Osteopetrosis with micro-lacunar resorption because of defective integrin organization

In vitro differentiated monocytes were used to characterize the cellular defect in a type of osteopetrosis with minimally functional osteoclasts, in which defects associated with common causes of osteopetrosis were excluded by gene sequencing. Monocytes from the blood of a 28-year-old patient were differentiated in media with RANKL and CSF-1. Cell fusion, acid compartments within cells, and tartrate resistant acid phosphatase (TRAP) activity were normal. However, the osteoclasts made abnormally small pits on the dentine. Phalloidin labeling showed that the cell attachments lacked the peripheral ring structure that supports lacunar resorption. Instead, the osteoclasts had clusters of podosomes near the center of cell attachments. Antibody to the alpha v beta 3 integrin pair or to the C-terminal of beta 3 did not label podosomes, but antibody to alpha v labeled them. Western blots using antibody to the N-terminal of beta 3 showed a protein of reduced size. Integrins beta 1 and beta 5 were upregulated, but, in contrast to observations in beta 3 defects, alpha 2 had not increased. The rho-GTP exchange protein Vav3, a key attachment organizing protein, did not localize normally with peripheral attachment structures. Vav3 forms of 70 kD and 90 kD were identified on western blots. However, the proteins beta 3 integrin, Vav3, Plekhm1, and Src, implicated in attachment defects, had normal exon sequences. In this new type of osteopetrosis, the integrin-organizing complex is dysfunctional, and at least two attachment proteins may be partially degraded. Laboratory Investigation (2009) 89, 1007-1017; doi:10.1038/labinvest.2009.58; published online 22 June 2009