Human Adipose-Derived Mesenchymal Stromal Cells Injected Systemically into GRMD Dogs without Immunosuppression are Able to Reach the Host Muscle and Express Human Dystrophin

Abstract

Duchenne muscular dystrophy (DMD), a lethal X-linked disorder, is the most common and severe form of muscular dystrophies, affecting 1 in 3,500 male births. Mutations in the DMD gene lead to the absence of muscle dystrophin and a progressive degeneration of skeletal muscle. The possibility to treat DMD through cell therapy has been widely investigated. We have previously shown that human adipose-derived stromal cells (hASCs) injected systemically in SJL mice are able to reach and engraft in the host muscle, express human muscle proteins, and ameliorate the functional performance of injected animals without any immunosuppression. However, before starting clinical trials in humans many questions still need to be addressed in preclinical studies, in particular in larger animal models, when available. The best animal model to address these questions is the golden retriever muscular dystrophy (GRMD) dog that reproduces the full spectrum of human DMD. Affected animals carry a mutation that predicts a premature termination codon in exon 8 and a peptide that is 5% the size of normal dystrophin. These dogs present clinical signs within the first weeks and most of them do not survive beyond age two. Here we show the results of local and intravenous injections of hASCs into GRMD dogs, without immunosuppression. We observed that hASCs injected systemically into the dog cephalic vein are able to reach, engraft, and express human dystrophin in the host GRMD dystrophic muscle up to 6 months after transplantation. Most importantly, we demonstrated that injecting a huge quantity of human mesenchymal cells in a large-animal model, without immunosuppression, is a safe procedure, which may have important applications for future therapy in patients with different forms of muscular dystrophies.

Iron Determination in Whole Blood Samples of Dystrophic Mouse Strains Using X-Ray Fluorescence Spectrometry

“…-1 ) corresponds to the animal model (Dmd mdx /J male) that reached the highest weight (increase of 12% in grams), compared to other dystrophic mice when they reach adulthood (3-month-old). Considering that the DMD muscle generally shows an advanced stage of dystrophy, with muscle fibers abnormally bigger and surrounded by adipose tissues and replacing a healthier muscle, this animal may be severely affected [18]. These data can be seen in Figure 4; they were arranged in ascending order of concentration.…”

Concise Review: Mesoangioblast and Mesenchymal Stem Cell Therapy for Muscular Dystrophy: Progress, Challenges, and Future Directions

“…They detected it in the forelimb and hind limb muscles by Western blotting and in approximately 50% of myofibers by immunofluorescence using an antibody specific to human dystrophin, indicating that the donor cells differentiated into myofibers in the dystrophic muscle of the recipient [52]. Dystrophin protein was also detected in another study in 11%-19% of myofibers in the GRMD dog model for DMD by immunofluorescent microscopy and Western blotting, using antibodies specific for human dystrophin [53]. Adult human adipose-derived MSCs also gave rise to human b2-microglobulin-positive cells in mdx muscle.…”

Mezenkimal kök hücreler ve veteriner hekimlikte kullanımı

“…Veteriner hekimlikte MKH tabanlı tedaviler atlarda ve köpeklerde tendon ve ligament yaralanmalarında kıkırdak ve eklem hasarlarında, muskuler distrofide (25), veteriner diş hekimliğinde pulpanın rejenerasyonu amacıyla (11), dermatolojide doku kayıplı yara ve yanıklarda deri iyileşmesi amacıyla (12) kullanım alanı bulmaktadır. Günümüzde veteriner hekimlik alanında çalışmalar genellikle araştırma tabanlı olarak deney hayvanlarında yapılmaktadır.…”