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Principal Investigator: Basak Icli
Organization: TUFTS MEDICAL CENTER
Fiscal Year: 2024
Award: $598,681
Funding agency: National Heart Lung and Blood Institute
PROJECT SUMMARY/ABSTRACT
White and brown adipose tissues are highly vascularized organs, capable of plasticity based on
metabolic demands and energy expenditure. However maladaptive regulation of these tissues can lead to
insulin resistance. Critical gaps remain in our understanding of how angiogenesis impacts adipose tissue
dysfunction and overall metabolism. MicroRNAs (miRs) are implicated in the regulation of the angiogenic
response to pathophysiological stimuli. The role of miRs in regulating the angiogenic response in diet-induced
insulin resistance is poorly understood.
Using a miRNA-Seq approach, we identified that miR-409-3p expression was significantly increased in
endothelial cells (ECs) of brown adipose tissue (BAT) of diet-induced obese (DIO) mice and in human diabetic
plasma samples compared to non-diabetic patients. Overexpression of miR-409-3p markedly inhibited EC
growth and migration, whereas miR-409-3p inhibition had the opposite effects. Preliminary studies indicate that
miR-409-3p targets the 3’UTRs of Zinc Finger E-box binding Homeobox 1 (ZEB1) and Mitogen-activated
protein kinase kinase kinase kinase 3 (MAP4K3). Overexpression of miR-409-3p decreased ZEB1 and
MAP4K3 expression in ECs, whereas inhibition had the opposite effect. SiRNA knockdown of ZEB1 or
MAP4K3 expression in ECs phenocopied the effects of miR-409-3p overexpression and significantly
decreased EC proliferation and migration. 3T3-L1 cells or human skin fat organoids co-cultured with
supernatant harvested from ECs overexpressing miR-409-3p had decreased expression of brown adipocyte
markers (UCP1, Cidea) by RT-qPCR and Western blot analyses, whereas supernatant harvested from ECs
deficient in miR-409-3p increased expression of brown adipocyte markers. Systemic intravenous delivery of
LNA-anti-miR-409-3p inhibitor to DIO mice significantly increased angiogenesis by CD31 staining,
accompanied by higher UCP-1 in BAT and sWAT by RT-qPCR, Western blot, and immunohistochemistry
analyses, while improving glucose and insulin tolerance and overall metabolism. Therefore, we hypothesize
that miR-409-3p serves as a critical regulator of EC growth and angiogenesis in adipose tissue and may
improve metabolic dysfunction in DIO. To explore this, we first propose in Aim1 to investigate the molecular
mechanisms by which miR-409-3p regulates EC growth and angiogenesis. In Aim2, we will delineate the
mechanisms by which miR-409-3p in ECs regulates browning in adipose tissues. Finally, in Aim3, we will
explore the effect of miR-409-3p neutralization in the vasculature of adipose tissues and development of DIO
and insulin resistance in mice. Successful completion of these studies will shed insights on the regulatory role
of miR-409-3p between impaired angiogenesis in diet-induced obesity and adipose tissue dysfunction, an
effect that could be exploited for therapeutic intervention in obesity-induced insulin resistance.
Terms: <3' Untranslated Regions><3'UTR><3T3-L1 Cells><Ablation><Abscission><Acceleration><Adipocytes><Adipose Cell><Adipose tissue><Adult-Onset Diabetes Mellitus><Age><Angiogenesis Antagonists><Angiogenesis Blockers><Angiogenesis Inhibition><Angiogenesis Inhibitors><Angiogenetic Antagonists><Angiogenetic Inhibitors><Angiogenic Antagonists><Angiogenic Inhibition><Angiogenic Inhibitors><Angiostatic Agents><Animal Model><Animal Models and Related Studies><Anti-Angiogenetic Agents><Anti-Angiogenic Agents><Anti-Angiogenic Drugs><Antiangiogenesis Agents><Antiangiogenic Agents><Antiangiogenic Drugs><Assay><Binding><Bioassay><Biological Assay><Blood Plasma><Body Tissues><Brown Adipose Tissue><Brown Fat><CD31><Cell Communication and Signaling><Cell Growth in Number><Cell Locomotion><Cell Migration><Cell Movement><Cell Multiplication><Cell Proliferation><Cell Signaling><Cellular Expansion><Cellular Growth><Cellular Migration><Cellular Motility><Cellular Proliferation><Co-culture><Cocultivation><Coculture><Coculture Techniques><Code><Coding System><Development><Diet><Dysfunction><Endocrine Gland Secretion><Endothelial Cells><Energy Expenditure><Energy Metabolism><Excision><Extirpation><Fat Cells><Fats><Fatty Tissue><Fatty acid glycerol esters><Functional RNA><Functional disorder><Gene Expression><Generalized Growth><Genes><Growth><Growth Agents><Growth Factor><Growth Substances><Harvest><Heat Production><Hibernating Gland><High Fat Diet><Homeo Boxes><Homeobox><Hormones><Human><Immunoblotting><Immunohistochemistry><Immunohistochemistry Cell/Tissue><Immunohistochemistry Staining Method><Impairment><Insulin Resistance><Intermediary Metabolism><Intracellular Communication and Signaling><Intravenous><Ketosis-Resistant Diabetes Mellitus><Kinases><Length><Lipocytes><MAP ERK Kinase Kinase 1><MAP Kinase Kinase Kinase 1><MAP3K1><MAP3K1 Protein><MAP3K1 gene><MAPK/ERK Kinase Kinase 1><MAPKKK1><MEK Kinase><MEK Kinase 1><MEKK-1 Protein Kinase><MEKK1><MEKK1 Protein><MEKK1 Protein Kinase><Mature Lipocyte><Mature fat cell><Maturity-Onset Diabetes Mellitus><Measures><Metabolic><Metabolic Diseases><Metabolic Disorder><Metabolic Processes><Metabolic dysfunction><Metabolism><Mice><Mice Mammals><Micro RNA><MicroRNA Expression Profiling><MicroRNAs><Mitogen-Activated Kinase Kinase Kinase 1><Mitogen-Activated Protein Kinase Kinase Kinase 1><Modern Man><Molecular><Molecular Interaction><Murine><Mus><NIDDM><Neovascularization Inhibitors><Non-Coding><Non-Coding RNA><Non-Insulin Dependent Diabetes><Non-Insulin-Dependent Diabetes Mellitus><Non-translated RNA><Noncoding RNA><Noninsulin Dependent Diabetes><Noninsulin Dependent Diabetes Mellitus><Nontranslated RNA><Nucleotides><Nutrient><Obese Mice><Obesity><Organ><Organoids><PECAM1><PECAM1 gene><Patients><Phenocopy><Phosphotransferase Gene><Phosphotransferases><Physiopathology><Plasma><Plasma Serum><Play><Proteins><Proteins Growth Factors><RNA Seq><RNA sequencing><RNAseq><Regulation><Removal><Reticuloendothelial System, Serum, Plasma><Role><Sampling><Short interfering RNA><Signal Pathway><Signal Transduction><Signal Transduction Systems><Signaling><Skin><Slow-Onset Diabetes Mellitus><Small Interfering RNA><Stable Diabetes Mellitus><Staining method><Stains><Stimulus><Surgical Removal><T2 DM><T2D><T2DM><Testing><Therapeutic Hormone><Therapeutic Intervention><Thermogenesis><Thesaurismosis><Tissue Growth><Tissues><Transcript><Transphosphorylases><Type 2 Diabetes Mellitus><Type 2 diabetes><Type II Diabetes Mellitus><Type II diabetes><Untranslated RNA><VEGF><VEGFs><Vascular Endothelial Growth Factors><Vascularization><Waste Products><Western Blotting><Western Immunoblotting><Zinc Finger Domain><Zinc Finger Motifs><Zinc Fingers><Zinc decreased><Zinc deficiency><Zinc low><Zn deficiency><Zn levels low><Zn++ low><adipose><adiposity><adult onset diabetes><ages><angiogenesis><antiangiogenic><biological signal transduction><cell growth><cell motility><corpulence><developmental><diabetic><diet-associated obesity><diet-induced obesity><diet-related obesity><diets><disease risk><disorder risk><endothelial dysfunction><global miRNA profiling><glucose tolerance><improved><in vivo><inhibitor><insight><insulin resistant><insulin sensitivity><insulin tolerance><intervention therapy><ketosis resistant diabetes><knock-down><knockdown><locked nucleic acid><low Zinc level><maturity onset diabetes><metabolism disorder><miRNA><miRNA expression profiling><miRNA sequencing><miRNA-seq><miRNAs><micro RNA expression profiling><microRNA profiling><microRNA sequencing><model of animal><non-diabetic><noncoding><nondiabetic><ob/ob mouse><obesity development><ontogeny><overexpress><overexpression><pathophysiology><programs><protein blotting><resection><response><siRNA><social role><subcutaneous><subdermal><transcriptome sequencing><transcriptomic sequencing><type 2 DM><type II DM><type two diabetes><white adipose tissue><yellow adipose tissue>