The goal of this proposal is to provide the applicant with additional mentored research experience and critical career development training. The additional experimental training proposed in this plan, wholly funded by NHBLI Division of Intramural Research, will provide him with the ability to perform cutting-edge cell biology research, while the career development training and instruction from mentors and advisors will provide him with the tools necessary to excel as an independent investigator. The candidate is an expert in non-muscle myosin 2, the major force-producing mechano-enzyme in non- muscle cells. Through his graduate and postdoctoral work, he has published 5 first-author, 5 middle-author and 1 review on myosin 2. By proposing to investigate the key kinases that regulate myosin 2, for which we are clearly lacking critical cellular information, the candidate will build upon his current expertise while establishing projects that unmistakably separate him from the work of his Ph.D. and postdoctoral advisors. This proposal seeks to investigate how these kinases are simultaneously regulating myosin 2 at steady-state (Aim 1) and during two dynamic morphology-altering processes: epithelial-mesenchymal transition (EMT) (Aim 2) and nuclear translocation (squeezing) during cell invasion (Aim 3). As both myosin 2 and the kinases that regulate myosin 2 are known to be involved in a wide-array of pathologies (e.g. cancer, diabetes, hypertension), and EMT and cell invasion are critical components of cancer cell metastasis, this research has clear clinical relevance. The additional technical training in this grant will enable the candidate to establish a lab performing state-of- the-art cell biology research. This includes training in super-resolution imaging with Dr. Eric Betzig (JRC), cell invasion using microfluidics with Dr. Jan Lammerding (Cornell), surface micropatterning with Dr. Ken Yamada (NIDCR/NIH), and in vitro characterization of myosin phosphorylation with Dr. James Sellers (NHLBI/NIH). In addition to the experimental training, the advisory committee assembled includes leading intramural and extramural scientists that were selected for their expertise in placing candidates in faculty positions, career development familiarity, relevant research expertise, and experience obtaining extramural funding. Furthermore, through the Office of Intramural Training at NIH, the training plan includes mechanisms to prepare the candidate for obtaining a position and excelling as an independent researcher. This includes counseling, coursework, workshops, and seminars that will provide mock interviews, practice job and chalk talks, grant writing experience, and effective teaching, mentoring, and lab management skills. Collectively, this tailored proposal will enable the candidate to build on his current expertise with new research and career skills to propel him to a successful career as an independent investigator.
The proposed work seeks to elucidate the mechanism by which myosin 2 is regulated. Myosin 2 forms the major contractile machinery in all non-muscle cells and is critical for cell movements during normal development and during various pathological states, including cancer metastasis. Completion of the proposed research will produce critical insights into how these processes occur and could potentially lead to novel strategies for therapeutic intervention.
