Uncovering the Role of Hyperglycemia Induced Long Non-Coding RNAs in Diabetic Vascular Complications

Das, Soumyodeep (2023) Uncovering the Role of Hyperglycemia Induced Long Non-Coding RNAs in Diabetic Vascular Complications. Masters thesis, Indian Institute of Science Education and Research Kolkata.

Text (MS dissertation of Soumyodeep Das (18MS105)) Thesis_18MS105.pdf - Submitted Version Restricted to Repository staff only Download (13MB)

Abstract

Background: Vascular smooth muscle cells (VSMCs) and macrophages undergo phenotype shifts in response to type 2 diabetes, leading to the development of vascular complications. Dysfunction of VSMCs leads to the establishment of disease phenotypes that reduce life expectancy, like Atherosclerosis. In contrast, loss of Macrophage polarisation to anti-inflammatory M2 phenotype leads to impaired Wound Healing resulting in ulceration and severe infection, leading to lower-leg amputation. Increasing evidence suggests that epigenetic regulators such as long non-coding RNAs (lncRNAs) potentially modulate M1-M2 macrophage phenotype switching and various VSMC functions such as phenotypic switching, proliferation, oxidative stress, and inflammation - all linked to accelerated Cardiovascular Diseases(CVDs), delayed wound resolution respectively, and other vascular complications related to Diabetes. It has been shown that high blood glucose, free fatty acids, proinflammatory cytokines, and growth factors could actually dysregulate lncRNAs in inflammatory, cardiac, vascular, and renal cells. This leads to an altered expression of important inflammatory and fibrotic genes that are related with diabetic vascular problems. Hypothesis: i) VSMC-specific lncRNA dysregulation is responsible for Phenotype switching in diabetic mice compared to non-diabetic mice. ii) Splenic Monocyte specific lncRNA dysregulation is responsible for Macrophage phenotype switching in diabetic mice. Methodology: Aorta and Spleen were harvested from dbdb(diabetic) and db/+(Non-diabetic, control) mice to culture primary cells from which RNA was isolated and sequenced. RNA-seq data were analyzed to identify differentially expressed coding and non-coding genes, including lncRNAs. Results: Using different bioinformatic tools, we identified several lncRNAs that are differentially expressed in diabetic conditions and are predicted to be linked to immune regulation and immune response pathways (GO analysis) with enrichment of pro or antiinflammatory transcription factors at their promoter region(Motif Analysis). Conclusion: Preliminary results identified several novel candidate lncRNAs that may regulate macrophage function under diabetic conditions (e.g., MSTRG.1653) or are responsible for VSMC dysfunction in Metabolic memory(e.g., 3110039M20Rik). Further experiments will elucidate these lncRNA targets' function and mechanistic roles in impaired Macrophage polarization and VSMC phenotype switching in Diabetes and associated vascular complications.

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