Generation of gene-corrected functional osteoclasts from osteopetrotic induced pluripotent stem cells
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RESEARCH
Open Access
Generation of gene-corrected functional osteoclasts from osteopetrotic induced pluripotent stem cells Xiaojie Xian1†, Roksana Moraghebi1†, Henrik Löfvall1,2, Anders Fasth3, Kim Henriksen2, Johan Richter1, Niels-Bjarne Woods1 and Ilana Moscatelli1*
Abstract Background: Infantile malignant osteopetrosis (IMO) is an autosomal recessive disorder characterized by nonfunctional osteoclasts and a fatal outcome early in childhood. About 50% of patients have mutations in the TCIRG1 gene. Methods: IMO iPSCs were generated from a patient carrying a homozygous c.11279G>A (IVS18+1) mutation in TCIRG1 and transduced with a lentiviral vector expressing human TCIRG1. Embryoid bodies were generated and differentiated into monocytes. Non-adherent cells were harvested and further differentiated into osteoclasts on bovine bone slices. Results: Release of the bone resorption biomarker CTX-I into the media of gene-corrected osteoclasts was 5-fold higher than that of the uncorrected osteoclasts and 35% of that of control osteoclasts. Bone resorption potential was confirmed by the presence of pits on the bones cultured with gene-corrected osteoclasts, absent in the uncorrected IMO osteoclasts. Conclusions: The disease phenotype was partially corrected in vitro, providing a valuable resource for therapy development for this form of severe osteopetrosis. Keywords: Infantile malignant osteopetrosis, iPSC generation, Gene transfer, Osteoclast differentiation
Background Infantile malignant osteopetrosis (IMO) is the most severe form of the genetically heterogeneous group of disorders named osteopetrosis [1]. IMO is characterized by defective functionality or differentiation of osteoclasts that leads to increased bone mass, dental abnormalities, and frequent bone fractures [2]. IMO patients also display a progressive reduction of marrow cavities leading to anemia, hepatosplenomegaly, and frequent infections * Correspondence: [email protected] † Xiaojie Xian and Roksana Moraghebi contributed equally to this work. 1 Department of Molecular Medicine and Gene Therapy, Lund Strategic Center for Stem Cell Biology, Lund University, BMC A12, 221 84 Lund, Sweden Full list of author information is available at the end of the article
[3, 4]. Furthermore, compression of the cranial nerves leads to impairment of neurologic functions with blindness and sometimes also deafness [3, 4]. More than 50% of the patients have mutations in the TCIRG1 gene encoding the a3 subunit of the V-ATPase proton pump, which is necessary for the acidification of the resorption lacunae [5]. The only curative treatment to date for IMO is allogeneic hematopoietic stem cell transplantation (HSCT), and IMO is usually fatal within the first 10 years of life if not treated [1]. Despite the greatly improved outcome overtime of HSCT for IMO [2, 6, 7], alternative autologous therapies such as HSC-targeted gene therapy [8, 9] have the potential to restore the resorptive function of osteoclasts without some of the
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