Control of multilineage hematopoiesis by 5' cap-proximal modification - PROJECT SUMMARY/ABSTRACT This proposal outlines a five-year plan to prepare the PI, Dr. Andrew Levine, for an academic career as a physician-scientist. Dr. Levine received his MD/PhD degrees from the Weill Cornell/Rockefeller/Sloan Kettering Tri-Institutional MD-PhD program and completed residency and fellowship training in Laboratory Medicine and Hematopathology at the University of California, San Francisco (UCSF), where he is currently a Clinical Instructor. Dr. Levine has developed a mentored research plan together with Dr. Davide Ruggero, a Professor of Urology at UCSF and an expert in the field of post-transcriptional RNA regulatory mechanisms at the organismal level. Dr. Ruggero has served as a mentor to numerous trainees, many of whom have obtained independent positions as faculty in academia or are running their own research groups in industry. The mentorship plan ensures Dr. Levine’s full access to the resources and expertise of UCSF, a world-class research institution, which will provide an outstanding environment for Dr. Levine to progress to scientific and career independence. Dr. Levine has also assembled a K08 advisory committee composed of mostly senior investigators and physician- scientists whose scientific expertise and mentorship will greatly assist Dr. Levine’s career development. They include Drs. Kevin Shannon, Samie Jaffrey, and Hani Goodarzi. In addition, Dr. Levine will participate in formal coursework, retreats, seminars, and workshops including through the UCSF Graduate Program, UCSF Office of Career Development, UCSF Clinical and Translational Sciences Institute, and UC Davis Genome Center. Finally, through the Ruggero laboratory and UCSF core facilities, Dr. Levine will have direct access to an extraordinary combination of scientific resources. Dr. Levine’s research proposal seeks to understand the role of post- transcriptional RNA regulatory mechanisms in hematopoiesis. Specifically, mRNA nucleotide modification is increasingly appreciated to constitute one major yet poorly understood RNA regulatory mechanism that is commonly affected in bone marrow failure syndromes and hematopoietic neoplasms. During Dr. Levine’s time as a mentee of Dr. Ruggero, he has discovered that RNA cap-proximal nucleotide methylation, mediated by Cap methyltransferase 2 (CMTR2), plays a critical role in balancing multilineage hematopoiesis. In Aim 1, through use of novel genetic models Dr. Levine will elucidate the precise developmental stage(s) at which CMTR2 is required for hematopoiesis. In Aim 2, through use of cell culture and in vivo models and cutting-edge sequencing technologies, Dr. Levine will explore the hypothesis that CMTR2 mediates its effects by regulating the translation and stability of key subsets of mRNAs within hematopoietic stem/progenitor cells to influence cell fate choices. These experiments will elucidate the in vivo workings of a poorly understood epitranscriptomic mark, and will elucidate how the presence of a single base modification on RNAs enforces complex cellular differentiation pathways during hematopoietic development. These studies will moreover provide an ideal platform for Dr. Levine to complete his training and launch his independent career.
