Search

Abstract 14039: The Cardioprotective and Anti-Inflammatory Effect of Inhaled Nitric Oxide During Fontan Surgery: A Prospective Randomized Study
- Jacek Kolcz,
- Elzbieta Karnas and
- Ewa K ZUBA SURMA
Circulation Volume 146, Issue Suppl_18 November 2022

Abstract Export Citations

Abstract
Introduction: Open heart surgery that requires cardiopulmonary by-pass (CPB) evokes tremendous stress and inflammatory responses, affecting postoperative organ function, morbidity, and mortality. Although part of this reaction triggers protective and anti-inflammatory phenomena, it is harmful, particularly in patients with single ventricle congenital heart defects during Fontan surgery. Unfortunately, there is no perfect anti-inflammatory and stress defence strategy despite decades of research.Hypothesis: This study aimed to investigate the influence of nitric oxide (NO) inhaled directly into the oxygenator during CPB on a proteome profile of inflammatory cytokines, metabolic response markers, heart and lung protection indicators, and the early clinical outcomes.Methods: The study involved 115 patients randomized to the Fontan-NO group, receiving NO inhalation (n=48) and the Fontan group, not receiving NO (n=49). Eighteen patients were excluded from the study. The clinical data were prospectively analyzed. Proteome profiler essays were used for repeated measurements of investigated analytes.Results: Fontan-NO patients had significantly lower levels of lactate (p=0.04) and glucose (p=0.02), lower pulmonary artery pressure (p=0.03) and catecholamine index (p=0.042), shorter ventilatory support time (p=0.03) and ICU stay (p=0.04). Plasma factors analysis has shown a significantly higher concentration of interleukin-10, tissue inhibitor of metalloproteinase, prolactin, and angiopoietin and a lower concentration of interleukin-6, interleukin-8, interleukin-1β, granulocyte-macrophage colony-stimulating factor, pentraxin, matrix metalloproteinase-8, troponin-I, CK-MB, and insulin in Fontan-NO group. Multiple regression analysis revealed the influence of Il-6 and Il-8 on respiratory support time and ICU stay and the relationship between catecholamine index, duration of effusions, and indicators of myocardial damage.Conclusions: NO inhaled into the oxygenator during CPB can alleviate the inflammatory response, improve myocardial and lung protection, and reduce metabolic stress in patients with a single ventricle undergoing Fontan surgery. The observed effects can influence short-term clinical outcomes.
Abstract 1276: Pluripotent Bone Marrow (BM)-Derived Very Small Embryonic-Like (VSEL) Stem Cells are Mobilized after Acute Myocardial Infarction in Mice
- Ewa K Zuba-Surma,
- Magdalena Kucia,
- Yiru Guo,
- Buddhadeb Dawn,
- Mariusz Z Ratajczak and
- Roberto Bolli
Circulation Volume 116, Issue suppl_1616 October 2007

Abstract Export Citations

Abstract
Recently, we have identified in bone marrow (BM) a population of SSEA-1+/Oct-4+/Sca-1+/lin-/CD45- pluripotent, very small embryonic-like stem cells (VSELs) with the ability to differentiate into cardiac lineage in vitro. The pathophysiologic role of these cells in acute myocardial infarction (AMI) is unknown. The aim of this study was to investigate if VSELs are mobilized into peripheral blood (PB) after AMI. C57BL/6 mice (6- or 15-wk-old) underwent a 30-min coronary occlusion followed by reperfusion and were euthanized at 24 h, 48 h, or 7 days after AMI. PB samples were collected for flow cytometric, confocal microscopic, and RQ-PCR analysis. Sham controls underwent sham surgery (1-h open-chest state) while controls did not. By flow cytometry, VSELs were barely detectable in PB under baseline conditions but increased significantly after AMI, peaking at 48 h post AMI both in young (6-wk-old) and older (15-wk-old) mice (3.33±0.37 and 7.73±1.02 cells/μl of PB, respectively) (Fig 1A). In contrast to hematopoietic stem cells (HSCs), sorted PB-derived VSELs were positive for Oct-4 and negative for CD45 (Fig 1B). Furthermore, RQ-PCR analysis revealed increased level of mRNA for markers of pluripotency, such as Oct-4, Nanog, Rex-1, Dppa1, and Rif1, in total PB cells of 6-wk-old mice at 48 h after AMI (9.50, 6.04, 5.28, 3.18 and 2.07-fold increase as compared with sham control, respectively). This is the first report that pluripotent stem cells (VSELs) are mobilized from the bone marrow into the peripheral blood after AMI in both young and older animals. Mobilization of VSELs with cardiogenic potential raises the possibility that these cells may play a role in repair of infarcted myocardium.
Abstract 15327: In vivo Ischemic Limb Injury Activates Proliferation and Mobilization of Pluripotent Bone Marrow- Derived Very Small Embryonic-Like Stem Cells
Circulation Volume 126, Issue suppl_2120 November 2012

Abstract Export Citations

Abstract
Very small embryonic-like stem cells (VSELs) were identified as rare quiescent, nonhematopoietic (CD45-/Lineage-) and pluripotent Oct-4+ stem cells in adult murine and human tissues including bone marrow (BM). Murine BM- derived VSELs were shown to differentiate into cells from all three germ layers and to participate in heart repair after injection into infarcted myocardium by enhancing perfusion and angiogenesis. However, their activation during physiological or pathological events in vivo have not been reported. In this study, we examined if acute ischemic injury may stimulate both proliferation of quiescent VSELs in BM and their mobilization into peripheral blood (PB). Wild-type C57BL/6 mice (9-week old) underwent hind limb ischemia by permanent proximal femoral artery occlusion. Mice were injeced with bromodeoxyuridine (BrdU, 1mg/mouse/i.p.)every two days and were scarified at 2, 7, 14 and 28 days following ischemia (N=5/group). PB and BM from non- and ischemic limbs were collected from individual animals. Healthy non-ischemic mice were used as control groups in each time point. The presence of proliferating (BrdU+) VSELs (CD45-/Lineage-/Sca-1+), endothelial progenitor cells (EPCs; CD45-dim/Lin-/Sca-1+/Flk-1+) and hematopoietic stem cells (HSCs; CD45+/Lineage-/Sca-1+) in PB and BM was evaluated by multiparameter flow cytometry and ImageStream system. The expression of genes related to the VSEL and EPC presence was examined by real time RT-PCR. We established that the content of VSELs was significantly increased in BM of ischemic mice after 7 days post injury. We observed elevated number of BrdU+ VSELs in BM and circulating in PB of the injured animals indicating vast impact of acute ischemia on activation of these cells. Similar results were obtained for BM- derived and circulating EPCs. We conclude that acute tissue injury such as limb ischemia may provide stimulatory agents to activate proliferation of quiescent VSELs in BM and their mobilization into PB. Thus, the ischemic injury may recruit the normally quiescent pluripotent stem cell pools to enhance endogenous mechanisms of tissue repair including stem/ progenitor cell- dependent angiogenesis which we currently investigate.
Abstract 937: Antigenically-Defined Subsets of Bone Marrow Mesenchymal Stem Cells Exhibit Differential Cardiomyogenic and Angiogenic Potential
- Ewa K Zuba-Surma,
- Ahmed Abdel-Latif,
- Robert J Vincent,
- Smita Ranjan,
- Sumit Tiwari,
- Mamdouh Khayat,
- Roberto Bolli and
- Buddhadeb Dawn
Circulation Volume 114, Issue suppl_1831 October 2006

Abstract Export Citations

Abstract
Bone marrow mesenchymal stem cells (BMMSCs) are typically isolated as adherent cells; the importance of antigen expression on cardiomyogenic potential of (BMMSCs) remains unclear. Although BMMSCs differentiate into a cardiomyocytic phenotype following 5-azacytidine treatment, the effect of non-toxic growth factors on differentiation induction remains unknown. We investigated the cardiomyogenic potential of two subpopulations of BMMSCs in medium containing growth factors crucial for cardiac development. Sca-1+/CD45-/c-kit-/Thy1+/ CD105+ (BMMSC+) and Sca-1+/CD45-/c-kit-/Thy1+/CD105- (BMMSC-) MSCs were obtained from the adherent fraction of BM from C57BL/6 mice using two-step FACS isolation. Cells were differentiated in DMEM with FBS, IGF-1, Dynorphin B, TGFβ-1 and FGF-2. After 30 days of culture, the expression of cardiac-specific transcription factors (TFs) and intracellular antigens (IAs) was quantitatively evaluated by confocal microscopy (Fig. 1A). BMMSC- cells exhibited greater cardiac differentiation potential compared with BMMSC+ and unfractionated BMMSC populations (29.1±1.7% vs. 15.7±1.6% vs. 15.8±0.9% of cells positive for cardiac markers, Fig. 1B). BMMSC- cells also exhibited greater angiogenic potential assessed by tube formation after 6 h of culture in Matrigel (Fig. 1C) when compared with BMMSC± and unfractionated BMMSC populations (364±11.4 vs. 268±14.4 vs. 293.7±33.6 of endotubules per field, Fig. 1D). These results indicate that selected non-toxic growth factors can effectively induce cardiac differentiation of BMMSCs and that BMMSCs lacking CD105 expression have greater proclivity to cardiac and endothelial differentiation.
Abstract 1142: Effects of Culture Conditions on Cardiomyogenic Differentiation and Expansion of Bone Marrow-Derived Myocardial Tissue-Committed Stem Cells
- Ewa K Zuba-Surma,
- Ahmed Abdel-Latif,
- Magda Kucia,
- Greg Hunt,
- Smita Ranjan,
- Mariusz Z Ratajczak,
- Roberto Bolli and
- Buddhadeb Dawn
Circulation Volume 114, Issue suppl_1831 October 2006

Abstract Export Citations

Abstract
The bone marrow harbors a population of Sca-1+/Lin-/CD45- tissue-committed stem cells (TCSCs) that express early cardiac markers. Given the rarity (approx. 0.01% of all cells) of these cells, optimization of culture conditions for cardiac differentiation and expansion is crucial for clinical translation. Thus, TCSCs were isolated from bone marrow of EGFP transgenic mice using FACS. Cardiac differentiation and expansion were induced on a feeder layer of bone marrow stromal cells in basic medium (BM, containing FBS, and IGF-1), BM supplemented with dynorphin B (DynB), oxytocin (OT), FGF-2 or insulin (for differentiation), or BM with different doses of FBS supplemented with combinations of LIF, SCF, EGF, PDGF-B and TPO (for expansion). After 30 days of culture, differentiation was quantitated by the expression of cardiac-specific transcription factors and intracellular antigens in EGFP+ cells by confocal microscopy (Fig. 1A). Expansion was assessed by counting the number of EGFP+ TCSCs after 30 days of culture. BM alone, BM+DynB, BM+insulin, BM+FGF-2, and BM+OT induced cardiac differentiation in 12.0±2.5%, 8.2±0.6%, 8.2±0.3%, 17.1±0.3% and 20.6±9.0% of TCSCs, respectively (Fig. 1B). Under expansion conditions, TCSCs survived only in media with low concentrations of FBS (2 and 5%) and in the presence of LIF alone or combinations of LIF, EGF and PDGF-B. Addition of SCF failed to enhance expansion (Fig.1C). These results indicate that TCSCs differentiate into a cardiac phenotype in vitro and that this process is profoundly affected by culture conditions. However, additional strategies need to be formulated to effectively expand these primitive cells.
Response to Letter Regarding Article, “Adult Bone Marrow Cell Therapy Improves Survival and Induces Long-Term Improvement in Cardiac Parameters: A Systematic Review and Meta-Analysis”
Circulation Volume 127, Issue 1516 April 2013

Export Citations PDF/EPUB

Abstract
Adult Bone Marrow Cell Therapy Improves Survival and Induces Long-Term Improvement in Cardiac Parameters: A Systematic Review and Meta-Analysis
- Vinodh Jeevanantham,
- Matthew Butler,
- Andre Saad,
- Ahmed Abdel-Latif,
- Ewa K. Zuba-Surma and
- Buddhadeb Dawn
Circulation Volume 126, Issue 531 July 2012

Abstract Export Citations PDF/EPUB

Abstract
Background—Despite rapid clinical translation and widespread enthusiasm, the therapeutic benefits of adult bone marrow cell (BMC) transplantation in patients with ischemic heart disease continue to remain controversial. A synthesis of the available data is critical to appreciate and underscore the true impact of this promising approach.Methods and Results—A total of 50 studies (enrolling 2625 patients) identified by database searches through January 2012 were included. Weighted mean differences for changes in left ventricular (LV) ejection fraction, infarct size, LV end-systolic volume, and LV end-diastolic volume were estimated with random-effects meta-analysis. Compared with control subjects, BMC-treated patients exhibited greater LV ejection fraction (3.96%; 95% confidence interval, 2.90–5.02; P<0.00001) and smaller infarct size (−4.03%, 95% confidence interval, −5.47 to −2.59; P<0.00001), LV end-systolic volume (−8.91 mL; 95% confidence interval, −11.57 to −6.25; P<0.00001), and LV end-diastolic volume (−5.23 mL; 95% confidence interval, −7.60 to −2.86; P<0.0001). These benefits were noted regardless of the study design (randomized controlled study versus cohort study) and the type of ischemic heart disease (acute myocardial infarction versus chronic ischemic heart disease) and persisted during long-term follow-up. Importantly, all-cause mortality, cardiac mortality, and the incidence of recurrent myocardial infarction and stent thrombosis were significantly lower in BMC-treated patients compared with control subjects.Conclusions—Transplantation of adult BMCs improves LV function, infarct size, and remodeling in patients with ischemic heart disease compared with standard therapy, and these benefits persist during long-term follow-up. BMC transplantation also reduces the incidence of death, recurrent myocardial infarction, and stent thrombosis in patients with ischemic heart disease.
Abstract 12650: Greater Antiapoptotic, Antiinflammatory, and Proangiogenic Attributes of a Defined Subpopulation of Bone Marrow Mesenchymal Stem Cells May Facilitate Cardiac Repair via Paracrine Effects
- Ewa K Zuba-Surma,
- Yu-Ting Xuan,
- Anna Labedz-Maslowska,
- Harold Elias,
- Elzbieta Kamycka,
- Ahmed Abdel-Latif,
- Yanqing Zhu,
- Ou-Li Wang,
- Arash Davani and
- Buddhadeb Dawn
Circulation Volume 126, Issue suppl_2120 November 2012

Abstract Export Citations

Abstract
Bone marrow mesenchymal stem cells (MSCs) isolated by adherence to plastic are highly heterogeneous. In this study, we tested the hypothesis that an antigenically-defined and homogenous CD45-/c-kit-/Sca-1+/CD90+/CD105- (CD105-) subpopulation of MSCs will exhibit greater angiogenic and antiapoptotic attributes, which may underlie superior cardiac repair by these cells. The CD105- MSCs and their precise antigenic control CD45-/c-kit-/Sca-1+/CD90+/CD105+ (CD105+) MSCs were obtained by two-step FACS isolation and used for comparative analysis of antiapoptotic, inflammatory and angiogenic factor expression. Moreover, the phenotype of sorted MSCs was evaluated by imaging cytometry (ImageStream system). We found that CD105- MSCs expressed higher mRNA levels of VGEF, HGF, and SDF-1, indicating greater ability to induce angiogenesis, when compared with unfractionated MSCs. Consistently, CD105- MSCs formed greater number of endotubules on Matrigel. Importantly, the CD105- MSCs also exhibited reduced expression of proinflammatory agents (TNF-a) and factors involved in adverse cardiac remodeling (MMP-2 and MMP-9) when compared with unfractionated MSCs (Fig.1A). The CD105- MSCs exhibited greater amounts of Bcl-XL, p-Bad, and HO-1, and resisted 2-h hypoxia/2-h reoxygenation-induced apoptosis. The CD105- MSCs also showed greater levels of p-STAT3 and p-ERK1/2, indicating innately higher activation level of cellular antiapoptotic machinery. Along with these functional differences, we also found striking phenotypic differences between CD105- and CD105+ MSCs (Fig.1B), confirming the distinct functional characteristics and a primitive phenotype of CD105- MSCs. We conclude that the CD105- MSCs represent a superior cellular substrate for infarct repair by virtue of their antiapoptotic, antiinflammatory, and angiogenic properties, which collectively contribute to differentiation potential and paracrine activities of these cells.
Abstract 14451: Bone Marrow Cell Therapy Improves Cardiac Function, Survival, and Clinical Outcomes in Patients With Chronic Ischemic Heart Disease
- Jane Ha,
- Muhammad R Afzal,
- Anweshan Samanta,
- Ewa Zuba-Surma,
- Ji Yoo and
- Buddhadeb Dawn
Circulation Volume 142, Issue Suppl_317 November 2020

Abstract Export Citations

Abstract
Introduction: The effects of bone marrow cell (BMC) therapy in patients with chronic ischemic heart disease (CIHD) remain controversial.Hypothesis: We hypothesized that injection of BMCs in patients with CIHD would improve left ventricular (LV) structure and function. We also hypothesized that BMC therapy would improve clinical outcomes in CIHD patients.Methods: We performed a systemic review and meta-analysis of pooled data from published randomized controlled trials (RCTs) that evaluated the efficacy of BMC administration in patients with CIHD. The effects of BMC injection on LV ejection fraction (LVEF), LV end-systolic volume (LVESV), LV end-diastolic volume (LVEDV), infarct size, and patient outcomes were analyzed using random-effects meta-analysis.Results: The literature search yielded 30 RCTs enrolling 1,548 patients. Transplantation of BMCs resulted in an improvement in LVEF in cell-treated patients compared with controls (2.89%; 95% confidence interval (CI): 1.80 to 3.97; P<0.001). There was a trend toward reduced LVESV (-4.96 ml; 95% CI: -11.64 to 1.71 ml; P= 0.14) and LVEDV (-5.95 ml; 95% CI: -12.09 to 0.18 ml; P=0.06). The improvement in LVEDV was more pronounced (-7.42 ml; 95% CI: -13.68 to -1.17ml; P=0.02) in patients with baseline LVEF <40%, indicating improved LV remodeling. BMC injection was also associated with marked reduction in the risk of all-cause mortality, rehospitalization due to heart failure, and ventricular arrhythmias in CIHD patients.Conclusions: BMC injection improves cardiac function and remodeling in patients with CIHD. These benefits are more pronounced in patients with LVEF <40% at baseline. Perhaps more importantly, BMC therapy also improves clinical outcomes that are critically important for this patient population with LV dysfunction, including survival, rehospitalization due to heart failure, and ventricular arrhythmias.
Abstract 1023: Transplantation Of Bone Marrow-derived Very Small Embryonic-like Stem Cells (VSELs) Improves Left Ventricular Function And Remodeling After Myocardial Infarction
- Ewa K Zuba-Surma,
- Hisham Taher,
- Magdalena Kucia,
- Yiru Guo,
- Santosh K SanganalMath,
- Greg Hunt,
- Robert J Vincent,
- Buddhadeb Dawn,
- Mariusz Z Ratajczak and
- Roberto Bolli
Circulation Volume 116, Issue suppl_1616 October 2007

Abstract Export Citations

Abstract
Adult bone marrow (BM) contains a rare population of pluripotent Sca-1+/Lin-/CD45- very small embryonic-like stem cells (VSELs) that differentiate into cardiomyocytes in vitro. The clinical usefulness of these primitive cells, however, is limited by their rarity, which makes it impossible to isolate a large number of VSELs for transplantation. Therefore, we examined if in vitro expansion followed by predifferentiation of VSELs in cardiomyogenic medium would enhance the cardiac reparative benefits of these cells. VSELs isolated from adult mouse BM were expanded in vitro; cardiac differentiation was induced by culturing expanded cells in a cardiomyogenic medium for 5 days. Mice underwent a 30-min coronary occlusion followed by reperfusion and, 48 h later, received intramyocardial injection of vehicle (group I, n = 11) or expanded VSELs (100,000 cells) without (group II, n = 8) or with (group III, n = 11) prediffer-entiation. At 35 d after MI, mice in group III exhibited improved LV ejection fraction (54.5 ± 3.3% vs. 41.5 ± 3.3% in group I, P<0.05) and infarct wall thickening fraction (51.5 ± 5.7% vs. 28.7 ± 4.5% in group I, P<0.05) (Fig). VSEL transplantation also ameliorated postinfarct LV dilation as evidenced by a smaller LV end-diastolic volume (60.6 ± 5.4 μl vs. 79.8 ± 8.1 μl in group I, P<0.05) (Fig). In contrast, transplantation of VSELs without predifferentiation (group II) failed to produce statistically significant benefits. We conclude that transplantation of expanded and predifferentiated VSELs improves LV remodeling and dysfunction after MI. Our results lay the groundwork for novel therapies based on the use of these pluripotent cells, which are present in normal adult BM.
Adult Bone Marrow Cell Therapy for Ischemic Heart Disease: Evidence and Insights From Randomized Controlled Trials
- Muhammad R. Afzal,
- Anweshan Samanta,
- Zubair I. Shah,
- Vinodh Jeevanantham,
- Ahmed Abdel-Latif,
- Ewa K. Zuba-Surma and
- Buddhadeb Dawn
Circulation Research Volume 117, Issue 628 August 2015

Abstract Export Citations PDF/EPUB

Abstract
Rationale:Notwithstanding the uncertainties about the outcomes of bone marrow cell (BMC) therapy for heart repair, further insights are critically needed to improve this promising approach.Objective:To delineate the true effect of BMC therapy for cardiac repair and gain insights for future trials through systematic review and meta-analysis of data from eligible randomized controlled trials.Methods and Results:Database searches through August 2014 identified 48 eligible randomized controlled trials (enrolling 2602 patients). Weighted mean differences for changes in left ventricular (LV) ejection fraction, infarct size, LV end-systolic volume, and LV end-diastolic volume were analyzed with random-effects meta-analysis. Compared with standard therapy, BMC transplantation improved LV ejection fraction (2.92%; 95% confidence interval, 1.91–3.92; P<0.00001), reduced infarct size (−2.25%; 95% confidence interval, −3.55 to −0.95; P=0.0007) and LV end-systolic volume (−6.37 mL; 95% confidence interval, −8.95 to −3.80; P<0.00001), and tended to reduce LV end-diastolic volume (−2.26 mL; 95% confidence interval, −4.59 to 0.07; P=0.06). Similar effects were noted when data were analyzed after excluding studies with discrepancies in reporting of outcomes. The benefits also persisted when cardiac catheterization was performed in control patients as well. Although imaging modalities partly influenced the outcomes, LV ejection fraction improved in BMC-treated patients when assessed by magnetic resonance imaging. Early (<48 hours) BMC injection after myocardial Infarction was more effective in reducing infarct size, whereas BMC injection between 3 and 10 days proved superior toward improving systolic function. A minimum of 50 million BMCs seemed to be necessary, with limited additional benefits seen with increasing cell numbers. BMC therapy was safe and improved clinical outcomes, including all-cause mortality, recurrent myocardial Infarction, ventricular arrhythmia, and cerebrovascular accident during follow-up, albeit with differences between acute myocardial Infarction and chronic ischemic heart disease subgroups.Conclusions:Transplantation of adult BMCs improves LV ejection fraction, reduces infarct size, and ameliorates remodeling in patients with ischemic heart disease. These effects are upheld in the analyses of studies using magnetic resonance imaging and also after excluding studies with discrepant reporting of outcomes. BMC transplantation may also reduce the incidence of death, recurrent myocardial Infarction, ventricular arrhythmia, and cerebrovascular accident during follow-up.
Abstract 11430: Transplantation of an Antigenically-defined MSC Subpopulation Induces Superior Cardiac Repair After a Reperfused Myocardial Infarction
- Guangming Cheng,
- Lin Zhao,
- Magdy Girgis,
- Arash Davani,
- Yanjuan Yang,
- Jeryl Hauptman,
- Robert J Vincent,
- Ewa K Zuba-Surma and
- Buddhadeb Dawn
Circulation Volume 140, Issue Suppl_119 November 2019

Abstract Export Citations

Abstract
Introduction: The outcomes of cardiac repair with bone marrow mesenchymal stem cells (MSCs) have been variable and generally suboptimal. Bone marrow MSCs are quite heterogeneous in nature.Hypothesis: We hypothesized that an antigenically-defined CD45-/c-kit-/Sca-1+/CD90+/CD105- (abbreviated as CD105- MSCs) homogeneous MSC subpolulation will induce superior repair after a reperfused myocardial infarct (MI).Methods: Adherent unfractionated MSCs from C57BL/6 mice were culture-expanded and CD105- and CD105+ subpopulations were isolated using a two-step FACS protocol. The characteristics of MSC subpopulations were tested in vitro. Then we tested the efficacy of these cells for cardiac repair in vivo. Vehicle (n=10), unfractionated (n=8), CD105+ (n=9) and CD105- MSCs (n=9) were injected into the infarct border zone 2 d after a reperfused MI in C57BL/6 mice. Serial echocardiograms were performed to assess LV structure and function at 4 d prior to MI (BSL1), and at 2 d (BSL2) and 35 d after cell injection. Infarct size, fibrosis, myocyte hypertrophy and apoptosis were assessed by morphometry and immunohistochemistry.Results: In vitro, compared with controls, CD105- MSCs exhibited greater angiogenic and pro-survival properties. In vivo, LVEF was similarly and significantly reduced among four groups of mice at 2 d after cell injection. After 35 d, compared with unfractionated MSC and CD105+ MSC groups, CD105- MSC-treated mice exhibited higher LVEF and smaller LVEDV (Figure), indicating that CD105- MSC injection induces superior infarct repair.Conclusions: These findings provide comprehensive evidence that a homogeneous CD105- MSC subpopulation is better suited for infarct repair compared with unfractionated MSCs or CD105+ MSCs.
Abstract 19198: Adult Human Heart Contains Two Populations of C-kit-Oct-4+SSEA-4+ Very Small Embryonic-Like Stem Cells (Isl-1- and Isl-1+)
- Wojciech Wojakowski,
- Ewa K Zuba-Surma,
- Marcin Syzdol,
- Adam Kowalowka,
- Elzbieta Kamycka,
- Tomasz Kargul,
- Justyna Drukala,
- Ryszard Bachowski,
- Stanislaw Wos,
- Mariusz Z Ratajczak and
- Michal Tendera
Circulation Volume 126, Issue suppl_2120 November 2012

Abstract Export Citations

Abstract
Very small embryonic-like cells (VSELs) express pluripotent stem markers (Oct-4,Nanog, SSEA-4). and in adult mice may be expanded and differentiatiated into three germ layers. In humans VSELs were found in cord blood and peripheral blood (PB). AIM: Comparison of VSELs isolated from human myocardium with cardiac stem cells (CSC) and to measure the mobilization into PB after cardiac surgery. METHODS: 32 patients undergoing CABG were enrolled. Right atrial appendage (RAA), left ventricle (LV) were collected. PB samples were drawn before and immediately after surgery, 24 hrs and 5-7 days after weaning from cardiopulmonary bypass. Tissue samples (0.5-1.0 cm3) were cut, digested in collagenase, passed through 70um cell strainer, centrifugated, resuspended and stained with antibodies CD34 FITC/CD133 APC/SSEA-4PE/CD45 APC-Cy7. Analyses were carried out on Image Stream System which is a combination of FACS and immunofluorescent microscope. RESULTS: VSELs and CSC were identified in LAA. VSELs had markedly smaller diameter (4.7 ± 0.2µm vs. 7.7 ± 0.4µm), were c-kit negative, expressed markers of pluripotent cells (Oct-4, SSEA-4) and CD133. Two populations of VSELs - both Isl-1 negative and positive were identified. Subpopulation of Oct-4+SSEA-4+Isl+ VSELs contained cells of intermediate size (5.7 ± 0.7µm) and might form more mature population of VSELs progenitors. VSELs undergo significant mobilization into PB following the CABG. CONCLUSION: Human myocardium in adults contains two subpopulations of Oct-4+SSEA4+ VSELs which are distinct from CSC. Cardiac surgery with cardiopulmonary bypass induced mobilization of VSELs into peripheral blood.
Abstract 5376: Evidence of Mobilization of Pluripotent and Very Small Embryonic-Like (VSEL) Stem Cells in Patients with Myocardial Ischemia- A Potential Therapeutic Target
- Ahmed Abdel-Latif,
- Ewa K Zuba-Surma,
- Magdalena Kucia,
- Khaled M Ziada,
- Mushabbar A Syed,
- Debabrata Mukherjee,
- Hussam Hamdalla,
- Susan S Smyth and
- Mariusz Z Ratajczak
Circulation Volume 118, Issue suppl_1828 October 2008

Abstract Export Citations

Abstract
Studies have shown that predestined bone marrow-derived stem cells are mobilized in acute myocardial infarction (MI). In contrast, the mobilization of pluripotent stem cells (PSCs), including Very Small Embryonic-Like stem cells (VSELs), in MI and other ischemic conditions has not been demonstrated. In this study, we sought to investigate whether PSCs such as VSELs are mobilized into peripheral blood (PB) in various clinical conditions of myocardial ischemia using a multidimensional approach employing flow cytometric, ImageStream system (ISS), and RQ-PCR analyses. PB was collected from patients with: chronic ischemic heart disease (IHD), non ST-segment elevation MI (NSTEMI), ST-segment elevation MI (STEMI), and controls. By flow cytometry, PSCs, hematopoetic stem cells and VSELs’ content was lowest in controls and highest in patients with STEMI particularly at the time of presentation (BSL) (Fig.1A). A similar pattern of expression of the pluripotency marker Oct-4 was observed by ISS (Fig. 1A and 1B). By RQ-PCR analyses, the expression of pluripotency markers (Oct-4 and Nanog), cardiac markers (Nkx-2.5 and GATA-4), endothelial markers (vWF and VE-Cadherin), and CXCR-4 was highest at BSL in STEMI patients (Fig. 1C). Mobilization of PSCs correlated positively with the extent of myocardial injury as estimated by cardiac enzymes (R=0.72, P=0.02) and negatively with age (R= −0.64, P=0.046). This is the first comprehensive and systematic report that demonstrates that PSCs are mobilized in patients with ischemic heart disease. Mobilization of PSCs with cardiogenic potential in myocardial ischemia suggests that these cells may play a role in repair of infarcted myocardium.Figure 1A. Absolute numbers of circulating pluripotent (SSEA-4+;Oct-4+) non-hematopoietic (CD45−) VSELs and hematopoietic (CD45+) cells in peripheral blood by flow cytometry and ISS (number of cells per 1 μl of blood).Figure 1B. Image of circulating Oct-4+ VSEL from the blood of STEMI patient (BSL) by ISS. Intranuclear Oct-4 shown in yellow (PE, Phycoerythrin); nucleus visualized with 7-aminoactinomycin D (7-AAD).Figure 1C. Expression of rnRNA for markers of pluripotent cells (Qct-4) and cardiac or endothelial progenitors (Nkx2.5/Csx and vWF, respectively) in peripheral blood-derived cells by RQ-PCR. Data are shown as fold difference when compared to controls (expressed as 1 for each gene).All data presented as Mean ± SEM; * P<005 vs controls.
Abstract 17476: Transplantation of an Antigenically-Defined Subpopulation of Bone Marrow Mesenchymal Stem Cells Results in Superior Cardiac Repair After a Reperfused Myocardial Infarction
- Guangming Cheng,
- Ewa K Zuba-Surma,
- Yu-Ting Xuan,
- Yanjuan Yang,
- Magdy Girgis,
- Ou-Li Wang,
- Yanqing Zhu,
- Santosh Sanganalmath,
- Shelley Lietzau-Mourer,
- Dong Xu and
- Buddhadeb Dawn
Circulation Volume 124, Issue suppl_2122 November 2011

Abstract Export Citations

Abstract
Although bone marrow mesenchymal stem cells (MSCs) have been used quite extensively for cardiac repair, the outcomes have been highly variable. We investigated whether antigenically-defined subpopulations of MSCs would promote superior repair after myocardial infarction (MI). Unfractionated MSCs were obtained from the adherent fraction of bone marrow cells from C57BL/6 mice using expansion culture, and subpopulations were obtained using two-step FACS isolation. Our in vitro studies identified a Sca-1+/CD45-/c-kit-/Thy1+/CD105- MSC population (CD105- MSCs) that exhibited greater mRNA expression of angiogenic molecules (VEGF, HGF), SDF-1, and reduced expression of deleterious molecules, including TNF-a (Figure). The CD105- MSCs also exhibited greater propensity toward cardiac differentiation. The efficacy of these cells for cardiac repair in vivo was tested in a mouse model of reperfused MI. Wild-type (C57BL/6) mice underwent a 30-min coronary occlusion followed by reperfusion, and 48 h later received intramyocardial injection of vehicle (Vehicle, n=8), unfractionated MSCs (MSCs, n=8), or CD105- MSCs (CD105- MSCs, n=6). Echocardiography was performed 4 d prior to coronary occlusion/reperfusion (BSL1) and at 48 h (BSL2) and 35 d after cell injection. Mice were sacrificed at 35 d. LV ejection fraction (EF) was significantly and similarly reduced in all groups at 48 h after MI (Figure). At 35 d after MI, mice injected with CD105- MSCs exhibited greater EF (Figure) and smaller LV end-diastolic volume compared with vehicle-treated and unfractionated MSC-treated mice. We conclude that compared with unfractionated MSCs, intramyocardial transplantation of CD105- MSCs after a reperfused MI results in improved cardiac repair. The use of this MSC subpopulation may potentially improve the outcomes of cell therapy in humans.
Abstract 15466: Bioactive Contents and Cardiovascular Regenerative Potential of Extracellular Vesicles Derived From Induced Pluripotent Stem Cells
- Marta Adamiak,
- Sylwia Bobis-Wozowicz,
- Elzbieta Karnas,
- Sylwia Kedracka-Krok,
- Michal Sarna,
- Guangming Cheng,
- Anweshan Samanta,
- Magdy Girgis,
- Zbigniew Madeja,
- Buddhadeb Dawn and
- Ewa Zuba-Surma
Circulation Volume 132, Issue suppl_310 November 2015

Abstract Export Citations

Abstract
Introduction: Extracellular vesicles (EVs) are tiny membrane-enclosed vesicles released by cells by membrane budding or exocytosis. The molecular profile of contents and regenerative potential of EVs derived from murine induced pluripotent stem cells (miPSCs) have not been investigated.Hypothesis: We hypothesized that miPSC-derived EVs carry bioactive substances, including mRNA and miRNA, which would influence the biology and fate of target cells.Methods: Murine iPSCs were generated using established protocols and EVs were isolated from culture supernatants by sequential centrifugation. Atomic force microscopy (AFM), flow cytometry, and real-time RT-PCR were used to characterize EV contents. Endothelial cells were exposed to miPSC-EVs in culture.Results: AFM and dynamic light scattering showed that miPSC-EVs are homogenous spherical vesicles smaller than 100 nm (Fig, A). High-sensitivity flow cytometry confirmed the presence of several iPSC- specific markers along with typical exosomal markers (CD9, CD63 and CD81) on miPSC-EVs. miPSC-EVs were enriched in mRNAs, miRNAs and proteins from donor iPSCs as shown by real-time RT-PCR and mass spectroscopy, respectively. Moreover, miPSC-EVs contained transcripts regulating pluripotency, self-renewal and differentiation, including Oct-4, Nanog, GATA-4 as well as miR290-295 cluster (Fig, B,C). Importantly, several miRNAs were found to be higher in EVs when compared with parental miPSCs (Fig, B,C). Endothelial cells treated with miPSC-EVs exhibited greater proliferative, metabolic and angiogenic activities, and were more resistant to apoptosis.Conclusions: Our data show, for the first time, that miPSC-derived EVs are natural nanocarriers capable of transferring bioactive contents to mature cells affecting target cell function as well as regenerative potential. We conclude that iPSC-EVs may represent safe therapeutic alternatives to whole cell-based therapy for cardiovascular repair.
Abstract 12638: The Activation of STAT3 via CamKII is Critical for the Induction of Cardiac Differentiation by Wnt11 in Bone Marrow Mesenchymal Stem Cells
- Lei Chen,
- Yu-Ting Xuan,
- Harold Elias,
- Amy Cantilena,
- Ewa Zuba-Surma,
- Arash Davani,
- Guangming Cheng,
- Ou-Li Wang,
- Robert J Vincent,
- Yanjuan Yang and
- Buddhadeb Dawn
Circulation Volume 126, Issue suppl_2120 November 2012

Abstract Export Citations

Abstract
Wnt11, a member of the evolutionarily conserved Wnt family of glycoproteins, plays important roles in cellular migration, proliferation, and differentiation during embryogenesis. Although Wnt11 induces cardiac differentiation in bone marrow cells, the underlying molecular mechanisms remain unclear. We hypothesized that Wnt11-induced cardiac differentiation of mesenchymal stem cells (MSCs) involves calcium-calmodulin dependent kinase II (CamKII), which is activated via the noncanonical Wnt-Calcium pathway. Murine MSCs were isolated by adhesion to plastic and propagated in culture. For Wnt11 exposure, MSCs were cultured on the bottom of plates with Wnt11-secreting 293 cells on inserts. Empty vector-bearing 293 cells served as controls. Exposure to Wnt11 increased the levels of both phosphorylated (activated) CamKII and p-STAT3 in MSCs (Fig. 1A). The addition of CamKII-bearing plasmids to culture increased p-STAT3 levels in MSCs in a dose-dependent fashion, indicating that CamKII can induce robust STAT3 activation in MSCs (Fig. 1B). Further, when Wnt11-exposed MSC homogenates were immunoprecipitated with anti-CamKII antibodies and immunoblotted for p-STAT3, the blots revealed physical association of CamKII with p-STAT3 (Fig. 1C), indicating a novel signaling paradigm in MSCs. Finally, the critical role of STAT3 in Wnt11-induced cardiac differentiation was confirmed with the use of a selective STAT3 inhibitor WP1066, which prevented the Wnt11-induced increase in cardiac transcription factor Nkx2.5 in MSCs (Fig. 1D). Consistently, in MSCs cultured in presence of Wnt11, the addition of STAT3 inhibitor WP1066 decreased the number of MSCs expressing cardiac-specific troponin T (Fig. 1E). We conclude that the activation of STAT3 via CamKII plays an essential role in the induction of cardiomyocytic differentiation of MSCs by Wnt11. This information may be utilized for effective cardiogenic priming of MSCs prior to transplantation for cardiac repair.
Abstract 212: Pharmacologic Late Preconditioning With Opioid δ1-agonist Confers Cardio- And Neuroprotetive Benefits During Cardiac Arrest
- Hisham Taher,
- Yiru Guo,
- Ralphiel S Payne,
- Greg Hunt,
- Robert J Vincent,
- Santosh K SanganalMath,
- Nathan J Reed,
- Ewa K Zuba-Surma,
- Smita Ranjan,
- Sumit Tiwari,
- Arash Rezazadeh,
- Rif S El-Mallakh,
- Roberto Bolli and
- Buddhadeb Dawn
Circulation Volume 116, Issue suppl_1616 October 2007

Abstract Export Citations

Abstract
Cardiac arrest affects thousands of patients every year and the outcome continues to be dismal. Administration of opioid delta1-receptor agonists induces pharmacologic late preconditioning against myocardial infarction. We hypothesized that administration of TAN 670, a selective opioid d1-agonist, 24 h prior to a resuscitated cardiac arrest would confer cardio- and neuroprotection in an analogous fashion. C57BL/6 mice received TAN 670 (60 mg/kg i.p.) or vehicle (n = 9/group) and 24 h later were subjected to cardiac arrest (pulseless electrical activity) by exsanguination via withdrawal of blood from LV cavity. Mice were resuscitated by reinfusion of normothermic blood and gentle chest compression. Sham controls underwent the same procedure without blood withdrawal (n = 9). Cardiac and neurologic function was assessed prior to and at serial time-points after cardiac arrest. Apoptosis was quantitated by immunostaining. Preconditioning with TAN 670 attenuated cardiac arrest-induced apoptosis in cardiomyocytes (Fig 1A) and neurons in the CA2 region of the hippocampus (Fig 1B) and improved neurologic function, evidenced by preservation of learning and memory assessed by modified Barnes maze (Fig 1C). Activity testing showed improvement in the margin time in TAN 670-treated mice. We conclude that pretreatment with an opioid d1-receptor agonist 24 h prior to a resuscitated cardiac arrest induces delayed protection against apoptosis in both the heart and brain. These findings reveal the existence of late preconditioning against cardiac arrest-induced cellular damage, a novel strategy that has therapeutic implications for organ protection during cardiac arrest in humans.
Abstract 9899: Wnt11-Pretreated Bone Marrow Mesenchymal Stem Cells Induce Superior Cardiac Repair After a Reperfused Myocardial Infarction
- Guangming Cheng,
- Yu-Ting Xuan,
- Carrie M Quinn,
- Santosh K Sanganalmath,
- Magdy Girgis,
- Ou-Li Wang,
- Yanqing Zhu,
- Yanjuan Yang,
- Michael P Flaherty,
- Shelley Lietzau-Mourer,
- Ewa K Zuba-Surma,
- Robert J Vincent,
- Dong Xu,
- Arash Davani and
- Buddhadeb Dawn
Circulation Volume 124, Issue suppl_2122 November 2011

Abstract Export Citations

Abstract
Wnt11, a member of the evolutionarily conserved Wnt family of glycoproteins, plays important roles in cellular migration, proliferation, and differentiation during embryogenesis. However, the impact of Wnt11 signaling in cardiac repair after an acute myocardial infarction (MI) remains largely unknown. Based on the ability of Wnt11 to induce cardiomyogenic differentiation in adult cells, we hypothesized that pretreatment of bone marrow mesenchymal stem cells (MSCs) with Wnt11 would improve the outcomes of infarct repair. In vitro, Wnt11 pretreatment upregulated the expression of antiapoptotic and angiogenic molecules in MSCs. Wnt11 also increased the expression of Nkx2.5 (Figure), a marker of cardiac commitment. To test the efficacy of Wnt11 pretreatment on MSC-induced cardiac repair in vivo, MSCs were cultured in presence of Wnt11-secreting 293 cells (on an insert) for 5 days and transplanted intramyocardially after a reperfused MI. Age-matched male wild-type (C57BL/6J) mice underwent a 30-min coronary occlusion followed by reperfusion, and 48 h later received intramyocardial injection of vehicle, untreated MSCs (1 million in 30 microL), or Wnt11-pretreated MSCs (1 million in 30 microL). Echocardiography was performed 4 d prior to coronary occlusion/reperfusion (BSL1) and at 48 h (BSL2), 21 d, and 35 d after cell injection. After 35 d, compared with mice that received vehicle or untreated MSCs, mice injected with Wnt11-pretreated MSCs exhibited greater LV ejection fraction (Figure) and smaller LV end-diastolic volume, indicating that Wnt11 pretreatment transforms MSCs into superior vehicles of infarct repair. We conclude that intramyocardial transplantation of Wnt11-pretreated MSCs after a reperfused MI results in improved myocardial repair. These results underscore that pretreatment with Wnt11 may potentially enhance the outcomes of therapeutic infarct repair in humans.
Abstract 13768: Intracoronary Infusion of Bone Marrow-Derived Mesenchymal Stromal Cells Expressing Heme Oxygenase-1 to Prevent Reperfusion Injury in Porcine Model of Myocardial Infarction
- Wojciech Wojakowski,
- Michał Tendera,
- Wiesław Cybulski,
- Ewa K Zuba-Surma,
- Krzysztof Szade,
- Urszula Florczyk,
- Magdalena Kozakowska,
- Agnieszka Szymula,
- ukasz Krzych,
- Urszula Pasławska,
- Robert Pasławski,
- Krzysztof Milewski,
- Piotr P Buszman,
- Adrian Janiszewski,
- Piotr Dziegiel,
- Paweł E Buszman,
- Alicja Jóozkowicz and
- Jóozef Dulak
Circulation Volume 124, Issue suppl_2122 November 2011

Abstract Export Citations

Abstract
Combination of mesenchymal stromal cells (MSC) with antioxidative, antiapoptotic and proangiogenic enzyme heme oxygenase-1 (HO-1) is a novel approach for prevention of reperfusion injury in acute MI.Aim was to evaluate the effects of intracoronary infusion of bone marrow (BM)-derived MSC overexpressing HO-1 administered immediately before reperfusion in porcine model of MI with regard to reduction of the infarct area (IA), improvement of left ventricular ejection fraction (LVEF), LV remodeling, diastolic function and expression of inflammation-related genes.Methods: MSC were isolated from porcine BM and transduced 48 h before infusion with 50 MOI/cell of adenoviruses with HO-1 or GFP genes. MI was produced by 60-minutes inflation of OTW balloon catheter located in medial LAD in 27 domestic pigs. Prior to reperfusion animals were randomized to receive either HO-1-MSC, unmodified MSC, GFP+ MSC or placebo (normal saline) by slow infusion through the OTW catheter distally to the occlusion. Echocardiography was done before procedure, 30 min. after the reperfusion and 14 days later.Results: Administration of HO-1 MSC improved LVEF after reperfusion significantly more than unmodified MSC. After 14 days there was a trend towards superiority of HO-1-MSC, however both types of MSC were comparably effective vs. placebo. There was no effect of cells on LV remodeling and diastolic function. Both types of MSC significantly reduced the infarct size. Expression of HIF-1 was significantly lower in the infarct border zone in pigs treated with cells than in placebo group. GFP-positive cells expressing PCNA were localized in the endothelium within the infarct border zone.Conclusion: Intracoronary infusion of MSC prior to reperfusion reduced the infarct size and improved LV contractility and there was a trend towards more effective prevention of reperfusion injury in animals receiving MSC expressing HO-1. Proliferating donor MSC were identified within recipients endothelium.

Search

Narrow Results

Journal Title

Article Type

Subject

Date Range

Author

Export Citation

Advanced

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract

Abstract