Role of SDF:CXCR4 in diet-induced cardiac injury
Introduction Diabetic cardiomyopathy (DCM) is a major cardiovascular complication in patients with diabetes and is defined as ventricular dysfunction (in diabetes) independent of coronary artery disease. In this study, we define a novel role for the SDF‐1: CXCR4 axis in diabetes‐associated myocardial dysfunction. Methods Wild‐type mice were randomly assigned to a high fat high sugar diet (HFHS) or control diet (LF) for 14 months. Serial echocardiography was used to assess cardiac function. The hSDF‐1 plasmid was injected into the LV wall of HFHS mice 7 months and 14 months after HFHS diet. The mRNA levels in the hearts were quantified by qPCR. Results HFHS‐fed mice (vs. control diet) showed significantly increased deceleration time (diastolic dysfunction) 7 months after HFHS diet and decreased Ejection Fraction (EF) (systolic dysfunction) 14 months after the HFHS diet. We observed a significant increase in cardiac myocytes surface area and a decrease in vascular density in the hearts of HFHS mice. HFHS‐fed mice showed decreased P‐selectin, E‐selectin expression and increased CXCR4, Resistin, and DDR2 expression in the heart. There is no significant improvement in diastolic function after the SDF‐1 injection at 7 month. However, direct myocardial injection of hSDF‐1 plasmid led to a significant improvement in EF compared to the control group. To determine the role of CM CXCR4 in hyperglycemia associated cardiac injury, we quantified cardiac function and survival rate in HFHS‐fed CM‐CXCR4 null mice. We observed a significantly increased mortality rate before 14 months on HFHS fed CM‐CXCR4 null mice compare to HFHS‐fed control mice. Conclusion HFHS diet induces diastolic dysfunction in the short term then systolic dysfunction after a long term. SDF‐1 treatment ameliorates systolic dysfunction but not diastolic dysfunction. Our data suggest that cardiac myocyte expression of CXCR4 has an important role in diabetes‐associated cardiac injury.
Dong F;Patel K;Kiedrowski M;Penn M
Faseb Journal
2020
2020-04
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
journalArticle
<a href="http://doi.org/10.1096/fasebj.2020.34.s1.01787" target="_blank" rel="noreferrer noopener">10.1096/fasebj.2020.34.s1.01787</a>
Molecular basis of takotsubo syndrome
Introduction Takotsubo syndrome (TTS), also known as the “Broken Heart Syndrome” or “Apical Ballooning Syndrome is defined by its characteristic anomaly: when the heart contracts during systole, the apex of the heart dilates as the base of the heart contracts. Severe TTS can lead to cardiogenic shock and death in 3–4% of patients. There is no standard medical therapy for TTS because the mechanism underlying the development of the syndrome is unknown. Our goal is to determine the molecular mechanisms of TTS as a first step towards better treatment plans and outcomes. Methods Our model of TTS is a Kv1.5 null mouse, with compromised coronary metabolic dilation, subjected to transaortic constriction (TAC). Two weeks after TAC, when Kv1.5 null mouse showed profound apical ballooning during ventricular contraction (echocardiography), myocardial blood flow (MBF) was measured by contrast echocardiography in the base and apex of the left ventricle of mice under control conditions and during acute administration of norepinephrine to increase cardiac work. Hearts were collected and gene expression (RNA deep sequencing) in both the apex and the base were performed and followed by bioinformatic analysis. Wild type (WT) and unstressed Kv1.5 null mice were used as controls. To increase the scientific rigor, we purposefully analyzed RNA expression from different animals with Real‐Time‐PCR, to confirm the sequencing data. Results A total of 3875 genes were identified by differentially expressed between TTS vs unstressed Kv1.5 null hearts. Gene Set Enrichment Analysis shows many families of genes downregulated (metabolism) and upregulated (inflammation, hypoxia, apoptosis) in the apical (ballooning) area of the heart in TTS (compared to the base of the heart). RT‐qPCR revealed significant upregulation of the genes for Postn, Lox, and C920009B18Rik (p<0.01) and down‐regulation of the genes Ucp3, Acaa2, Pfkb1, Mir133a‐2 (p<0.05). These significant changes in expression were also shown in the apex of TTS hearts as compared to the apex of control Kv1.5 null hearts (p<0.01). We also found that in the mice with TTS, MBF was lower in the apex than in the base. Conclusion Our results suggest that the apex of the heart in TTS is receiving insufficient perfusion compared to the base, which may be one of the root problems in TTS. There is significant upregulation of structural genes in the apex of TTS hearts versus their bases and the apexes of normal hearts, whereas there is a significant downregulation of genes playing roles in metabolism. This confirms the importance of these specific genes in the development of the anomaly shown in TTS.
Chandler S;Joshi H;Hoff E;Patel K;Ohanyan V;Kiedrowski M;Chilian W;Dong F
Faseb Journal
2020
2020-04
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
journalArticle
<a href="http://doi.org/10.1096/fasebj.2020.34.s1.02623" target="_blank" rel="noreferrer noopener">10.1096/fasebj.2020.34.s1.02623</a>
Early Up-regulation Of Myocardial Cxcr4 Expression Is Critical For Cardiac Improvement With Chemical Preconditioning In Acute Myocardial Infarction
Cardiac regeneration; Cardiovascular System & Cardiology; heart failure; Myocardial infarction
Kamath M; Kiedrowski M; Weber K; Forudi F; Penn M S; Dong F
Circulation
2013
2013-11
Journal Article or Conference Abstract Publication
n/a
Cardiac Pressure Overload Initiates A Systemic Stem Cell Response
& Experimental Medicine; acute myocardial-infarction; Biotechnology & Applied Microbiology; bone marrow; bone marrow; cardiac stem cells; cardiomyocytes; Cell Biology; endogenous stem cells; endothelial; endothelial progenitor cells; heart; Hematology; hypertrophy; identification; murine; peripheral-blood; progenitor cells; regeneration; Research; spleen; SSEA-1; transaortic constriction; transplantation
Finan A; Kiedrowski M; Turturice B A; Sopko N A; Penn M S
Cytotherapy
2012
2012-09
Journal Article or Conference Abstract Publication
<a href="http://doi.org/10.3109/14653249.2012.684380" target="_blank" rel="noreferrer noopener">10.3109/14653249.2012.684380</a>
Rat Mesenchymal Stem Cell Secretome Promotes Elastogenesis And Facilitates Recovery From Simulated Childbirth Injury
Cell Biology; Elastin; Electromyography (EMG); expression; External urethral sphincter (EUS); female; female rats; mice; Paracrine factors; protection; Research & Experimental Medicine; Stress urinary incontinence (SUI); stress urinary-incontinence; stromal cells; tissue; transplantation; transplantation; Urethra; urethral sphincter; vaginal distension
Dissaranan C; Cruz M A; Kiedrowski M; Balog B M; Gill B C; Penn M S; Goldman H B; Damaser M S
Cell Transplantation
2014
2014
Journal Article or Conference Abstract Publication
<a href="http://doi.org/10.3727/096368913x670921" target="_blank" rel="noreferrer noopener">10.3727/096368913x670921</a>
Mesenchymal Stem Cells Facilitate Pudendal Nerve Recovery From Simulated Childbirth Injury
Urology & Nephrology
Damaser M; Lin D L; Hanzlicek B; Kiedrowski M; Balog B; Penn M; Goldman H
Neurourology and Urodynamics
2012
2012-08
Journal Article or Conference Abstract Publication
n/a
Mesenchymal Stem Cells Or Their Secretome Promote Recovery And Alter Urethral Elastin In An Animal Model Of Childbirth Injuries
Biotechnology & Applied Microbiology; Cell Biology; Engineering
Damaser M; Deng K; Lin D L; Hanzlicek B; Balog B; Penn M; Kiedrowski M; Zhu H
Journal of Tissue Engineering and Regenerative Medicine
2014
2014-06
Journal Article or Conference Abstract Publication
n/a
IMPACT OF RHOGDI GENE TRANSFECTION OF BLADDER SMOOTH MUSCLE CONTRACTILITY IN A VALIDATED EX-VIVO MURINE MODEL
Urology & Nephrology
19th Annual Fall Scientific Meeting of SMSNA
Introduction Plasmid-based gene therapy is an intriguing option for treating malignant and bladder pathologies. The RhoA pathway is involved in bladder smooth muscle regulation, cancer invasion and metastasis. Rho GDP-dissociation inhibitor (RhoGDI) is an inhibitor of the RhoA pathway. We validated ex-vivo bladder gene transfer to facilitate assessment of gene targets for treating bladder pathology. Methods Basic Local Alignment Search Tool was used to identify human RhoGDI coding sequences and to compare between rats and humans, which were cloned into a eCMV-based expression vector. NBTII rat bladder cancer cell lines were transfected using FuGENE (Promega, USA) and human protein expression and interaction with endogenous RhoA were tested using flow cytometry, immunofluorescence, and RNA expression analysis. Bladders were harvested from female Lewis Rats (∼250g) and sectioned and cultured for 72-hours following transfection with RhoGDI and FuGENE. Transfected bladder tissues were analyzed as described above. Non-transfected cultured bladder segments were analyzed using myography for viability and intact smooth muscle physiology in response to 120 mM KCl and 30uM carbachol. Results Human and rodent RhoGDI protein homology is 96%. Human RhoGDI was successfully detected exclusively in transfected NBTII cells with a top efficiency of 26%. qPCR analysis demonstrated rodent RhoA and RhoGDI levels were not impacted but ROCK1 and ROCK2 mRNA were significantly decreased by 23.6% (p=0.034) and 40.0% (p=0.015), respectively following human RhoGDI transfection. Human RhoGDI was detected in transfected ex-vivo cultured bladder segments in both FuGENE and microinjection experiments. Similar to NBTII cells, qPCR analysis demonstrated rodent RhoA and RhoGDI levels were not impacted but ROCK1 and ROCK2 mRNA were significantly decreased by 15.0% (p=0.035) and 22.4% (p=0.010) after FuGENE transfection and 20.5% (p=0.024) and 21.4% (p=0.015) after microinjections. Ex-vivo cultured bladder strips successfully contracted to KCl (mean 0.88+/-0.48 mN/mg tissue) and carbachol (1.82+/-0.97 mN/mg tissue) stimulation. After microinjection, RhoGDI caused a significant reduction in KCl mediated constriction although this was not observed in FuGENE experiments. Conclusion Ex-vivo bladder culture, transfection, and physiological assessment are feasible and may provide a high-throughput method to test novel gene transfer technologies before in-vivo testing. RhoGDI plasmid microinjection transfection appears to decrease contractility of ex-vivo bladder smooth muscle.
Joice G; Bell J M; La Favor J; Yoshida T; Torga G; Harris K; Liu X; Kiedrowski M; Penn M; Bivalacqua T
Journal of Sexual Medicine
2019
2019-04
Article information provided for research and reference use only. All rights are retained by the journal listed under publisher and/or the creator(s).
<a href="http://doi.org/10.1016/j.jsxm.2019.01.194" target="_blank" rel="noreferrer noopener">10.1016/j.jsxm.2019.01.194</a>
185 Impact of RhoGDI Gene Transfection of Bladder Smooth Muscle Contractility in a Validated Ex-vivo Murine Model
Joice G; Bell J M; La Favor J; Yoshida T; Torga G; Harris K; Liu X; Kiedrowski M; Penn M; Bivalacqua T
The Journal of Sexual Medicine
2019
2019-04
<a href="https://doi.org/10.1016/j.jsxm.2019.01.194">https://doi.org/10.1016/j.jsxm.2019.01.194</a>