The Experts below are selected from a list of 552 Experts worldwide ranked by ideXlab platform
Giulio Cossu - One of the best experts on this subject based on the ideXlab platform.
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Oxidative stress preconditioning of mouse perivascular myogenic progenitors selects a subpopulation of cells with a distinct survival advantage in vitro and in vivo
Cell Death & Disease, 2018Co-Authors: Cesare Gargioli, Giulio Cossu, Giuseppina Turturici, Maria M. Barreca, Walter Spinello, Claudia Fuoco, Stefano Testa, Salvatore Feo, Stefano M. Cannata, Gabriella SconzoAbstract:Cell engraftment, survival and integration during transplantation procedures represent the crux of cell-based therapies. Thus, there have been many studies focused on improving cell viability upon implantation. We used severe oxidative stress to select for a mouse Mesoangioblast subpopulation in vitro and found that this subpopulation retained self-renewal and myogenic differentiation capacities while notably enhancing cell survival, proliferation and migration relative to unselected cells. Additionally, this subpopulation of cells presented different resistance and recovery properties upon oxidative stress treatment, demonstrating select advantages over parental Mesoangioblasts in our experimental analysis. Specifically, the cells were resistant to oxidative environments, demonstrating survival, continuous self-renewal and improved migration capability. The primary outcome of the selected cells was determined in in vivo experiments in which immunocompromised dystrophic mice were injected intramuscularly in the tibialis anterior with selected or non-selected Mesoangioblasts. Resistant Mesoangioblasts exhibited markedly enhanced survival and integration into the host skeletal muscle, accounting for a more than 70% increase in engraftment compared with that of the unselected Mesoangioblast cell population and leading to remarkable muscle recovery. Thus, the positive effects of sorting on Mesoangioblast cell behaviour in vitro and in vivo suggest that a selection step involving oxidative stress preconditioning may provide a novel methodology to select for resistant cells for use in regenerative tissue applications to prevent high mortality rates upon transplantation.
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Skeletal Muscle Differentiation on a Chip Shows Human Donor Mesoangioblasts' Efficiency in Restoring Dystrophin in a Duchenne Muscular Dystrophy Model
Stem cells translational medicine, 2016Co-Authors: Elena Serena, Giulio Cossu, Susi Zatti, Alice Zoso, Francesca Lo Verso, F. Saverio Tedesco, Nicola ElvassoreAbstract:Acknowledgements Restoration of the protein dystrophin on muscle membrane is the goal of many research lines aimed at curing Duchenne muscular dystrophy (DMD). Results of ongoing preclinical and clinical trials suggest that partial restoration of dystrophin might be sufficient to significantly reduce muscle damage. Different myogenic progenitors are candidates for cell therapy of muscular dystrophies, but only satellite cells and pericytes have already entered clinical experimentation. This study aimed to provide in vitro quantitative evidence of the ability of Mesoangioblasts to restore dystrophin, in terms of protein accumulation and distribution, within myotubes derived from DMD patients, using a microengineered model. We designed an ad hoc experimental strategy to miniaturize on a chip the standard process of muscle regeneration independent of variables such as inflammation and fibrosis. It is based on the coculture, at different ratios, of human dystrophin-positive myogenic progenitors and dystrophin-negative myoblasts in a substrate with muscle-like physiological stiffness and cell micropatterns. Results showed that both healthy myoblasts and Mesoangioblasts restored dystrophin expression in DMD myotubes. However, Mesoangioblasts showed unexpected efficiency with respect to myoblasts in dystrophin production in terms of the amount of protein produced (40% vs. 15%) and length of the dystrophin membrane domain (210–240 µm vs. 40–70 µm). These results show that our microscaled in vitro model of human DMD skeletal muscle validated previous in vivo preclinical work and may be used to predict efficacy of new methods aimed at enhancing dystrophin accumulation and distribution before they are tested in vivo, reducing time, costs, and variability of clinical experimentation. This study aimed to provide in vitro quantitative evidence of the ability of human Mesoangioblasts to restore dystrophin, in terms of protein accumulation and distribution, within myotubes derived from patients with Duchenne muscular dystrophy (DMD), using a microengineered model. An ad hoc experimental strategy was designed to miniaturize on a chip the standard process of muscle regeneration independent of variables such as inflammation and fibrosis. This microscaled in vitro model, which validated previous in vivo preclinical work, revealed that Mesoangioblasts showed unexpected efficiency as compared with myoblasts in dystrophin production. Consequently, this model may be used to predict efficacy of new drugs or therapies aimed at enhancing dystrophin accumulation and distribution before they are tested in vivo.
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Intra‐arterial transplantation of HLA‐matched donor Mesoangioblasts in Duchenne muscular dystrophy
EMBO molecular medicine, 2015Co-Authors: Giulio Cossu, Francesco Saverio Tedesco, S.c. Previtali, S. Napolitano, M.p. Cicalese, F. Nicastro, M. Noviello, U. Roostalu, M.g. Natali Sora, M. ScarlatoAbstract:Intra-arterial transplantation of Mesoangioblasts proved safe and partially efficacious in preclinical models of muscular dystrophy. We now report the first-in-human, exploratory, non-randomized open-label phase I-IIa clinical trial of intra-arterial HLA-matched donor cell transplantation in 5 Duchenne patients. We administered escalating doses of donor-derived Mesoangioblasts in limb arteries under immunosuppressive therapy (tacrolimus). Four consecutive infusions were performed at 2-month intervals, preceded and followed by clinical, laboratory, and muscular MRI analyses. Two months after the last infusion, a muscle biopsy was performed. Safety was the primary endpoint. The study was relatively safe: One patient developed a thalamic stroke with no clinical consequences and whose correlation with Mesoangioblast infusion remained unclear. MRI documented the progression of the disease in 4/5 patients. Functional measures were transiently stabilized in 2/3 ambulant patients, but no functional improvements were observed. Low level of donor DNA was detected in muscle biopsies of 4/5 patients and donor-derived dystrophin in 1. Intra-arterial transplantation of donor Mesoangioblasts in human proved to be feasible and relatively safe. Future implementation of the protocol, together with a younger age of patients, will be needed to approach efficacy.
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Research Article Intra-arterial transplantation of HLA-matched donor Mesoangioblasts in Duchenne muscular dystrophy
2015Co-Authors: Giulio Cossu, Francesco Saverio Tedesco, S.c. Previtali, S. Napolitano, M.p. Cicalese, F. Nicastro, M. Noviello, U. Roostalu, Maria Grazia, Natali SoraAbstract:Intra-arterial transplantation of Mesoangioblasts proved safe and partially efficacious in preclinical models of muscular dystrophy. We now report the first-in-human, exploratory, non-randomized open-label phase I–IIa clinical trial of intra-arterial HLA-matched donor cell transplantation in 5 Duchenne patients. We administered escalating doses of donor-derived Mesoangioblasts in limb arteries under immunosuppressive therapy (tacrolimus). Four consecutive infusions were performed at 2-month intervals, preceded and followed by clinical, laboratory, and muscular MRI analyses. Two months after the last infusion, a muscle biopsy was performed. Safety was the primary endpoint. The study was relatively safe: One patient developed a thalamic stroke with no clinical consequences and whose correlation with Mesoangioblast infusion remained unclear. MRI documented the progression of the disease in 4/
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hemogenic endothelium generates Mesoangioblasts that contribute to several mesodermal lineages in vivo
Development, 2014Co-Authors: Emanuele Azzoni, Valentina Conti, Arianna Dellavalle, Ralf H. Adams, Giulio Cossu, Lara Campana, Silvia BrunelliAbstract:The embryonic endothelium is a known source of hematopoietic stem cells. Moreover, vessel-associated progenitors/stem cells with multilineage mesodermal differentiation potential, such as the ‘embryonic Mesoangioblasts’, originate in vitro from the endothelium. Using a genetic lineage tracing approach, we show that early extra-embryonic endothelium generates, in a narrow time-window and prior to the hemogenic endothelium in the major embryonic arteries, hematopoietic cells that migrate to the embryo proper, and are subsequently found within the mesenchyme. A subpopulation of these cells, distinct from embryonic macrophages, co-expresses mesenchymal and hematopoietic markers. In addition, hemogenic endothelium-derived cells contribute to skeletal and smooth muscle, and to other mesodermal cells in vivo , and display features of embryonic Mesoangioblasts in vitro . Therefore, we provide new insights on the distinctive characteristics of the extra-embryonic and embryonic hemogenic endothelium, and we identify the putative in vivo counterpart of embryonic Mesoangioblasts, suggesting their identity and developmental ontogeny.
Francesco Saverio Tedesco - One of the best experts on this subject based on the ideXlab platform.
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628. Transposons Expressing Full-Length Human Dystrophin Enable Genetic Correction of Dystrophic Mesoangioblasts and iPS-Derived Mesoangioblast-Like Cells
Molecular Therapy, 2016Co-Authors: Mariana Loperfido, Susan Jarmin, Sumitava Dastidar, Mario Di Matteo, Ilaria Perini, Marc Moore, Nisha Nair, Ermira Samara-kuko, Takis Athanasopoulos, Francesco Saverio TedescoAbstract:Duchenne muscular dystrophy (DMD) is a genetic neuromuscular disorder caused by the absence of dystrophin. We developed a novel gene therapy approach based on the use of the piggyBac (PB) transposon system to deliver the coding DNA sequence (CDS) of either full-length human dystrophin (DYS: 11.1 kb) or truncated microdystrophins (MD1: 3.6 kb; MD2: 4 kb). PB transposons encoding microdystrophins were transfected in C2C12 myoblasts, yielding 65±2% MD1 and 66±2% MD2 expression in differentiated multinucleated myotubes. A hyperactive PB (hyPB) transposase was then deployed to enable transposition of the large-size PB transposon (17 kb) encoding the full-length DYS and green fluorescence protein (GFP). Stable GFP expression attaining 78±3% could be achieved in the C2C12 myoblasts that had undergone transposition. Western blot analysis demonstrated expression of the full-length human DYS protein in myotubes. Subsequently, dystrophic Mesoangioblasts from a Golden Retriever muscular dystrophy dog were transfected with the large-size PB transposon resulting in 50±5% GFP-expressing cells after stable transposition. This was consistent with correction of the differentiated dystrophic Mesoangioblasts following expression of full-length human DYS. Alternatively, dystrophic Mesoangioblast-like cells were generated from iPS of DMD patients. These iPS-derived Mesoangioblasts, constitute an essentially unlimited supply of stem/progenitor cells that could be genetically corrected using PB transposons expressing dystrophin. These results pave the way toward a novel non-viral gene therapy approach for DMD using PB transposons underscoring their potential to deliver large therapeutic genes.
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piggyBac transposons expressing full-length human dystrophin enable genetic correction of dystrophic Mesoangioblasts.
Nucleic acids research, 2015Co-Authors: Mariana Loperfido, Susan Jarmin, Sumitava Dastidar, Mario Di Matteo, Ilaria Perini, Marc Moore, Nisha Nair, Ermira Samara-kuko, Takis Athanasopoulos, Francesco Saverio TedescoAbstract:Duchenne muscular dystrophy (DMD) is a genetic neuromuscular disorder caused by the absence of dystrophin. We developed a novel gene therapy approach based on the use of the piggyBac (PB) transposon system to deliver the coding DNA sequence (CDS) of either full-length human dystrophin (DYS: 11.1 kb) or truncated microdystrophins (MD1: 3.6 kb; MD2: 4 kb). PB transposons encoding microdystrophins were transfected in C2C12 myoblasts, yielding 65±2% MD1 and 66±2% MD2 expression in differentiated multinucleated myotubes. A hyperactive PB (hyPB) transposase was then deployed to enable transposition of the large-size PB transposon (17 kb) encoding the full-length DYS and green fluorescence protein (GFP). Stable GFP expression attaining 78±3% could be achieved in the C2C12 myoblasts that had undergone transposition. Western blot analysis demonstrated expression of the full-length human DYS protein in myotubes. Subsequently, dystrophic Mesoangioblasts from a Golden Retriever muscular dystrophy dog were transfected with the large-size PB transposon resulting in 50±5% GFP-expressing cells after stable transposition. This was consistent with correction of the differentiated dystrophic Mesoangioblasts following expression of full-length human DYS. These results pave the way toward a novel non-viral gene therapy approach for DMD using PB transposons underscoring their potential to deliver large therapeutic genes.
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Intra‐arterial transplantation of HLA‐matched donor Mesoangioblasts in Duchenne muscular dystrophy
EMBO molecular medicine, 2015Co-Authors: Giulio Cossu, Francesco Saverio Tedesco, S.c. Previtali, S. Napolitano, M.p. Cicalese, F. Nicastro, M. Noviello, U. Roostalu, M.g. Natali Sora, M. ScarlatoAbstract:Intra-arterial transplantation of Mesoangioblasts proved safe and partially efficacious in preclinical models of muscular dystrophy. We now report the first-in-human, exploratory, non-randomized open-label phase I-IIa clinical trial of intra-arterial HLA-matched donor cell transplantation in 5 Duchenne patients. We administered escalating doses of donor-derived Mesoangioblasts in limb arteries under immunosuppressive therapy (tacrolimus). Four consecutive infusions were performed at 2-month intervals, preceded and followed by clinical, laboratory, and muscular MRI analyses. Two months after the last infusion, a muscle biopsy was performed. Safety was the primary endpoint. The study was relatively safe: One patient developed a thalamic stroke with no clinical consequences and whose correlation with Mesoangioblast infusion remained unclear. MRI documented the progression of the disease in 4/5 patients. Functional measures were transiently stabilized in 2/3 ambulant patients, but no functional improvements were observed. Low level of donor DNA was detected in muscle biopsies of 4/5 patients and donor-derived dystrophin in 1. Intra-arterial transplantation of donor Mesoangioblasts in human proved to be feasible and relatively safe. Future implementation of the protocol, together with a younger age of patients, will be needed to approach efficacy.
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Efficient derivation and inducible differentiation of expandable skeletal myogenic cells from human ES and patient-specific iPS cells
Nature Protocols, 2015Co-Authors: Sara M Maffioletti, Mattia F M Gerli, Sara Benedetti, Mariana Loperfido, Sumitava Dastidar, Martina Ragazzi, Thierry Vandendriessche, Marinee K Chuah, Francesco Saverio TedescoAbstract:Skeletal muscle is the most abundant human tissue; therefore, an unlimited availability of myogenic cells has applications in regenerative medicine and drug development. Here we detail a protocol to derive myogenic cells from human embryonic stem (ES) and induced pluripotent stem (iPS) cells, and we also provide evidence for its extension to human iPS cells cultured without feeder cells. The procedure, which does not require the generation of embryoid bodies or prospective cell isolation, entails four stages with different culture densities, media and surface coating. Pluripotent stem cells are disaggregated to single cells and then differentiated into expandable cells resembling human Mesoangioblasts. Subsequently, transient Myod1 induction efficiently drives myogenic differentiation into multinucleated myotubes. Cells derived from patients with muscular dystrophy and differentiated using this protocol have been genetically corrected, and they were proven to have therapeutic potential in dystrophic mice. Thus, this platform has been demonstrated to be amenable to gene and cell therapy, and it could be extended to muscle tissue engineering and disease modeling. In this protocol, pluripotent stem cells are first differentiated into expandable cells resembling human Mesoangioblasts; subsequently, transient MyoD induction drives differentiation into multinucleated myotubes.
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pw1 peg3 expression regulates key properties that determine Mesoangioblast stem cell competence
Nature Communications, 2015Co-Authors: Chiara Bonfanti, Stefania Antonini, Sara Benedetti, Francesco Saverio Tedesco, Rossana Tonlorenzi, Giuliana Rossi, Monica Giannotta, Giovanna Marazzi, Elisabetta Dejana, David SassoonAbstract:Mesoangioblasts are vessel-associated progenitor cells that show therapeutic promise for the treatment of muscular dystrophy. Mesoangioblasts have the ability to undergo skeletal muscle differentiation and cross the blood vessel wall regardless of the developmental stage at which they are isolated. Here we show that PW1/Peg3 is expressed at high levels in Mesoangioblasts obtained from mouse, dog and human tissues and its level of expression correlates with their myogenic competence. Silencing PW1/Peg3 markedly inhibits myogenic potential of Mesoangioblasts in vitro through MyoD degradation. Moreover, lack of PW1/Peg3 abrogates Mesoangioblast ability to cross the vessel wall and to engraft into damaged myofibres through the modulation of the junctional adhesion molecule-A. We conclude that PW1/Peg3 function is essential for conferring proper Mesoangioblast competence and that the determination of PW1/Peg3 levels in human Mesoangioblasts may serve as a biomarker to identify the best donor populations for therapeutic application in muscular dystrophies.
Rossana Tonlorenzi - One of the best experts on this subject based on the ideXlab platform.
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PW1/Peg3 expression regulates key properties that determine Mesoangioblast stem cell competence
Nature Communications, 2015Co-Authors: Chiara Bonfanti, Stefania Antonini, Sara Benedetti, Rossana Tonlorenzi, Giuliana Rossi, Francesco Saverio-tedesco, Monica Giannotta, Giovanna Marazzi, Elisabetta Dejana, David SassoonAbstract:Mesoangioblasts are vessel-associated progenitor cells that show therapeutic promise for the treatment of muscular dystrophy. Mesoangioblasts have the ability to undergo skeletal muscle differentiation and cross the blood vessel wall regardless of the developmental stage at which they are isolated. Here we show that PW1/Peg3 is expressed at high levels in Mesoangioblasts obtained from mouse, dog and human tissues and its level of expression correlates with their myogenic competence. Silencing PW1/Peg3 markedly inhibits myogenic potential of Mesoangioblasts in vitro through MyoD degradation. Moreover, lack of PW1/Peg3 abrogates Mesoangioblast ability to cross the vessel wall and to engraft into damaged myofibres through the modulation of the junctional adhesion molecule-A. We conclude that PW1/Peg3 function is essential for conferring proper Mesoangioblast competence and that the determination of PW1/Peg3 levels in human Mesoangioblasts may serve as a biomarker to identify the best donor populations for therapeutic application in muscular dystrophies.
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pw1 peg3 expression regulates key properties that determine Mesoangioblast stem cell competence
Nature Communications, 2015Co-Authors: Chiara Bonfanti, Stefania Antonini, Sara Benedetti, Francesco Saverio Tedesco, Rossana Tonlorenzi, Giuliana Rossi, Monica Giannotta, Giovanna Marazzi, Elisabetta Dejana, David SassoonAbstract:Mesoangioblasts are vessel-associated progenitor cells that show therapeutic promise for the treatment of muscular dystrophy. Mesoangioblasts have the ability to undergo skeletal muscle differentiation and cross the blood vessel wall regardless of the developmental stage at which they are isolated. Here we show that PW1/Peg3 is expressed at high levels in Mesoangioblasts obtained from mouse, dog and human tissues and its level of expression correlates with their myogenic competence. Silencing PW1/Peg3 markedly inhibits myogenic potential of Mesoangioblasts in vitro through MyoD degradation. Moreover, lack of PW1/Peg3 abrogates Mesoangioblast ability to cross the vessel wall and to engraft into damaged myofibres through the modulation of the junctional adhesion molecule-A. We conclude that PW1/Peg3 function is essential for conferring proper Mesoangioblast competence and that the determination of PW1/Peg3 levels in human Mesoangioblasts may serve as a biomarker to identify the best donor populations for therapeutic application in muscular dystrophies.
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Additional file 3: Figure S3. of Mesoangioblast delivery of miniagrin ameliorates murine model of merosin-deficient congenital muscular dystrophy type 1A
2015Co-Authors: Teuta Domi, Rossana Tonlorenzi, Emanuela Porrello, Daniele Velardo, Alessia Capotondo, Alessandra Biffi, Stefano Amadio, Alessandro Ambrosi, Yuko Miyagoe-suzuki, Shin’ichi TakedaAbstract:Expression of integrin α6 in treated and not-treated dy2J mice. Cryosections of the tibialis anterior muscle from Wt, dy2J mice untreated (saline solution) or treated with MABs + mMAG stained with anti α6 integrin antibody. The immunofluorescence shows positive staining limited to the nerves and blood vessels in Wt skeletal muscle. In dy2J mice, faint and dotted staining for α6 integrin in the skeletal muscle was observed, as it was after Mesoangioblast treatment (either MABs alone or MABs + mMAG). DAPI staining identifies nuclei. Scale bar = 100 μm. (TIFF 1.05 mb
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Inflammation Converts Human Mesoangioblasts Into Targets of Alloreactive Immune Responses: Implications for Allogeneic Cell Therapy of DMD
Molecular therapy : the journal of the American Society of Gene Therapy, 2014Co-Authors: M. Noviello, Mattia F M Gerli, Francesco Saverio Tedesco, Rossana Tonlorenzi, S. Napolitano, M.p. Cicalese, Attilio Bondanza, Maria Rosaria Carbone, Sarah Marktel, Fabio CiceriAbstract:Stem cell therapy is a promising approach to regenerate healthy tissues starting from a limited amount of self-renewing cells. Immunological rejection of cell therapy products might represent a major limitation. In this study, we investigated the immunological functional profile of Mesoangioblasts, vessel-associated myogenic stem cells, currently tested in a phase 1–2a trial, active in our Institute, for the treatment of Duchenne muscular dystrophy. We report that in resting conditions, human Mesoangioblasts are poorly immunogenic, inefficient in promoting the expansion of alloreactive T cells and intrinsically resistant to T-cell killing. However, upon exposure to interferon-γ or differentiation into myotubes, Mesoangioblasts acquire the ability to promote the expansion of alloreactive T cells and acquire sensitivity to T-cell killing. Resistance of Mesoangioblasts to T-cell killing is largely due to the expression of the intracellular serine protease inhibitor-9 and represents a relevant mechanism of stem cell immune evasion.
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Human iPSC-derived Mesoangioblasts, like their tissue-derived counterparts, suppress T cell proliferation through IDO- and PGE-2-dependent pathways (v1; ref status: approved 1,
2013Co-Authors: Karen English, Giulio Cossu, Rossana Tonlorenzi, Francesco SaverioAbstract:Human Mesoangioblasts are currently in a phase I/II clinical trial for the treatment of patients with Duchenne muscular dystrophy. However, limitations associated with the finite life span of these cells combined with the significant numbers of Mesoangioblasts required to treat all of the skeletal muscles in these patients restricts their therapeutic potential. Induced pluripotent stem cell (iPSC)-derived Mesoangioblasts may provide the solution to this problem. Although, the idea of using iPSC-derived cell therapies has been proposed for quite some time, our understanding of how the immune system interacts with these cells is inadequate. Herein, we show that iPSC-derived Mesoangioblasts (HIDEMs) from healthy donors and, importantly, limb-girdle muscular dystrophy 2D patients exert immunosuppressive effects on T cell proliferation. Interferon gamma (IFN-γ) and tumour necrosis factor alpha (TNF-α) play crucial roles in the initial activation of HIDEMs and importantly indoleamine 2,3 dioxygenase (IDO) and prostaglandin E2 (PGE-2) were identified as key mechanisms involved in HIDEM suppression of T cell proliferation. Together with recent studies confirming the myogenic function and regenerative potential of these cells, we suggest that HIDEMs could provide an unlimited alternative source for Mesoangioblast-based therapies.
Silvia Brunelli - One of the best experts on this subject based on the ideXlab platform.
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Vessel-associated myogenic precursors control macrophage activation and clearance of apoptotic cells
Clinical and experimental immunology, 2014Co-Authors: Lidia Bosurgi, Silvia Brunelli, Angelo A. Manfredi, Elena Rigamonti, Antonella Monno, Patrizia Rovere-queriniAbstract:Swift and regulated clearance of apoptotic cells prevents the accumulation of cell remnants in injured tissues and contributes to the shift of macrophages towards alternatively activated reparatory cells that sustain wound healing. Environmental signals, most of which are unknown, in turn control the efficiency of the clearance of apoptotic cells and as such determine whether tissues eventually heal. In this study we show that vessel-associated stem cells (Mesoangioblasts) specifically modulate the expression of genes involved in the clearance of apoptotic cells and in macrophage alternative activation, including those of scavenger receptors and of molecules that bridge dying cells and phagocytes. Mesoangioblasts, but not immortalized myoblasts or neural precursor cells, enhance CD163 membrane expression in vitro as assessed by flow cytometry, indicating that the effect is specific. Mesoangioblasts transplanted in acutely or chronically injured skeletal muscles determine the expansion of the population of CD163+ infiltrating macrophages and increase the extent of CD163 expression. Conversely, macrophages challenged with Mesoangioblasts engulf significantly better apoptotic cells in vitro. Collectively, the data reveal a feed-forward loop between macrophages and vessel-associated stem cells, which has implications for the skeletal muscle homeostatic response to sterile injury and for diseases in which homeostasis is jeopardized, including muscle dystrophies and inflammatory myopathies.
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hemogenic endothelium generates Mesoangioblasts that contribute to several mesodermal lineages in vivo
Development, 2014Co-Authors: Emanuele Azzoni, Valentina Conti, Arianna Dellavalle, Ralf H. Adams, Giulio Cossu, Lara Campana, Silvia BrunelliAbstract:The embryonic endothelium is a known source of hematopoietic stem cells. Moreover, vessel-associated progenitors/stem cells with multilineage mesodermal differentiation potential, such as the ‘embryonic Mesoangioblasts’, originate in vitro from the endothelium. Using a genetic lineage tracing approach, we show that early extra-embryonic endothelium generates, in a narrow time-window and prior to the hemogenic endothelium in the major embryonic arteries, hematopoietic cells that migrate to the embryo proper, and are subsequently found within the mesenchyme. A subpopulation of these cells, distinct from embryonic macrophages, co-expresses mesenchymal and hematopoietic markers. In addition, hemogenic endothelium-derived cells contribute to skeletal and smooth muscle, and to other mesodermal cells in vivo , and display features of embryonic Mesoangioblasts in vitro . Therefore, we provide new insights on the distinctive characteristics of the extra-embryonic and embryonic hemogenic endothelium, and we identify the putative in vivo counterpart of embryonic Mesoangioblasts, suggesting their identity and developmental ontogeny.
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Transplanted Mesoangioblasts Require Macrophage IL-10 for Survival in a Mouse Model of Muscle Injury
Journal of immunology (Baltimore Md. : 1950), 2012Co-Authors: Lidia Bosurgi, Silvia Brunelli, Giulio Cossu, Gianfranca Corna, Michela Vezzoli, Thierry Touvier, Angelo A. Manfredi, Patrizia Rovere-queriniAbstract:The aim of this study was to verify whether macrophages influence the fate of transplanted Mesoangioblasts--vessel-associated myogenic precursors--in a model of sterile toxin-induced skeletal muscle injury. We have observed that in the absence of macrophages, transplanted Mesoangioblasts do not yield novel fibers. Macrophages retrieved from skeletal muscles at various times after injury display features that resemble those of immunoregulatory macrophages. Indeed, they secrete IL-10 and express CD206 and CD163 membrane receptors and high amounts of arginase I. We have reconstituted the muscle-associated macrophage population by injecting polarized macrophages before Mesoangioblast injection: alternatively activated, immunoregulatory macrophages only support Mesoangioblast survival and function. This action depends on the secretion of IL-10 in the tissue. Our results reveal an unanticipated role for tissue macrophages in Mesoangioblast function. Consequently, the treatment of muscle disorders with Mesoangioblasts should take into consideration coexisting inflammatory pathways, whose activation may prove crucial for its success.
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Sphingosine 1-phosphate induces differentiation of Mesoangioblasts towards smooth muscle. A role for GATA6.
PloS one, 2011Co-Authors: Chiara Donati, Silvia Brunelli, Giuseppina Marseglia, Alberto Magi, Simona Serratì, Francesca Cencetti, Caterina Bernacchioni, Genni Nannetti, Matteo Benelli, Francesca TorricelliAbstract:Different cells can contribute to repair following vascular injury by differentiating into smooth muscle (SM) cells; however the extracellular signals involved are presently poorly characterized. Mesoangioblasts are progenitor cells capable of differentiating into various mesoderm cell types including SM cells. In this study the biological action exerted by the pleiotropic sphingolipid sphingosine 1-phosphate (S1P) in human Mesoangioblasts has been initially investigated by cDNA microarray analysis. Obtained data confirmed the anti-apoptotic action of this sphingolipid and identified for the first time a strong differentiating action toward SM cells. Quantitative mRNA and protein analysis corroborated the microarray results demonstrating enhanced expression of myogenic marker proteins and regulation of the expression of transcription factor GATA6 and its co-regulator, LMCD1. Importantly, GATA6 up-regulation induced by S1P was responsible for the enhanced expression of SM-specific contractile proteins. Moreover, by specific gene silencing experiments GATA6 was critical in the pro-differentiating activity of the cytokine TGFβ. Finally, the pharmacological inhibition of endogenous S1P formation in response to TGFβ abrogated GATA6 up-regulation, supporting the view that the S1P pathway plays a physiological role in mediating the pro-myogenic effect of TGFβ. This study individuates GATA6 as novel player in the complex transcriptional regulation of Mesoangioblast differentiation into SM cells and highlights a role for S1P to favour vascular regeneration.
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Human cardiac Mesoangioblasts isolated from hypertrophic cardiomyopathies are greatly reduced in proliferation and differentiation potency
Cardiovascular research, 2009Co-Authors: Beatriz G. Gálvez, Arianna Dellavalle, Silvia Brunelli, Diego Covarello, Rosanna Tolorenzi, Stefania Crippa, Salman Afroze Azmi Mohammed, Ludovica Scialla, Ivan Cuccovillo, Fabiola MollaAbstract:AIMS: Our objective was to test whether progenitor cell proliferation and differentiation potential may vary depending upon the disease of the donor. METHODS AND RESULTS: Human cardiac Mesoangioblasts were isolated from cardiac muscle biopsies of patients undergoing open heart surgery for correction of mitral regurgitation following an acute myocardial infarction (MR-MI) or correction of mitral and aortic regurgitation with ensuing left ventricular hypertrophy (MAR-LVH). The cells express surface markers and cardiac genes similar to mouse cardiac Mesoangioblasts; they have limited self-renewing and clonogenic activity and are committed mainly to cardiogenesis. Although cardiac differentiation can be induced by 5-azacytidine or by co-culture with rat neonatal cardiomyocytes, human cells do not contract spontaneously like their mouse counterparts. When locally injected in the infarcted myocardium of immunodeficient mice, cardiac Mesoangioblasts generate a chimeric heart that contains human myocytes and some capillaries; likewise, they colonize chick embryo hearts when transplanted in ovo. At variance with cells from patients with MR-MI, when isolation was performed on biopsies from MAR-LVH, cells could be isolated in much lower numbers, proliferated less extensively and failed to differentiate. CONCLUSION: Cardiac Mesoangioblasts are present in the human heart but this endogenous progenitor population is progressively exhausted, possibly by continuous and inefficient regeneration attempts.
Suzanne E. Berry - One of the best experts on this subject based on the ideXlab platform.
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Injection of Vessel-Derived Stem Cells Prevents Dilated Cardiomyopathy and Promotes Angiogenesis and Endogenous Cardiac Stem Cell Proliferation in mdx/utrn−/− but Not Aged mdx Mouse Models for Duchenne Muscular Dystrophy
Stem cells translational medicine, 2012Co-Authors: Ju Lan Chun, Robert T. O'brien, Minho Song, Blake F. Wondrasch, Suzanne E. BerryAbstract:Duchenne muscular dystrophy (DMD) is the most common form of muscular dystrophy. DMD patients lack dystrophin protein and develop skeletal muscle pathology and dilated cardiomyopathy (DCM). Approximately 20% succumb to cardiac involvement. We hypothesized that Mesoangioblast stem cells (aorta-derived Mesoangioblasts [ADMs]) would restore dystrophin and alleviate or prevent DCM in animal models of DMD. ADMs can be induced to express cardiac markers, including Nkx2.5, cardiac tropomyosin, cardiac troponin I, and α-actinin, and adopt cardiomyocyte morphology. Transplantation of ADMs into the heart of mdx/utrn(-/-) mice prior to development of DCM prevented onset of cardiomyopathy, as measured by echocardiography, and resulted in significantly higher CD31 expression, consistent with new vessel formation. Dystrophin-positive cardiomyocytes and increased proliferation of endogenous Nestin(+) cardiac stem cells were detected in ADM-injected heart. Nestin(+) striated cells were also detected in four of five mdx/utrn(-/-) hearts injected with ADMs. In contrast, when ADMs were injected into the heart of aged mdx mice with advanced fibrosis, no functional improvement was detected by echocardiography. Instead, ADMs exacerbated some features of DCM. No dystrophin protein, increase in CD31 expression, or increase in Nestin(+) cell proliferation was detected following ADM injection in aged mdx heart. Dystrophin was observed following transplantation of ADMs into the hearts of young mdx mice, however, suggesting that pathology in aged mdx heart may alter the fate of donor stem cells. In summary, ADMs delay or prevent development of DCM in dystrophin-deficient heart, but timing of stem cell transplantation may be critical for achieving benefit with cell therapy in DMD cardiac muscle.
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14 1 t whole body mri for detection of Mesoangioblast stem cells in a murine model of duchenne muscular dystrophy
Magnetic Resonance in Medicine, 2011Co-Authors: Boris Odintsov, Ju Lan Chun, James A. Mulligan, Suzanne E. BerryAbstract:Noninvasive imaging procedures will be important for stem cell therapy for muscular dystrophy (MD). Mesoangioblasts regenerate muscle in animal models of muscular dystrophy. In this study, superparamagnetic iron oxide nanoparticles were used to visualize Mesoangioblasts in vivo with MRI. Mesoangioblasts incorporated superparamagnetic iron oxide without transfection reagents, and cell differentiation was not negatively impacted. A custom-built radiofrequency coil with an adjustable field of view and 14.1 T magnet were used for whole-body MRI of mice. High-resolution images of Mesoangioblasts in skeletal and cardiac muscle of Mdx mice were obtained following local delivery. Labeled cells were verified by Prussian blue staining and dystrophin expression, indicating that the wild-type Mesoangioblasts survived and differentiated in muscle. Iron-labeled cells were detected with MRI in vivo 6 months following intracardiac injection but were determined to be activated macrophages. Iron-labeled cells were not detected by MRI following systemic delivery but were present in skeletal and cardiac muscle, visualized by Prussian blue staining. Systemically delivered Mesoangioblasts were detected in lungs by Prussian blue staining and DiI but not by MRI in our study. MRI may be useful for short-term tracking of Mesoangioblasts delivered locally but not for long-term monitoring or detection after systemic delivery.
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Aorta-derived Mesoangioblasts differentiate into the oligodendrocytes by inhibition of the Rho kinase signaling pathway
Stem cells and development, 2011Co-Authors: Lei Wang, Ju Lan Chun, Anant Kamath, Janie Frye, Gary A. Iwamoto, Suzanne E. BerryAbstract:Mesoangioblasts are vessel-derived stem cells that differentiate into mesodermal derivatives. We have isolated postnatal aorta-derived Mesoangioblasts (ADMs) that differentiate into smooth, skeletal, and cardiac muscle, and adipocytes, and regenerate damaged skeletal muscle in a murine model for Duchenne muscular dystrophy. We report that the marker profile of ADM is similar to that of Mesoangioblasts isolated from embryonic dorsal aorta, postnatal bone marrow, and heart, but distinct from Mesoangioblasts derived from skeletal muscle. We also demonstrate that ADM differentiate into myelinating glial cells. ADM localize to peripheral nerve bundles in regenerating muscles and exhibit morphology and marker expression of mature Schwann cells, and myelinate axons. In vitro, ADM spontaneously express markers of oligodendrocyte progenitors, including the chondroitin sulphate proteoglycan NG2, nestin, platelet-derived growth factor (PDGF) receptor α, the A2B5 antigen, thyroid hormone nuclear receptor α, and O4. P...
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14.1 T whole body MRI for detection of Mesoangioblast stem cells in a murine model of duchenne muscular dystrophy
Magnetic Resonance in Medicine, 2011Co-Authors: Boris Odintsov, Ju Lan Chun, James A. Mulligan, Suzanne E. BerryAbstract:Noninvasive imaging procedures will be important for stem cell therapy for muscular dystrophy (MD). Mesoangioblasts regenerate muscle in animal models of muscular dystrophy. In this study, superparamagnetic iron oxide nanoparticles were used to visualize Mesoangioblasts in vivo with MRI. Mesoangioblasts incorporated superparamagnetic iron oxide without transfection reagents, and cell differentiation was not negatively impacted. A custom-built radiofrequency coil with an adjustable field of view and 14.1 T magnet were used for whole-body MRI of mice. High-resolution images of Mesoangioblasts in skeletal and cardiac muscle of Mdx mice were obtained following local delivery. Labeled cells were verified by Prussian blue staining and dystrophin expression, indicating that the wild-type Mesoangioblasts survived and differentiated in muscle. Iron-labeled cells were detected with MRI in vivo 6 months following intracardiac injection but were determined to be activated macrophages. Iron-labeled cells were not detected by MRI following systemic delivery but were present in skeletal and cardiac muscle, visualized by Prussian blue staining. Systemically delivered Mesoangioblasts were detected in lungs by Prussian blue staining and DiI but not by MRI in our study. MRI may be useful for short-term tracking of Mesoangioblasts delivered locally but not for long-term monitoring or detection after systemic delivery. Magn Reson Med, 2011. © 2011 Wiley Periodicals, Inc.
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multipotential Mesoangioblast stem cell therapy in the mdx utrn mouse model for duchenne muscular dystrophy
Regenerative Medicine, 2007Co-Authors: Suzanne E. Berry, Jianming Liu, Eric J. Chaney, Stephen J. KaufmanAbstract: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 α-smooth muscle act...