FAPESP Week /21/2017

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1 Texas Tech University Dr Naima Moustaid-Moussa Texas Tech University FAPESP Week /21/2017 University of Sao Paulo Dr Sonia Jancar ICB-USP Theresa Ramalho MS PhD candidate, Immunology ICB-USP Dr Latha Ramalingam Texas Tech University Dr Luciano Filgueiras ICB-USP Anti-Inflammatory Effects of Omega 3 Fatty Acids in Metabolic Disorders: Role of Lipid Mediators and MicroRNAs

2 FAPESP Week /21/2017 Presenters Texas Tech University University of Sao Paulo Dr Naima Moustaid-Moussa Texas Tech University Theresa Ramalho MS PhD candidate, Immunology ICB-USP

3 Metabolic Disorders Immune System Metabolism Sterile Inflammation o Atherosclerosis o Obesity o Metabolic syndrome o Diabetes

4 Diabetes Incidence

5 Diabetes Physiology IFNγ Hyperglycemia o Insulin resistance o Steatosis Lipid Mediators Sterile TNFα Co-morbidities o Retinopathy o Neuropathy o Defective wound healing IL-1β Inflammation IL-1α IL = Interleukin; TNF: Tumor Necrosis Factor; IFN: Interferon

6 Lipid Mediators (PAF) Metabolic Syndrome (PAFR KO) Cancer Platelet Activating Fator (PAF) LPAT Arachdonic Acid Membrane Phospholipid Carcinoma (mouth and uterus) Melanoma Type 1 Diabetes Streptozotocin model LTB4 5-LO. Leukotrienes COX TXAS.. Prostaglandins Thromboxanes

7 PAFR and Metabolic Homeostasis

8 PAFR and Metabolic Homeostasis Macrophage Inflammatory macrophage infiltration in adipose tissue in PAFR KO mice M1 PAF receptor Anti-inflammatory phenotype Steatosis in PAFR KO Insulin Resistance in PAFR KO mice Filgueiras LR et al., Clinical Science, 2016 Ishizuka Eket al, Sci Rep, 2016

9 PAFR and Metabolic Homeostasis Macrophage MyD88 NFkB Nucleus IL-12 IL-6 - Metabolic Syndrome Filgueiras LR et al., Clinical Science, 2016 Ishizuka Eket al, Sci Rep, 2016

10 LTB4 in Type 1 Diabetes

11 Lipid Mediators (LTB4) BLT1 Platelet Activating Fator (PAF) LPAT Arachdonic Acid 5-LO Membrane Phospholipid Type 1 Diabetes Streptozotocin model LTB4. Leukotrienes COX TXAS.. Prostaglandins Thromboxanes

12 LTB4 in Type 1 Diabetes LTB4 promotes sterile inflammation Macrophage BLT1 receptor Pro-inflammatory phenotype Filgueiras LR et al., Sci Sign, 2015 Ramalho TR et al, Unpublished, 2017

13 LTB4 in Type 1 Diabetes LTB 4 Macrophage BLT1 G αi MyD88 NFkB Nucleus Epigenetic effects IL-6 IL-1β TNFγ Susceptibility to sepsis Filgueiras LR et al., Sci Sign, 2015 Filgueiras LR et al., J Diabetes Comp, 2016 Ramalho TR et al, Unpublished, 2017

14 Progress & Accomplishments 1. TTU and USP team met in Summer 2016 at the International Immunology meeting in Australia 2. Jancar's Postdoc Luciano attended the 2016 FAPESP meeting at TTU, presented the SPRINT project and visited Moustaid-Moussa s lab 3. TTU Team visited USP in March 2017 and Moustaid-Moussa gave an invited lecture in the dept. of Immunology, USP 4. Visiting scholar (Theresa) at TTU for one year (funded by CAPES, Brazil) 5. Invited presentation by AFMR, April 2017, and invited peer-reviewed manuscript in press 6. Poster presentations at the Immuno-metabolism and Chronic Disease conference, Fiji, Aug 2017

15 Obesity, a global & complex disease

16 Obesity Complex disease 1/3 of American adults Medical costs Per capita medical spending $2,741 higher Increases risk for metabolic diseases Health Disparities Linked to changes in metabolism/inflammation (sterile inflammation) 17.9% Obesity 52.5% Overweight Ministry of Health Brazil cdc.gov 16

17

18 Obesity and the Metabolic Syndrome Hyperglycemia Abdominal Obesity Type 2 Diabetes Reduced HDL CVD Elevated Triglycerides Elevated Blood Pressure Prevalence: 35-40% (AHA/NHLBI, IDF, 2005) Alberti et al., Circulation, 2009

19 Obesity and inflammation LEAN OBESE Diet (ω 3-PUFAs), genetics, pharmacology IL-10 TNFa IL-6 Ang II Adiponectin PAI-1 Stem cells/ preadipocytes Adipocyte Blood Vessel T n 1 T n 2 IFNY MCP-1 Resistin M1 macrophage M2 macrophage T reg Effector T cell Adapted from: Kalupahana and Moustaid-Moussa. Obesity Reviews, 2011

20 Pathogenesis of obesity-associated insulin resistance Liver Adipose Tissue Inflammation Glycogenolysis Gluconeogenesis Cytokines Lipolysis Glucose Pancreas Insulin Free fatty acids Ectopic fat Insulin resistance Intestine Skeletal muscle Kalupahana, Moustaid-Moussa and Claycombe. Molecular Aspects of Medicine 2012

21 Dietary/genetic regulation of adipose tissue inflammation Food and Plant Bioactives (Omega 3 s, tart cherry anthocyanins, tocotrienols, Switchgrass extracts) Cells, animals, model organisms, bariatric patients Endocrine systems (Renin Angiotensin System) Small Adipocytes Obesity Large inflamed Adipocytes Metabolic Syndrome Breast Cancer Mechanisms linking adipocyte inflammation to metabolic disorders

22 Omega 3 fatty acids prevent dietinduced obesity and inflammation

23 Lipid Mediators Metabolic Syndrome (PAFR KO) Platelet Activating Fator (PAF) LPAT Arachdonic Acid 5-LO Membrane Phospholipid Omega 3 fatty acids (EPA) BLT1? LTB4. Leukotrienes COX TXAS T1 Diabetes.. Prostaglandins Thromboxanes Anti-inflammatory mediators Omega 3 fatty acids: hypotriglyceridemic, cardioprotective and anti-inflammatory

24 Research Hypothesis High fat diet OBESITY/T2 Diabetes INFLAMMATION Lipogenesis Adipogenesis Adipocyte Hypertrophy Lipid mediators? MCP-1 TNFα IL-6 MACROPHAGE RECRUITMENT T1 Diabetes 24

25 Study design Low-fat diet (10% fat, 70% CHO, 20%proteins) Male C57BL/6J mice LF HF HF-EPA-P HF-EPA-R High-fat diet (45% fat, 35% CHO, 20%proteins) 11 Weeks EPA diet (HF with 36g/kg EPA) Tissue collection and analyses High-fat diet EPA diet 6 weeks 5 weeks Metabolic Changes Gene Expression Inflammation Cell Culture Experiments Microarray qrt-pcr Macrophage infiltration Adipokines secretion

26 26 EPA reduces body weight and restores glucose tolerance 600 Body Weight (g) LF HF EPA-R HF-EPA-P HF-R c Age (weeks) Kalupahana NS et al. J Nutr 2010, 140: a b Blood glucose (mg/dl) Plasma insulin (ng/ml) LF HF-EPA-P HF HF-EPA-R Time (minutes) a a LF HF-EPA-P HF HF-EPA-R b a

27 EPA Lowers TG Deposition in Liver 44% 70% 11% 44% Kalupahana et al. J Nutr

28 EPA reduces adipose macrophages independent of adiposity LF HF-EPA-P Macrophage Area (μm 2 ) x* a a b a HF HF-EPA-R 0 LF HF-EPA-P HF HF-EPA-R Mechanisms mediating these anti-inflammatory effects? LeMieux, M.J. et al. J Nutr Labeled means without a common letter differ, P <0.05

29 29 Summary of Results Omega 3 fatty acids target specific mirnas and genes related to inflammation and lipid mediators (unpublished) We thank members of Prof. Jancar and Prof Moustaid-Moussa s lab for their contributions to this SPRINT project

30 30 Future studies Validation of gene and mirna findings Effects of EPA on LTB4 in adipose tissue Inhibition of LTB4/BLT1 and PAFR in wild type, obese and diabetic animals to determine effects on: Inflammation Target genes and mirnas Research Manuscript Funding after research publication: NIH- FAPESP and other funding agencies

31 Nutritional Sciences

32 Prof Sonia Jancar Laboratory of Immunopharmacology Department of Immunology Institute of Biomedical Sciences University of Sao Paulo Sonia Jancar

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