TUG1 enhances high gluose impaired endothelial progenitor cell function via miR-29c-3p/PDGF-BB/Wnt signaling

Abstract Background Diabetes is associated with the dysfunction of endothelial progenitor cells (EPCs), characterized as impaired angiogenesis, a phenomenon thought to be involved in the development of diabetic foot. LncRNA plays an essential role in microvascular dysfunction and signaling pathways in patients with diabetes. LncRNA taurine upregulated gene 1 (TUG1) participates in angiogenesis in various cells. However, the mechanisms of TUG1 activity in EPCs have not been elucidated. Methods We isolated and then characterized EPCs from the peripheral blood of mice using immunofluorescence and flow cytometry. Western blot detected the wnt/β-catenin pathway in high glucose treated EPCs. Bioinformatics analysis predicted a putative binding site for TUG1 on miR-29c-3p. The interactions among TUG1, platelet-derived growth factor-BB (PDGF-BB) and miR-29c-3p were analyzed by luciferase assays. In vivo, diabetic mouse ischemic limb were treated with normal saline or TUG1 overexpression lentiviruses. Results We found that EPC migration, invasion and tube formation declined after treatment with high glucose, but improved with TUG1 overexpression. Mechanically, wnt/β-catenin pathway and autophagy were involved in the function of TUG1 overexpression in high glucose treated EPCs. Moreover, TUG1 regulates the PDGF-BB/wnt pathway and function of high glucose treated EPCs via miR-29c-3p. In vivo, injection of TUG1 lentivirus in a diabetic mouse ischemic limb model stimulated angiogenesis. Conclusions Our findings suggest that TUG1 restores high glucose treated EPC function by regulating miR-29c-3p/PDGF-BB/Wnt signaling.

Download Full-text

TUG1 enhances high gluose impaired endothelial progenitor cell function via miR-29c-3p/PDGF-BB/Wnt signaling

10.21203/rs.3.rs-33078/v2 ◽

2020 ◽

Author(s):

Yang Li ◽

Kangkang Zhi ◽

Shilong Han ◽

Xue Li ◽

Maoquan Li ◽

...

Keyword(s):

Wnt Signaling ◽

High Glucose ◽

Cell Function ◽

Microvascular Dysfunction ◽

Diabetic Mouse ◽

Tube Formation ◽

Endothelial Progenitor ◽

Ischemic Limb ◽

Patients With Diabetes

Abstract Background: Diabetes is associated with the dysfunction of endothelial progenitor cells (EPCs), characterized as impaired angiogenesis, a phenomenon thought to be involved in the development of diabetic foot. LncRNA plays an essential role in microvascular dysfunction and signaling pathways in patients with diabetes. LncRNA taurine upregulated gene 1 (TUG1) participates in angiogenesis in various cells. However, the mechanisms of TUG1 activity in EPCs have not been elucidated. Methods: We isolated and then characterized EPCs from the peripheral blood of mice using immunofluorescence and flow cytometry. Western blot detected the wnt/β-catenin pathway in high glucose treated EPCs. Bioinformatics analysis predicted a putative binding site for TUG1 on miR-29c-3p. The interactions among TUG1, platelet-derived growth factor-BB (PDGF-BB) and miR-29c-3p were analyzed by luciferase assays. In vivo, diabetic mouse ischemic limb were treated with normal saline or TUG1 overexpression lentiviruses. Results: We found that EPC migration, invasion and tube-formation declined after treatment with high-glucose, but improved with TUG1 overexpression. Mechanically, wnt/β-catenin pathway and autophagy were involved in the function of TUG1 overexpression in high glucose treated EPCs. Moreover, TUG1 regulates the PDGF-BB/wnt pathway and function of high glucose treated EPCs via miR-29c-3p. In vivo, injection of TUG1 lentivirus in a diabetic mouse ischemic limb model stimulated angiogenesis. Conclusions: Our findings suggest that TUG1 restores high glucose treated EPC function by regulating miR-29c-3p/PDGF-BB/Wnt signaling.

Download Full-text

TUG1 enhances high glucose-impaired endothelial progenitor cell function via miR-29c-3p/PDGF-BB/Wnt signaling

Stem Cell Research & Therapy ◽

10.1186/s13287-020-01958-3 ◽

2020 ◽

Vol 11 (1) ◽

Author(s):

Yang Li ◽

Kangkang Zhi ◽

Shilong Han ◽

Xue Li ◽

Maoquan Li ◽

...

Keyword(s):

Wnt Signaling ◽

High Glucose ◽

Cell Function ◽

Microvascular Dysfunction ◽

Diabetic Mouse ◽

Tube Formation ◽

Endothelial Progenitor ◽

Ischemic Limb ◽

Patients With Diabetes

Abstract Background Diabetes is associated with the dysfunction of endothelial progenitor cells (EPCs), characterized as impaired angiogenesis, a phenomenon thought to be involved in the development of diabetic foot. lncRNA plays an essential role in microvascular dysfunction and signaling pathways in patients with diabetes. lncRNA taurine upregulated gene 1 (TUG1) participates in angiogenesis in various cells. However, the mechanisms of TUG1 activity in EPCs have not been elucidated. Methods We isolated and then characterized EPCs from the peripheral blood of mice using immunofluorescence and flow cytometry. Western blot detected the wnt/β-catenin pathway in high glucose-treated EPCs. Bioinformatics analysis predicted a putative binding site for TUG1 on miR-29c-3p. The interactions among TUG1, platelet-derived growth factor-BB (PDGF-BB), and miR-29c-3p were analyzed by luciferase assays. In vivo, diabetic mouse ischemic limb was treated with normal saline or TUG1 overexpression lentiviruses. Results We found that EPC migration, invasion, and tube formation declined after treatment with high glucose, but improved with TUG1 overexpression. Mechanically, wnt/β-catenin pathway and autophagy were involved in the function of TUG1 overexpression in high glucose-treated EPCs. Moreover, TUG1 regulates the PDGF-BB/wnt pathway and function of high glucose-treated EPCs via miR-29c-3p. In vivo, injection of TUG1 lentivirus in a diabetic mouse ischemic limb model stimulated angiogenesis. Conclusions Our findings suggest that TUG1 restores high glucose-treated EPC function by regulating miR-29c-3p/PDGF-BB/Wnt signaling.

Download Full-text

STK35 Gene Therapy Attenuates Endothelial Dysfunction and Improves Cardiac Function in Diabetes

Frontiers in Cardiovascular Medicine ◽

10.3389/fcvm.2021.798091 ◽

2022 ◽

Vol 8 ◽

Author(s):

Darukeshwara Joladarashi ◽

Yanan Zhu ◽

Matthew Willman ◽

Kevin Nash ◽

Maria Cimini ◽

...

Keyword(s):

High Glucose ◽

Interstitial Fibrosis ◽

Microvascular Dysfunction ◽

Diabetic Mouse ◽

Tube Formation ◽

Left Ventricular ◽

Vascular Density ◽

Viral Particles ◽

Microvascular Pathology ◽

Angiogenic Genes

Diabetic cardiomyopathy (DCM) is characterized by microvascular pathology and interstitial fibrosis that leads to progressive heart failure. The mechanisms underlying DCM pathogenesis remain obscure, and no effective treatments for the disease have been available. In the present study, we observed that STK35, a novel kinase, is decreased in the diabetic human heart. High glucose treatment, mimicking hyperglycemia in diabetes, downregulated STK35 expression in mouse cardiac endothelial cells (MCEC). Knockdown of STK35 attenuated MCEC proliferation, migration, and tube formation, whereas STK35 overexpression restored the high glucose-suppressed MCEC migration and tube formation. Angiogenesis gene PCR array analysis revealed that HG downregulated the expression of several angiogenic genes, and this suppression was fully restored by STK35 overexpression. Intravenous injection of AAV9-STK35 viral particles successfully overexpressed STK35 in diabetic mouse hearts, leading to increased vascular density, suppression of fibrosis in the heart, and amelioration of left ventricular function. Altogether, our results suggest that hyperglycemia downregulates endothelial STK35 expression, leading to microvascular dysfunction in diabetic hearts, representing a novel mechanism underlying DCM pathogenesis. Our study also emerges STK35 is a novel gene therapeutic target for preventing and treating DCM.

Download Full-text

Loss of endothelial glucocorticoid receptor promotes angiogenesis via upregulation of Wnt/β-catenin pathway

Angiogenesis ◽

10.1007/s10456-021-09773-x ◽

2021 ◽

Author(s):

Bing Liu ◽

Han Zhou ◽

Tiening Zhang ◽

Xixiang Gao ◽

Bo Tao ◽

...

Keyword(s):

Endothelial Cells ◽

Glucocorticoid Receptor ◽

Wnt Signaling ◽

Limb Ischemia ◽

Tube Formation ◽

Autophagy Flux ◽

Nuclear Receptor Family ◽

Canonical Wnt

Abstract Objective The glucocorticoid receptor (GR) is a member of the nuclear receptor family that controls key biological processes in the cardiovascular system and has recently been shown to modulate Wnt signaling in endothelial cells. Wnt/β-catenin signaling has been demonstrated to be crucial in the process of angiogenesis. In the current study, we studied whether GR could regulate angiogenesis via the Wnt/β-catenin pathway. Approach and Resultsa Key components of the Wnt/β-catenin pathway were evaluated using quantitative PCR and Western blot in the presence or absence of GR. Enhanced angiogenesis was found in GR deficiency in vitro and confirmed with cell viability assays, proliferation assays and tube formation assays. Consistent with these in vitro findings, endothelial cell-specific GR loss GR in vivo promoted angiogenesis in both a hind limb ischemia model and sponge implantation assay. Results were further verified in a novel mouse model lacking endothelial LRP5/6, a key receptor in canonical Wnt signaling, and showed substantially suppressed angiogenesis using these same in vitro and in vivo assays. To further investigate the mechanism of GR regulation of Wnt signaling, autophagy flux was investigated in endothelial cells by visualizing auto phagolysosomes as well as by assessing P62 degradation and LC3B conversion. Results indicated that potentiated autophagy flux participated in GR-Wnt regulation. Conclusions Lack of endothelial GR triggers autophagy flux, leads to activation of Wnt/β-catenin signaling and promotes angiogenesis. There may also be a synergistic interaction between autophagy and Wnt/β-catenin signaling.

Download Full-text

miR-126 modulates angiogenic growth parameters of peripheral blood endothelial progenitor cells

Biological Chemistry ◽

10.1515/hsz-2014-0259 ◽

2015 ◽

Vol 396 (3) ◽

pp. 245-252 ◽

Cited By ~ 14

Author(s):

Sebastian M. Goerke ◽

Lena S. Kiefer ◽

G. Björn Stark ◽

Filip Simunovic ◽

Günter Finkenzeller

Keyword(s):

Tissue Engineering ◽

Progenitor Cells ◽

Endothelial Progenitor Cells ◽

Peripheral Blood ◽

Growth Parameters ◽

Therapeutic Option ◽

Tube Formation ◽

Loss Of Function ◽

Endothelial Progenitor

Abstract Vascularization plays an important role in tissue engineering applications. It is known that implantation of differentiated endothelial cells or endothelial progenitor cells (EPCs) from cord blood (cbEPCs) gives rise to the formation of a complex functional neovasculature, whereas EPCs isolated from peripheral blood (pbEPCs) have a limited capability to form blood vessels upon implantation. MicroRNA-126 (miR-126) has been shown to have pro-angiogenic effects in vivo. In this study, we investigated whether modulation of miR-126 expression in pbEPCs may alter their angiogenic properties. Gain of function and loss of function experiments revealed that miR-126 has anti-angiogenic effects in pbEPCs. Overexpression of miR-126 resulted in decreased proliferation, migration, invasion and tube formation, while inhibition of miR-126 induced the opposite effects. However, modulation of miR-126 expression did not influence apoptotic susceptibility of pbEPCs. This study provides evidence that inhibition of miR-126 improves angiogenesis-related growth parameters in pbEPCs and may represent a therapeutic option to ameliorate the angiogenic and vasculogenic properties of pbEPCs.

Download Full-text

High Glucose Aggravates the Detrimental Effects of Pancreatic Stellate Cells on Beta-Cell Function

International Journal of Endocrinology ◽

10.1155/2014/165612 ◽

2014 ◽

Vol 2014 ◽

pp. 1-8 ◽

Cited By ~ 9

Author(s):

Min Zha ◽

Wei Xu ◽

Qing Zhai ◽

Fengfei Li ◽

Bijun Chen ◽

...

Keyword(s):

Insulin Secretion ◽

High Glucose ◽

Cell Function ◽

Stellate Cells ◽

Pancreatic Stellate Cells ◽

High Concentration ◽

Gk Rats ◽

Chop Expression

Background and Aims. We here assess the effects of PSCs onβ-cell function and apoptosisin vivoandin vitro.Materials and Methods.PSCs were transplanted into Wistar and Goto-Kakizaki (GK) rats. Sixteen weeks after transplantation,β-cell function, apoptosis, and islet fibrosis were assessed.In vitrothe effects of PSCs conditioned medium (PSCs-CM) and/or high concentration of glucose on INS-1 cell function was assessed by measuring insulin secretion, INS-1 cell survival, apoptosis, and endoplasmic reticulum stress (ER stress) associated CHOP expression.Results. PSCs transplantation exacerbated the impairedβ-cell function in GK rats, but had no significant effects in Wistar rats.In vitro, PSCs-CM caused impaired INS-1 cell viability and insulin secretion and increased apoptosis, which were more pronounced in the presence of high glucose.Conclusion.Our study demonstrates that PSCs induceβ-cell failurein vitroandin vivo.

Download Full-text