Integrated microRNA–protein plasma biomarkers improve discrimination of multiple system atrophy from Parkinson’s disease

Multiple system atrophy (MSA) is a fatal, rapidly progressive atypical parkinsonism that responds poorly to Parkinson’s disease (PD) medications, and diagnostic precision remains a critical unmet need. Earlier, accurate diagnosis would give patients realistic prognostic expectations and enable timely clinical-trial enrollment. We sought plasma biomarkers discriminating MSA from PD.

Plasma small RNA sequencing and extracellular vesicle proteomics in discovery cohorts, filtered by the Biomedical Oriented Logistic Dantzig (BOLD) selector, nominated two microRNAs (hsa-miR-520a-5p, hsa-miR-22-3p) and two proteins (LCAT, kallistatin). Candidates were quantified in independent cohorts by XENO-Q qPCR and sandwich ELISA, and multivariate logistic regression models including all pairwise interactions were trained (N = 65) and validated in an independently recruited testing cohort (MSA N = 22; PD N = 16). The combined microRNA–protein model achieved test-set AUC 0.813 (95% CI 0.654–0.971), with sensitivity 0.818 and specificity 0.750, outperforming microRNA-only (AUC 0.599) and protein-only (AUC 0.747) models and indicating complementary rather than redundant information.

The markers converge on pathways of glycerophospholipid metabolism, cholesterol homeostasis, myelin integrity, and inflammatory regulation, consistent with the oligodendrocyte pathology central to MSA. These exploratory findings show that a non-invasive plasma multi-analyte panel can differentiate MSA from PD with clinically meaningful accuracy, and support prospective validation in larger cohorts. The authors would like to express their sincere appreciation toward Mr.

Christian Hanson from Biophysics and Bioanalytical Technology Lab, Academia Sinica, for the consolidation and interpretation of EV characterization results; Mr. Yan-Ru Ju from the Institute of Biotechnology, National Taiwan University, for their support in statistical analysis and optimization of BOLD selector; Prof. Tang-Long Shen from Department of Plant Pathology and Microbiology, National Taiwan University, for his insightful discussion about the consolidation and interpretation of EV proteomics profiling; and Prof.

Chi-An Cheng for providing the high-throughput SIMOA platform and analysis. Teh-Cheng Wang, and the staff of the Fifth Core Lab, Department of Medical Research, National Taiwan University Hospital, for their technical support and insightful discussion throughout the study and for the tremendous help from members of the “Center for Parkinson & Movement Disorders” in assisting with patient assessment and inclusion/exclusion of sample collection. This work was supported by the National Science and Technology Council, Taiwan (grant numbers 109-2314-B-002-120-MY3 and 112-2314-B-002-163- to R.M.W.; 109-2311-B-002-024, 110-2311-B-002-032, and 111-2311-B-002-001 to S.P.L.; and MOST-111-2118-M-001-007-MY2 to F.K.H.P.); National Taiwan University (grant numbers NTU-108L880304, 107L880305, and NTU-AS-112L104306 to S.P.L.); National Taiwan University Hospital (grant number 111-UN0007 to R.M.W. and S.P.L.); Academia Sinica (grant number AS-IA-112-M03 to F.K.H.P.); and Mission MSA (grant number MSAC-2023-12-002 to S.P.L. and R.M.W.).

S.W.Y. was supported by the Ministry of Science and ICT, Ministry of Trade, Industry and Energy, Ministry of Health and Welfare, and Ministry of Food and Drug Safety, Republic of Korea (grant number RS-2022-00141618). The funders had no role in the study design; participant recruitment; data collection, analysis, or interpretation; preparation of the manuscript; or the decision to submit the manuscript for publication. Institute of Biotechnology, National Taiwan University, Taipei, Taiwan Hsiang-Hsuan Lin-Wang, Yi-Tzang Tsai, Chia-Chen Lu & Shau-Ping Lin Institute of Statistical Science, Academia Sinica, Taipei, Taiwan Jing-Wen Huang, Yan-Hong Chen & Frederick Kin Hing Phoa Department of Medicine, National Taiwan University Cancer Center, Taipei, Taiwan Graduate Institute of Clinical Medicine, College of Medicine, National Taiwan University, Taipei, Taiwan Department of Neurology, Center of Excellence for Parkinson Disease and Movement Disorders, National Taiwan University Hospital, Taipei, Taiwan Genomics and Systems Biology Degree Program, National Taiwan University and Academia Sinica, Taipei, Taiwan Yung-Tsai Chu, Pin-Jui Kung & Shau-Ping Lin Institute of Atomic and Molecular Sciences, Academia Sinica, Taipei, Taiwan Elisha Wei-Chun Ho & Charles Pin-Kuang Lai Department of Neurology, Taipei Medical University – Shuang Ho Hospital, New Taipei City, Taiwan Department of Neurology, School of Medicine, College of Medicine, Taipei Medical University, Taipei, Taiwan Department of Systems Biology, College of Life Science and Biotechnology, Yonsei University, Seoul, Korea Department of Cell Biology and Anatomy, National Cheng Kung University, Tainan, Taiwan Cancer Precision Medicine Center, Japanese Foundation for Cancer Research, Tokyo, Japan Department of Molecular and Cellular Medicine, Tokyo Medical University, Tokyo, Japan NTU Health Science and Wellness Research Center, Taipei, Taiwan Research Center for Developmental Biology and Regenerative Medicine, National Taiwan University, Taipei, Taiwan Agricultural Biotechnology Research Center, Academia Sinica, Taipei, Taiwan Correspondence to Ruey-Meei Wu or Shau-Ping Lin.