Cell Therapy For Cardiovascular Diseases Research Articles

Stem cell therapy for cardiac regeneration: key points

Cardiovascular diseases are a leading cause of disability and mortality among the working population, necessitating the development and implementation of new, more effective treatment and rehabilitation methods for such patients, including the use of modern cellular and tissue technologies. In this review, an analysis and summary of research results over the past decades regarding the effectiveness of various types of stem cells in heart pathology were conducted. Priority directions include the search for more accessible and safe sources of stem cells, the development of new effective methods for their cultivation, and the use of scaffolds for transplantation. Given the pathogenetic mechanisms of cardiovascular pathology and myocardial regeneration, particular interest for clinical application is drawn to tissue-specific progenitors from the myocardium for replacing injured cardiomyocytes, endothelial progenitors for correcting accompanying endothelial dysfunction, and multipotent mesenchymal cells of various origins with low immunogenicity that exhibit trophic, anti-inflammatory, immunomodulatory, and anti-apoptotic effects. Questions regarding the optimal method, dose, and frequency of stem cell administration, as well as the standardization of criteria for their quality and effectiveness, remain open. Additionally, the combined use of different types of stem cells may serve as a new priority strategy in assessing the effectiveness and safety of cell therapy, especially considering the paradigm of repeated transplantation. The successful translation of obtained preclinical results into further large, well-planned, placebo-controlled clinical trials will enhance the safety and effectiveness of cell therapy for cardiovascular diseases, which is one of the current challenges of modern regenerative medicine.

Human Adipose-Derived Mesenchymal Stem Cell-Based Microspheres Ameliorate Atherosclerosis Progression In Vitro.

Atherosclerosis (AS) is a chronic inflammatory disease associated with lipid deposition, which could be converted into acute clinical events by thrombosis or plaque rupture. Adipose-derived mesenchymal stem cell (ADSC)-encapsulated repair units could be an effective cure for the treatment of AS patients. In this study, we encapsulate human adipose-derived mesenchymal stem cells (hADSCs) in collagen microspheres to fabricate stem cell repair units. Besides, we show that encapsulation in collagen microspheres and cultured in vitro for 14 days maintain the viability and stemness of hADSCs. Moreover, we generate AS progression model and niche in vitro by combining hyperlipemia serum of AS patients with AS cell models. We further systematically demonstrate that hADSC-based microspheres could ameliorate AS progression by inhibiting oxidative stress injury, cell apoptosis, endothelial dysfunction, inflammation, and lipid accumulation. In addition, we perform transcriptomic analysis and functional studies to demonstrate how hADSCs (three dimensional cultured in microspheres) respond to AS niche compared with healthy microenvironment. These findings reveal a role for ADSC-based microspheres in the treatment of AS and provide new ideas for stem cell therapy in cardiovascular disease. The results may have implications for improving the efficiency of hADSC therapies by illuminating the mechanisms of hADSCs exposed in special pathological niche.

CRISPR-Cas9 editing of TLR4 to improve the outcome of cardiac cell therapy

Inflammation and fibrosis limit the reparative properties of human mesenchymal stromal cells (hMSCs). We hypothesized that disrupting the toll-like receptor 4 (TLR4) gene would switch hMSCs toward a reparative phenotype and improve the outcome of cell therapy for infarct repair. We developed and optimized an improved electroporation protocol for CRISPR-Cas9 gene editing. This protocol achieved a 68% success rate when applied to isolated hMSCs from the heart and epicardial fat of patients with ischemic heart disease. While cell editing lowered TLR4 expression in hMSCs, it did not affect classical markers of hMSCs, proliferation, and migration rate. Protein mass spectrometry analysis revealed that edited cells secreted fewer proteins involved in inflammation. Analysis of biological processes revealed that TLR4 editing reduced processes linked to inflammation and extracellular organization. Furthermore, edited cells expressed less NF-ƙB and secreted lower amounts of extracellular vesicles and pro-inflammatory and pro-fibrotic cytokines than unedited hMSCs. Cell therapy with both edited and unedited hMSCs improved survival, left ventricular remodeling, and cardiac function after myocardial infarction (MI) in mice. Postmortem histologic analysis revealed clusters of edited cells that survived in the scar tissue 28 days after MI. Morphometric analysis showed that implantation of edited cells increased the area of myocardial islands in the scar tissue, reduced the occurrence of transmural scar, increased scar thickness, and decreased expansion index. We show, for the first time, that CRISPR-Cas9-based disruption of the TLR4-gene reduces pro-inflammatory polarization of hMSCs and improves infarct healing and remodeling in mice. Our results provide a new approach to improving the outcomes of cell therapy for cardiovascular diseases.

Human umbilical cord mesenchymal stem cells derived exosomes exert antiapoptosis effect via activating PI3K/Akt/mTOR pathway on H9C2 cells.

In recent years, as an alternative to stem cell therapy for cardiovascular diseases (CVD), exsomes have attracted wide attention among researchers. The present study aimed to investigate the role of human umbilical cord mesenchymal stem cells (UC-MSCs) derived exosomes play on H9C2 cells apoptosis and possible mechanisms. Exosomes were isolated from normal UC-MSCs culture media and hypoxic preconditioning culture media. Transmission electron microscopy was used to observe the morphology of exosomes. Nanoparticle tracking analysis was used to detect the size distribution and concentration of exosomes. Western blot analysis was used to analyzed the surface marker CD63 of exosomes. H9C2 cells were induced apoptosis by hypoxia and serum deprivation (H/SD) and then were treated respectively by group. Cell Counting Kit-8 assay was used to detect viability of H9C2 cells. Apoptosis was detected by Hochest staining and annexin V-FITC/PI. The expression levels of related proteins of apoptosis, autophagy, and PI3K/Akt/mTOR pathway were analyzed by Western blot analysis. Immunofluorescence was used to analyze LC3B expression. Hypoxic preconditioning increased the exosomes secretion of UC-MSCs. UC-MSCs derived exosomes could inhibit H/SD-induced H9C2 cells apoptosis. Hypoxic preconditioning strengthened this antiapoptosis effect of UC-MSCs. Hypoxic preconditioning UC-MSCs derived exosomes (H-Exo) downregulated LC3B-II/I and beclin-1 and upregulated P62, p-Akt/Akt and p-mTOR/mTOR. The antiapoptotic effect of H-Exo could be attenuated by treatment with LY294002 and rapamycin. UC-MSCs derived exosomes could inhibit H9C2 cells apoptosis induced by H/SD through regulating autophagy via PI3K/Akt/mTOR pathway. Hypoxia preconditioning could enhance above effects through increasing exosomes secretion of UC-MSCs.

Meet the First Authors.

Meet the First Authors.

The Biological Mechanisms of Action of Cardiac Progenitor Cell Therapy.

Cell therapy for cardiovascular diseases is regarded as a rapidly growing field within regenerative medicine. Different cellular populations enriched for cardiac progenitor cells (CPCs), or derivate a-cellular products, are currently under preclinical and clinical evaluation. Here, we have reviewed the described mechanisms whereby resident post-natal CPCs, isolated in different ways, act as a therapeutic product on the damaged myocardium. Several biological mechanisms of action have been described which can explain the multiple therapeutic effects of CPC treatment observed on cardiac function and remodelling. These mechanisms span from direct cardiovascular differentiation, through induction of resident progenitor proliferation, to paracrine effects on cardiac and non-cardiac cells mediated by exosomes and non-coding RNAs. All the reported mechanisms of action support an integrated view including cardiomyogenesis, cardioprotection, and anti-fibrotic effects. Moreover, future developments of CPC therapy approaches may support cell-free strategies, exploiting effective pleiotropic cell-derived products, such as exosomes.

Meet the First Authors.

Meet the First Authors.

Single-Cell Sequencing Technologies for Cardiac Stem Cell Studies.

Today with the rapid advancements in stem cell studies and the promising potential of using stem cells in clinical therapy, there is an increasing demand for in-depth comprehensive analysis on individual cell transcriptome and epigenome, as they play critical roles in a number of cell functions such as cell differentiation, growth, and reprogramming. The development of single-cell sequencing technologies has helped in revealing some exciting new perspectives in stem cells and regenerative medicine research. Among the various potential applications, single-cell analysis for cardiac stem cells (CSCs) holds tremendous promises in understanding the mechanisms of heart development and regeneration, which might light up the path toward cell therapy for cardiovascular diseases. This review briefly highlights the recent progresses in single-cell sequencing analysis technologies and their applications in CSC research.

Paracrine-Mediated Systemic Anti-Inflammatory Activity of Intravenously Administered Mesenchymal Stem Cells: A Transformative Strategy for Cardiac Stem Cell Therapeutics.

Stem cell therapy as a treatment option for acute myocardial infarction or heart failure caused by ischemic or nonischemic cardiomyopathy has focused on direct cardiac delivery of stem cells to facilitate cardiac engraftment. Once engrafted, it was believed that these cells would either transdifferentiate into new functioning myocytes or stimulate the expansion of resident myocardial stem cells. However, a sea change in thinking about cell therapy in cardiovascular disease has occurred as mounting evidence indicates that (1) inflammation is a major mechanism contributing to the progressive myocardial dysfunction seen in patients post-acute myocardial infarction and in patients with cardiomyopathy, and (2) systemic paracrine-mediated anti-inflammatory effects of stem cells can drive beneficial cardiac effects in these diseases. These concepts lead to a potentially transformative strategy that intravenous delivery of stem cells, through systemic anti-inflammatory mechanisms, improves myocardial function and thereby obviates the need for invasive methods of stem cell delivery. Until recently, the prevailing view of the mechanism responsible for any potential benefit of stem cells in patients with acute myocardial infarction (AMI) or with heart failure (HF) caused by ischemic or nonischemic cardiomyopathy (ICM/NICM) was that benefit derived from local effects—once engrafted in damaged myocardium, the stem cells either transdifferentiate into functional myocardium, stimulate resident myocardial stem cells to expand, and repopulate the heart with functioning myocytes or secrete substances leading to myocardial healing. This mechanistic perspective implied that the greater the number of engrafted cells in the myocardium, the greater the cardiac benefit. Because few intravenously administered stem cells engraft in injured myocardium, invasive strategies providing direct delivery of stem cells to the heart were uniformly adopted. This necessarily involved either catheter-based delivery (intracoronary or transendocardial injection) or surgical delivery (direct intramyocardial injection). A transformative concept relating to stem cell treatment of patients with AMI or ICM/NICM has recently …

GW28-e0659 Regenerative rehabilitation: a hybrid strategy for heart disease

The aim of this study is to anticipate the role of cardiac rehabilitation in enhancement of efficacy of cell therapy in cardiovascular disease. In this narrative study search several data base using key words including cell therapy, stem cell, regeneration rehabilitation, cardiac rehabilitation and

Forging the Fate of Cellular Therapies for Cardiovascular Disease.

The field of cell therapy for cardiovascular disease has progressed at an uneven rate and will likely follow the translational paths of other biological therapies. The field will require novel public–private partnerships to guide it to its ultimate applications. The long process of developing new therapeutics for cardiovascular disease may often seem like a guideless journey into the unknown. The process begins with definition of an unmet clinical need and is combined with insights into the pathophysiologic nature of disease. Whether the proposed therapy is a device, small molecule, or biological each requires extensive preclinical and clinical investigation prior to approval. While each new treatment must create its own path to approval, outcomes may be predicted by prior experiences. Cell-based therapies for cardiac and vascular disease have progressed at an uneven pace. High-profile publications suggested plasticity and potency of bone marrow–derived cells when delivered into infarcted myocardium and spurred clinical investigation seemingly before the ink dried on the original papers.1 These human studies identified a promise of therapy without harmful effects when data were aggregated.2 However, these uniformly positive small studies have not been confirmed by larger studies. This has prompted a reexamination of the underlying premise on which the original studies were based.3,4 The path by which cell-based therapeutics is being developed is illuminated by the development of other biological approaches for cardiovascular disease. The similarities allow for educated guesses as to the outcome of this therapeutic journey. Will cells ever be widely used as first-line therapy for cardiac and vascular diseases? Which diseases will be the first approved targets for cell-based therapies? How might the development process be improved? I think the answers to these questions may be informed by the fates of other biologics, including antibodies, gene therapy, and peptides. Few academic …

Optimizing stem cells for cardiac repair: Current status and new frontiers in regenerative cardiology.

Cell therapy has the potential to improve healing of ischemic heart, repopulate injured myocardium and restore cardiac function. The tremendous hope and potential of stem cell therapy is well understood, yet recent trials involving cell therapy for cardiovascular diseases have yielded mixed results with inconsistent data thereby readdressing controversies and unresolved questions regarding stem cell efficacy for ischemic cardiac disease treatment. These controversies are believed to arise by the lack of uniformity of the clinical trial methodologies, uncertainty regarding the underlying reparative mechanisms of stem cells, questions concerning the most appropriate cell population to use, the proper delivery method and timing in relation to the moment of infarction, as well as the poor stem cell survival and engraftment especially in a diseased microenvironment which is collectively acknowledged as a major hindrance to any form of cell therapy. Indeed, the microenvironment of the failing heart exhibits pathological hypoxic, oxidative and inflammatory stressors impairing the survival of transplanted cells. Therefore, in order to observe any significant therapeutic benefit there is a need to increase resilience of stem cells to death in the transplant microenvironment while preserving or better yet improving their reparative functionality. Although stem cell differentiation into cardiomyocytes has been observed in some instance, the prevailing reparative benefits are afforded through paracrine mechanisms that promote angiogenesis, cell survival, transdifferentiate host cells and modulate immune responses. Therefore, to maximize their reparative functionality, ex vivo manipulation of stem cells through physical, genetic and pharmacological means have shown promise to enable cells to thrive in the post-ischemic transplant microenvironment. In the present work, we will overview the current status of stem cell therapy for ischemic heart disease, discuss the most recurring cell populations employed, the mechanisms by which stem cells deliver a therapeutic benefit and strategies that have been used to optimize and increase survival and functionality of stem cells including ex vivo preconditioning with drugs and a novel “pharmaco-optimizer” as well as genetic modifications.

Stem cell therapy for cardiovascular diseases

Stem cell therapy for cardiovascular diseases

Adult Stem Cell Therapy and Heart Failure, 2000 to 2016: A Systematic Review.

Stem cell therapy is a promising treatment strategy for patients with heart failure, which accounts for more than 10% of deaths in the United States annually. Despite more than a decade of research, further investigation is still needed to determine whether stem cell regenerative therapy is an effective treatment strategy and can be routinely implemented in clinical practice. To describe the progress in cardiac stem cell regenerative therapy using adult stem cells and to highlight the merits and limitations of clinical trials performed to date. Information for this review was obtained through a search of PubMed and the Cochrane database for English-language studies published between January 1, 2000, and July 26, 2016. Twenty-nine randomized clinical trials and 7 systematic reviews and meta-analyses were included in this review. Although adult stem cells were once believed to have the ability to create new heart tissue, preclinical studies suggest that these cells release cardioprotective paracrine factors that activate endogenous pathways, leading to myocardial repair. Subsequent randomized clinical trials, most of which used autologous bone marrow mononuclear cells, have found only a modest benefit in patients receiving stem cell therapy. The lack of a significant benefit may result from variations in trial methods, discrepancies in reporting, and an overreliance on surrogate end points. Although stem cell therapy for cardiovascular disease is not yet ready for routine clinical application, significant progress continues to be made. Physicians should be aware of the current status of this treatment so that they can better inform their patients who may be in search of alternative therapies.

Advances in stem cell therapy for cardiovascular disease (Review).

Cardiovascular disease constitutes the primary cause of mortality and morbidity worldwide, and represents a group of disorders associated with the loss of cardiac function. Despite considerable advances in the understanding of the pathologic mechanisms of the disease, the majority of the currently available therapies remain at best palliative, since the problem of cardiac tissue loss has not yet been addressed. Indeed, few therapeutic approaches offer direct tissue repair and regeneration, whereas the majority of treatment options aim to limit scar formation and adverse remodeling, while improving myocardial function. Of all the existing therapeutic approaches, the problem of cardiac tissue loss is addressed uniquely by heart transplantation. Nevertheless, alternative options, particularly stem cell therapy, has emerged as a novel and promising approach. This approach involves the transplantation of healthy and functional cells to promote the renewal of damaged cells and repair injured tissue. Bone marrow precursor cells were the first cell type used in clinical studies, and subsequently, preclinical and clinical investigations have been extended to the use of various populations of stem cells. This review addresses the present state of research as regards stem cell therapy for cardiovascular disease.

Stem Cell Therapy for Cardiovascular Diseases and its Major Setback

Stem cells have inherent property to proliferate and differentiate into varieties of cells thus have significant importance in cell based therapy. However, significant number of cells die once transplanted into ischemic myocardium because of unfavourable environment inside heart. Preconditioning with pharmacological drugs or with short cycles of hypoxia/anoxia followed by reoxygenation evokes endogenous protective mechanism against ischemic stress both in vitro and in vivo models and it may be therefore a necessary step to enhance their survival and proliferation upon transplantation into the infarcted heart.

Stem cell therapy for cardiovascular diseases: a progressive journey.

This review presents the findings from a select group of clinical trials of stem cell therapy for both acute and chronic heart disease. These studies are used to highlight the insight gained through such investigations and the remaining challenges. Clinical trials assessing the use of adult stem cells for treating patients with heart disease have produced encouraging results that suggest these therapies are well tolerated and could potentially improve clinical outcomes, including left ventricular function, infarct size and the occurrence of future adverse clinical events. However, most have shown only modest benefits, and the results often appear to be inconsistent across trials. Nevertheless, analyses from these trials have provided useful information regarding factors that affect the potency of these therapies, the mechanisms underlying their effects and the patient populations most likely to derive benefits. Stem cell therapy has the potential to overcome the limited regenerative capacity of the heart and induce cardiac repair and regeneration. The challenge lies in finding the most effective approach, which can only be determined through larger rigorously conducted clinical trials.

Transient Expression of Telomerase Reverse Transcriptase in Aged Human Stromal Vascular Fraction Improves Function and Regenerative Capacity

Objective: Autologous adult stem cell therapy for cardiovascular disease has shown minimal or no benefit in clinical trials. This may in part be explained by aging-related alteration of secretory function and loss of replicative capacity of autologous cell preparations. Previously we have shown that functional and replicative capacity of human aortic endothelial cells can be increased by retroviral overexpression of telomerase. However, there is a concern for immortalization and insertional mutagenesis with this approach. More recently, we have developed methods for transient expression of human telomerase reverse transcriptase (TERT). We hypothesize that this method will increase telomere length (TL) sufficiently to enhance the regenerative capacity of adult stem cells. In the current study we assessed the effects of this approach on adipose-derived stromal vascular fraction (SVF).

Current Concepts in Stem Cell Therapy for Cardiovascular Diseases: What We Know and Don't Know

Medical therapies and mechanical interventions for the treatment of myocardial infarction (MI) and ischemic heart failure have seen great progress. However, current therapies only slow the progression to heart failure, but do not stimulate regeneration to recover the loss of functional myocytes. Stem cell-based therapy is a novel modality that can potentially be used for the treatment of ischemic cardiac injury and heart failure wherein cardiac tissue is regenerated thereby improving cardiac function and reducing the morbidity and mortality of patients. Many preclinical and clinical trials have employed various types of stem cells including bone marrow-derived mononuclear cells, skeletal myoblasts, resident cardiac stem cells, mesenchymal stem cells, and endothelial progenitor cells, and these studies have suggested great potential for the clinical application of stem cell therapy for cardiac disease. Here we will review and summarize the current evidence indicating the potential of stem cell therapy, focusing on the clinical trials conducted to date.

FUNCTIONAL ENHANCEMENT OF CIRCULATING ANGIOGENIC CELLS CULTURED ON A COLLAGEN BIOMATERIAL USING THE CCN1 MATRICELLULAR PROTEIN

Circulating angiogenic cells (CACs) play an important role in vascular homeostasis and hold tremendous promise for cell therapy for cardiovascular diseases. The regenerative potential of these cells can be improved by culture on a natural-based collagen matrix through the up-regulation of key integrin proteins. Further improvements are needed however, as the effects remain minimal or transient. The aim of this study was to evaluate whether the incorporation of CCN1, a secreted pro-angiogenic matricellular protein of the CCN (CYR61/CTGF/NOV) family, could further enhance the revascularization potential of CACs. Human CACs were cultured on fibronectin or a type I collagen matrix. Using RT-qPCR, CCN1 target integrins (Itg) αV and Itgβ3 were up-regulated 13.5±4.3 and 37.8±5.2 fold respectively, in CACs cultured on matrix compared to fibronectin. Using a CCN1-GFP protein, the association of CCN1 was 1.9±0.1 fold greater in CACs from matrix compared to fibronectin and this association was reduced by 41.5% and 42.6%, when an ItgαV or Itgβ3 blocking antibody was applied, respectively (p=0.008). Direct integrin interaction was confirmed by protein pull-down using a His-tagged CCN1 protein. Functionally, CCN1-matrix treatment increased proliferating Ki67+ cells 2.2± 0.2 fold over CACs on matrix alone (p=0.001) and 8.7±2.7 over fibronectin-cultured CACs (p=0.0002). When used in a migration assay, CCN1-matrix cells exhibited a 5.7±0.4 and 1.8±0.2 fold increase in migration towards vascular endothelial growth factor (VEGF) compared to fibronectin (p=0.001) and matrix (p=0.02) cultured cells, respectively. In a HUVEC/CAC angiogenic co-culture assay, CACs treated with the CCN1-matrix demonstrated a 4.1±1.6 and 7.3±1.4 fold increase in cell incorporation into capillary-like structures compared to matrix (p=0.03) and fibronectin (p=0.02) culture CACs. Tubule network formation during the angiogenic assay was also greater by 5.1±0.6 fold (p=0.007) for the CCN1-matrix treated CACs compared to matrix-cultured cells. In vivo, intramuscular injection of CCN1-matrix cultured CACs to ischemic hindlimbs of mice resulted in blood flow recovery to 80±1.8% of baseline, which was greater than the recovery observed for matrix-cultured CACs and PBS treatments (66±1.6% and 35.5±2.3%, respectively; p≤0.02). Matrix-cultured CACs were more responsive to CCN1 and exhibited greater angiogenic properties, suggesting that the specific integrin up-regulation may make the cells more responsive to extracellular signals. We also showed that pre-treatment of matrix-cultured CACs with CCN1 significantly improved their ability to restore perfusion in ischemic tissue. These findings demonstrate a novel mechanism which may be used to promote tissue neovascularization.