miR-10a-3p modulates adiposity and suppresses adipose inflammation through TGF-β1/Smad3 signaling pathway

Background Obesity is a multifactorial disease characterized by an enhanced amount of fat and energy storage in adipose tissue (AT). Obesity appears to promote and maintain low-grade chronic inflammation by activating a subset of inflammatory T cells, macrophages, and other immune cells that infiltrate the AT. Maintenance of AT inflammation during obesity involves regulation by microRNAs (miRs), which also regulate the expression of genes implicated in adipocyte differentiation. This study aims to use ex vivo and in vitro approaches to evaluate the role and mechanism of miR-10a-3p in adipose inflammation and adipogenesis. Methods Wild-type BL/6 mice were placed on normal (ND) and high-fat diet (HFD) for 12 weeks and their obesity phenotype, inflammatory genes, and miRs expression were examined in the AT. We also used differentiated 3T3-L1 adipocytes for mechanistic in vitro studies. Results Microarray analysis allowed us to identify an altered set of miRs in the AT immune cells and Ingenuity pathway analysis (IPA) prediction demonstrated that miR-10a-3p expression was downregulated in AT immune cells in the HFD group as compared to ND. A molecular mimic of miR-10a-3p reduced expression of inflammatory M1 macrophages, cytokines, and chemokines, including transforming growth factor-beta 1 (TGF-β1), transcription factor Krüppel-like factor 4 (KLF4), and interleukin 17F (IL-17F) and induced expression of forkhead box P3 (FoxP3) in the immune cells isolated from AT of HFD-fed mice as compared to ND. In differentiated 3T3-L1 adipocytes, the miR-10a-3p mimics also reduced expression of proinflammatory genes and lipid accumulation, which plays a role in the dysregulation of AT function. In these cells, overexpression of miR-10a-3p reduced the expression of TGF-β1, Smad3, CHOP-10, and fatty acid synthase (FASN), relative to the control scramble miRs. Conclusion Our findings suggest that miR-10a-3p mimic mediates the TGF-β1/Smad3 signaling to improve metabolic markers and adipose inflammation. This study provides a new opportunity for the development of miR-10a-3p as a novel therapeutic for adipose inflammation, and its associated metabolic disorders. Graphical Abstract

[1]  F. Park,et al.  Piceatannol induces regulatory T cells and modulates the inflammatory response and adipogenesis , 2023, Biomedicine & pharmacotherapy = Biomedecine & pharmacotherapie.

[2]  S. Kiran,et al.  MicroRNAs: a crossroad that connects obesity to immunity and aging , 2022, Immunity & ageing : I & A.

[3]  Kepeng Wang,et al.  Interleukin-17 Family Cytokines in Metabolic Disorders and Cancer , 2022, Genes.

[4]  J. Hong,et al.  Integrative understanding of immune-metabolic interaction , 2022, BMB reports.

[5]  Jian-wei Tian,et al.  MicroRNA-10a/b inhibit TGF-β/Smad-induced renal fibrosis by targeting TGF-β receptor 1 in diabetic kidney disease , 2022, Molecular therapy. Nucleic acids.

[6]  S. Kiran,et al.  High-Fat Diet-Induced Dysregulation of Immune Cells Correlates with Macrophage Phenotypes and Chronic Inflammation in Adipose Tissue , 2022, Cells.

[7]  A. Piwowar,et al.  Molecular Mechanism of Lipotoxicity as an Interesting Aspect in the Development of Pathological States—Current View of Knowledge , 2022, Cells.

[8]  P. Hsieh,et al.  The Chemokine Systems at the Crossroads of Inflammation and Energy Metabolism in the Development of Obesity , 2021, International journal of molecular sciences.

[9]  G. Hotamisligil,et al.  Metabolic Messengers: tumour necrosis factor , 2021, Nature Metabolism.

[10]  Mohammad Shoaib Prince,et al.  Role of Inflammatory Cytokines, Growth Factors and Adipokines in Adipogenesis and Insulin Resistance , 2021, Inflammation.

[11]  L. Rochette,et al.  The emerging role of miRNA-132/212 cluster in neurologic and cardiovascular diseases: Neuroprotective role in cells with prolonged longevity , 2021, Mechanisms of Ageing and Development.

[12]  Y. Cong,et al.  MicroRNA-10a Negatively Regulates CD4+ T Cell IL-10 Production through Suppression of Blimp1 , 2021, The Journal of Immunology.

[13]  R. El Bekay,et al.  miR-21 mimic blocks obesity in mice: A novel therapeutic option , 2021, Molecular therapy. Nucleic acids.

[14]  S. Kiran,et al.  High Fat Diet-Induced CD8+ T Cells in Adipose Tissue Mediate Macrophages to Sustain Low-Grade Chronic Inflammation , 2021, Frontiers in Immunology.

[15]  Guoqing Yang,et al.  STAT3 phosphorylation in central leptin resistance , 2021, Nutrition & Metabolism.

[16]  Youhua Liu,et al.  MicroRNA-10 negatively regulates inflammation in diabetic kidney via targeting activation of NLRP3 inflammasome. , 2021, Molecular therapy : the journal of the American Society of Gene Therapy.

[17]  E. Taylor.  The complex role of adipokines in obesity, inflammation, and autoimmunity , 2021, Clinical science.

[18]  Nancie J. MacIver,et al.  The Role of the Adipokine Leptin in Immune Cell Function in Health and Disease , 2021, Frontiers in Immunology.

[19]  S. Ramakrishnan,et al.  Tregs facilitate obesity and insulin resistance via a Blimp-1/IL-10 axis , 2020, JCI insight.

[20]  Gongshe Yang,et al.  Elevated miR-10a-5p facilitates cell cycle and restrains adipogenic differentiation via targeting Map2k6 and Fasn, respectively. , 2020, Acta biochimica et biophysica Sinica.

[21]  Ashish Ranjan Sharma,et al.  Therapeutic advances of miRNAs: A preclinical and clinical update , 2020, Journal of advanced research.

[22]  L. Kane,et al.  Noncanonical STAT3 activity sustains pathogenic Th17 proliferation and cytokine response to antigen , 2020, The Journal of experimental medicine.

[23]  Xiaohui Guo,et al.  Aberrant expression of miR-214 is associated with obesity-induced insulin resistance as a biomarker and therapeutic , 2020, Diagnostic Pathology.

[24]  F. Beguinot,et al.  Chronic Adipose Tissue Inflammation Linking Obesity to Insulin Resistance and Type 2 Diabetes , 2020, Frontiers in Physiology.

[25]  J. Granneman,et al.  MicroRNA-10a-5p regulates macrophage polarization and promotes therapeutic adipose tissue remodeling , 2019, Molecular metabolism.

[26]  J. Tuomilehto,et al.  Reduced miR-181d level in obesity and its role in lipid metabolism via regulation of ANGPTL3 , 2019, Scientific Reports.

[27]  J. D. Cortés-García,et al.  Increased levels of adipose tissue-resident Th17 cells in obesity associated with miR-326. , 2019, Immunology letters.

[28]  B. Liu,et al.  A simplified system for the effective expression and delivery of functional mature microRNAs in mammalian cells , 2019, Cancer Gene Therapy.

[29]  A. Roy,et al.  Association of obesity and type 2 diabetes , 2019 .

[30]  D. Kabelitz,et al.  An update on immune dysregulation in obesity‐related insulin resistance , 2019, Scandinavian journal of immunology.

[31]  L. Niu,et al.  miR-10b-5p regulates 3T3-L1 cells differentiation by targeting Apol6. , 2019, Gene.

[32]  A. Xu,et al.  Adipocyte-secreted exosomal microRNA-34a inhibits M2 macrophage polarization to promote obesity-induced adipose inflammation , 2019, The Journal of clinical investigation.

[33]  L. Niu,et al.  miR-144-3p Promotes Adipogenesis Through Releasing C/EBPα From Klf3 and CtBP2 , 2018, Front. Genet..

[34]  S. Gogg,et al.  Impaired Adipogenesis and Dysfunctional Adipose Tissue in Human Hypertrophic Obesity. , 2018, Physiological reviews.

[35]  M. González-Gay,et al.  Obesity, Fat Mass and Immune System: Role for Leptin , 2018, Front. Physiol..

[36]  Jiao Li,et al.  miR-10a rejuvenates aged human mesenchymal stem cells and improves heart function after myocardial infarction through KLF4 , 2018, Stem Cell Research & Therapy.

[37]  S. Fruh.  Obesity: Risk factors, complications, and strategies for sustainable long‐term weight management , 2017, Journal of the American Association of Nurse Practitioners.

[38]  A. Siani,et al.  Role of microRNAs in obesity and obesity-related diseases , 2017, Genes & Nutrition.

[39]  Wai-Yee Chan,et al.  Conserved miR-10 family represses proliferation and induces apoptosis in ovarian granulosa cells , 2017, Scientific Reports.

[40]  A. Hata,et al.  TGF-β Signaling from Receptors to Smads. , 2016, Cold Spring Harbor perspectives in biology.

[41]  Zhiqiu Hu,et al.  MiR-10a improves hepatic fibrosis by regulating the TGFβl/Smads signal transduction pathway , 2016, Experimental and therapeutic medicine.

[42]  Vikas Gupta,et al.  MicroRNA therapeutics: Discovering novel targets and developing specific therapy , 2016, Perspectives in clinical research.

[43]  Y. Abed,et al.  Obesity and inflammation: the linking mechanism and the complications , 2016, Archives of medical science : AMS.

[44]  Dawei Sun,et al.  FOSL2 Positively Regulates TGF-β1 Signalling in Non-Small Cell Lung Cancer , 2014, PloS one.

[45]  I. Pinchuk,et al.  miR-10a inhibits dendritic cell activation and Th1/Th17 cell immune responses in IBD , 2014, Gut.

[46]  R. Kelishadi,et al.  Insulin and leptin levels in overweight and normal-weight Iranian adolescents: The CASPIAN-III study , 2014, Journal of research in medical sciences : the official journal of Isfahan University of Medical Sciences.

[47]  P. Sethupathy,et al.  miR-182 and miR-10a Are Key Regulators of Treg Specialisation and Stability during Schistosome and Leishmania-associated Inflammation , 2013, PLoS pathogens.

[48]  Chia‐cheng Chang,et al.  Leptin differentially regulate STAT3 activation in ob/ob mouse adipose mesenchymal stem cells , 2012, Nutrition & Metabolism.

[49]  J. Gugenheim,et al.  Identification of Adipose Tissue Dendritic Cells Correlated With Obesity-Associated Insulin-Resistance and Inducing Th17 Responses in Mice and Patients , 2012, Diabetes.

[50]  J. Bluestone,et al.  MicroRNA 10a Marks Regulatory T Cells , 2012, PloS one.

[51]  Apostolos Zaravinos,et al.  Differential Expression of MicroRNAs in Adipose Tissue after Long-Term High-Fat Diet-Induced Obesity in Mice , 2012, PloS one.

[52]  N. Tan,et al.  Getting ‘Smad' about obesity and diabetes , 2012, Nutrition & Diabetes.

[53]  E. Wagner,et al.  FOSL2 promotes leptin gene expression in human and mouse adipocytes. , 2012, The Journal of clinical investigation.

[54]  P. Sun,et al.  Protection from obesity and diabetes by blockade of TGF-β/Smad3 signaling. , 2011, Cell metabolism.

[55]  K. Clément,et al.  Krüppel-like factor 4 regulates macrophage polarization. , 2011, The Journal of clinical investigation.

[56]  C. Deltas,et al.  microRNAs: a newly described class of encoded molecules that play a role in health and disease. , 2010, Hippokratia.

[57]  M. Civelek,et al.  MicroRNA-10a regulation of proinflammatory phenotype in athero-susceptible endothelium in vivo and in vitro , 2010, Proceedings of the National Academy of Sciences.

[58]  K. Flegal,et al.  Prevalence and trends in obesity among US adults, 1999-2008. , 2010, JAMA.

[59]  Zhihua Liu,et al.  MicroRNA-10b Promotes Migration and Invasion through KLF4 in Human Esophageal Cancer Cell Lines* , 2010, The Journal of Biological Chemistry.

[60]  S. Gogg,et al.  Inflammation and impaired adipogenesis in hypertrophic obesity in man. , 2009, American Journal of Physiology. Endocrinology and Metabolism.

[61]  N. Rothwell,et al.  Leptin induces interleukin‐1β release from rat microglial cells through a caspase 1 independent mechanism , 2007, Journal of neurochemistry.

[62]  Chih-Hsin Tang,et al.  Leptin-Induced IL-6 Production Is Mediated by Leptin Receptor, Insulin Receptor Substrate-1, Phosphatidylinositol 3-Kinase, Akt, NF-κB, and p300 Pathway in Microglia1 , 2007, The Journal of Immunology.

[63]  M. Fasshauer,et al.  Fatty acid synthase gene expression in human adipose tissue: association with obesity and type 2 diabetes , 2007, Diabetologia.

[64]  A. Tuzcu,et al.  The correlation between adiposity and adiponectin, tumor necrosis factor α, interleukin-6 and high sensitivity C-reactive protein levels. Is adipocyte size associated with inflammation in adults? , 2007, Journal of endocrinological investigation.

[65]  C. Herder,et al.  Relationship between adipocyte size and adipokine expression and secretion. , 2007, The Journal of clinical endocrinology and metabolism.

[66]  Arya M. Sharma,et al.  Peroxisome Proliferator-Activated Receptor γ and Adipose Tissue—Understanding Obesity-Related Changes in Regulation of Lipid and Glucose Metabolism , 2007 .

[67]  U. Smith,et al.  Cytokines Promote Wnt Signaling and Inflammation and Impair the Normal Differentiation and Lipid Accumulation in 3T3-L1 Preadipocytes* , 2006, Journal of Biological Chemistry.

[68]  V. Giusti,et al.  Adipose tissue is a regulated source of interleukin-10. , 2005, Cytokine.

[69]  R. Vettor,et al.  Resistin and adiponectin expression in visceral fat of obese rats: effect of weight loss. , 2002, Obesity research.

[70]  M. Lane,et al.  Role of C/EBP homologous protein (CHOP-10) in the programmed activation of CCAAT/enhancer-binding protein-beta during adipogenesis. , 2000, Proceedings of the National Academy of Sciences of the United States of America.

[71]  R. Derynck,et al.  Roles of Autocrine TGF-β Receptor and Smad Signaling in Adipocyte Differentiation , 2000, The Journal of cell biology.

[72]  B. Spiegelman,et al.  Cross-Regulation of C/EBPα and PPARγ Controls the Transcriptional Pathway of Adipogenesis and Insulin Sensitivity , 1999 .

[73]  Z. Wu,et al.  Induction of peroxisome proliferator-activated receptor gamma during the conversion of 3T3 fibroblasts into adipocytes is mediated by C/EBPbeta, C/EBPdelta, and glucocorticoids , 1996, Molecular and cellular biology.

[74]  B. Spiegelman,et al.  Tumor Necrosis Factor α: A Key Component of the Obesity-Diabetes Link , 1994, Diabetes.

[75]  W. Hsueh,et al.  Adipose Tissue T Regulatory Cells: Implications for Health and Disease. , 2021, Advances in experimental medicine and biology.

[76]  U. Senarath,et al.  Worldwide epidemic of obesity , 2020, Obesity and Obstetrics.

[77]  F. Hu,et al.  Obesity , 2017, Nature Reviews Disease Primers.

[78]  Allison,et al.  Obesity and Obstetrics , 2010 .

[79]  B. Spiegelman,et al.  C/EBPalpha induces adipogenesis through PPARgamma: a unified pathway. , 2002, Genes & development.

[80]  D. de Silva.  Tumour necrosis factor , 1990, The Ceylon medical journal.