Changes in gene expression in human skeletal stem cells transduced with constitutively active Gsα correlates with hallmark histopathological changes seen in fibrous dysplastic bone

In order to dissect pathways enchained in skeletal stem/progenitors by Fibrous Dysplasia mutations, we engineered human skeletal stem/progenitors with the mutation and performed transcriptomic analysis. FD mutation profoundly alters the properties of skeletal stem/progenitors by pushing hBMSCs towards formation of disorganized bone with a concomitant lack of fat development. In addition, the mutation created an altered in trans environment that pushed the overall system towards neovascularization, inflammation and osteoclastogenesis. We used microarrays to detail the global programme of gene expressionc of human skeletal progenitors containing the Gsα activating mutation R201C Human bone marrow stromal cells (hBMSCs) (derived from bone marrow aspirates) from three independent healthy donors were isolated. Lentiviral vectors (LV-GSαR201C and LV-ctr) were generated, produced and titrated. The LV-vector integrated copy number was calculated by Q-PCR as described and was established as ~1 copy of integrated lentiviral sequence per transduced cell. hBMSCs were transduced with LV-GSαR201C and LV-ctr or mock treated.