Circulating plasma miR-23b-3p as a biomarker target for Idiopathic Parkinson’s disease: comparison with small extracellular vesicle miRNA

Sanskriti Rai¹ Prahalad S. Bharti¹ Rishabh Singh¹ Simran Rastogi¹ Komal Rani² Gyan P. Modi³ Krishna K. Inampudi¹ Hem C. Pandey⁴ Sanjay Yadav⁵ Roopa Rajan⁶ Fredrik Nikolajeff⁷ Saroj Kumar^{1, 8*}

• ¹Department of Biophysics, All India Institute of Medical Sciences, India
• ²Department of Pathology & Laboratory Medicine, All india institute of medical sciences, Bibinagar, India
• ³Department of Pharmaceutical Engineering & Technology, Indian Institute of Technology (BHU), India
• ⁴Department of Transfusion Medicine, All India Institute of Medical Sciences, India
• ⁵Department of Biochemistry, All India Institute of Medical Sciences Raebareli, India
• ⁶Department of Neurology, All India Institute of Medical Sciences, India
• ⁷Department of Health, Education and Technology, Luleå University of Technology, Sweden
• ⁸Luleå University of Technology, Sweden

Background: Parkinson’s disease (PD) is an increasingly common neurodegenerative condition, which causes movement dysfunction and a broad range of nonmotor symptoms. There is no molecular or biochemical diagnosis test for PD. The miRNAs are a class of small non-coding RNAs and are extensively studied owing to their altered expression in pathological states and facile harvesting and analysis techniques.
Methods: A total of 48 samples (16 each of PD, aged-matched and young controls) were recruited. The small extracellular vesicles (sEV) were isolated and validated using western blot, transmission electron microscope, and nanoparticle tracking analysis. Small RNA isolation, library preparation, and small RNA sequencing followed by differential expression and targeted prediction of miRNA were performed. The real-time PCR was performed with the targeted miRNA on PD, aged-matched, and young healthy control of plasma and plasma-derived sEV to demonstrate their potential as a diagnostic biomarker.
Results: In RNA sequencing, we identified 14.89% up-regulated (fold change 1.11 to 11.04, p<0.05) and 16.54% downregulated (fold change -1.04 to -7.28, p<0.05) miRNAs in PD and controls. Four differentially expressed miRNAs (miR-23b-3p, miR-29a-3p, miR-19b-3p, and miR-150-3p) were selected. The expression of miR-23b-3p was “upregulated” (p=0.002) in plasma whereas “downregulated” (p=0.0284) in plasma-derived sEVs in PD than age-matched controls. The ROC analysis of miR-23b-3p revealed better AUC values in plasma (AUC= 0.8086, p=0.0029) and plasma-derived sEVs (AUC= 0.7278, p= 0.0483) of PD and age-matched controls.
Conclusion: We observed an opposite expression profile of miR-23b-3p in PD and aged-matched healthy control in plasma and plasma-derived sEV fractions, where the expression of miR-23b-3p is increased in PD plasma while decreased in plasma-derived sEVs fraction. We further observed the different miR-23b-3p expression profiles in the young and aged-matched healthy control.