032,China [15]Department of Neurology,Rui Jin Hospital Affiliated to Shanghai Jiao Tong University School of Medicine,Shanghai 200025,China

出　　处：《Translational Neurodegeneration》2024年第1期603-618,共16页转化神经变性病(英文)

基　　金：supported by National Natural Science Foundation of China(81870856,81870992,82071416,82071417,82188101,32171236);Central Government Guiding Local Science and Technology Development Projects(ZYYD2022C17);Key Research and Development Program of Guangzhou(2023B03J0631);Municipal University(Faculty)Joint Funding Project(202102010010);Municipal and University(Institute)Jointly Funded Project for Basic and Applied Basic Research(202201020472);Guangdong Basic and Applied Basic Research Foundation(2021B1515140026,2022B1515230004);the Science and Technology Commission of Shanghai Municipality(STCSM)(20XD1425000,2019SHZDZX02 and 22JC1410400).

摘　　要：Background Seed amplification assays(SAA)enable the amplification of pathological misfolded proteins,includ‑ingα-synuclein(αSyn),in both tissue homogenates and body fluids of Parkinson’s disease(PD)patients.SAA involves repeated cycles of shaking or sonication coupled with incubation periods.However,this amplification scheme has limitations in tracking protein propagation due to repeated fragmentation.Methods We introduced a modified form of SAA,known as Quiescent SAA(QSAA),and evaluated biopsy and autopsy samples from individuals clinically diagnosed with PD and those without synucleinopathies(control group).Brain biopsy samples were obtained from 14 PD patients and 6 controls without synucleinopathies.Addition‑ally,skin samples were collected from 214 PD patients and 208 control subjects.Data were analyzed from April 2019 to May 2023.Results QSAA successfully amplifiedαSyn aggregates in brain tissue sections from mice inoculated with pre-formed fibrils.In the skin samples from 214 PD cases and 208 non-PD cases,QSAA demonstrated high sensitivity(90.2%)and specificity(91.4%)in differentiating between PD and non-PD cases.Notably,moreαSyn aggregates were detected by QSAA compared to immunofluorescence with the pS129-αSyn antibody in consecutive slices of both brain and skin samples.Conclusion We introduced the new QSAA method tailored for in situ amplification ofαSyn aggregates in brain and skin samples while maintaining tissue integrity,providing a streamlined approach to diagnosing PD with individual variability.The integration of seeding activities with the location of deposition ofαSyn seeds advances our understanding of the mechanism underlyingαSyn misfolding in PD.

关 键 词：Α-SYNUCLEIN Seed amplification assay Quiescent SAA Parkinson’s disease Seeding activity 

分 类 号：R742.5[医药卫生—神经病学与精神病学]