Cell therapy of a patient with type III <i>osteogenesis imperfecta</i> caused by mutation in <i>COL1A2</i> gene and unstable collagen type I

Abstract

The allogenic bone marrow derived mesenchymal stem cells transplantation was given to the newborn girl diagnosed with osteogenesis imperfecta type III, with multiple bone fractures, extreme shortness and limbs deformities. The treatment was performed at the age of 4 and 6 weeks. The clinical diagnosis was supported by biochemical analysis of collagen type I recovered from culture medium of cultivated patient's skin fibroblast, which revealed its triple helix instability at temperature about 2˚C lower than normal. Sequencing of both genes encoding procollagen type I revealed heterozygous substitution G23569A in COL1A2 gene causing change of glycine at position 517 to aspartate. The donor of mesenchymal stem cells was the girl's father. She received two intravenous infusions of suspended cultured mesenchymal cells in 16 days apart without any side effects. An analysis of procollagen type I secreted to the culture medium by bone marrow-derived mesenchymal stem cells obtained from the patient, 3 months following transplantation revealed its normal triple helix stability. During the subsequent two years of follow up two new bone fractures were noted. Currently a twoyear-old girl's presents extreme growth and weight deficiency. The motoric development is also retarded, but the patient constantly improves and makes progresses.

Osteogenesis Imperfecta: Current and Prospective Therapies

“…Moreover, no alloreactivity to donor hfMSCs or possible toxic reactions to the procedure were observed [48]. Other clinical trials with HLA-matched family members were conducted [49]. Successful application of stem cell therapy in paediatric patients, both prenatally (in utero) and postnatally, has been also reported in other studies [28,39].…”

Mutations in COL1A1 and COL1A2 Genes Associated with Osteogenesis Imperfecta (OI) Types I or III.

“…The PCRs were conducted using FastStartTaq DNA Polymerase kit (Roche, Germany) and the amplicons were verified. The reaction mixtures were prepared as described elsewhere (Witecka et al, 2008;Majka et al, 2013). The amplicons were verified following electrophoresis in 1% agarose gel using 5 µl of the obtained PCR products.…”

“…Sanger's enzymatic method was used for DNA sequencing, therefore, 20 ng of each purified PCR product was mixed with 5 pM of the appropriate sequencing primer, for SpI: 5'-TCT-GGG-GAG-CCG-CTA-GCG-CGG-3'; for PvuII: 5'-TTT-CAT-CCG-TGG-CAG-CAT-CAT-AAG-C-3' with Big Dye Terminator v.3.1. The sequencing PCR was conducted according to a protocol described elsewhere (Witecka et al, 2008;Majka et al, 2013).…”

Analysis of miRNAs in Osteogenesis imperfecta Caused by Mutations in COL1A1 and COL1A2: Insights into Molecular Mechanisms and Potential Therapeutic Targets

“…The DNA amplification reactions were conducted using FastStartTaq DNA polymerase kit (Roche, Basel, Switzerland). The composition of the reaction mixture and conditions used are described elsewhere [84,85]. Briefly, for each gene, 10 amplification reactions were conducted, and the DNA fragments were visualized in 1% agarose gel (Agarose, LE Analytical Grade, Promega, Madison, WI, USA) in TAE buffer.…”