Single-fiber morphometry and spatial transcriptomics reveal selective oxidative muscle fiber atrophy in...

Cancer-related fatigue is the most common and persistent symptom in breast cancer, with fatigue reported up to 10 years post-diagnosis. Unlike many cancers, fatigue in breast cancer often arises during early-stage disease in the absence of cachexia. While many factors contribute to fatigue, the direct contribution of cancer-associated skeletal muscle pathology remains poorly understood. Here we analyzed pectoralis major muscle biopsies from individuals with non-metastatic breast cancer and non-cancer controls using single-fiber morphometry and spatial transcriptomics. We identified fiber-type-specific structural alterations and spatially localized transcriptional reprogramming within the muscle microenvironment. Single-fiber morphometry revealed selective atrophy of oxidative type I and type IIa muscle fibers, while glycolytic type IIx fibers were relatively preserved. Concordant spatial transcriptomic profiling revealed suppression of oxidative metabolic programs, evidence of mitochondrial dysfunction, and spatially localized catabolic signaling originating from intramuscular adipocytes. This study introduces an integrated framework for profiling skeletal muscle architecture and spatially localized gene expression in surgically obtained muscle biopsies and represents the first application of spatial transcriptomics to human skeletal muscle from individuals with cancer. These findings demonstrate structural and metabolic remodeling of skeletal muscle in non-metastatic breast cancer and suggest targeting muscle metabolism represents a promising therapeutic strategy for cancer-related fatigue.