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Gcn5 Impacts FGF Signaling at Multiple Levels and Activates C- Myc Target Genes During Early Differentiation of Embryoid Bodies

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Gcn5 Impacts FGF Signaling at Multiple Levels and Activates C- Myc Target Genes During Early Differentiation of Embryoid Bodies The Texas Medical Center Library DigitalCommons@TMC The University of Texas MD Anderson Cancer Center UTHealth Graduate School of The University of Texas MD Anderson Cancer Biomedical Sciences Dissertations and Theses Center UTHealth Graduate School of (Open Access) Biomedical Sciences 8-2017 Gcn5 Impacts FGF Signaling At Multiple Levels And Activates C- Myc Target Genes During Early Differentiation Of Embryoid Bodies Li Wang Follow this and additional works at: https://digitalcommons.library.tmc.edu/utgsbs_dissertations Part of the Biological Factors Commons, Cell Biology Commons, Developmental Biology Commons, and the Molecular Biology Commons Recommended Citation Wang, Li, "Gcn5 Impacts FGF Signaling At Multiple Levels And Activates C-Myc Target Genes During Early Differentiation Of Embryoid Bodies" (2017). The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Dissertations and Theses (Open Access). 799. https://digitalcommons.library.tmc.edu/utgsbs_dissertations/799 This Dissertation (PhD) is brought to you for free and open access by the The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences at DigitalCommons@TMC. It has been accepted for inclusion in The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Dissertations and Theses (Open Access) by an authorized administrator of DigitalCommons@TMC. For more information, please contact [email protected]. GCN5 IMPACTS FGF SIGNALING AT MULTIPLE LEVELS AND ACTIVATES C-MYC TARGET GENES DURING EARLY DIFFERENTIATION OF EMBRYOID BODIES by Li Wang, B. Sc. APPROVED: ______________________________ Sharon Y.R. Dent, Ph.D. Advisory Professor ______________________________ Mark T. Bedford, Ph.D. ______________________________ David G. Johnson, Ph.D. ______________________________ Xiaobing Shi, Ph.D. ______________________________ Pierre D. McCrea, Ph.D. APPROVED: ____________________________ Dean, The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences GCN5 IMPACTS FGF SIGNALING AT MULTIPLE LEVELS AND ACTIVATES C-MYC TARGET GENES DURING EARLY DIFFERENTIATION OF EMBRYOID BODIES A DISSERTATION Presented to the Faculty of The University of Texas MD Anderson Cancer Center UTHealth And Graduate School of Biomedical Sciences in Partial Fulfillment of the Requirements for the Degree of DOCTOR OF PHILOSOPHY by Li Wang, B. Sc. Houston, Texas August 2017 ii ACKNOWLEDGMENTS Ph.D. graduation marks a milestone in life, and it reminds me of everyone who has touched my life during this long, yet extremely rewarding and never-short-of-surprise journey. I would not be here today without the help and support from those who had stood by my side along this path. I would like to express my sincere gratitude to my advisory professor, Dr. Sharon Dent, for her continuous support of my Ph.D. study and research. Sharon has always encouraged me to think outside the box and given me the freedom to pursue my scientific interests, yet she has kept me on track through her constructive criticisms. Despite her hectic schedule as the director of Science Park, she has always made herself available to help me, from evaluating detailed experimental results to insightful discussions over career plans. During my preparation for graduation, she has been an instrumental force to support and motivate me in crafting a quality research article and a scholarly dissertation, all in less than two months. It is because of her that my graduate life has been such a rewarding experience filled with enlightenment and positivity. I am proud, as I am grateful to have Sharon as my Ph.D. mentor. Besides my advisor, I would like to thank the rest of my advisory committee, Dr. Mark Bedford, Dr. David Johnson, Dr. Pierre McCrea, Dr. Xiaobing Shi (2012-2017) and Dr. Michelle Barton (2011-2012). They have iii offered insightful suggestions and comments to help shape my project. They have also asked hard questions, driving me to delve further in the literature and think more in order to refine my project. Although I have had members located both in Smithville and Houston, my committee has made it easy for me to arrange meetings timely in compliance with GSBS requirements. I sincerely appreciate their flexibility and willingness to help all these years. The Dent Lab has been a nourishing environment for me to grow in for the past 6 years. This is a lab that appreciates and promotes hard work, creative ideas, critical thinking, individuality and teamwork. I have always felt supported and encouraged during my time in the lab. In particular, Dr. Calley Hirsch, a previous lab member, introduced me to stem cell biology, which eventually set the stage for me to build a project exploring gene regulation mechanisms in stem cell differentiation. I will always appreciate her for opening my eyes to this exciting research field. Dr. Andria Schibler (NIH) had been my lab “buddy” while she was here getting her Ph.D. We worked through countless late night hours together. Her creative ideas, resourcefulness in problem solving, and her humorous personality had been and will always be part of the fond memories of my grad-school life. Dr. Evangelia Koutelou has been my go-to person ever since I started diving into the world of stem cell differentiation (or embryonic development). She is well versed in many disciplines and techniques ranging from biochemistry to cell biology and to embryonic development. I learned many skills from her that iv proved to be instrumental for my research, for instance, isolating stem cells from early stage mouse embryos, differentiating stem cells into specific lineages (we made neurons and vessels from stem cells!), preserving the EBs for cryosections/imaging, basic techniques to create stable cell lines, and so on. But more importantly, I find her enthusiasm toward science and her passionate, positive attitude toward life to be the most inspiring and encouraging. I feel so fortunate to have come to know Andria and Lia as great companions at work, also wonderful friends in life. I would like to also extend my appreciation to our lab manager, Mr. Andrew Salinger, and our mouse expert, Amanda Martin. They have provided critical logistic and material support for my research. Throughout the years, I had the opportunity to collaborate with scientists outside our lab. For instance, I worked with Dr. Ryan McCarthy (Barton Lab) for mass cytometry experiments, also collaborated with Dr. Junya Tomida (Wood Lab) to create the ES cells expressing GCN5 tagged with Flag-HA. These collaborations proved to be very educational and helpful. The core facilities of MD Anderson Cancer Center, particularly, at Science Park have been very important for me to carry out my research. We have an amazing next generation sequencing (NGS) team and strong bioinformatics support here in Science Park, also a very capable confocal imaging core. And I had the opportunity to work with the CyTOF team (mass cytometry) in Houston. Their quality and timely services made it possible for v me to generate reliable data fundamental for my publication and this dissertation. Science Park, a reclusive research institute surrounded by beautiful pine forests, has been a wonderful place for me to experience graduate life. I was able to concentrate on my study and research in the quiet, safe campus, yet also disengage anytime by simply stepping into wonderful natural world, or to explore city life close by. My overall grad-school experience proved that I had made the absolutely right decision to come here in the first place. Finally, the love and support of my family has been always assuring and comforting all these years. Although I wish my father were alive to witness this achievement in my career, he would have been very proud. As Nobel Laureate Dr. Bruce Beutler pointed out, it is important for scientists, especially the rising scientists, to develop an area of research, make progressive discoveries, and earn reputation for solid work. Graduate study/research made a great start on this course for me, I am excited and enthusiastically anticipate the journey ahead. vi GCN5 IMPACTS FGF SIGNALING AT MULTIPLE LEVELS AND ACTIVATES C-MYC TARGET GENES DURING EARLY DIFFERENTIATION OF EMBRYOID BODIES Li Wang, B.Sc. Advisory Professor: Sharon Y.R. Dent, Ph.D. Precise control of gene expression during development is orchestrated by transcription factors, signaling pathways and co-regulators, with complex cross-regulatory events often occurring. Growing evidence has identified chromatin modifiers as important regulators for development as well, yet how particular chromatin modifying enzymes affect specific developmental processes remains largely unclear. Embryonic stem cells (ESCs) are self- renewing, pluripotent, and have the abilities to generate almost all cell types in adult tissues. The dual capacity of ESCs to self-renew and differentiate offers unlimited potential for studying gene regulation events at specific developmental stages in vitro that parallel developmental events during embryogenesis in vivo. In this dissertation project, we use a murine ESC aggregation assay (embryoid body formation, EB) to model the early development stages that proceed gastrulation, and report that GCN5, a histone acetyltransferase vii (HAT) essential for embryonic development, is required for proper expression of multiple genes encoding components of the FGF signaling pathway, and for normal activation of ERK and p38
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