Exercise-associated changes of leukocyte gene expression in Stratin-associated myopathy - A case study

Biological Relevance and Intent of the Experiment Statin-associated muscle symptoms (SAMS) are among the most common adverse effects limiting the use of statin therapy, yet their underlying biological mechanisms remain poorly defined. While metabolic and structural causes have been proposed, increasing attention is being given to the possibility of immune system involvement in SAMS pathogenesis. Understanding whether and how immunological pathways contribute to SAMS is essential for improving diagnostic, preventive, and therapeutic strategies. The present study explores the immunological component of SAMS using a longitudinal, case-based transcriptomic approach. By examining gene expression in peripheral blood mononuclear cells (PBMCs) during controlled exercise challenges before, during, and after the development of SAMS, the study investigates how the immune response to physical stress is altered in the presence of statin-induced myopathy. The intent of this experiment is to: Identify immune-related gene expression changes associated with SAMS, Determine how these changes evolve across disease onset and recovery, Uncover potential biomarkers or immune pathways involved in the development and resolution of SAMS, Provide mechanistic insight into the role of the immune system in exercise-induced symptoms exacerbated by statin use. Experimental Workflow Participant Recruitment and Baseline Testing At baseline (Visit 1, prior to statin therapy), the participant underwent a standardized cardiopulmonary exercise test (CPX). Blood samples were collected at three timepoints around exercise: TP1: Pre-exercise TP2: Peak exercise TP3: One hour post-exercise Statin Therapy and Symptom Development After Visit 1, while the participant was on Statin medication, SAMS developed subsequently, marked by muscle discomfort and weakness. Follow-Up Testing During Symptom Phases The participant underwent repeat CPX testing and blood sampling: Visit 2: During active SAMS (symptomatic phase) Visit 3: After partial symptom resolution (recovery phase) For the same three timepoints (TP1–TP3) blood samples were collected at each visit. RNA Isolation and Sequencing PBMCs were isolated from all blood samples. RNA was extracted and subjected to next-generation RNA sequencing to profile transcriptomic changes. Data Analysis Differential Gene Expression Analysis was used to identify genes with altered expression between visits and timepoints. Weighted Gene Co-expression Network Analysis (WGCNA) was applied to detect co-regulated gene modules. Pathway and Gene Ontology Enrichment Analysis (e.g., GO, Reactome) was performed to interpret the biological functions of key modules and differentially expressed genes. Key Findings 39 co-expression modules were identified from WGCNA analysis. Several modules that responded to peak exercise in the healthy state (V1) showed blunted responses during SAMS (V2) and partial restoration during recovery (V3). 16 key genes that may contribute to the immune pathways associated with SAMS were identified.