Davis Wachtell

The TRPV4 skeletal dysplasias are characterized by short stature, short limbs with prominent large joints, and progressive scoliosis. They result from dominant missense mutations that activate the TRPV4 calcium permeable ion channel. As a platform to understand the mechanism of disease and to test the hypothesis that channel inhibition could treat these disorders, we developed a knock-in mouse that conditionally expresses the p.R594H Trpv4 mutation. Embryonic, chondrocyte-specific induction of the mutation using Col2a1-Cre resulted in a skeletal dysplasia affecting the long bones, spine, and craniofacial skeletal elements, consistent with the human skeletal dysplasia phenotypes produced by TRPV4 mutations. Cartilage growth plate histological abnormalities included disorganized proliferating chondrocyte columns and reduced hypertrophic chondrocyte development, reflecting abnormal endochondral ossification. In vivo treatment with the TRPV4-specific inhibitor GSK2798745 markedly improved the radiographic skeletal phenotype and rescued the growth plate histological abnormalities. ScRNA-Seq of chondrocyte transcripts from affected mice identified calcium-mediated effects on multiple signaling pathways as potential mechanisms underlying the defects in linear and cartilage appositional growth observed in both mutant mice and patients. These results provide preclinical evidence demonstrating TRPV4 inhibition as a rational, mechanism-based therapeutic strategy to ameliorate disease progression and severity in the TRPV4 skeletal dysplasias.

Osteogenesis imperfecta, a genetically heterogeneous disorder, is characterized by brittle bones with recurrent fractures as well as short stature. A recent study investigating cellular signaling in growth plate chondrocytes identified an increased expression of FGF receptors and elevated FGF signaling. We inhibited FGF receptor activation with the small molecule infigratinib in growing Aga2/+ mice and then characterized linear growth, cartilage growth plate histology, and bone morphometrics. Infigratinib treatment resulted in increased female femur length, increased growth plate lengths in both sexes, specifically in the proliferative zone, and reduced phospho-ERK1/2 as well as SOX9 expression in lower hypertrophic chondrocytes. Infigratinib had no impact on cortical bone but reduced male trabecular bone parameters in Aga2/+ mice. Three point bending biomechanical tests showed that Aga2/+-treated males had increased bone strength for some parameters, but reduced bone strength in treated WT males, correlating with the negative impact on trabecular bone parameters. WT and Aga2-treated male mice also showed increased serum testosterone levels. Together, these data establish FGFR as a therapeutic target for short stature in osteogenesis imperfecta and identify potential negative effects of pan-FGFR inhibition, particularly in the male skeleton.

Osteogenesis imperfecta (OI), or brittle bone disease, is a disorder characterized by bone fragility and increased fracture incidence. All forms of OI also feature short stature, implying an effect on endochondral ossification. Using the Aga2 ^+/– mouse, which has a mutation in type I collagen, we show an affected growth plate primarily due to a shortened proliferative zone. We used single-cell RNA-Seq analysis of tibial and femoral growth plate tissues to understand transcriptional consequences on growth plate cell types. We show that perichondrial cells, which express abundant type I procollagen, and growth plate chondrocytes, which were found to express low amounts of type I procollagen, had ER stress and dysregulation of the same unfolded protein response pathway as previously demonstrated in osteoblasts. Aga2 ^+/– proliferating chondrocytes showed increased FGF and MAPK signaling, findings consistent with accelerated differentiation. There was also increased Sox9 expression throughout the growth plate, which is expected to accelerate early chondrocyte differentiation but reduce late hypertrophic differentiation. These data reveal that mutant type I collagen expression in OI has an impact on the cartilage growth plate. These effects on endochondral ossification indicate that OI is a biologically complex phenotype going beyond its known impacts on bone to negatively affect linear growth.

Keeping track of other people’s gaze is an essential task in social cognition and key for successfully reading other people’s intentions and beliefs (theory of mind). Recent behavioral evidence suggests that we construct an implicit model of other people’s gaze, which may incorporate physically incoherent attributes such as a construct of force-carrying beams that emanate from the eyes. Here, we used functional magnetic resonance imaging and multivoxel pattern analysis to test the prediction that the brain encodes gaze as implied motion streaming from an agent toward a gazed-upon object. We found that a classifier, trained to discriminate the direction of visual motion, significantly decoded the gaze direction in static images depicting a sighted face, but not a blindfolded one, from brain activity patterns in the human motion-sensitive middle temporal complex (MT+) and temporo-parietal junction (TPJ). Our results demonstrate a link between the visual motion system and social brain mechanisms, in which the TPJ, a key node in theory of mind, works in concert with MT+ to encode gaze as implied motion. This model may be a fundamental aspect of social cognition that allows us to efficiently connect agents with the objects of their attention. It is as if the brain draws a quick visual sketch with moving arrows to help keep track of who is attending to what. This implicit, fluid-flow model of other people’s gaze may help explain culturally universal myths about the mind as an energy-like, flowing essence.