Increased Osteoclast Activity Contributes to Bone Resorption and Osteopenia in a Rett Syndrome Mouse Model

ABSTRACT Rett syndrome is a severe neurodevelopmental disorder caused predominantly by loss-of-function mutations in the X-linked gene MECP2 . Besides a vast array of neurological and physiological impairments, patients also frequently develop severe osteopenia with increased fracture risk, however, the mechanisms underlying these skeletal defects are not completely understood. Previous work in Mecp2 -null mouse models has suggested that osteopenia is mainly due to impaired osteoblast function and reduced bone formation. Here, we examined bone mass, microarchitecture, and remodeling parameters in a Mecp2 -null mouse model during postnatal development, with a particular focus on osteoclast involvement. Micro-computed tomography and histomorphometric analyses showed reduced bone mineral density and trabecular bone volume, associated with increased trabecular separation and cortical thinning. These structural alterations were accompanied by increased osteoclast number per bone surface, elevated urinary deoxypyridinoline, and higher expression of osteoclast-associated genes, including Cathepsin K . Furthermore, gene expression analysis revealed an age-dependent shift in bone remodeling. At postnatal day 35, mutant mice showed reduced expression of Dlx5 and Dlx6 , consistent with low bone turnover. By postnatal day 55, Rankl and Cathepsin K were markedly upregulated, suggesting an increase in osteoclast resorptive activity, while key osteoblast markers and the RANKL/OPG ratio did not change significantly. A potential cell-autonomous contribution of Mecp2 to osteoclast maturation is also suggested by the analysis of public transcriptomic datasets on human osteoclast differentiation. Together, our findings identify increased osteoclast activity as a significant contributor to Rett-associated osteopenia and suggest that skeletal pathology in Mecp2 deficiency progresses from an early low-turnover state to a later phase of increased osteoclast resorption. HIGHLIGHTS What are the main findings. Mecp2 -null mice display reduced bone mass and altered bone microarchitecture during postnatal development, associated not only with reduced osteoblast activity, but also with increased osteoclast number, elevated urinary deoxypyridinoline, and increased expression of osteoclast-associated genes. Bone remodelling shows an age-dependent shift in Mecp2 deficiency, from an early low-turnover state at postnatal day 35 to increased osteoclast resorptive activity at postnatal day 55. What are the implications of the main findings? Rett-associated osteopenia is not explained solely by impaired osteoblast function, but also involves a significant osteoclast contribution to skeletal deterioration. These findings refine the pathophysiological model of bone involvement in Rett syndrome and support the idea that skeletal alterations evolve dynamically during disease progression.