Hyperactive BMP and Mechanosignaling Remodel Chromatin to Drive Aberrant Osteogenesis in FOP

Abstract Fibrodysplasia ossificans progressiva (FOP) is a rare genetic disorder in which a recurrent ACVR1 (R206H) mutation drives progressive heterotopic ossification (HO). While aberrant BMP hypersensitivity has been studied, how this mutation enforces a persistent pro-osteogenic state remains unclear. Here, we combined super-resolution stochastic optical reconstruction microscopy (STORM), transposase-accessible chromatin with sequencing (ATAC-Seq), and RNA sequencing (RNA-Seq) to investigate how Acvr1 R206H remodels chromatin to promote osteogenic transcriptional programs. Mutant mouse embryonic fibroblasts (MEFs) exhibited globally decondensed chromatin and increased accessibility at developmental and osteogenic loci enriched for HOX, TEAD, and RUNX motifs. Integration of ATAC-Seq and RNA-Seq data identified transcriptional networks primed for osteochondrogenic gene expression, including ossification, extracellular matrix organization, and cell adhesion pathways, consistent with enhanced BMP–SMAD and mechanotransduction activity. Time-course experiments revealed heightened responses to BMP ligands in Acvr1 R206H/+ MEFs compared to wild-type, highlighting ligand hypersensitivity. Importantly, pharmacological modulation showed that chromatin alterations were dynamic and reversible: activation of Rho/ROCK in wild-type cells reproduced the mutant chromatin state, while inhibition of Rho/ROCK or BMP–SMAD signaling restored condensation to wild-type levels in mutant cells. Together, these findings establish that Acvr1 R206H enforces a pro-osteogenic chromatin landscape through convergent BMP–SMAD and Rho/ROCK signaling, predisposing progenitors to aberrant differentiation trajectories. Our study reframes FOP as a disorder of persistent, but reversible, chromatin states and identifies novel therapeutic opportunities to restore mesenchymal cell homeostasis and prevent pathological bone formation. Significance Statement Fibrodysplasia ossificans progressiva (FOP) is a rare genetic disorder in which a mutation in ACVR1/ALK2 drives progressive heterotopic ossification. However, how this mutation enforces a persistent pro-osteogenic state is unclear. Here, we show that the Acvr1 R206H mutation remodels chromatin architecture and accessibility through hyperactive BMP–SMAD and Rho/ROCK signaling, activating transcription factor networks that drive osteochondrogenic gene expression. These chromatin changes are dynamic and reversible with targeted pathway inhibition, revealing therapeutic potential to restore mesenchymal cell plasticity and prevent pathological bone formation.