Identification and validation of anti-protein arginine methyltransferase 5 (PRMT5) antibody as a novel biomarker for systemic sclerosis (SSc)

Minrui Liang; Lingbiao Wang; Xiaolong Tian; Kun Wang; Xiaoyi Zhu; Linlin Huang; Qing Li; Wenjing Ye; Chen Chen; Haihua Yang; Wanqing Wu; Xiangjun Chen; Xiaoxia Zhu; Yu Xue; Weiguo Wan; Yanling Wu; Liwei Lu; Jiucun Wang; Hejian Zou; Tianlei Ying; Feng Zhou

doi:10.1136/ard-2024-225596

Identification and validation of anti-protein arginine methyltransferase 5 (PRMT5) antibody as a novel biomarker for systemic sclerosis (SSc)

Ann Rheum Dis. 2024 May 8:ard-2024-225596. doi: 10.1136/ard-2024-225596. Online ahead of print.

Authors

Minrui Liang^#^{1

2

3}, Lingbiao Wang^#^{4

2

3}, Xiaolong Tian^#⁵, Kun Wang^#^{6

7}, Xiaoyi Zhu^#⁵, Linlin Huang^{4

2

3}, Qing Li^{6

7}, Wenjing Ye^{4

2

3}, Chen Chen^{4

2

8}, Haihua Yang⁹, Wanqing Wu¹⁰, Xiangjun Chen¹⁰, Xiaoxia Zhu^{4

2

3}, Yu Xue^{4

2

3}, Weiguo Wan^{4

2

3}, Yanling Wu⁵, Liwei Lu¹¹, Jiucun Wang¹², Hejian Zou^{4

2

3}, Tianlei Ying¹³, Feng Zhou^{14

7}

Affiliations

¹ Department of Rheumatology, Huashan Hospital, Fudan University, Shanghai, China mliang10@fudan.edu.cn tlying@fudan.edu.cn zhou_feng@fudan.edu.cn.
² Institute of Rheumatology, Immunology and Allergy, Fudan University, Shanghai, China.
³ Huashan Rare Disease Center, Huashan Hospital, Fudan University, Shanghai, China.
⁴ Department of Rheumatology, Huashan Hospital, Fudan University, Shanghai, China.
⁵ Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS) and Shanghai Institute of Infectious Disease and Biosecurity, Shanghai Frontiers Science Center of Pathogenic Microorganisms and Infection, Shanghai Engineering Research Center for Synthetic Immunology, School of Basic Medical Sciences, Fudan University, Shanghai, China.
⁶ Shanghai Key Laboratory of Medical Epigenetics, International Co-laboratory of Medical Epigenetics and Metabolism, Ministry of Science and Technology, Institutes of Biomedical Sciences, Fudan University, Shanghai, China.
⁷ Key Laboratory of Carcinogenesis and Cancer Invasion, Ministry of Education, Liver Cancer Institute, Zhongshan Hospital, Fudan University, Shanghai, China.
⁸ Department of Emergency Medicine, Zhongshan Hospital, Fudan University, Shanghai, China.
⁹ Department of Respiratory and Critical Care Medicine, Huashan Hospital, Fudan University, Shanghai, China.
¹⁰ Department of Neurology, Huashan Hospital, Fudan University, Shanghai, China.
¹¹ Department of Pathology, The University of Hong Kong, Hong Kong, China.
¹² State Key Laboratory of Genetic Engineering, Collaborative Innovation Center for Genetics and Development, School of Life Sciences, and Human Phenome Institute, Fudan University, Shanghai, China.
¹³ Key Laboratory of Medical Molecular Virology (MOE/NHC/CAMS) and Shanghai Institute of Infectious Disease and Biosecurity, Shanghai Frontiers Science Center of Pathogenic Microorganisms and Infection, Shanghai Engineering Research Center for Synthetic Immunology, School of Basic Medical Sciences, Fudan University, Shanghai, China mliang10@fudan.edu.cn tlying@fudan.edu.cn zhou_feng@fudan.edu.cn.
¹⁴ Shanghai Key Laboratory of Medical Epigenetics, International Co-laboratory of Medical Epigenetics and Metabolism, Ministry of Science and Technology, Institutes of Biomedical Sciences, Fudan University, Shanghai, China mliang10@fudan.edu.cn tlying@fudan.edu.cn zhou_feng@fudan.edu.cn.

^# Contributed equally.

PMID: 38684324
DOI: 10.1136/ard-2024-225596

Abstract

Objectives: In the complex panorama of autoimmune diseases, the characterisation of pivotal contributing autoantibodies that are involved in disease progression remains challenging. This study aimed to employ a global antibody profiling strategy to identify novel antibodies and investigate their association with systemic sclerosis (SSc).

Methods: We implemented this strategy by conducting immunoprecipitation (IP) following on-bead digestion with the sera of patients with SSc or healthy donors, using antigen pools derived from cell lysates. The enriched antigen-antibody complex was proceeded with mass spectrometry (MS)-based quantitative proteomics and over-represented by bioinformatics analysis. The candidate antibodies were then orthogonally validated in two independent groups of patients with SSc. Mice were immunised with the target antigen, which was subsequently evaluated by histological examination and RNA sequencing.

Results: The IP-MS analysis, followed by validation in patients with SSc, revealed a significant elevation in anti-PRMT5 antibodies among patients with SSc. These antibodies exhibited robust diagnostic accuracy in distinguishing SSc from healthy controls and other autoimmune conditions, including systemic lupus erythematosus and Sjögren's syndrome, with an area under the curve ranging from 0.900 to 0.988. The elevation of anti-PRMT5 antibodies was verified in a subsequent independent group with SSc using an additional method, microarray. Notably, 31.11% of patients with SSc exhibited seropositivity for anti-PRMT5 antibodies. Furthermore, the titres of anti-PRMT5 antibodies demonstrated a correlation with the progression or regression trajectory in SSc. PRMT5 immunisation displayed significant inflammation and fibrosis in both the skin and lungs of mice. This was concomitant with the upregulation of multiple proinflammatory and profibrotic pathways, thereby underscoring a potentially pivotal role of anti-PRMT5 antibodies in SSc.

Conclusions: This study has identified anti-PRMT5 antibodies as a novel biomarker for SSc.

Keywords: Autoantibodies; Autoimmune Diseases; Scleroderma, Systemic.