Milan Unit, Istituto di Ricerca Genetica e Biomedica, CNR, 20138 Milan, Italy; Humanitas Clinical and Research Center, via Manzoni 56, 20089 Rozzano (Mi), Italy.
Mouse & Animal Pathology Laboratory, Fondazione Filarete, 20139 Milano, Italy.
Stem Cell Reports. 2015 Oct 13;5(4):558-68. doi: 10.1016/j.stemcr.2015.08.005. Epub 2015 Sep 3.
Autosomal recessive osteopetrosis is a human bone disease mainly caused by TCIRG1 gene mutations that prevent osteoclasts resorbing activity, recapitulated by the oc/oc mouse model. Bone marrow transplantation is the only available treatment, limited by the need for a matched donor. The use of induced pluripotent stem cells (iPSCs) as an unlimited source of autologous cells to generate gene corrected osteoclasts might represent a powerful alternative. We generated iPSCs from oc/oc mice, corrected the mutation using a BAC carrying the entire Tcirg1 gene locus as a template for homologous recombination, and induced hematopoietic differentiation. Similarly to physiologic fetal hematopoiesis, iPSC-derived CD41(+) cells gradually gave rise to CD45(+) cells, which comprised both mature myeloid cells and high proliferative potential colony-forming cells. Finally, we differentiated the gene corrected iPSC-derived myeloid cells into osteoclasts with rescued bone resorbing activity. These results are promising for a future translation into the human clinical setting.
常染色体隐性遗传骨质硬化症是一种人类骨骼疾病，主要由 TCIRG1 基因突变引起，该突变阻止破骨细胞的吸收活性，oc/oc 小鼠模型对此进行了重现。骨髓移植是唯一可用的治疗方法，但受到需要匹配供体的限制。利用诱导多能干细胞（iPSCs）作为无限的自体细胞来源，以产生基因校正的破骨细胞，可能代表一种强大的替代方法。我们从 oc/oc 小鼠中生成 iPSCs，使用携带整个 Tcirg1 基因座的 BAC 作为同源重组模板来校正突变，并诱导造血分化。与生理胎儿造血一样，iPSC 衍生的 CD41(+)细胞逐渐产生 CD45(+)细胞，其中包括成熟的髓样细胞和高增殖潜能的集落形成细胞。最后，我们将基因校正的 iPSC 衍生的髓样细胞分化为具有挽救骨吸收活性的破骨细胞。这些结果为未来在人类临床环境中的转化提供了希望。
