Stem cells help mice with muscular dystrophy: study

[mj23]

http://news.yahoo.com/s/nm/20080120/...y_stemcells_dc

[Wise Young]

Originally Posted by mj23

http://news.yahoo.com/s/nm/20080120/...y_stemcells_dc

So, the headline news of this study, published in Nature Medicine, is that this is the first time muscular dystrophy has been successfully treated with embryonic stem cells. The lead scientist, Rita Perlingeiro, turned on a gene called Pax6 to differentiate mouse embryonic cells to begin differentiating in the direction of muscle cells and then transplanted these cells into transgenic mice that have the gene dystrophin (known to cause muscular dystrophy). The cells turned into tumor cells. So, they carefully selected the cells that they injected from various surface markers and found that if they transplant only the selected cells, they did not get tumors.

What the article does not and should have said is that many other studies have shown that stem cells from fetuses, umbilical cord blood stem cells, and bone marrow stem cells have all been reported to improve MD mice. Furthermore, cord blood cells have been reported to improve kids of muscular dystrophy.

Here are some of the references

1. Liu Y, Yan X, Sun Z, Chen B, Han Q, Li J and Zhao RC (2007). Flk-1+ adipose-derived mesenchymal stem cells differentiate into skeletal muscle satellite cells and ameliorate muscular dystrophy in mdx mice. Stem Cells Dev. 16: 695-706. Institute of Basic Medical Sciences & School of Basic Medicine, Center of Excellence in Tissue Engineering, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, PR China. Duchenne muscular dystrophy (DMD) is a severe hereditary disease characterized by the absence of dystrophin on the sarcolemma of muscle fiber. This absence results in widespread muscle damage and satellite cell activation. After depletion of the satellite cell pool, skeletal muscle is then invariably replaced by connective tissue, leading to progressive muscle weakness. Herein, we isolated Flk-1(+) mesenchymal stem cells (MSCs) from adult adipose tissue and induced them to differentiate into skeletal muscle cells in culture. Within mdx mice, an animal model of DMD, adipose tissue-derived Flk-1(+) MSCs (AD-MSCs) homed to and differentiated into cells that repaired injured muscle tissue. This repair correlated with reconstitution of dystrophin expression on the damaged fibers. Flk-1(+) AD-MSCs also differentiated into muscle satellite cells. This differentiation may have accounted for long-term reconstitution. These cells also differentiated into endothelial cells, thereby possibly improving fiber regeneration as a result of the induced angiogenesis. Therefore, Flk-1(+) AD-MSC transplants may repair muscular dystrophy.
2. Vieira NM, Brandalise V, Zucconi E, Jazedje T, Secco M, Nunes VA, Strauss BE, Vainzof M and Zatz M (2007). Human multipotent adipose derived stem cells restore dystrophin expression of Duchenne skeletal muscle cells in vitro. Biol Cell. BACKGROUND INFORMATION: Duchenne muscular dystrophy (DMD) is a devastating X-linked disorder characterized by progressive muscle degeneration and weakness. The use of cell therapy for the repair of defective muscle is being pursued as a possible treatment for DMD. Mesenchymal stem cells have the potential to differentiate and display a myogenic phenotype in vitro. Since liposuctioned human fat is available in large quantities, it may be an ideal source of stem cells for therapeutic applications. Adipose-derived stem cells (ASCs) are able to restore dystrophin expression in the muscles of mdx mice. However, the outcome when these cells interact with human dystrophic muscle is still unknown. RESULTS: We show here that ASCs participate in myotube formation when cultured together with differentiating human DMD myoblasts, resulting in the restoration of dystrophin expression. Similarly, dystrophin was induced when ASCs were cocultivated with DMD myotubes. Experiments with GFP-positive ASCs and DAPI stained DMD myoblasts indicated that ASCs participate in human myogenesis through cellular fusion. CONCLUSIONS: These results show that ASCs have the potential to interact with dystrophic muscle cells restoring dystrophin expression of DMD cells in vitro. The possibility of using adipose tissue as a source for stem cell therapies for muscular diseases is extremely exciting.
3. Asakura A, Hirai H, Kablar B, Morita S, Ishibashi J, Piras BA, Christ AJ, Verma M, Vineretsky KA and Rudnicki MA (2007). Increased survival of muscle stem cells lacking the MyoD gene after transplantation into regenerating skeletal muscle. Proc Natl Acad Sci U S A. 104: 16552-7. Stem Cell Institute, Paul and Sheila Wellstone Muscular Dystrophy Center, University of Minnesota Medical School, Minneapolis, MN 55455, USA. asakura@umn.edu. MyoD is a myogenic master transcription factor that plays an essential role in muscle satellite cell (muscle stem cell) differentiation. To further investigate the function of MyoD in satellite cells, we examined the transplantation of satellite cell-derived myoblasts lacking the MyoD gene into regenerating skeletal muscle. After injection into injured muscle, MyoD(-/-) myoblasts engrafted with significantly higher efficiency compared with wild-type myoblasts. In addition, MyoD(-/-) myoblast-derived satellite cells were detected underneath the basal lamina of muscle fibers, indicating the self-renewal property of MyoD(-/-) myoblasts. To gain insights into MyoD gene deficiency in muscle stem cells, we investigated the pathways regulated by MyoD by GeneChip microarray analysis of gene expression in wild-type and MyoD(-/-) myoblasts. MyoD deficiency led to down-regulation of many muscle-specific genes and up-regulation of some stem cell markers. Importantly, in MyoD(-/-) myoblasts, many antiapoptotic genes were up-regulated, whereas genes known to execute apoptosis were down-regulated. Consistent with these gene expression profiles, MyoD(-/-) myoblasts were revealed to possess remarkable resistance to apoptosis and increased survival compared with wild-type myoblasts. Forced expression of MyoD or the proapoptotic protein Puma increased cell death in MyoD(-/-) myoblasts. Therefore, MyoD(-/-) myoblasts may preserve stem cell characteristics, including their resistance to apoptosis, expression of stem cell markers, and efficient engraftment and contribution to satellite cells after transplantation. Furthermore, our data offer evidence for improved therapeutic stem cell transplantation for muscular dystrophy, in which suppression of MyoD in myogenic progenitors would be beneficial to therapy by providing a selective advantage for the expansion of stem cells.
4. Torrente Y, Belicchi M, Marchesi C, Dantona G, Cogiamanian F, Pisati F, Gavina M, Giordano R, Tonlorenzi R, Fagiolari G, Lamperti C, Porretti L, Lopa R, Sampaolesi M, Vicentini L, Grimoldi N, Tiberio F, Songa V, Baratta P, Prelle A, Forzenigo L, Guglieri M, Pansarasa O, Rinaldi C, Mouly V, Butler-Browne GS, Comi GP, Biondetti P, Moggio M, Gaini SM, Stocchetti N, Priori A, D'Angelo MG, Turconi A, Bottinelli R, Cossu G, Rebulla P and Bresolin N (2007). Autologous transplantation of muscle-derived CD133+ stem cells in Duchenne muscle patients. Cell Transplant. 16: 563-77. Fondazione IRCCS Ospedale Maggiore Policlinico of Milan, Department of Neurological Sciences, Dino Ferrari Center, University of Milan, Italy. torrenteyvan@hotmail.com. Duchenne muscular dystrophy (DMD) is a lethal X-linked recessive muscle disease due to defect on the gene encoding dystrophin. The lack of a functional dystrophin in muscles results in the fragility of the muscle fiber membrane with progressive muscle weakness and premature death. There is no cure for DMD and current treatment options focus primarily on respiratory assistance, comfort care, and delaying the loss of ambulation. Recent works support the idea that stem cells can contribute to muscle repair as well as to replenishment of the satellite cell pool. Here we tested the safety of autologous transplantation of muscle-derived CD133+ cells in eight boys with Duchenne muscular dystrophy in a 7-month, double-blind phase I clinical trial. Stem cell safety was tested by measuring muscle strength and evaluating muscle structures with MRI and histological analysis. Timed cardiac and pulmonary function tests were secondary outcome measures. No local or systemic side effects were observed in all treated DMD patients. Treated patients had an increased ratio of capillary per muscle fibers with a switch from slow to fast myosin-positive myofibers.
5. Berry SE, Liu J, Chaney EJ and Kaufman SJ (2007). Multipotential mesoangioblast stem cell therapy in the mdx/utrn-/- mouse model for Duchenne muscular dystrophy. Regen Med. 2: 275-88. University of Illinois, Department of Cell and Developmental Biology, 601 South Goodwin Avenue, Urbana, IL 61801, USA. BACKGROUND: Duchenne muscular dystrophy is a progressive, lethal muscle-wasting disease for which there is no treatment. MATERIALS & METHODS: We have isolated wild-type mesoangioblasts from aorta and tested their effectiveness in alleviating severe muscle disease in the dystrophin/utrophin knockout (mdx/utrn-/-) mouse model for Duchenne muscular dystrophy. RESULTS: Mesoangioblast clones express Sca-1 and Flk-1 and differentiate into smooth and skeletal muscle, glial cells and adipocytes in vitro. Mesoangioblasts proliferate in vivo, incorporate into muscle fibers, form new fibers, and promote synthesis of dystrophin and utrophin. Muscle fibers that have incorporated mesoangioblasts, as well as surrounding fibers, are protected from damage, with approximately 50-fold less damage than fibers in muscle injected with saline. Some mesoangioblasts localize beneath the basal lamina and express c-met, whereas others differentiate into smooth muscle cells at the periphery of vessels and express alpha-smooth muscle actin. In mdx/utrn-/- muscle, some mesoangioblasts also form Schwann cells. DISCUSSION & CONCLUSION: Mesoangioblasts differentiate into multiple cell types damaged during the progression of severe muscle disease and protect fibers from damage. As such, they are good candidates for therapy of Duchenne muscular dystrophy and perhaps other neuromuscular diseases.
6. Liu YN, Yan X, Sun Z, Han Q and Zhao RC (2007). [Mice adipose derived Flk-1+ mesenchymal stem cells can ameliorate Duchenne's muscular dystrophy in Mdx mice for their multilineage potential]. Zhongguo Shi Yan Xue Ye Xue Za Zhi. 15: 306-12. Center of Tissue Engineering, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing 100005, China. Duchenne muscular dystrophy (DMD) is a common X-linked disease characterized by widespread muscle damage that invariably leads to paralysis and death. There is currently no therapy for this disease. This study was purposed to investigate the feasibility to use adult adipose-derived mesenchymal stem cells (AD-MSCs) in the therapy of DMD. The Flk-1(+) MSCs were isolated from adipose tissue of adult GFP mice; the phenotype and cell cycle of MSCs were analyzed by flow cytometry; the AD-MSCs were directionally differentiated by myoblast and endotheliablast induction system in vitro and were identified by immumofluorecence staining and RT-PCR; the AD-MSCs were transplanted into CTX-injured mice model or mdx mice (DMD animal model) through tail vein; the distribution and differentiation of AD-MSCs were detected by immunofluorescence staining and RT-PCR respectively, and statistic analysis was performed. The results showed that the Flk-1(+) AD-MSCs could be induced to differentiate into myoblasts and endothelial cells in vitro. After transplanted into CTX-injured mice model or mdx mice, GFP-positive cells could be detected in damaged muscle, and these donor-derived cells were also positive for MHC, vWF, or Pax7. Flk-1(+) AD-MSC transplantation also partly reconstituted the expression of dystrophin, and reduced the percentage of centronucleated myofibers in mdx mice. It is concluded that Flk-1(+) AD-MSCs represent a possible tool for future cell therapy applications in DMD disease, as they can be delivered through the circulation for their potential of muscle homing. And Flk-1(+) AD-MSCs also show the ability to contribute to muscle repair, improvement of blood supply and long term reconstitution of dystrophy muscle.
7. Israeli D, Ziaei S, Gjata B, Benchaouir R, Rameau P, Marais T, Fukada S, Segawa M, Yamamoto H, Gonin P, Danos O and Garcia L (2007). Expression of mdr1 is required for efficient long term regeneration of dystrophic muscle. Exp Cell Res. 313: 2438-50. Genethon, CNRS UMR 8115, Evry, France. israeli@genethon.fr. The mouse mdr1a and mdr1b genes are expressed in skeletal muscle, though their precise role in muscle is unknown. Dystrophic muscle is characterized by repeated cycles of degeneration and regeneration. To explore the role of the mdr1 genes during muscle regeneration, we have created a triple knockout mouse lacking the mdr1a, mdr1b, and the dystrophin genes. The resulting ReX mice developed normally and were fertile. However, as adults, ReX had a higher proportion of degenerating muscle fibers and greater long-term loss of muscle mass than mdx. ReX muscles were also characterized by a reduced proportion of muscle side population (mSP) cells, of myogenic cells, and a reduced capacity for muscle regeneration. We found too that mSP cells derived from dystrophic muscle are more myogenic than those from normal muscle. Thus, in dystrophic muscle, the mdr1 gene plays an important role in the preservation of the mSP and of the myogenic regenerative potential. Moreover, our results suggest a hitherto unappreciated role of mdr1 in precursor cells of regenerating tissue; they therefore provide an important clue to the physiological significance of mdr1 expression in stem cells.
8. Peault B, Rudnicki M, Torrente Y, Cossu G, Tremblay JP, Partridge T, Gussoni E, Kunkel LM and Huard J (2007). Stem and progenitor cells in skeletal muscle development, maintenance, and therapy. Mol Ther. 15: 867-77. Stem Cell Research Center, Children's Hospital of Pittsburgh, Department of Orthopaedic Surgery, University of Pittsburgh School of Medicine, Pittsburgh, PA 15213, USA. Satellite cells are dormant progenitors located at the periphery of skeletal myofibers that can be triggered to proliferate for both self-renewal and differentiation into myogenic cells. In addition to anatomic location, satellite cells are typified by markers such as M-cadherin, Pax7, Myf5, and neural cell adhesion molecule-1. The Pax3 and Pax7 transcription factors play essential roles in the early specification, migration, and myogenic differentiation of satellite cells. In addition to muscle-committed satellite cells, multi-lineage stem cells encountered in embryonic, as well as adult, tissues exhibit myogenic potential in experimental conditions. These multi-lineage stem cells include side-population cells, muscle-derived stem cells (MDSCs), and mesoangioblasts. Although the ontogenic derivation, identity, and localization of these non-conventional myogenic cells remain elusive, recent results suggest their ultimate origin in blood vessel walls. Indeed, purified pericytes and endothelium-related cells demonstrate high myogenic potential in culture and in vivo. Allogeneic myoblasts transplanted into Duchenne muscular dystrophy (DMD) patients have been, in early trials, largely inefficient owing to immune rejection, rapid death, and limited intramuscular migration--all obstacles that are now being alleviated, at least in part, by more efficient immunosuppression and escalated cell doses. As an alternative to myoblast transplantation, stem cells such as mesoangioblasts and CD133+ progenitors administered through blood circulation have recently shown great potential to regenerate dystrophic muscle.
9. Yu M, Zhang C, Zhang Y, Feng S, Yao X and Lu X (2007). BM stem cell transplantation rescues pathophysiologic features of aged dystrophic mdx muscle. Cytotherapy. 9: 44-52. Department of Neurology, The First Affiliated Hospital of Sun Yat-Sen University, Guangzhou, China. BACKGROUND: The value of transplantation of BM stem cells in aged (12-month-old) mdx was evaluated because it is thought to be a more ideal model for studying the praxiology of Duchenne muscular dystrophy (DMD). The possible mechanisms of stem cell differentiation were then discussed. METHODS: BM was isolated from 8-10-week-old male C57 BL/10 mice. After injecting BM cells into 12-month-old female mdx mice through the tail vein, the expression of dystrophin and MyoD was detected at different time points by immunofluorescence staining, RT-PCR and Western blot. RESULTS: The C57 male mice donor-specific and Y-chromosome-specific sequence could be detected in all female aged mdx mice, implying the success of the transplantation. Expression of dystrophin and MyoD was detected and increased over time. DISCUSSION: BM cells were recruited to the muscle and partially restored specific pathophysiologic features of the dystrophic muscle in aged mdx mice. Muscle differentiation of BM cells recapitulated embryonic myogenesis.
10. Chan J, Waddington SN, O'Donoghue K, Kurata H, Guillot PV, Gotherstrom C, Themis M, Morgan JE and Fisk NM (2007). Widespread distribution and muscle differentiation of human fetal mesenchymal stem cells after intrauterine transplantation in dystrophic mdx mouse. Stem Cells. 25: 875-84. Experimental Fetal Medicine Group, Institute of Reproductive and Developmental Biology, Imperial College London, Hammersmith Campus, Du Cane Road, London, United Kingdom. jerrychan@nus.edu.sg. Duchenne muscular dystrophy (DMD) is a common X-linked disease resulting from the absence of dystrophin in muscle. Affected boys suffer from incurable progressive muscle weakness, leading to premature death. Stem cell transplantation may be curative, but is hampered by the need for systemic delivery and immune rejection. To address these barriers to stem cell therapy in DMD, we investigated a fetal-to-fetal transplantation strategy. We investigated intramuscular, intravascular, and intraperitoneal delivery of human fetal mesenchymal stem cells (hfMSCs) into embryonic day (E) 14-16 MF1 mice to determine the most appropriate route for systemic delivery. Intramuscular injections resulted in local engraftment, whereas both intraperitoneal and intravascular delivery led to systemic spread. However, intravascular delivery led to unexpected demise of transplanted mice. Transplantation of hfMSCs into E14-16 mdx mice resulted in widespread long-term engraftment (19 weeks) in multiple organs, with a predilection for muscle compared with nonmuscle tissues (0.71% vs. 0.15%, p < .01), and evidence of myogenic differentiation of hfMSCs in skeletal and myocardial muscle. This is the first report of intrauterine transplantation of ontologically relevant hfMSCs into fully immunocompetent dystrophic fetal mice, with systemic spread across endothelial barriers leading to widespread long-term engraftment in multiple organ compartments. Although the low-level of chimerism achieved is not curative for DMD, this approach may be useful in other severe mesenchymal or enzyme deficiency syndromes, where low-level protein expression may ameliorate disease pathology.
11. Nunes VA, Cavacana N, Canovas M, Strauss BE and Zatz M (2007). Stem cells from umbilical cord blood differentiate into myotubes and express dystrophin in vitro only after exposure to in vivo muscle environment. Biol Cell. 99: 185-96. Human Genome Research Center, Department of Genetics and Evolutionary Biology, University of Sao Paulo, Sao Paulo, SP, Brazil. vanunes@ib.usp.br. BACKGROUND INFORMATION: Duchenne muscular dystrophy is a disease characterized by progressive and irreversible muscle degeneration for which there is no therapy. HUCB (human umbilical cord blood) has been considered as an important source of haematopoietic and mesenchymal stem cells, each having been shown to differentiate into distinct cell types. However, it remains unclear if these cells are able to differentiate into muscle cells. RESULTS: We have showed that stem cells from HUCB did not differentiate into myotubes or express dystrophin when cultured in muscle-conditioned medium or with human muscle cells. However, delivery of GFP (green fluorescent protein)-transduced mononucleated cells from HUCB, which comprises both haematopoietic and mesenchymal populations, into quadriceps muscle of mdx (mouse dystrophy X-chromosome linked) mice resulted in the expression of human myogenic markers. After recovery of these cells from mdx muscle and in vitro cultivation, they were able to fuse and form GFP-positive myotubes that expressed dystrophin. CONCLUSIONS: These results indicate that chemical factors and cell-to-cell contact provided by in vitro conditions were not enough to trigger the differentiation of stem cells into muscle cells. Nevertheless, we showed that the HUCB-derived stem cells were capable of acquiring a muscle phenotype after exposure to an in vivo muscle environment, which was required to activate the differentiation programme.
12. Sampaolesi M, Blot S, D'Antona G, Granger N, Tonlorenzi R, Innocenzi A, Mognol P, Thibaud JL, Galvez BG, Barthelemy I, Perani L, Mantero S, Guttinger M, Pansarasa O, Rinaldi C, Cusella De Angelis MG, Torrente Y, Bordignon C, Bottinelli R and Cossu G (2006). Mesoangioblast stem cells ameliorate muscle function in dystrophic dogs. Nature. 444: 574-9. San Raffaele Scientific Institute, Universita Vita e Salute, Stem Cell Research Institute, Via Olgettina 58, 20132 Milan, Italy. Duchenne muscular dystrophy remains an untreatable genetic disease that severely limits motility and life expectancy in affected children. The only animal model specifically reproducing the alterations in the dystrophin gene and the full spectrum of human pathology is the golden retriever dog model. Affected animals present a single mutation in intron 6, resulting in complete absence of the dystrophin protein, and early and severe muscle degeneration with nearly complete loss of motility and walking ability. Death usually occurs at about 1 year of age as a result of failure of respiratory muscles. Here we report that intra-arterial delivery of wild-type canine mesoangioblasts (vessel-associated stem cells) results in an extensive recovery of dystrophin expression, normal muscle morphology and function (confirmed by measurement of contraction force on single fibres). The outcome is a remarkable clinical amelioration and preservation of active motility. These data qualify mesoangioblasts as candidates for future stem cell therapy for Duchenne patients.
13. Galvez BG, Sampaolesi M, Brunelli S, Covarello D, Gavina M, Rossi B, Constantin G, Torrente Y and Cossu G (2006). Complete repair of dystrophic skeletal muscle by mesoangioblasts with enhanced migration ability. J Cell Biol. 174: 231-43. Stem Cell Research Institute, San Raffaele Hospital, 20132 Milan, and Department of Experimental Medicine, Human Anatomy Institute, University of Pavia, Italy. Efficient delivery of cells to target tissues is a major problem in cell therapy. We report that enhancing delivery of mesoangioblasts leads to a complete reconstitution of downstream skeletal muscles in a mouse model of severe muscular dystrophy (alpha-sarcoglycan ko). Mesoangioblasts, vessel-associated stem cells, were exposed to several cytokines, among which stromal- derived factor (SDF) 1 or tumor necrosis factor (TNF) alpha were the most potent in enhancing transmigration in vitro and migration into dystrophic muscle in vivo. Transient expression of alpha4 integrins or L-selectin also increased several fold migration both in vitro and in vivo. Therefore, combined pretreatment with SDF-1 or TNF-alpha and expression of alpha4 integrin leads to massive colonization (>50%) followed by reconstitution of >80% of alpha-sarcoglycan-expressing fibers, with a fivefold increase in efficiency in comparison with control cells. This study defines the requirements for efficient engraftment of mesoangioblasts and offers a new potent tool to optimize future cell therapy protocols for muscular dystrophies.
14. Bachrach E, Perez AL, Choi YH, Illigens BM, Jun SJ, del Nido P, McGowan FX, Li S, Flint A, Chamberlain J and Kunkel LM (2006). Muscle engraftment of myogenic progenitor cells following intraarterial transplantation. Muscle Nerve. 34: 44-52. Howard Hughes Medical Institute, Program in Genomics, Children's Hospital, 300 Longwood Avenue, Boston, Massachusetts 02115, USA. Cell-based therapy continues to be a promising avenue for the treatment of Duchenne muscular dystrophy (DMD), an X-linked skeletal muscle-wasting disease. Recently, we demonstrated that freshly isolated myogenic progenitors contained within the adult skeletal muscle side population (SP) can engraft into dystrophic fibers of nonirradiated mdx(5cv) mice after intravenous transplantation. Engraftment rates, however, have not been therapeutically significant, achieving at most 1% of skeletal muscle myofibers expressing protein from donor-derived nuclei. To enhance the engraftment of transplanted myogenic progenitors, an intraarterial delivery method was adapted from a previously described procedure. Cultured, lentivirus-transduced skeletal muscle SP cells, derived from mdx(5cv) mice, were transplanted into the femoral artery of noninjured mdx(5cv) mice. Based on the expression of microdystrophin or green fluorescent protein (GFP) transgenes in host muscle, sections of the recipient muscles exhibited 5%-8% of skeletal muscle fibers expressing donor-derived transgenes. Further, donor muscle SP cells, which did not express any myogenic markers prior to transplant, expressed the satellite cell transcription factor, Pax7, and the muscle-specific intermediate filament, desmin, after extravasation into host muscle. The expression of these muscle-specific markers indicates that progenitors within the side population can differentiate along the myogenic lineage after intraarterial transplantation and extravasation into host muscle. Given that femoral artery catheterization is a common, safe clinical procedure and that the transplantation of cultured adult muscle progenitor cells has proven to be safe in mice, our data may represent a step toward the improvement of cell-based therapies for DMD and other myogenic disorders.

[tim34]

Dear Dr. Wise, one of the researcher is going to conduct human trials with adipose tissue mesenchymal stem cell from donor to treat muscular dystrophy. (refer 1, 2 & 6 from above). He proposed IV allogenic AD-MSC and then The stem cells will be subjected to a one time treatment with proprietory formula (also very harmless and not involving gene transfer) to signal them to differentiate into skeletal muscle cells. Also they will be made availabel in large numbers at very early passage so that they have retained full differentiation potential with minimal immunosuppressive properties


Will the above said improve the condition of MD ? What is your opinion, if all that said is correct ? awaiting reply.

[Wise Young]

Originally Posted by tim34

Dear Dr. Wise, one of the researcher is going to conduct human trials with adipose tissue mesenchymal stem cell from donor to treat muscular dystrophy. (refer 1, 2 & 6 from above). He proposed IV allogenic AD-MSC and then The stem cells will be subjected to a one time treatment with proprietory formula (also very harmless and not involving gene transfer) to signal them to differentiate into skeletal muscle cells. Also they will be made availabel in large numbers at very early passage so that they have retained full differentiation potential with minimal immunosuppressive properties


Will the above said improve the condition of MD ? What is your opinion, if all that said is correct ? awaiting reply.

Tim,

There are two claims being made here. The first is that adipose tissue mesenchymal stem cells (or any mesenchymal stem cells, for that matter) will be beneficial for muscular dystrophy. The second is that the cells obtained from an allogeneic donor will not be rejected quickly. Let me provide some of the evidence behind each of these claims.

Mesenchymal stem cell therapy of muscular dystrophy. Mesenchymal stem cells are the cells for muscular dystrophy since they give rise to myoblasts that make muscles. Mari Dezawa (one of the top scientists in the field using mesenchymal stem cells and who just moved from Kyoto University to Tohoku University in Senda) said:

1. Dezawa M (2006). Insights into autotransplantation: the unexpected discovery of specific induction systems in bone marrow stromal cells. Cell Mol Life Sci. 63: 2764-72. Department of Anatomy and Neurobiology, Kyoto University Graduate School of Medicine, Kyoto, Japan. dezawa@anat2.med.kyoto-u.ac.jp. Many kinds of cells, including embryonic stem cells and tissue stem cells, have been considered candidates for transplantation therapy for neuro- and muscle-degenerative diseases. Bone marrow stromal cells (MSCs) also have great potential as therapeutic agents since they are easily isolated and can be expanded from patients without serious ethical or technical problems. Recently, new methods for the highly efficient and specific induction of functional neurons and skeletal muscle cells have been developed for MSCs. These induced cells were transplanted into animal models of stroke, Parkinson's disease and muscle degeneration, resulting in the successful integration of transplanted cells and improvement in the behavior of the transplanted animals. Here I describe the discovery of these induction systems and focus on the potential use of MSC-derived cells for 'auto-cell transplantation therapy' in neuro- and muscle-degenerative diseases.

.

In the above article, Mari Dezawa was talking about autografts which would not have any immune-rejection problems. However, it makes no sense to use autografts (to transplant one's own cells to oneself) when there is a genetic disorder because it must recapitulates the genetic problem. Allografts (grafts from another individual) would be necessary or one can genetically modify autografts to express the normal dystrophin gene. But, to my knowledge, this is not yet available.

Most of the studies of mesenchymal stem cell therapies of animal models of muscular dystrophy, however, are from fetuses. Others have focussed o mesoangioblasts, a muscle progenitor cell.

1. Nunes VA, Cavacana N, Canovas M, Strauss BE and Zatz M (2007). Stem cells from umbilical cord blood differentiate into myotubes and express dystrophin in vitro only after exposure to in vivo muscle environment. Biol Cell. 99: 185-96. Human Genome Research Center, Department of Genetics and Evolutionary Biology, University of Sao Paulo, Sao Paulo, SP, Brazil. vanunes@ib.usp.br. BACKGROUND INFORMATION: Duchenne muscular dystrophy is a disease characterized by progressive and irreversible muscle degeneration for which there is no therapy. HUCB (human umbilical cord blood) has been considered as an important source of haematopoietic and mesenchymal stem cells, each having been shown to differentiate into distinct cell types. However, it remains unclear if these cells are able to differentiate into muscle cells. RESULTS: We have showed that stem cells from HUCB did not differentiate into myotubes or express dystrophin when cultured in muscle-conditioned medium or with human muscle cells. However, delivery of GFP (green fluorescent protein)-transduced mononucleated cells from HUCB, which comprises both haematopoietic and mesenchymal populations, into quadriceps muscle of mdx (mouse dystrophy X-chromosome linked) mice resulted in the expression of human myogenic markers. After recovery of these cells from mdx muscle and in vitro cultivation, they were able to fuse and form GFP-positive myotubes that expressed dystrophin. CONCLUSIONS: These results indicate that chemical factors and cell-to-cell contact provided by in vitro conditions were not enough to trigger the differentiation of stem cells into muscle cells. Nevertheless, we showed that the HUCB-derived stem cells were capable of acquiring a muscle phenotype after exposure to an in vivo muscle environment, which was required to activate the differentiation programme.

Bone marrow is a rich and proven source of mesenchymal stem cells. The following group in China has substantial experience with both umbilical cord blood and bone marrow stem cell treatments of Duchenne muscular dystrophy.

1. Feng SW, Lu XL, Liu ZS, Zhang YN, Liu TY, Li JL, Yu MJ, Zeng Y and Zhang C (2008). Dynamic distribution of bone marrow-derived mesenchymal stromal cells and change of pathology after infusing into mdx mice. Cytotherapy. 10: 254-64. Department of Neurology, First Affiliated Hospital, Sun Yat-sen University, Guangzhou, PR China. BACKGROUND: Mesenchymal stromal cells (MSC) are attractive candidates for the treatment of Duchenne muscular dystrophy (DMD) but how the donor MSC distribute in multiple organs and whether the increased dystrophin leads to a change in the pathology of mdx mice is still uncertain. In this research we detected the distribution of MSC and the pathology of mdx mice after MSC infusion. METHODS: MSC were isolated from rat bone marrow (BM) and expanded in proliferation medium. MSC of the fifth passage were delivered intravenously into irradiated mdx mice. The distribution of MSC labeled by [3H]TdR into a recipient's organs was calculated by radioactivity. The expression of dystrophin was detected at weeks 4, 8, 12 and 16 after MSC transplantation by immunofluorescence staining, RT-PCR and Western blot. Serum creatine kinase (CK) and centrally nucleated fiber (CNF) were also detected to assess the change in pathology. RESULTS: 24-48 h after transplantation, MSC were mainly found in the BM, liver and lung. The radioactivity in these organs decreased, whereas skeletal and myocardial muscle radioactivity increased gradually over time. In accordance with the increased radioactivity in skeletal muscle, the amount of dystrophin-positive myofibers increased. Furthermore, serum CK and CNF decreased slightly, suggesting specific pathophysiologic features of the dystrophic muscle were partially restored. DISCUSSION: Upon certification of the distribution of transplanted MSC in irradiated mdx mice, we found evidence of myogenic differentiation of MSC in skeletal muscle. This research may help us understand the mechanism of therapy of MSC transplantation.

Liu, et al. (2007) used adipose-derived mesenchymal stem cells to treat mdx mice:

1. Liu Y, Yan X, Sun Z, Chen B, Han Q, Li J and Zhao RC (2007). Flk-1+ adipose-derived mesenchymal stem cells differentiate into skeletal muscle satellite cells and ameliorate muscular dystrophy in mdx mice. Stem Cells Dev. 16: 695-706. Institute of Basic Medical Sciences & School of Basic Medicine, Center of Excellence in Tissue Engineering, Chinese Academy of Medical Sciences & Peking Union Medical College, Beijing, PR China. Duchenne muscular dystrophy (DMD) is a severe hereditary disease characterized by the absence of dystrophin on the sarcolemma of muscle fiber. This absence results in widespread muscle damage and satellite cell activation. After depletion of the satellite cell pool, skeletal muscle is then invariably replaced by connective tissue, leading to progressive muscle weakness. Herein, we isolated Flk-1(+) mesenchymal stem cells (MSCs) from adult adipose tissue and induced them to differentiate into skeletal muscle cells in culture. Within mdx mice, an animal model of DMD, adipose tissue-derived Flk-1(+) MSCs (AD-MSCs) homed to and differentiated into cells that repaired injured muscle tissue. This repair correlated with reconstitution of dystrophin expression on the damaged fibers. Flk-1(+) AD-MSCs also differentiated into muscle satellite cells. This differentiation may have accounted for long-term reconstitution. These cells also differentiated into endothelial cells, thereby possibly improving fiber regeneration as a result of the induced angiogenesis. Therefore, Flk-1(+) AD-MSC transplants may repair muscular dystrophy.

Goncalves, et al. (2006) reported success in getting human mesenchymal stem cells to fuse into Duchenne muscular dsytrophy myotubes:

1. Goncalves MA, de Vries AA, Holkers M, van de Watering MJ, van der Velde I, van Nierop GP, Valerio D and Knaan-Shanzer S (2006). Human mesenchymal stem cells ectopically expressing full-length dystrophin can complement Duchenne muscular dystrophy myotubes by cell fusion. Hum Mol Genet. 15: 213-21. Department of Molecular Cell Biology, Leiden University Medical Center, Wassenaarseweg 72, 2333 AL Leiden, The Netherlands. Duchenne muscular dystrophy (DMD) is the most prevalent inheritable muscle disease. It is caused by mutations in the approximately 2.5-megabase dystrophin (Dys) encoding gene. Therapeutic attempts at DMD have relied on injection of allogeneic Dys-positive myoblasts. The immune rejection of these cells and their limited availability have prompted the search for alternative therapies and sources of myogenic cells. Stem cell-based gene therapy aims to restore tissue function by the transplantation of gene-corrected autologous cells. It depends on (i) the capacity of stem cells to participate in tissue regeneration and (ii) the efficient genetic correction of defective autologous stem cells. We explored the potential of bone marrow-derived human mesenchymal stem cells (hMSCs) genetically modified with the full-length Dys-coding sequence to engage in myogenesis. By tagging hMSCs with enhanced green fluorescent protein (EGFP) or the membrane dye PKH26, we demonstrated that they could participate in myotube formation when cultured together with differentiating human myoblasts. Experiments performed with EGFP-marked hMSCs and DsRed-labeled DMD myoblasts revealed that the EGFP-positive DMD myotubes were also DsRed-positive indicating that hMSCs participate in human myogenesis through cellular fusion. Finally, we showed that hMSCs transduced with a tropism-modified high-capacity hybrid viral vector encoding full-length Dys could complement the genetic defect of DMD myotubes.

So, I think that mesenchymal stem cells are a reasonable cell treatment. The problem is the other assumption, that the cells and their progeny are immune-privileged. It is true that mesenchymal stem cells are “anti-immune” to the extent that they see to escape the host immune system for a time, perhaps because they are not expressing all the HLA-antigens. However, there is no guarantee that they will not be rejected. Moreover, it is not clear that these cells will have any place to “engraft” (other than muscle) where they can continually produce stem cells that can contribute to muscle formation.

However, I believe that umbilical cord blood cells would be a better option than adipose-derived mesenchymal stem cells. First, there is a lot of experience transplanting umbilical cord blood cells and having them engraft in the bone marrow. Second, the cells can be HLA-matched and are readily available. Third, the cells, once they are engrafted in the bone marrow, will continuously shower the muscles with normal mesenchymal stem cells rather than a one shot deal where they may fuse with the muscle and then go away.

Wise.

[Scott Buxton]

Thank you for sharing this encyclopedic knowledge. Scott.

[tim34]

Originally Posted by tim34

Dear Dr. Wise, one of the researcher is going to conduct human trials with adipose tissue mesenchymal stem cell from donor to treat muscular dystrophy. (refer 1, 2 & 6 from above). He proposed IV allogenic AD-MSC and then The stem cells will be subjected to a one time treatment with proprietory formula (also very harmless and not involving gene transfer) to signal them to differentiate into skeletal muscle cells. Also they will be made availabel in large numbers at very early passage so that they have retained full differentiation potential with minimal immunosuppressive properties


Will the above said improve the condition of MD ? What is your opinion, if all that said is correct ? awaiting reply.

Dr. Wise, thank you for such informative explaination of various comparative stem cell treatments but If I volunteer the above trial, is their a possibility to improve the present condition using specifically "Allogenic" "Adipose Tissue" "mesenchymal stem cell" with the "signal to differentiate into skeletal muscles" ? How much improvement can you bet? ... Sir, as a layman I am not able to co-relate the mouse mdel strudy with the trial that is being done, so I would like to understand the result, if above trial is done on humans.thank you again.. tim

[Wise Young]

Originally Posted by tim34

Dr. Wise, thank you for such informative explaination of various comparative stem cell treatments but If I volunteer the above trial, is their a possibility to improve the present condition using specifically "Allogenic" "Adipose Tissue" "mesenchymal stem cell" with the "signal to differentiate into skeletal muscles" ? How much improvement can you bet? ... Sir, as a layman I am not able to co-relate the mouse mdel strudy with the trial that is being done, so I would like to understand the result, if above trial is done on humans.thank you again.. tim

Let me try to put all the information together. I am sorry it is so long and scattered.

The bottom line is that adipose mesenchymal stem cell is one of several options that have now been reported to be beneficial in animal models of muscular dystrophy. The animal models are reasonable because they are mice that express the muscular dystrophy gene (dystrophin) and develop many of the symptoms of the disease. There is no guarantee that a therapy that works in these mice will work in humans but the animal models is one of the reasons why the field is making significant progress.

I said that mesenchymal stem cells are a good source of cells for treating muscular dystrophy because they actually should produce the progenitor cells for muscle, called myoblasts. I also pointed out that I believe it is necessary for the cells to be allografts (i.e. coming from another individual) because if the muscular dystrophy is a genetic disease, transplanting cells with the same genes should not solve the problem. However, allograts are a problem because they will be immune-rejected unless they are HLA-matched.

Some scientists claim that mesenchymal stem cells are immune-privileged and although I think that there is some data to support the possibility that immune-rejection of these cells are slowed down and take several months, I believe that the cells may be rejected eventually. Therefore, I suggested the use of umbilical cord blood cells. There is lot of clinical experience engrafting umbilical cord blood cells into the bone marrow. Over 6000 such transplants are done every year, using allogeneic umbilical cord blood to replace stem cells in the bone marrow in people with leukemia, anemia, and other hematopoietic diseases.

Once engrafted, the new cells in the bone marrow will be sending out mesenchymal stem cells in the blood stream of the individual, "showering" the muscles with such cells, rather than a one-shot deal where mesenchymal cells would fuse with the muscle and help for a period of time and repeated treatments would be necessary. That is just my opinion and the kind of cells that I would look for. By the way, a group in Guangzhou Medical Center in China had published two studies reporting successful treatment of children with muscular dystrophy, using umblical cord blood cells. I had reviewed this before. That is probably what I would do if I or a loved one has muscular dystrophy.

Wise.

[smirking1]

Lucky damn mice :-)

[tim34]

Thank you dr. Wise. It is me who is a little dumb while you take all your sincere efforts for people like me (who are lost) to help understand a bit of modern medicine. I have gone through many of the articles on this forum where your kindheartedness have gone far to explain smallest of matter which might help the sufferer. The forum success is a reflection and many rely on your comments and take it very seriously. Thank you sir for being available for us.

[tim34]

Dear Dr. Wise,
Related to our previous discussion I found this article in support of adipose tissue derived mesnchymal stem cell and its ability to differentiate when induced.
Myogenic potential of adipose-tissue-derived cells.
http://www.ncbi.nlm.nih.gov/pubmed/16825428

Quote from the article "Finally, we show that uncultured adipose-tissue-associated cells have a high regenerative capacity in vivo since they can be incorporated into muscle fibers following ischemia and can restore significantly dystrophin expression in mdx mice."

This has sugessted a regenerating capacity of AD-MSC. Your comment sir. Does that now looks more optimistic in Humans ?