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 8-2019 Investigating the role of CD109 in Pancreatic Ductal Adenocarcinoma MennatAllah Shaheen Follow this and additional works at: https://digitalcommons.library.tmc.edu/utgsbs_dissertations Part of the Biology Commons, Genetics Commons, Genomics Commons, and the Medicine and Health Sciences Commons Recommended Citation Shaheen, MennatAllah, "Investigating the role of CD109 in Pancreatic Ductal Adenocarcinoma" (2019). The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences Dissertations and Theses (Open Access). 971. https://digitalcommons.library.tmc.edu/utgsbs_dissertations/971 This Thesis (MS) 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 digitalcommons@library.tmc.edu. INVESTIGATING THE ROLE OF CD109 IN PANCREATIC DUCTAL ADENOCARCINOMA By: MennatAllah Shaheen, B.Sc. Approval Page APPROVED: ______________________________ Giulio F. Draetta M.D. Ph.D. Advisory Professor ______________________________ Swathi Arur Ph.D. ______________________________ Richard R. Behringer Ph.D. ______________________________ George T. Eisenhoffer Jr. Ph.D. ______________________________ Haoqiang Ying Ph.D. APPROVED: ____________________________ Dean, The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences INVESTIGATING THE ROLE OF CD109 IN PANCREATIC DUCTAL ADENOCARCINOMA Title Page A Thesis Presented to the Faculty of The University of Texas MD Anderson Cancer Center UTHealth Graduate School of Biomedical Sciences In Partial Fulfillment of the Requirements of the Degree of MASTER OF SCIENCE By: MennatAllah Shaheen, B.Sc. Houston, Texas August, 2018 ii Abstract Investigating the Role of CD109 in Pancreatic Ductal Adenocarcinoma MennatAllah Shaheen, M.S. Advisory Professor: Giulio F. Draetta, M.D., Ph.D. Pancreatic Ductal Adenocarcinoma (PDAC) is the 3rd leading cause of cancer death in the US. We performed loss of function genomic screening on a cohort of four patient derived PDAC cell populations and our data shows a cell surface receptor CD109 to be a common vulnerability, the biologic role of which in PDAC is yet unstudied and largely unknown. We hypothesized that CD109 expression provides PDAC cells with a survival advantage, and promotes cancer progression through activation of downstream signaling. We believe therefore that targeting CD109 could improve PDAC patients’ survival. Here we report that CD109 plays a role in cell proliferation, viability, and clonogenicity in vitro. We also find that it promotes tumor formation and progression in nude mice, therefore decreasing their survival. We revealed an association between CD109 expression and YAP/TAZ signaling through RPPA and RNA Sequencing data. This data establishes CD109 as a cell surface protein exclusively expressed in PDAC rather than healthy pancreatic tissue, demonstrating pro-oncogenic behavior and tumor initiation potential in vitro and in vivo. This helps us understand more about PDAC and provides insights into a relatively unknown protein with a therapeutic potential. iii Acknowledgements I feel I can’t thank my mentor Dr Giulio Draetta enough for giving me the opportunity to be a part of his laboratory, to be under his kind knowing supervision, and most importantly making me feel welcomed that I am part of the lab family. It proved to be the best environment to learn, wrok, and make progress. Whole hearted thanks to my advisory committee, Dr swathi Arur, Dr Richard Behringer, Dr George Eisenhoffer, and Dr Haoqiang Yin, who were extremely supportive throughout my masters work, and until the very end were things were crazy. Their time, feedback, expertise, and flexibility were critical to the success of this project. Thank you to Dr Draetta’s lab, especially Johnathon Rose who essentially taught me from scratch and directly mentored me even though he had his own projects to attend to. Thank you Sanjana Srinivisan, and Melinda Seoung, for your help, support, and friendship. I am very happy to know you all. Finally, thank you Dr Wantong Yao. You have been the voice of logic, and practicality. I am honored you have been on my oral defense committee. I believe the best way to repay mentors is by benefiting from what they taught you, and passing on this knowledge. This way the circle of giving goes on forever. iv Dedication First, and foremost, thank You Allah for blessing me with this experience. I pray my knowledge will eventually benefit patients and can help make lives better. This work, small as it is, is dedicated to my family, my parents Ahmed Shaheen, and Sahar Sayed, and my siblings, Tuqa, Ziad, and Zeinab, who have sacrificed beyond imagination for what is best for us. I can write hundreds of pages expressing how thankful I am but it will never do justice to your efforts. Without you we wouldn’t be where we are. My utmost love, and gratefulness, to my dear husband, Mustafa Hussein, who believed in me so much as to cross oceans and continents so I can become a better person. He made this an enjoyable adventure with his unconditional support, and love through good and hard times. I hope I made you proud, and I ma forever thankful to you. Thank you to my friend, Menna Khedr for providing encouragement and advice from day one to graduation. You don’t know how much I appreciate it. Finally, thanks to my Reema. You are our little family away from home. Our spark of happiness, and joy. And our motivation to be better people to be your parents. v Table of Contents Approval Page ................................................................................................................................ i Title Page ...................................................................................................................................... ii Abstract ........................................................................................................................................ iii Acknowledgements ...................................................................................................................... iv Dedication ..................................................................................................................................... v Table of Contents ......................................................................................................................... vi List of Figures ............................................................................................................................... ix List of Tables ................................................................................................................................. x 1. Introduction: .......................................................................................................................... 1 1.1 Loss of Function Genomic Screening ................................................................................. 1 1.2 CD109: A History ................................................................................................................ 2 1.3 CD109: Structure ................................................................................................................ 3 1.4 CD109 in Physiological Conditions ..................................................................................... 5 1.5 CD109 in Cancer ................................................................................................................ 5 1.6 CD109 Mechanism of Action .............................................................................................. 7 1.7 Project Summary and Hypothesis ...................................................................................... 8 2. Materials and Methods ............................................................................................................. 9 2.1 Cell Lines ............................................................................................................................ 9 2.2 Plasmid Production ........................................................................................................... 11 vi 2.3 Lentivirus Production ........................................................................................................ 11 2.4 Rescue Model ................................................................................................................... 12 2.5 Inducible system ............................................................................................................... 12 2.6 Western Blot ..................................................................................................................... 13 2.7 Colony Formation Assay ................................................................................................... 13 2.8 Cell Cycle Analysis ..........................................................................................................