Generation of an immunodeficient mouse model of tcirg1-deficient autosomal recessive osteopetrosis.

Palagano, Eleonora; Muggeo, Sharon; Crisafulli, Laura; Tourkova, Irina L; Strina, Dario; Mantero, Stefano; Fontana, Elena; Locatelli, Silvia L et al. · Bone Rep · 2020

basic_science · Level V

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Abstract

Autosomal recessive osteopetrosis is a rare skeletal disorder with increased bone density due to a failure in osteoclast bone resorption. In most cases, the defect is cell-autonomous, and >50% of patients bear mutations in the <i>TCIRG1</i> gene, encoding for a subunit of the vacuolar proton pump essential for osteoclast resorptive activity. The only cure is hematopoietic stem cell transplantation, which corrects the bone pathology by allowing the formation of donor-derived functional osteoclasts. Therapeutic approaches using patient-derived cells corrected <i>ex vivo</i> through viral transduction or gene editing can be considered, but to date functional rescue cannot be demonstrated <i>in vivo</i> because a relevant animal model for xenotransplant is missing. We generated a new mouse model, which we named NSG oc/oc, presenting severe autosomal recessive osteopetrosis owing to the <i>Tcirg1</i> <sup><i>oc</i></sup> mutation, and profound immunodeficiency caused by the NSG background. We performed neonatal murine bone marrow transplantation and xenotransplantation with human CD34<sup>+</sup> cells. We demonstrated that neonatal murine bone marrow transplantation rescued NSG oc/oc mice, in line with previous findings in the oc/oc parental strain and with evidence from clinical practice in humans. Importantly, we also demonstrated human cell chimerism in the bone marrow of NSG oc/oc mice transplanted with human CD34<sup>+</sup> cells. The severity and rapid progression of the disease in the mouse model prevented amelioration of the bone pathology; nevertheless, we cannot completely exclude that minor early modifications of the bone tissue might have occurred. Our work paves the way to generating an improved xenograft model for <i>in vivo</i> evaluation of functional rescue of patient-derived corrected cells. Further refinement of the newly generated mouse model will allow capitalizing on it for an optimized exploitation in the path to novel cell therapies.