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A Role for TNMD in Adipocyte Differentiation and Adipose Tissue Function: a Dissertation

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A Role for TNMD in Adipocyte Differentiation and Adipose Tissue Function: a Dissertation University of Massachusetts Medical School eScholarship@UMMS GSBS Dissertations and Theses Graduate School of Biomedical Sciences 2016-06-30 A Role for TNMD in Adipocyte Differentiation and Adipose Tissue Function: A Dissertation Ozlem Senol-Cosar University of Massachusetts Medical School Let us know how access to this document benefits ou.y Follow this and additional works at: https://escholarship.umassmed.edu/gsbs_diss Part of the Cell Biology Commons, Cellular and Molecular Physiology Commons, and the Developmental Biology Commons Repository Citation Senol-Cosar O. (2016). A Role for TNMD in Adipocyte Differentiation and Adipose Tissue Function: A Dissertation. GSBS Dissertations and Theses. https://doi.org/10.13028/M2PC7S. Retrieved from https://escholarship.umassmed.edu/gsbs_diss/837 This material is brought to you by eScholarship@UMMS. It has been accepted for inclusion in GSBS Dissertations and Theses by an authorized administrator of eScholarship@UMMS. For more information, please contact [email protected]. A ROLE FOR TNMD IN ADIPOCYTE DIFFERENTIATION AND ADIPOSE TISSUE FUNCTION A Dissertation Presented By OZLEM SENOL-COSAR Submitted to the Faculty of the University of Massachusetts Graduate School of Biomedical Sciences, Worcester In partial fulfillment of the requirements for the degree of DOCTOR OF PHILOSOPHY JUNE 30, 2016 Interdisciplinary Graduate Program A ROLE FOR TNMD IN ADIPOCYTE DIFFERENTIATION AND ADIPOSE TISSUE FUNCTION A Dissertation Presented By OZLEM SENOL-COSAR This work was undertaken in the Graduate School of Biomedical Sciences Interdisciplinary Graduate Program The signature of the Thesis Advisor signifies validation of Dissertation content __________________________________ Michael P. Czech, Ph.D., Thesis Advisor The signatures of the Dissertation Defense Committee signify completion and approval as to style and content of the Dissertation __________________________________ David A. Guertin, Ph.D., Member of Committee __________________________________ Yong -Xu Wang, Ph.D., Member of Committee ________________________________ Anthony N. Imbalzano, Ph.D., Member of Committee ___________________________________ Evan D. Rosen, Ph.D., Member of Committee The signature of the Chair of the Committee signifies that the written dissertation meets the requirements of the Dissertation Committee ___________________________________ Dale L. Greiner, Ph.D. Chair of Committee The signature of the Dean of the Graduate School of Biomedical Sciences signifies that the student has met all graduation requirements of the school. ___________________________________ Anthony Carruthers, Ph.D. Dean of the Graduate School of Biomedical Sciences June 30, 2016 ii Dedication I would like to dedicate this thesis to Dr. Guldal Kirkali, my beloved advisor for my Master’s thesis. I thank her always for teaching me innumerable lessons, especially how important being influential in other people’s life. I thank her for the countless hours of conversations, fun times and valuable mentorship. Meeting her has been one of the luckiest things that ever happened to me. iii Acknowledgements I thank everyone who has supported me throughout my educational life. First and foremost, I would like to thank my mentor Dr. Michael Czech whose endless encouragement and guidance helped me in every step of my graduate work. I thank him for being a great inspiration in my life, for allowing me to pursue my scientific interests in the lab and for letting me to grow as an independent scientist. I feel so lucky to be part of the Czech lab where I have known many amazing people. I would like to thank all the past and present members. I would particularly like to thank Dr. Rachel Roth Flach for her support, advice, ever- lasting encouragement, guidance and friendship. Her innumerable hours spent helping me were critical for my success. I cannot thank Marina DiStefano enough for always being there for me to share ideas, help me with my experiments, listen, understand and provide sound advice. I would like to thank Dr. Joseph Virbasius for valuable discussions, suggestions, edits and very fun conversations. I would like to thank Laura Danai for her contagious positive energy. I will always miss our fun times. I also want thank Drs. Adilson Guilherme and David Pedersen for being great office mates and scientific resources. I sincerely thank to my TRAC committee for their insightful suggestions. I would especially like to thank my committee chair Dr. Dale Greiner for his patience and support. I would like to thank Drs. Yong-Xu Wang, David Guertin, Marcus Cooper, Stephen Jones and Amy Walker for attending my TRAC meetings and providing me thoughtful suggestions and interesting directions for iv my research. I would also like to Drs. Anthony Imbalzano and Evan Rosen for being in my dissertation committee and putting in time to evaluate my thesis. Special thanks go to my friends outside of the lab who made Worcester a very fun place to live. I would like to thank Ebru for being a great friend over the last six years, always making time for me and for brightening my day when I most needed it. I also thank Sezin, Aysegul, Arda and Alper for all the fun times, long lunches and for listening to my long political comments. I would like to express my sincere appreciation and gratitude to my parents, my mother Sengul and my father Abbas for their unwavering support, motivation and faith that they have shown throughout my education. I am the luckiest person for being their daughter and I can only hope that, one day, I will be as dedicated and caring for my own children. I would like to thank my sister, Cigdem for her unconditional love, support and for being a great example for me. Her pride in my accomplishments has always kept me going, particularly during tough moments in graduate school. Most importantly, I would like to thank my loving husband Ozgur who has been extremely supportive, patient and encouraging ever since we met. Thank you for all those hours that you have spent studying with me in libraries, waiting in front of labs for me to finish an experiment, listening to me and being always with me. Last but not least, I would like to thank my daughter Ada for being such a sweet and easy baby when her mommy was writing her thesis. I hope the time that I spent writing instead of cuddling you will pay off. Mommy loves you so much! v Abstract Adipose tissue is one of the most dynamic tissues in the body and is vital for metabolic homeostasis. In the case of excess nutrient uptake, adipose tissue expands to store excess energy in the form of lipids, and in the case of reduced nutrient intake, adipose tissue can shrink and release this energy. Adipocytes are most functional when the balance between these two processes is intact. To understand the molecular mechanisms that drive insulin resistance or conversely preserve the metabolically healthy state in obese individuals, our laboratory performed a screen for differentially regulated adipocyte genes in insulin resistant versus insulin sensitive subjects who had been matched for BMI. From this screen, we identified the type II transmembrane protein tenomodulin (TNMD), which had been previously implicated in glucose tolerance in gene association studies. TNMD was upregulated in omental fat samples isolated from the insulin resistant patient group compared to insulin sensitive individuals. TNMD was predominantly expressed in primary adipocytes compared to the stromal vascular fraction from this adipose tissue. Furthermore, TNMD expression was greatly increased in human preadipocytes by differentiation, and silencing TNMD blocked adipogenic gene induction and adipogenesis, suggesting its role in adipose tissue expansion. Upon high fat diet feeding, transgenic mice overexpressing Tnmd specifically in adipose tissue developed increased epididymal adipose tissue (eWAT) mass without a difference in mean cell size, consistent with elevated in vi vitro adipogenesis. Moreover, preadipocytes isolated from transgenic epididymal adipose tissue demonstrated higher BrdU incorporation than control littermates, suggesting elevated preadipocyte proliferation. In TNMD overexpressing mice, lipogenic genes PPARG, FASN, SREBP1c and ACLY were upregulated in eWAT as was UCP-1 in brown fat, while liver triglyceride content was reduced. Transgenic animals displayed improved systemic insulin sensitivity, as demonstrated by decreased inflammation and collagen accumulation and increased Akt phosphorylation in eWAT. Thus, the data we present here suggest that TNMD plays a protective role during visceral adipose tissue expansion by promoting adipogenesis and inhibiting inflammation and tissue fibrosis. vii Table of Content Signature Page ..................................................................................................... ii Dedication ........................................................................................................... iii Acknowledgements ............................................................................................ iv Abstract ............................................................................................................... vi List of Tables ........................................................................................................ x List of Figures ..................................................................................................... xi List of Frequently Used Abbreviations ........................................................... xiii CHAPTER
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