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Pyocyanin, a Virulence Factor Produced by Sepsis- Causing Pseudomonas Aeruginosa, Promotes Adipose Wasting and Cachexia

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Pyocyanin, a Virulence Factor Produced by Sepsis- Causing Pseudomonas Aeruginosa, Promotes Adipose Wasting and Cachexia University of Kentucky UKnowledge Theses and Dissertations--Pharmacology and Nutritional Sciences Pharmacology and Nutritional Sciences 2019 PYOCYANIN, A VIRULENCE FACTOR PRODUCED BY SEPSIS- CAUSING PSEUDOMONAS AERUGINOSA, PROMOTES ADIPOSE WASTING AND CACHEXIA Nika Larian University of Kentucky, [email protected] Digital Object Identifier: https://doi.org/10.13023/etd.2019.238 Right click to open a feedback form in a new tab to let us know how this document benefits ou.y Recommended Citation Larian, Nika, "PYOCYANIN, A VIRULENCE FACTOR PRODUCED BY SEPSIS-CAUSING PSEUDOMONAS AERUGINOSA, PROMOTES ADIPOSE WASTING AND CACHEXIA" (2019). Theses and Dissertations-- Pharmacology and Nutritional Sciences. 31. https://uknowledge.uky.edu/pharmacol_etds/31 This Doctoral Dissertation is brought to you for free and open access by the Pharmacology and Nutritional Sciences at UKnowledge. It has been accepted for inclusion in Theses and Dissertations--Pharmacology and Nutritional Sciences by an authorized administrator of UKnowledge. For more information, please contact [email protected]. STUDENT AGREEMENT: I represent that my thesis or dissertation and abstract are my original work. Proper attribution has been given to all outside sources. I understand that I am solely responsible for obtaining any needed copyright permissions. I have obtained needed written permission statement(s) from the owner(s) of each third-party copyrighted matter to be included in my work, allowing electronic distribution (if such use is not permitted by the fair use doctrine) which will be submitted to UKnowledge as Additional File. I hereby grant to The University of Kentucky and its agents the irrevocable, non-exclusive, and royalty-free license to archive and make accessible my work in whole or in part in all forms of media, now or hereafter known. I agree that the document mentioned above may be made available immediately for worldwide access unless an embargo applies. I retain all other ownership rights to the copyright of my work. I also retain the right to use in future works (such as articles or books) all or part of my work. I understand that I am free to register the copyright to my work. REVIEW, APPROVAL AND ACCEPTANCE The document mentioned above has been reviewed and accepted by the student’s advisor, on behalf of the advisory committee, and by the Director of Graduate Studies (DGS), on behalf of the program; we verify that this is the final, approved version of the student’s thesis including all changes required by the advisory committee. The undersigned agree to abide by the statements above. Nika Larian, Student Dr. Lisa Cassis, Major Professor Dr. Howard Glauert, Director of Graduate Studies PYOCYANIN, A VIRULENCE FACTOR PRODUCED BY SEPSIS-CAUSING PSEUDOMONAS AERUGINOSA, PROMOTES ADIPOSE WASTING AND CACHEXIA _____________________________________ DISSERTATION _____________________________________ A dissertation submitted in partial fulfillment of the requirements for the degree of Doctor of Philosophy in the College of Medicine at the University of Kentucky By Nika Larian Lexington, KY Director: Dr. Lisa Cassis, Professor of Nutritional Sciences and Pharmacology, Vice President for Research Lexington, KY 2019 Copyright © Nika Larian 2019 ABSTRACT OF DISSERTATION PYOCYANIN, A VIRULENCE FACTOR PRODUCED BY SEPSIS-CAUSING PSEUDOMONAS AERUGINOSA, PROMOTES ADIPOSE WASTING AND CACHEXIA Sepsis is a leading cause of death among critically ill patients that results in metabolic alterations including hypercatabolism, lipoatrophy, and muscle wasting, contributing to the development of cachexia. Septic cachexia is associated with loss of body weight, fat mass, and lean mass and dysregulated immune function. There are currently no efficacious treatment strategies for septic cachexia, and nutritional interventions have limited success in preventing hypercatabolic wasting. Pyocyanin is a virulence factor produced by sepsis-causing Pseudomonas aeruginosa that has been shown to activate the aryl hydrocarbon receptor (AhR), increase inflammation, and produce reactive oxygen species. Thus, pyocyanin represents a novel mechanistic target in the development of septic cachexia. In Aim 1, we hypothesized that pyocyanin reduces adipocyte differentiation and activates AhR in vitro and in vivo. In vitro, pyocyanin reduced differentiation of 3T3-L1 cells to adipocytes and promoted expression of proinflammatory cytokines. These effects were associated with activation of AhR. We established an in vivo model of pyocyanin-induced cachexia using repeat intraperitoneal exposure to pyocyanin in male and female C57BL/6J mice. Acutely, pyocyanin reduced differentiation of stem cells isolated from adipose stromal vascular tissue and augmented expression of proinflammatory cytokines. Chronically, pyocyanin reduced body weight and fat mass, which was associated with adipose-specific AhR activation. Pyocyanin had sexually dimorphic effects on lipolysis and adipocyte inflammation. These data suggest a role of pyocyanin in adipose cachexia associated with sepsis. In Aim 2, we hypothesized that pyocyanin activates adipocyte AhR to promote adipose tissue wasting and cachexia. To test this hypothesis, we used a mouse model of adipocyte-specific deficiency of AhR and chronically administered pyocyanin to male and female mice. In male mice with adipocyte AhR deficiency, effects of pyocyanin to promote adipose wasting and cachexia were attenuated. In contrast, female adipocyte AhR deficient mice had an augmented response to pyocyanin to decrease body weight. Results suggest divergent mechanisms of pyocyanin to regulate adiposity and body weight through adipocyte AhR between male and female mice. These data support a role for pyocyanin in the development of adipose cachexia associated with Pseudomonas aeruginosa sepsis that is partially regulated by adipocyte AhR. Targeting pyocyanin’s effects on adipocytes represents a potentially novel therapeutic approach for septic cachexia that could mitigate septic cachexia, a condition associated with increased risk of mortality in this population. KEYWORDS: Pyocyanin, Sepsis, Cachexia, Aryl Hydrocarbon Receptor, Adipocyte, Pseudomonas aeruginosa Nika Larian May 9, 2019 PYOCYANIN, A VIRULENCE FACTOR PRODUCED BY SEPSIS-CAUSING PSEUDOMONAS AERUGINOSA, PROMOTES ADIPOSE WASTING AND CACHEXIA By Nika Larian Dr. Lisa Cassis Director of Dissertation Dr. Howard Glauert Director of Graduate Studies May 9, 2019 Date ACKNOWLEDGMENTS A dissertation is not just a publication, it is a personal story. Woven between these many lines of text are the people, places, and experiences that have crafted my story as a little girl who dreamed so fiercely of growing up and becoming a doctor, a scientist, and a woman with the power to create change. I have many characters to thank in this story. First, I would first like to thank my mentor Dr. Lisa Cassis for her endless support of my research and professional development. You are the reason I came to UK to pursue a PhD, and you are the model of a strong woman in science. Next, I want to thank the other members of the Cassis Lab, who have truly been a family over the past several years. Dr. Mark Ensor for somehow making injecting hundreds of mice bearable and for constant help and support- I truly couldn’t have survived this last year without you; Dr. Robin Shoemaker for teaching me basically every technique I know; Dr. Sean Thatcher for always having an answer to my questions and always offering a helping hand; Victoria English for being the lab queen and answering my dog emergency calls; Dr. Yasir Al-Siraj for setting an excellent example of a graduate student; Heba Ali for being the most caring and selfless person I’ve ever met. I would also like to thank my other committee members, Dr. Kevin Pearson, Dr. Xi- ang Li, Dr. Dawn Brewer, and my outside examiner Dr. Nathan Vanderford. Your support, guidance, and perspective have been invaluable in this process. Importantly, I would like to thank the friends who made this a story worth living. Jessie, I cannot even imagine graduate school without you by my side every step of the way. I didn’t anticipate making close friends in graduate school, iii and I certainly didn’t anticipate meeting someone I would consider a sister for life. Through the highs and lows, you have carried me (sometimes kicking and screaming) to the finish line. To my other friends at UK- Kaia, Peter, Jared, Adam, Madison, and all the students in the Department of Pharmacology and Nutritional Sciences, thank you for the community you created. To my friends outside of the department, Megan, Kim, Natalie, and Savanna, thank you for reminding me that I am more than a scientist. Importantly, I would like to thank my family, particularly my mom and dad. Dad, thank you for making me tough and fiery, for urging me to find independence, and for instilling in me that education is the most powerful asset. This will be better than an MD, I promise. Mom, there aren’t enough words to express my love and gratitude. Thank you for your unconditional love, for being the hardest working woman I know, for being the most selfless woman I know, for being everything to me. Mela, you may not be able to read, but you also deserve this PhD for being by my side through every late night and early morning, for being my guardian angel since my first semester. Finally, I would like to thank God for showing me that strength is built in adversity, that my story is ultimately written by You, and that I should enjoy each page instead
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