Impact of anti-apoptotic and anti-oxidative effects of bone marrow mesenchymal stem cells with transient overexpression of heme oxygenase-1 on myocardial ischemia. Am J Physiol Heart Circ Physiol 298(5):H1320-9

Division of Cardiovascular Medicine, Kanazawa University Graduate School of Medicine, Takara-machi 13-1, Kanazawa, Ishikawa, Japan.
AJP Heart and Circulatory Physiology (Impact Factor: 3.84). 02/2010; 298(5):H1320-9. DOI: 10.1152/ajpheart.01330.2008
Source: PubMed


Although mesenchymal stem cells (MSCs) have therapeutic potential for tissue injury, intolerance and poor cell viability limit their reparative capability. Therefore, we examined the impact of bone marrow-derived MSCs, in which heme oxygenase-1 (HO-1) was transiently overexpressed, on the repair of an ischemic myocardial injury. When MSCs and HO-1-overexpressed MSCs (MSC(HO-1)) were exposed to serum deprivation/hypoxia or H(2)O(2)-induced oxidative stress, MSC(HO-1) exhibited increased resistance to cell apoptosis compared with MSCs (17 +/- 1 vs. 30 +/- 2%, P < 0.05) and were markedly resistant to cell death (2 +/- 1 vs. 32 +/- 2%, P < 0.05). Under these conditions, vascular endothelial growth factor (VEGF) production was 2.1-fold greater in MSC(HO-1) than in MSCs. Pretreatment of MSCs and MSC(HO-1) with phosphatidylinositol 3-kinase (PI 3-kinase)/protein kinase B (Akt) pathway inhibitors such as LY-294002 (50 muM) or wortmannin (100 nM) significantly decreased VEGF production. In a rat infarction model with MSCs or MSC(HO-1) (5 x 10(6) +/- 0.1 x 10(6) cells/rat) transplantation, the number of TdT-mediated dUTP nick end-labeling-positive cells was significantly lower in the MSC(HO-1) group than in the MSC group (12.1 +/- 1.0 cells/field vs. 26.5 +/- 2.6, P < 0.05) on the 4th day after cell transplantation. On the 28th day, increased capillary density associated with decreased infarction size was observed in the MSC(HO-1) group (1,415 +/- 47/mm(2) with 21.6 +/- 2.3%) compared with those in the MSCs group (1,215 +/- 43/mm(2) with 28.2 +/- 2.3%, P < 0.05), although infarction size relative to area at risk was not different in each group at 24 h after transplantation. These results demonstrate that MSC(HO-1) exhibit markedly enhanced anti-apoptotic and anti-oxidative capabilities compared with MSCs, thus contributing to improved repair of ischemic myocardial injury through cell survival and VEGF production associated with the PI 3-kinase/Akt pathway.

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    • "Our results showed that the number of apoptotic MSC-HO-1 cells were lower than the control cells. Supporting our study, it has been shown recently that the MSC-HO-1s had better anti-oxidative and antiapoptotic properties than unmodified MSCs (Tsubokawa et al. 2010). It should be noted that in the Tsubokawa study, a nonviral vector was used for expression of HO-1 while in the present study we transiently over-expressed the human HO-1 gene in the MSCs using an adenoviral expression system. "
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    ABSTRACT: The capacity of mesenchymal stem cells (MSCs) to survive and engraft in the target tissue may lead to promising therapeutic effects. However, the fact that the majority of MSCs die during the first few days following transplantation complicates cell therapy. Hence, it is necessary to strengthen the stem cells to withstand the rigors of the microenvironment to improve the efficacy of cell therapy. In this study, we manipulated MSCs to express a cytoprotective factor, heme oxygenase-1 (HO-1), to address this issue. Full-length cDNA of human HO-1 was isolated and cloned into TOPO vector by TOPO cloning reaction. Then, the construct was ligated to gateway adapted adenovirus expression vector by LR recombination reaction. Afterwards, the recombinant virus expressing HO-1 was produced in appropriate mammalian cell line and used to infect MSCs. The HO-1 engineered MSCs were exposed to hypoxic and oxidative stress conditions fol-lowed by evaluation of the cells' viability and apoptosis. Transient expression of HO-1 was detected within MSCs. It was observed that HO-1 expression could protect MSCs against cell death and the apoptosis triggered by hypoxic and oxidative stress conditions. The MSCs-HO-1 retained their ability to differentiate into adipogenic, chondrogenic, or osteogenic lineages. These findings could be applied as a strategy for prevention of graft cell death in MSCs-based cell therapy and is a good demonstration of how an understanding of cellular stress responses can be used for practical applications.
    Cell Stress and Chaperones 04/2012; 17(2). DOI:10.1007/s12192-011-0298-y · 3.16 Impact Factor
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    • "This result has since been shown to be reproducible in a large animal model of myocardial ischemia.[89] Other genetic means to enhance the therapeutic effect of MSCs in the cardiovascular setting include the overexpression of; VEGF,[30] eNOS,[90] HGF,[31] BCL-2,[91] heme-oxygenase-1,[9293] Integrin-linked kinase,[94] angiopoietin-1,[95] PI3K-C2a,[96] GATA-4,[97] SDF-1,[98] CCR1,[99] CXCR1/CXCR2,[100] CXCR4,[101] and Cx43.[102] Our group has also demonstrated that MSCs can be retrovirally engineered to secrete high levels of Epo allows for a more robust host-derived angiogenic response, enhanced MSC survival, better preserves myocardial contractility in a murine model of MI and can reduce neutrophilic cellular infiltration.[39] "
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    ABSTRACT: The fields of regenerative medicine and cellular therapy have been the subject of tremendous hype and hope. In particular, the perceived usage of somatic cells like mesenchymal stromal cells (MSCs) has captured the imagination of many. MSCs are a rare population of cells found in multiple regions within the body that can be readily expanded ex vivo and utilized clinically. Originally, it was hypothesized that transplantation of MSCs to sites of injury would lead to de novo tissue-specific differentiation and thereby replace damaged tissue. Now, it is generally agreed that MSC home to sites of injury and direct positive remodeling via the secretion of paracrine factors. Consequently, their clinical utilization has largely revolved around their abilities to promote neovascularization for ischemic disorders and modulate overly exuberant inflammatory responses for autoimmune and alloimmune conditions. One of the major issues surrounding the development of somatic cell therapies like MSCs is that despite evoking a positive response, long-term engraftment and persistence of these cells is rare. Consequently, very large cell doses need be administered for raising production, delivery, and efficacy issues. In this review, we will outline the field of MSC in the context of ischemia and discuss causes for their lack of persistence. In addition, some of the methodologies be used to enhance their therapeutic potential will be highlighted.
    Journal of cardiovascular disease research 03/2011; 2(1):3-13. DOI:10.4103/0975-3583.78581
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    • "In addition, decreased expression of the proapoptotic protein Bax and reductions in the levels of proinflammatory molecules tumor necrosis factor (TNF)-a, interleukin (IL)-1b, and IL-6 were also observed (Zeng et al., 2008a,b). Moreover, MSCs in which HO-1 was transiently overexpressed also showed enhanced antiapoptotic and antioxidative properties leading to enhanced repair of the myocardium (Tsubokawa et al., 2010). Additional experiments have taken the idea of HO-1 overexpression further by producing MSCs that overexpress both HO-1 and inducible nitric oxide synthase (iNOS). "
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    ABSTRACT: The use of stem cells for tissue regeneration and repair is advancing both at the bench and bedside. Stem cells isolated from bone marrow are currently being tested for their therapeutic potential in a variety of clinical conditions including cardiovascular injury, kidney failure, cancer, and neurological and bone disorders. Despite the advantages, stem cell therapy is still limited by low survival, engraftment, and homing to damage area as well as inefficiencies in differentiating into fully functional tissues. Genetic engineering of mesenchymal stem cells is being explored as a means to circumvent some of these problems. This review presents the current understanding of the use of genetically engineered mesenchymal stem cells in human disease therapy with emphasis on genetic modifications aimed to improve survival, homing, angiogenesis, and heart function after myocardial infarction. Advancements in other disease areas are also discussed.
    Human gene therapy 11/2010; 21(11):1513-26. DOI:10.1089/hum.2010.165 · 3.76 Impact Factor
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