Molecular Mechanisms Underlying the Metastasis Suppressor Activity of NME1 in Malignant Melanoma Item Type dissertation Authors Pamidimukkala, Nidhi Vela Publication Date 2019 Abstract Metastatic melanoma is exceedingly lethal and is responsible for the majority of skin cancer related deaths. Impactful research on melanoma metastasis is crucial to improving prognosis. The discovery of metastasis suppressor genes, genes that inhibit... Keywords ALDOC; NME1; Metastasis; Fructose-Bisphosphate Aldolase; Melanoma; Transcription, Genetic Download date 26/09/2021 16:56:05 Link to Item http://hdl.handle.net/10713/9611 Curriculum Vitae Nidhi V. Pamidimukkala [email protected] https://www.linkedin.com/in/nidhi-pamidimukkala Doctor of Philosophy, 2019 EDUCATION 2012-2019 Ph.D., Graduate Program in Molecular Medicine, Cancer Biology Tack University of Maryland School of Medicine – Baltimore, MD 2005-2009 B.S., Biological Sciences, Mellon College of Science Carnegie Mellon University - Pittsburgh, PA 2005-2009 Minor, Chemistry, Mellon College of Science Carnegie Mellon University - Pittsburgh, PA RESEARCH EXPERIENCE 2012-2019 Graduate Research Assistant, Mentor: David M. Kaetzel, Ph.D University of Maryland School of Medicine-Baltimore, MD 2009-2012 Biochemist-MBV Bioprocessing Group Merck & Co, Inc.-Union, NJ 2008-2008 Future Talent Intern-Drug Metabolism Group Merck & Co., Inc- Rahway, NJ 2007-2009 Undergraduate Research Assistant, Mentor: Gordon S. Rule, Ph.D Carnegie Mellon University-Pittsburgh, PA RESEARCH PUBLICATIONS Pamidimukkala, N.V , Leonard, M.K, Snyder, D., McCorkle, J.R., Kaetzel, D.M. (2018). Metastasis Suppressor NME1 Directly Activates Transcription of the ALDOC Gene in Melanoma Cells. Anticancer Research,38(11), 6059-6068. PMID: 30396920 Leonard, M.K, Novak, M., Snyder, D., Snow, G., Pamidimukkala, N.V , McCorkle, J.R., Yang, X.H., Kaetzel, D.M. (2019). The Metastasis Suppressor NME1 Inhibits Melanoma Cell Motility via Direct Transcriptional Induction of the Integrin beta-3 Gene. Experimental Cell Research, 374(1), 86-93. PMID: 30458180 REVIEW PUBLICATIONS Puts, G.S., Leonard, M.K, Pamidimukkala, N.V, Snyder, D., Kaetzel, D.M. (2018). Nuclear Functions of NME Proteins. Lab Inverstigation,98 (2), 211-218. PMID: 29058704 Leonard, M.K, Pamidimukkala, N.V, Puts, G.S., Snyder, D., Slominski, A.T., Kaetzel, D.M. (2017). The HGF/SF Mouse Model of UV-Induced Melanoma as an In Vivo Sensor for Metastasis-Regulating Gene. International Journal of Molecular Science, 18 (8), E1647 PMID: 28788083 MANUSCRIPTS IN PREPARATION Leonard, M.K*, Pamidimukkala, N.V*, Puts, G.S*., Manojlovic, Z., Jarrett, S.G., Snyder, D., Novak, M., Dabernat, S., DeFabo, E.C., Noonan, F.P., Slominski, A., Merlino, G., Carpten, J., Kaetzel, D.M., Comprehensive Molecular Profiling of UV- induced Metastatic Melanoma in Nme1/Nme2-deficient Mice Reveals Genomic and Transcriptomic Alterations Associated with Poor Survival in Human Melanoma Patients. In preparation. *Authors contributed equally Leonard, M.K., Pamidimukkala, N.V ., Novak, M., Snyder, D., Snow, G., McCorkle, J.R., Yang, X.H., Kaetzel, D.M., Impacts of NME1 Expression in the Tumor Microenvironment on Growth and Metastatic Potential of Melanoma Cells. In preparation Puts, G.C., Leonard, M.K., Jarrett, S.G., Snyder, D., Vincent, R., Yang, Y., Pamidimukkala, N.V. , Portney, B., Rassool, F., Zalzman, M., Kaetzel, D.M., Recruitment of the Metastasis Suppressor of NME1 to Double-Stranded Breaks in DNA Promotes the Non-Homologous End-Joining Pathway of Repair. In Preparation SELECTED PRESENTATIONS 8th Annual Biochemistry Retreat Baltimore, MD University of Maryland School of Medicine January 2019 Poster Presentation: Metastasis Suppressor NME1 Activates Transcription of the ALDOC Gene in Melanoma 9th Annual Cancer Biology Research Retreat Baltimore, MD University of Maryland School of Medicine May 2018 Oral Presentation: Transcriptional Regulation of the Aldolase C (ALDOC) Gene as a Potential Mechanism for the Metastasis Suppressor Activity of NME1 7th Annual Biochemistry Retreat Baltimore, MD University of Maryland School of Medicine January 2018 Poster Presentation: NME1 Regulation of Aldolase C (ALDOC) as a Potential Mechanism for the Metastasis Suppressor Activity in Melanoma Molecular Medicine Seminar Baltimore, MD University of Maryland School of Medicine November 2017 Oral Presentation: NME1 Regulation of Aldolase C (ALDOC) as a Potential Mechanism of Metastasis Suppression in Melanoma Cancer Focus Group Meeting Baltimore, MD University of Maryland School of Medicine June 2017 Oral Presentation: Induction of ALDOC Expression by NME1 May Mediate Metastasis Suppressor Activity in Malignant Melanoma 8th Annual Cancer Biology Research Retreat Baltimore, MD University of Maryland School of Medicine May 2017 Poster Presentation: Induction of ALDOC Expression by NME1 may Mediate Metastasis Suppressor Activity in Malignant Melanoma (Presentation Winner) AACR Annual Meeting Washington, D.C. Walter E. Washington Convention Center April 2017 Poster Presentation: Induction of Aldolase C by NME1 may Mediate Metastasis Suppressor Activity in Malignant Melanoma 6th Annual Biochemistry Retreat Baltimore, MD University of Maryland School of Medicine January 2017 Poster Presentation: Induction of ALDOC Expression by NME1 may Mediate Metastasis Suppressor Activity in Malignant Melanoma Pan American Society for Pigment Cell Research Baltimore, MD Royal Sonesta Harbor Court October 2016 Poster Presentation: Identifying Key Effectors in NME1-Mediated Metastasis Suppression 5th Annual Biochemistry Retreat Baltimore, MD University of Maryland School of Medicine January 2016 Poster Presentation: Identifying Key Effectors in NME1-Mediated Metastasis Suppression 6th Annual Cancer Biology Research Retreat Baltimore, MD University of Maryland School of Medicine May 2015 Poster Presentation: Identifying Key Effectors in NME1-Mediated Metastasis Suppression (Presentation Winner) Cancer Focus Group Meeting Baltimore, MD University of Maryland School of Medicine September 2014 Oral Presentation: The Role of NME1 in Mediating Metastasis Suppression in Malignant Melanoma Abstract Title of Dissertation: Molecular Mechanisms Underlying the Metastasis Suppressor Activity of NME1 in Malignant Melanoma Nidhi V. Pamidimukkala, Doctor of Philosophy, 2019 Dissertation Directed by: David M. Kaetzel, Ph.D, Professor, Biochemistry and Molecular Biology Metastatic melanoma is exceedingly lethal and is responsible for the majority of skin cancer related deaths. Impactful research on melanoma metastasis is crucial to improving prognosis. The discovery of metastasis suppressor genes, genes that inhibit metastasis but do not affect tumor growth, have advanced the field of metastasis research. NME1 (also referred to as NDPK-A or NM23-H1) was the first identified metastasis suppressor gene. NME1 is a multifaceted molecule, executing numerous cellular functions which are attributable to the suppressor phenotype. Our lab previously identified a signature of NME1-regualted genes which have prognostic value in human melanoma. As such, we hypothesize that NME1 possess an understudied transcriptional activity to regulate one or more of these signature genes. In the present study, we identified Aldolase C ( ALDOC ) from the gene signature as an ideal candidate to assess NME1 transcriptional activity. NME1 induced mRNA and protein expression of ALDOC in multiple melanoma cell lines. Transcriptional regulation was evidenced by elevated expression of ALDOC pre-mRNA and activation of the ALDOC promoter in NME1- expressing melanoma cells. Employing chromatin immunoprecipitation, we demonstrated NME1 localizes to the ALDOC gene and enriches the presence of active transcription indicators at the ALDOC promoter. Together, we provide novel systematic evidence for NME1 transcriptional activity, which may further be explored as a mechanism for the metastasis suppressor function. Furthermore, we establish unequivocal metastasis suppressor capability of NME1 using a transgenic mouse model susceptible to developing melanoma upon exposure to ultraviolet (UV) radiation. Mice deficient in NME1 expression experienced significantly increased melanoma metastases to the lung and lymph nodes compared to wild-type mice. Taken together, our studies demonstrate NME1 as a robust metastasis suppressor of UV-induced melanoma, and provide evidence for novel transcription factor activity of NME1. We aim to enhance our understanding of melanoma metastasis through our continued research of the metastasis suppressor gene, NME1. Molecular Mechanisms Underlying the Metastasis Suppressor Activity of NME1 in Malignant Melanoma by Nidhi V. Pamidimukkala Dissertation submitted to the Faculty of the Graduate School of the University of Maryland, Baltimore in partial fulfillment of the requirements for the degree of Doctor of Philosophy 2019 Acknowledgments I sincerely thank my mentor, Dr. David M. Kaetzel. I am so appreciative of everything you have taught me. I know you have mentored several students, but I am happy to have been your first graduate student at UMB. Thank you to my thesis committee, Drs. Thomas Hornyak, Shawn Lupold, Stuart Martin, Tony Passaniti, and Dudley Strickland, for all their valuable advice on my dissertation project. Thank you to all the past and present members of the Kaetzel lab, particularly, Drs. Katie Leonard and Gemma Puts. You both were there to offer support and advice in the lab and outside of it. Thank you for teaching me so much and providing such an