Hypothyroidism Reduces Myogenic Diversity and Impairs Skeletal Muscle Regeneration After Injury by Altering Myogenic and Non-Myogenic Pathways

Thyroid hormone signaling is an essential regulator of skeletal muscle development, function, and metabolism, yet the specific signaling pathways required for muscle regeneration are not yet defined. We utilized scRNA-seq and a cell cycle reporter mouse model (FUCCI, Fluorescent Ubiquitination-based Cell Cycle Indicator) to examine how hypothyroidism impacts repair processes after cardiotoxin-induced injury in mice. During regeneration, and up to 2 months after injury, hypothyroid muscles displayed smaller myofibers and a shift to slower oxidative fiber types.scRNA-seq of tibialis anterior muscle during regeneration revealed that hypothyroidism reduces myogenic lineage diversity. Cell cycle analysis confirmed delayed cell cycle progression at 5 and14 days after injury, with skeletal muscle stem cells stalled at the G1/S transition, hindering differentiation. Transcriptomic data revealed altered non-myogenic dynamics, including elevatedactivated fibro-adipogenic progenitors (FAPs) early in repair and persistent pro-inflammatory macrophages. Integrative regulon and ligand-receptor analysis further demonstrated that T3 acts through dual modes: a direct transcriptional control of myogenic cell cycle and oxidative programs and an indirect paracrine remodeling mediated by FAP and immune signaling networks. This study identifies novel effects of hypothyroidism on myogenic heterogeneity and reduced tissue repair, offering insights into muscle-wasting mechanisms relevant to hypothyroidism-associated myopathy and sarcopenia.