Thm2 interacts with paralog, Thm1 , and sensitizes to Hedgehog signaling in postnatal skeletogenesis

Mutations in the intraflagellar transport-A (IFT-A) gene, THM1 , have been identified in skeletal ciliopathies. Here, we report a genetic interaction between Thm1 , and its paralog, Thm2, in postnatal skeletogenesis. THM2 localizes to primary cilia, but Thm2 deficiency does not affect ciliogenesis and Thm2- null mice survive into adulthood. However, by postnatal day 14, Thm2 −/− ; Thm1 aln/ + mice exhibit small stature and small mandible. Radiography and microcomputed tomography reveal Thm2 −/− ; Thm1 aln/ + tibia are less opaque and have reduced cortical and trabecular bone mineral density. In the mutant tibial growth plate, the proliferation zone is expanded and the hypertrophic zone is diminished, indicating impaired chondrocyte differentiation. Additionally, mutant growth plate chondrocytes show increased Hedgehog signaling. Yet deletion of one allele of Gli2 , a major transcriptional activator of the Hedgehog pathway, exacerbated the Thm2 −/− ; Thm1 aln/ + small phenotype, and further revealed that Thm2 −/− ; Gli2 +/- mice have small stature. In Thm2 −/− ; Thm1 aln/ + primary osteoblasts, a Hedgehog signaling defect was not detected, but bone nodule formation was markedly impaired. This indicates a signaling pathway is altered, and we propose that this pathway may potentially interact with Gli2 . Together, our data reveal that loss of Thm2 with one allele of Thm1, Gli2, or both, present new IFT mouse models of osteochondrodysplasia. Our data also suggest Thm2 as a modifier of Hedgehog signaling in postnatal skeletal development.