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Human umbilical cord blood-derived mononuclear cells improve murine ventricular function upon intramyocardial delivery in right ventricular chronic pressure overload.

Oommen S, Yamada S, Cantero Peral S, Campbell KA, Bruinsma ES, Terzic A, Nelson TJ - Stem Cell Res Ther (2015)

Bottom Line: The RV volume load in PAB-only mice was 24.09±3.9 compared to 11.05±2.09 in the cell group (mm3, P-value<0.005).The analysis of pathogenic gene expression (BNP, ANP, Acta1, Myh7) in the cell-transplanted group showed a significant reversal with respect to the diseased PAB mice with a robust increase in cardiac progenitor gene expression such as GATA4, Kdr, Mef2c and Nkx2.5.Histological analysis indicated significant fibrosis in the RV in response to PAB that was reduced following UCB-MNC's transplantation along with concomitant increased Ki-67 expression and CD31 positive vessels as a marker of angiogenesis within the myocardium.

View Article: PubMed Central - PubMed

Affiliation: General Internal Medicine and Transplant Center, Mayo Clinic, Rochester, MN, USA. oommen.saji@mayo.edu.

ABSTRACT

Introduction: Stem cell therapy has emerged as potential therapeutic strategy for damaged heart muscles. Umbilical cord blood (UCB) cells are the most prevalent stem cell source available, yet have not been fully tested in cardiac regeneration. Herein, studies were performed to evaluate the cardiovascular safety and beneficial effect of mononuclear cells (MNCs) isolated from human umbilical cord blood upon intramyocardial delivery in a murine model of right ventricle (RV) heart failure due to pressure overload.

Methods: UCB-derived MNCs were delivered into the myocardium of a diseased RV cardiac model. Pulmonary artery banding (PAB) was used to produce pressure overload in athymic nude mice that were then injected intramyocardially with UCB-MNCs (0.4×10^6 cells/heart). Cardiac functions were then monitored by telemetry, echocardiography, magnetic resonance imaging (MRI) and pathologic analysis of heart samples to determine the ability for cell-based repair.

Results: The cardio-toxicity studies provided evidence that UCB cell transplantation has a safe therapeutic window between 0.4 to 0.8 million cells/heart without altering QT or ST-segments or the morphology of electrocardiograph waves. The PAB cohort demonstrated significant changes in RV chamber dilation and functional defects consistent with severe pressure overload. Using cardiac MRI analysis, UCB-MNC transplantation in the setting of PAB demonstrated an improvement in RV structure and function in this surgical mouse model. The RV volume load in PAB-only mice was 24.09±3.9 compared to 11.05±2.09 in the cell group (mm3, P-value<0.005). The analysis of pathogenic gene expression (BNP, ANP, Acta1, Myh7) in the cell-transplanted group showed a significant reversal with respect to the diseased PAB mice with a robust increase in cardiac progenitor gene expression such as GATA4, Kdr, Mef2c and Nkx2.5. Histological analysis indicated significant fibrosis in the RV in response to PAB that was reduced following UCB-MNC's transplantation along with concomitant increased Ki-67 expression and CD31 positive vessels as a marker of angiogenesis within the myocardium.

Conclusions: These findings indicate that human UCB-derived MNCs promote an adaptive regenerative response in the right ventricle upon intramyocardial transplantation in the setting of chronic pressure overload heart failure.

No MeSH data available.


Related in: MedlinePlus

UCB mononuclear cells suppressed PAB-induced increase in pathogenic gene expression and upregulated cardiac transcription factors. (A through E) Right ventricular free wall was used for quantitative real-time PCR analysis of cardiac pathogenic markers, such as BNP, ANP, Acta1, Myh7 and Myh6. Transcript levels of these markers were measured in heart tissue of the PAB-only group and compared with that of the UCB-MNC transplanted group. (F) Cardiac transcription factors (GATA4, Kdr, Mef2c and Nkx2.5) were significantly upregulated in the UCB-MNC transplanted heart tissue in comparison with PAB-only heart tissue (n = 4, ** P-value <0.05 versus PAB). Data are expressed as fold change. All data reported are representative of four independent experiments performed in triplicate and are normalized to those of GAPDH. Data are presented as mean ± SEM. PAB, pulmonary artery banding; SEM, standard error of the mean; UCB-MNCs, umbilical cord blood-mononuclear cells.
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Fig6: UCB mononuclear cells suppressed PAB-induced increase in pathogenic gene expression and upregulated cardiac transcription factors. (A through E) Right ventricular free wall was used for quantitative real-time PCR analysis of cardiac pathogenic markers, such as BNP, ANP, Acta1, Myh7 and Myh6. Transcript levels of these markers were measured in heart tissue of the PAB-only group and compared with that of the UCB-MNC transplanted group. (F) Cardiac transcription factors (GATA4, Kdr, Mef2c and Nkx2.5) were significantly upregulated in the UCB-MNC transplanted heart tissue in comparison with PAB-only heart tissue (n = 4, ** P-value <0.05 versus PAB). Data are expressed as fold change. All data reported are representative of four independent experiments performed in triplicate and are normalized to those of GAPDH. Data are presented as mean ± SEM. PAB, pulmonary artery banding; SEM, standard error of the mean; UCB-MNCs, umbilical cord blood-mononuclear cells.

Mentions: Specific cardiac pathogenic markers were used for the diagnosis of heart failure and recovery. The dysfunctional heart tissue of PAB mice showed an elevated level of pathogenic gene expression (BNP, ANP, Acta1and Myh7) compared to a significant reversal of these genes in the group of animals receiving cell transplantation (Figure 6A- D). The expression of Myh6 was not significantly changed under these tested conditions (Figure 6E). Transcriptional regulation by multiple cardiac transcription factors, such as GATA4, Kdr, Mef2c, and Nkx2.5, is associated with an adaptive cardiac regenerative process [20]. After PAB, the transcript levels of cardiac progenitors GATA4, Kdr, Mef2c, and Nkx2.5 were significantly lower than those of control tissue. The embryonic gene expression profile was reversed upon transplantation of UCB-MNCs (Figure 6F).Figure 6


Human umbilical cord blood-derived mononuclear cells improve murine ventricular function upon intramyocardial delivery in right ventricular chronic pressure overload.

Oommen S, Yamada S, Cantero Peral S, Campbell KA, Bruinsma ES, Terzic A, Nelson TJ - Stem Cell Res Ther (2015)

UCB mononuclear cells suppressed PAB-induced increase in pathogenic gene expression and upregulated cardiac transcription factors. (A through E) Right ventricular free wall was used for quantitative real-time PCR analysis of cardiac pathogenic markers, such as BNP, ANP, Acta1, Myh7 and Myh6. Transcript levels of these markers were measured in heart tissue of the PAB-only group and compared with that of the UCB-MNC transplanted group. (F) Cardiac transcription factors (GATA4, Kdr, Mef2c and Nkx2.5) were significantly upregulated in the UCB-MNC transplanted heart tissue in comparison with PAB-only heart tissue (n = 4, ** P-value <0.05 versus PAB). Data are expressed as fold change. All data reported are representative of four independent experiments performed in triplicate and are normalized to those of GAPDH. Data are presented as mean ± SEM. PAB, pulmonary artery banding; SEM, standard error of the mean; UCB-MNCs, umbilical cord blood-mononuclear cells.
© Copyright Policy - open-access
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4416353&req=5

Fig6: UCB mononuclear cells suppressed PAB-induced increase in pathogenic gene expression and upregulated cardiac transcription factors. (A through E) Right ventricular free wall was used for quantitative real-time PCR analysis of cardiac pathogenic markers, such as BNP, ANP, Acta1, Myh7 and Myh6. Transcript levels of these markers were measured in heart tissue of the PAB-only group and compared with that of the UCB-MNC transplanted group. (F) Cardiac transcription factors (GATA4, Kdr, Mef2c and Nkx2.5) were significantly upregulated in the UCB-MNC transplanted heart tissue in comparison with PAB-only heart tissue (n = 4, ** P-value <0.05 versus PAB). Data are expressed as fold change. All data reported are representative of four independent experiments performed in triplicate and are normalized to those of GAPDH. Data are presented as mean ± SEM. PAB, pulmonary artery banding; SEM, standard error of the mean; UCB-MNCs, umbilical cord blood-mononuclear cells.
Mentions: Specific cardiac pathogenic markers were used for the diagnosis of heart failure and recovery. The dysfunctional heart tissue of PAB mice showed an elevated level of pathogenic gene expression (BNP, ANP, Acta1and Myh7) compared to a significant reversal of these genes in the group of animals receiving cell transplantation (Figure 6A- D). The expression of Myh6 was not significantly changed under these tested conditions (Figure 6E). Transcriptional regulation by multiple cardiac transcription factors, such as GATA4, Kdr, Mef2c, and Nkx2.5, is associated with an adaptive cardiac regenerative process [20]. After PAB, the transcript levels of cardiac progenitors GATA4, Kdr, Mef2c, and Nkx2.5 were significantly lower than those of control tissue. The embryonic gene expression profile was reversed upon transplantation of UCB-MNCs (Figure 6F).Figure 6

Bottom Line: The RV volume load in PAB-only mice was 24.09±3.9 compared to 11.05±2.09 in the cell group (mm3, P-value<0.005).The analysis of pathogenic gene expression (BNP, ANP, Acta1, Myh7) in the cell-transplanted group showed a significant reversal with respect to the diseased PAB mice with a robust increase in cardiac progenitor gene expression such as GATA4, Kdr, Mef2c and Nkx2.5.Histological analysis indicated significant fibrosis in the RV in response to PAB that was reduced following UCB-MNC's transplantation along with concomitant increased Ki-67 expression and CD31 positive vessels as a marker of angiogenesis within the myocardium.

View Article: PubMed Central - PubMed

Affiliation: General Internal Medicine and Transplant Center, Mayo Clinic, Rochester, MN, USA. oommen.saji@mayo.edu.

ABSTRACT

Introduction: Stem cell therapy has emerged as potential therapeutic strategy for damaged heart muscles. Umbilical cord blood (UCB) cells are the most prevalent stem cell source available, yet have not been fully tested in cardiac regeneration. Herein, studies were performed to evaluate the cardiovascular safety and beneficial effect of mononuclear cells (MNCs) isolated from human umbilical cord blood upon intramyocardial delivery in a murine model of right ventricle (RV) heart failure due to pressure overload.

Methods: UCB-derived MNCs were delivered into the myocardium of a diseased RV cardiac model. Pulmonary artery banding (PAB) was used to produce pressure overload in athymic nude mice that were then injected intramyocardially with UCB-MNCs (0.4×10^6 cells/heart). Cardiac functions were then monitored by telemetry, echocardiography, magnetic resonance imaging (MRI) and pathologic analysis of heart samples to determine the ability for cell-based repair.

Results: The cardio-toxicity studies provided evidence that UCB cell transplantation has a safe therapeutic window between 0.4 to 0.8 million cells/heart without altering QT or ST-segments or the morphology of electrocardiograph waves. The PAB cohort demonstrated significant changes in RV chamber dilation and functional defects consistent with severe pressure overload. Using cardiac MRI analysis, UCB-MNC transplantation in the setting of PAB demonstrated an improvement in RV structure and function in this surgical mouse model. The RV volume load in PAB-only mice was 24.09±3.9 compared to 11.05±2.09 in the cell group (mm3, P-value<0.005). The analysis of pathogenic gene expression (BNP, ANP, Acta1, Myh7) in the cell-transplanted group showed a significant reversal with respect to the diseased PAB mice with a robust increase in cardiac progenitor gene expression such as GATA4, Kdr, Mef2c and Nkx2.5. Histological analysis indicated significant fibrosis in the RV in response to PAB that was reduced following UCB-MNC's transplantation along with concomitant increased Ki-67 expression and CD31 positive vessels as a marker of angiogenesis within the myocardium.

Conclusions: These findings indicate that human UCB-derived MNCs promote an adaptive regenerative response in the right ventricle upon intramyocardial transplantation in the setting of chronic pressure overload heart failure.

No MeSH data available.


Related in: MedlinePlus