Secretion of miRNA-326-3p by senescent adipose exacerbates myocardial metabolism in diabetic mice

Abstract

Background Adipose tissue homeostasis is at the heart of many metabolic syndromes such as diabetes. Previously it has been demonstrated that adipose tissues from diabetic patients are senescent but whether this contributes to diabetic cardiomyopathy (DCM) remains to be elucidated. Methods The streptozotocin (STZ) type 1 diabetic mice were established as animal model, and adult mouse ventricular myocytes (AMVMs) isolated by langendorff perfusion as well as neonatal mouse ventricular myocytes (NMVMs) were used as cell models. Senescent associated β galactosidase (SA-β-gal) staining and RT-qPCR were used to identify the presence of adipose senescence in diabetic adipose tissue. Senescent adipose were removed either by surgery or by senolytic treatment. Large extracellular vesicles (LEVs) derived from adipose tissue and circulation were separated by ultracentrifugation. Cardiac systolic and diastolic function was evaluated through cardiac ultrasound. Cardiomyocytes contraction function was evaluated by the Ionoptix HTS system and live cell imaging, mitochondrial morphology and functions were evaluated by transmission electron microscope, live cell fluorescent probe and seahorse analysis. RNA-seq for AMVMs and miRNA-seq for LEVs were performed, and bioinformatic analysis combined with RT-qPCR and Western blot were used to elucidate underlying mechanism that senescent adipose derives LEVs exacerbates myocardial metabolism. Results SA-β-gal staining and RT-qPCR identified the presence of adipose tissue senescence in STZ mice. Through surgical as well as pharmacological means we show that senescent adipose tissue participates in the pathogenesis of DCM in STZ mice by exacerbates myocardial metabolism through secretion of LEVs. Specifically, expression of miRNA-326-3p was up-regulated in LEVs isolated from senescent adipose tissue, circulation, and cardiomyocytes of STZ mice. Up-regulation of miRNA-326-3p coincided with myocardial transcriptomic changes in metabolism. Functionally, we demonstrate that miRNA-326-3p inhibited the expression of Rictor and resulted in impaired mitochondrial and contractile function in cardiomyocytes. Conclusion We demonstrate for the first time that senescent adipose derived LEVs exacerbates myocardial metabolism through up-regulated miRNA-326-3p which inhibits Rictor in cardiomyocytes. Furthermore, reducing senescence burden in adipose tissue is capable of relieving myocardial metabolism disorder in diabetes mellitus.

Human umbilical cord-derived mesenchymal stromal cells improve myocardial fibrosis and restore miRNA-133a expression in diabetic cardiomyopathy

“…Current research con rms that miRNAs play important roles in pathological processes in the cardiovascular system, including arrhythmia, cardiac hypertrophy, cardiac brosis and myocardial infarction [44]. For this reason, miRNAs have emerged as novel targets for DCM treatment and important tools for disease diagnosis, prevention, and treatment [45]. It is known that MSCs release large amounts of extracellular vesicles (EVs) containing various bioactive molecular including a range of miRNAs and may, therefore, be a potent sources of transferable miRNAs in disease settings [46].…”

The role of ncRNAs‐mediated pyroptosis in diabetes and its vascular complications

“…Similarly, gene sequencing analysis of serum samples from type 1 diabetic patients with and without microvascular complications revealed that miR‐518d‐3p and miR‐618 were significantly upregulated among 21 differentially expressed miRNAs in patients with microvascular complications 68 . In addition, miRNA‐326‐3p inhibited the expression of Rictor, leading to myocardial mitochondria damage, systolic dysfunction, and progression of DCM in the STZ‐induced type 1 diabetic mouse model 69 . circRNA sequencing analysis of proliferative vascular membranes in non‐DR and DR subjects revealed nearly 200 dysregulated circRNAs, and KEGG enrichment analysis displayed that the differentially expressed circRNAs were from the inflammatory response‐related pathways 70 .…”

“…68 In addition, miRNA-326-3p inhibited the expression of Rictor, leading to myocardial mitochondria damage, systolic dysfunction, and progression of DCM in the STZ-induced type 1 diabetic mouse model. 69 circRNA sequencing analysis of proliferative vascular membranes in non-DR and DR subjects revealed nearly 200 dysregulated circRNAs, and KEGG enrichment analysis displayed that the differentially expressed circRNAs were from the inflammatory response-related pathways. 70 In conclusion, ncRNAs regulate diabetes and VCD via cell proliferation, apoptosis, autophagy, pyroptosis, and inflammation.…”

Insight into extracellular vesicles in vascular diseases: intercellular communication role and clinical application potential

“…Senescent cells influence exosomal biogenesis and regulate autophagy flux, ultimately driving the progression of vascular diseases [ 122 ]. A study demonstrated that EVs mediated the crosstalk between senescent adipose and cardiomyocytes, and removing senescent adipose alleviated DCM symptoms in streptozotocin type 1 diabetic mice [ 123 ]. Mechanistically, inhibiting Rictor expression in cardiomyocytes is how miR-326-3p from senescent adipocyte-derived EVs leads to mitochondrial dysfunction and cardiac diastolic failure [ 123 ].…”

“…A study demonstrated that EVs mediated the crosstalk between senescent adipose and cardiomyocytes, and removing senescent adipose alleviated DCM symptoms in streptozotocin type 1 diabetic mice [ 123 ]. Mechanistically, inhibiting Rictor expression in cardiomyocytes is how miR-326-3p from senescent adipocyte-derived EVs leads to mitochondrial dysfunction and cardiac diastolic failure [ 123 ]. However, this study does not clarify the specific subtypes of senescent cells, and more accurate targets of interaction might be discovered in future research.…”