High Glucose Causes Human Cardiac Progenitor Cell Dysfunction by Promoting Mitochondrial Fission: Role of a GLUT1 Blocker

( He Yun Choi ),( Ji Hye Park ),( Woong Bi Jang ),( Seung Taek Ji ),( Seok Yun Jung ),( Da Yeon Kim ),( Songhwa Kang ),( Yeon Ju Kim ),( Jisoo Yun ),( Jae Ho Kim ),( Sang Hong Baek ),( Sang Mo Kwon ) 한국응용약물학회 2016 Biomolecules & Therapeutics(구 응용약물학회지) Vol.24 No.4

Cardiovascular disease is the most common cause of death in diabetic patients. Hyperglycemia is the primary characteristic of diabetes and is associated with many complications. The role of hyperglycemia in the dysfunction of human cardiac progenitor cells that can regenerate damaged cardiac tissue has been investigated, but the exact mechanism underlying this association is not clear. Thus, we examined whether hyperglycemia could regulate mitochondrial dynamics and lead to cardiac progenitor cell dysfunction, and whether blocking glucose uptake could rescue this dysfunction. High glucose in cardiac progenitor cells results in reduced cell viability and decreased expression of cell cycle-related molecules, including CDK2 and cyclin E. A tube formation assay revealed that hyperglycemia led to a significant decrease in the tube-forming ability of cardiac progenitor cells. Fluorescent labeling of cardiac progenitor cell mitochondria revealed that hyperglycemia alters mitochondrial dynamics and increases expression of fission-related proteins, including Fis1 and Drp1. Moreover, we showed that specific blockage of GLUT1 improved cell viability, tube formation, and regulation of mitochondrial dynamics in cardiac progenitor cells. To our knowledge, this study is the first to demonstrate that high glucose leads to cardiac progenitor cell dysfunction through an increase in mitochondrial fission, and that a GLUT1 blocker can rescue cardiac progenitor cell dysfunction and downregulation of mitochondrial fission. Combined therapy with cardiac progenitor cells and a GLUT1 blocker may provide a novel strategy for cardiac progenitor cell therapy in cardiovascular disease patients with diabetes.

High Glucose Causes Human Cardiac Progenitor Cell Dysfunction by Promoting Mitochondrial Fission: Role of a GLUT1 Blocker

Choi, He Yun,Park, Ji Hye,Jang, Woong Bi,Ji, Seung Taek,Jung, Seok Yun,Kim, Da Yeon,Kang, Songhwa,Kim, Yeon Ju,Yun, Jisoo,Kim, Jae Ho,Baek, Sang Hong,Kwon, Sang-Mo The Korean Society of Applied Pharmacology 2016 Biomolecules & Therapeutics(구 응용약물학회지) Vol.24 No.4

Cardiovascular disease is the most common cause of death in diabetic patients. Hyperglycemia is the primary characteristic of diabetes and is associated with many complications. The role of hyperglycemia in the dysfunction of human cardiac progenitor cells that can regenerate damaged cardiac tissue has been investigated, but the exact mechanism underlying this association is not clear. Thus, we examined whether hyperglycemia could regulate mitochondrial dynamics and lead to cardiac progenitor cell dysfunction, and whether blocking glucose uptake could rescue this dysfunction. High glucose in cardiac progenitor cells results in reduced cell viability and decreased expression of cell cycle-related molecules, including CDK2 and cyclin E. A tube formation assay revealed that hyperglycemia led to a significant decrease in the tube-forming ability of cardiac progenitor cells. Fluorescent labeling of cardiac progenitor cell mitochondria revealed that hyperglycemia alters mitochondrial dynamics and increases expression of fission-related proteins, including Fis1 and Drp1. Moreover, we showed that specific blockage of GLUT1 improved cell viability, tube formation, and regulation of mitochondrial dynamics in cardiac progenitor cells. To our knowledge, this study is the first to demonstrate that high glucose leads to cardiac progenitor cell dysfunction through an increase in mitochondrial fission, and that a GLUT1 blocker can rescue cardiac progenitor cell dysfunction and downregulation of mitochondrial fission. Combined therapy with cardiac progenitor cells and a GLUT1 blocker may provide a novel strategy for cardiac progenitor cell therapy in cardiovascular disease patients with diabetes.

The Calcineurin-Drp1-Mediated Mitochondrial Fragmentation is Aligned with the Differentiation of c-Kit Cardiac Progenitor Cells

Attaur Rahman,Yuhao Li,Nur Izzah Ismail,To-Kiu Chan,Yuzhen Li,Dachun Xu,Hao Zhou,Sang-Bing Ong Korean Society for Stem Cell Research 2023 International journal of stem cells Vol.16 No.2

Objective: The heart contains a pool of c-kit⁺ progenitor cells which is believed to be able to regenerate. The differentiation of these progenitor cells is reliant on different physiological cues. Unraveling the underlying signals to direct differentiation of progenitor cells will be beneficial in controlling progenitor cell fate. In this regard, the role of the mitochondria in mediating cardiac progenitor cell fate remains unclear. Specifically, the association between changes in mitochondrial morphology with the differentiation status of c-kit⁺ CPCs remains elusive. In this study, we investigated the relationship between mitochondrial morphology and the differentiation status of c-kit⁺ progenitor cells. Methods and Results: c-kit⁺ CPCs were isolated from 2-month-old male wild-type FVB mice. To activate differentiation, CPCs were incubated in α-minimal essential medium containing 10 nM dexamethasone for up to 7 days. To inhibit Drp1-mediated mitochondrial fragmentation, either 10 μM or 50 μM mdivi-1 was administered once at Day 0 and again at Day 2 of differentiation. To inhibit calcineurin, either 1 μM or 5 μM ciclosporin-A (CsA) was administered once at Day 0 and again at Day 2 of differentiation. Dexamethasone-induced differentiation of c-kit⁺ progenitor cells is aligned with fragmentation of the mitochondria via a calcineurin-Drp1 pathway. Pharmacologically inhibiting mitochondrial fragmentation retains the undifferentiated state of the c-kit⁺ progenitor cells. Conclusions: The findings from this study provide an alternative view of the role of mitochondrial fusion-fission in the differentiation of cardiac progenitor cells and the potential of pharmacologically manipulating the mitochondria to direct progenitor cell fate.

Engineered M13 Peptide Carrier Promotes Angiogenic Potential of Patient-Derived Human Cardiac Progenitor Cells and In Vivo Engraftment

장웅비,지승택,박지혜,Kim Yeon-Ju,Kang Songhwa,김다연,이나경,김진수,Lim Hye Ji,최재우,LE THI HONG VAN,LY THANH TRUONG GIANG,비누스,김동환,하종성,윤지수,Baek Sang Hong,권상모 한국조직공학과 재생의학회 2020 조직공학과 재생의학 Vol.17 No.3

BACKGROUND: Despite promising advances in stem cell-based therapy, the treatment of ischemic cardiovascular diseases remains a big challenge due to both the insufficient in vivo viability of transplanted cells and poor angiogenic potential of stem cells. The goal of this study was to develop therapeutic human cardiac progenitor cells (hCPCs) for ischemic cardiovascular diseases with a novel M13 peptide carrier. METHOD: In this study, an engineered M13 peptide carrier was successfully generated using a QuikChange Kit. The cellular function of M13 peptide carrier-treated hCPCs was assessed using a tube formation assay and scratch wound healing assay. The in vivo engraftment and cell survival bioactivities of transplanted cells were demonstrated by immunohistochemistry after hCPC transplantation into a myocardial infarction animal model. RESULTS: The engineered M13RGD?SDKP peptide carrier, which expressed RGD peptide on PIII site and SDKP peptide on PVIII site, did not affect morphologic change and proliferation ability in hCPCs. In contrast, hCPCs treated with M13RGD?SDKP showed enhanced angiogenic capacity, including tube formation and migration capacity. Moreover, transplanted hCPCs with M13RGD?SDKP were engrafted into the ischemic region and promoted in vivo cell survival. CONCLUSION: Our present data provides a promising protocol for CPC-based cell therapy via short-term cell priming of hCPCs with engineered M13RGD?SDKP before cell transplantation for treatment of cardiovascular disease.

Amine-enriched surface modification facilitates expansion, attachment, and maintenance of human cardiac-derived c-kit positive progenitor cells

Choi, S.H.,Jung, S.Y.,Yoo, S.M.,Asahara, T.,Suh, W.,Kwon, S.M.,Baek, S.H. Elsevier/North-Holland Biomedical Press 2013 INTERNATIONAL JOURNAL OF CARDIOLOGY Vol.168 No.1

Background: Stem cells have a low expansion rate and are difficult to maintain in vitro. To overcome the problems of cardiovascular regeneration, we developed a novel method of stem cell cultivation in culture vessels with amine and carboxyl coatings. Methods and results: We isolated cardiac stem/progenitor cells from infant-derived heart tissue by using c-kit antibody (human cardiac-derived c-kit positive progenitor cells; hCPC<SUP>c-kit+</SUP>); the cells differentiated into endothelial cells, smooth muscle cells, and cardiomyocytes. To characterize the effect of surface modification on hCPC<SUP>c-kit+</SUP> expansion, cellular attachment, c-kit expression maintenance, and cardiomyocyte differentiation, we tested hCPC<SUP>c-kit+</SUP> cultured on non-coated (control), amine-coated (amine), and carboxyl-coated (carboxyl) vessels. Ex vivo proliferation, c-kit maintenance, and cellular attachment were significantly enhanced in the amine group. The amine coating also increased procollagen type I (pro-COL1) expression and increased phosphorylation signals, such as focal adhesion kinase (FAK) and cytosolic Src, as well as enhanced ERK/CDK2 signaling. In addition, there was significant downregulation of the stress signal transducer, JNK, in the amine group. However, cardiomyogenesis remained unchanged in the control, amine, and carboxyl groups. Conclusions: Although surface modifications had no effect on early induction cardiomyogenesis, amine-enriched surface modification may increase hCPC<SUP>c-kit+</SUP> expansion. The amine-enriched surface improved cellular proliferation and attachment during ex vivo hCPC<SUP>c-kit+</SUP> expansion, possibly by modulating intracellular signal transducers.

Doxorubicin Regulates Autophagy Signals via Accumulation of Cytosolic Ca ²⁺ in Human Cardiac Progenitor Cells

Park, Ji Hye,Choi, Sung Hyun,Kim, Hyungtae,Ji, Seung Taek,Jang, Woong Bi,Kim, Jae Ho,Baek, Sang Hong,Kwon, Sang Mo MDPI 2016 INTERNATIONAL JOURNAL OF MOLECULAR SCIENCES Vol.17 No.10

<P>Doxorubicin (DOXO) is widely used to treat solid tumors. However, its clinical use is limited by side effects including serious cardiotoxicity due to cardiomyocyte damage. Resident cardiac progenitor cells (hCPCs) act as key regulators of homeostasis in myocardial cells. However, little is known about the function of hCPCs in DOXO-induced cardiotoxicity. In this study, we found that DOXO-mediated hCPC toxicity is closely related to calcium-related autophagy signaling and was significantly attenuated by blocking mTOR signaling in human hCPCs. DOXO induced hCPC apoptosis with reduction of SMP30 (regucalcin) and autophagosome marker LC3, as well as remarkable induction of the autophagy-related markers, Beclin-1, APG7, and P62/SQSTM1 and induction of calcium-related molecules, CaM (Calmodulin) and CaMKII (Calmodulin kinase II). The results of an LC3 puncta assay further indicated that DOXO reduced autophagosome formation via accumulation of cytosolic Ca<SUP>2+</SUP>. Additionally, DOXO significantly induced mTOR expression in hCPCs, and inhibition of mTOR signaling by rapamycin, a specific inhibitor, rescued DOXO-mediated autophagosome depletion in hCPCs with significant reduction of DOXO-mediated cytosolic Ca<SUP>2+</SUP> accumulation in hCPCs, and restored SMP30 and mTOR expression. Thus, DOXO-mediated hCPC toxicity is linked to Ca<SUP>2+</SUP>-related autophagy signaling, and inhibition of mTOR signaling may provide a cardio-protective effect against DOXO-mediated hCPC toxicity.</P>

Spatial Allocation and Specification of Cardiomyocytes during Zebrafish Embryogenesis

Hajime Fukui,Ayano Chiba,Takahiro Miyazaki,Haruko Takano,Hiroyuki Ishikawa,Naoki Mochiuzki,Toyonori Omori 대한심장학회 2017 Korean Circulation Journal Vol.47 No.2

Incomplete development and severe malformation of the heart result in miscarriage of embryos because of its malfunction as a pump for circulation. During cardiogenesis, development of the heart is precisely coordinated by the genetically-primed program that is revealed by the sequential expression of transcription factors. It is important to investigate how spatial allocation of the heart containing cardiomyocytes and other mesoderm-derived cells is determined. In addition, the molecular mechanism underlying cardiomyocyte differentiation still remains elusive. The location of ectoderm-, mesoderm-, and endoderm-derived organs is determined by their initial allocation and subsequent mutual cell-cell interactions or paracrine-based regulation. In the present work, we provide an overview of cardiac development controlled by the germ layers and discuss the points that should be uncovered in future for understanding cardiogenesis.

심근유래의 줄기세포 프라이밍 및 조직공학 기술에 의한 심혈관 재생 전략

박지혜(Ji Hye Park),김희정(Hee Jung Kim),정한나(Hanna Jung),박해리(Haeri Park),황혜원(Hyewon Hwang),권상모(Sang Mo Kwon) 한국생물공학회 2018 KSBB Journal Vol.33 No.3

The prevalence and mortality rate of cardiovascular disease continue to increase despite the development of new drugs and treatments. Stem cell therapy is a promising strategy to repair and/or regenerate damaged cardiac tissue. Cardiac progenitor cells may be stronger candidates than other stem cells for cardiac cell therapy. Cardiac progenitor cells (CPCs) residing in the heart (also referred to as cardiosphere-derived cells (CDCs) exert a therapeutic potential in repairing heart damage. Numerous studies have identified candidate CPC cell surface markers, including c-kit, stem cell antigen 1 (Sca1), and platelet-derived growth factor receptor α (PDGFRα). CPCs are capable of differentiating into multiple mature cardiac cell types, including cardiomyocytes (CM), smooth muscle cells (SMC), and endothelial cells (EC). Owing to their unique pluripotency, various pre-clinical and clinical studies are being conducted using cardiac progenitor cells to regenerate damaged heart tissues. However, poor stem cell survival and low engraftment efficiency underlie significant reductions in therapeutic efficacy. To overcome current limitations that reduce stem cell therapeutic efficacy, many methods have been implemented that aim to improve stem cell survival and engraftment. Recently, stem cell priming and tissue engineering technologies have been suggested to enhance stem cell therapeutic efficacy. It is possible to increase survival and engraftment of cardiac progenitor cells in injured cardiac tissue by cell priming and tissue engineering technologies before use. In this review, we provide an understanding of multiple therapeutic approaches including cell priming, scaffolds, tissue engineering, 3D printing techniques and clinical prospect for cardiovascular regeneration using patient-derived CPCs.

Characteristics and Cardiomyogenic Potential of Rat Fetal Cardiac Progenitor Cells at Different Developmental Stage

Tung Nguyen Thanh,신힘차,김활란,박소라,김지영,최병현 한국조직공학과 재생의학회 2017 조직공학과 재생의학 Vol.14 No.3

In recent years, several kinds of cardiac progenitor cells have been identified and isolated from heart tissue. These cells showed differentiation potential into cardiomyocytes, smooth muscle cells, and endothelial cells in vitro and in vivo. Morphogenetic events are tightly regulated during development to determine cell destiny and reshape the embryonic lineage. In this study, we directly compared the characteristics of rat fetal cardiac progenitor cells (rFCPCs) isolated from the chamber formation stage at embryonic day 12 (E12) and at the septation stage of E15. Both kinds of rFCPCs expressed mesenchymal stem cell markers (CD105, CD73, and CD29) but not CD34 and CD45. The E12 rFCPCs expressed a high level of Oct4 compared to E15 until passage 5 and showed a steep decline of Nkx2.5 expression at passage 5. However, Nkx2.5 expression at E15 was maintained until passage 5 and Oct4 expression slightly increased at passage 5. We also detected an intense staining for Oct4 antibody in E12 heart tissue sections. The average doubling time of the E12 rFCPCs from passage 3 to passage 15 was about 5 hours longer than E15. These cells could also be induced into cardiomyocytes expressing a-MHC, cTnT, cTnC, and Cx43 under cardiomyogenic culture conditions and rFCPCs at E15 showed more intense staining of a-MHC than cells at E12 by immunocytochemistry. Taken together, our results show that developmental differences between E12 and E15 may influence their properties and differentiation. Furthermore those differences should be considered when deciding on the optimal cell source for cell replacement therapy in cardiovascular regeneration.

Regulation of ROS-independent ERK signaling rescues replicative cellular senescence in ex vivo expanded human c-kit-positive cardiac progenitor cells

Choi, S.H.,Jung, S.Y.,Yoo, S.Y.,Yoo, S.M.,Kim, D.Y.,Kang, S.,Baek, S.H.,Kwon, S.M. Elsevier/North-Holland Biomedical Press 2013 INTERNATIONAL JOURNAL OF CARDIOLOGY Vol.169 No.1

Backgrounds: Although the rescue of cellular senescence during ex vivo expansion of human-derived cardiac progenitor cells (hCPC) is critical for the application of autologous stem cell therapy in cardiovascular disease, the underlying molecular pathways during replicative senescence in hCPC have not been fully defined. Thus, we examined whether the regulation of mitogen-activated protein kinases activation could facilitate the recovery of human c-kit-positive hCPCs (hCPC<SUP>c-kit+</SUP>) and whether senescence is reactive oxygen species (ROS)-dependent or -independent. Methods and results: To investigate the molecular pathways of replicative cellular senescence, we first evaluated cellular senescence in ex vivo-expanded hCPC<SUP>c-kit+</SUP> by using senescence-associated β-galactosidase (SA-β-gal) activity with enlarged cytoplasm and observed increased expression of cell senescence-related pivotal molecules, including TP53, cleavage Mdm2 (cMdm2), and Mdm2. Unexpectedly, we found that the extracellular signal-regulated kinase (ERK) was markedly activated in aged hCPC<SUP>c-kit+</SUP>, with reduced proliferative activity. SA-β-gal activity and cytoplasm size in senescent hCPC<SUP>c-kit+</SUP> were significantly reduced, with reduced TP53 and cMdm2 expression after treatment with a specific ERK inhibitor (U0126). We examined whether the signaling in ERK inhibitory rescue of hCPC<SUP>c-kit+</SUP> senescence is ROS-dependent. Interestingly, the increased ROS level was not changed after treatment with a specific ERK inhibitor. Similarly, the increased expression levels of endogenous antioxidant enzymes, e.g., peroxiredoxin (Prdx)-1 and 2, in senescent hCPC<SUP>c-kit+</SUP> were not changed after treatment with a specific ERK inhibitor. Conclusions: From the above results, we conclude that the specific inhibition of ERK during cellular senescence might rescue bioactivities of senescent hCPC<SUP>c-kit+</SUP> in a ROS-independent manner.