Robin Miskimins - US grants

DESCRIPTION: The applicant s aim is to gain understanding of how oligodendrocyte development and myelination are regulated in the CNS, with a view to design therapies for demyelinating disorders. Dr. Miskimins presents preliminary data indicating that the differentiation of oligodendrocyte progenitor cells is accompanied by a marked increase in the cyclin dependent kinase inhibitor p27kip1 (and a decrease in cyclin E). She proposes that the increase in p27kip1 is necessary for development and differentiation of oligodendrocytes. Specifically, the increase in p27kip1 would account for the cessation of proliferation prior to differentiation and could also play a role in the activation of the differentiation program. An initial goal is to define the specific stage of oligodendrocytes development at which p27kip1 levels rise. For this study Dr. Miskimins will utilize the oligodendrocyte progenitor (O-2A cell), as well as the CG-4 cell line which she reports retains many of the properties of the O-2A cell. Another set of studies is to define the effects of growth factors and signaling molecules (bFGF, TGFB and cAMP) known to moderate oligodendrocyte development on the expression of p27kip1. The applicant predicts that basic fibroblasts growth factor (bFGF) will interfere with the elevation of p27kip1 while transforming growth factor b (TGF b) and cAMP should increase p27kip1 levels. For these studies she will utilize O-2A cells and monitor p27kip1 by immunofluorescence in Western blots and she will quantitate mRNA by Northern blotting. The third aim is to determine if expression of p27kip1 is sufficient to promote differentiation of O-2A cells. Constructs for expression of p27kip1 under the control of an inducible promoter will be transfected into either CG-4 or proliferating O-2A cells. The applicant predicts that induction of p27kip1 should bring proliferation to a halt and promote differentiation. Antisense technology will be utilized to determine if p27kip1 is required for oligodendrocyte development. An addendum provides evidence suggesting that cyclin E levels should also be monitored and this is proposed.

[unreadable] DESCRIPTION (provided by applicant): The broad, long term goal of this proposal is to understand the mechanisms that control the growth, survival, and differentiation of oligodendrocytes, the myelin producing cells of the central nervous system. The project is therefore directly relevant to demyelinating diseases such as multiple sclerosis. The proposed experiments focus on the rote of the regulatory subunit of PI3 kinase, p85, in expression of the MBP gene. p85 greatly enhances MBP promoter activity. This function requires only the SH2 domains and is independent of PI3 kinase activity. The MBP gene is only expressed at high levels in mature oligodendrocytes. Its expression is sensitive to inputs from multiple signaling pathways that regulate the progression from growing progenitors to terminally differentiated myelin-producing cells. Although many of the growth factors and hormones that control this process have been characterized, much less is known about the intracellular signals that they generate, the way these signals are integrated, and their molecular targets. The proposed experiments should provide significant insight into some of these processes. In aim 1 we will seek to identify and characterize the protein(s) with which p85 interacts. The mechanism by which p85 modulates MBP expression will be explored by determining p85 molecular interactions at various stages of oligodendrocyte development. This will be pursued using GST pull downs and co-immunoprecipitation experiments. Aim 2 seeks to determine the molecular basis through which p85 activates the MBP gene by defining the target sequences in the MBP promoter, and by characterizing the proteins that interact with these elements. We will use mutagenesis to define the element(s) in the MBP promoter that is required for the response to p85. This will serve as the basis for identifying and characterizing the transcription regulatory factors involved in this process. Aim 3 is to investigate how p85 integrates with other signaling pathways in leading to increased MBP expression. Chemical inhibitors as well as interfering expression constructs for components of signaling pathways known to be involved in MBP expression will be used to determine whether p85 integrates with these pathways in leading to increased MBP promoter activity. Completion of the studies described will provide significant insight into the role of p85 in regulating oligodendrocyte differentiation. [unreadable] [unreadable]

Abstract Not Provided.

[unreadable] DESCRIPTION (provided by applicant): The Sanford School of Medicine at the University of South Dakota (USDSM) is planning a symposium with the focus of "Emerging strategies: Bridging Basic and Clinical Science" to be held June 13-15, 2008. USDSM has recently made great progress in growing its research mission, approximately doubling the level of extramural grant support in the past five years. A majority of this growth has occurred in the basic sciences. We believe that the next step in the progression of our research program is to increase the level of translational research. Indeed, we feel this area of research holds a tremendous potential for growth in our region due to some of the unique populations (e.g. Native American, rural) present in the region. Accordingly, a major goal of the conference is to allow researchers from USDSM and the greater Midwest region an opportunity to interact with speakers of international stature that have been successful in bridging basic and clinical sciences. We also recognize that accelerating our progress in research will require the formation of strategic partnerships. Thus a second goal of the symposium, which will be regional in scope with invitations sent to the greater Midwest region, will be to initiate such partnerships with neighboring institutions. We anticipate approximately 300 attendees which would include basic science faculty, clinical faculty, postdoctoral fellows, graduate students and undergraduate students in research tracks from the greater Midwest region. Invited talks will also be open to the public. In addition to the invited talks, a poster session will be held for attendees to present their current research. Ample discussion periods and an open forum panel discussion involving the invited speakers will allow for extensive interaction between participants. [unreadable] [unreadable] [unreadable] [unreadable]

Project Summary Providing medical students with meaningful research experiences during their training is an important piece in generating physicians who incorporate research into their careers. There is a strong need for research aware clinicians in rural areas in the Upper Midwest. Having a cadre that focuses its efforts in the area of pediatric medicine will also be important in addressing the health disparities seen in rural and American Indian populations. The ultimate goal of the DRPMS is to help meet the future needs of health-related research by contributing to the development of physicians who will be well- prepared to use evidence-based medicine in practice and contribute to translational research. The Northern Plains have a large number of health disparities among the children and adolescents. Thus there is a compelling need to support and promote the training of the physicians in training in developmental research. Faculty members at the Sanford School of Medicine are strong in research focused on development: prenatal development, postnatal development, diseases and issues of children and adolescents. The Developmental Research Program for Medical Students (DRPMS) proposed here will help provide medical students with exposure to research areas of development in order to kindle their interest in pursuing this as part of their careers. The Sanford School of Medicine has recently integrated its curriculum. As an extension of this, the school is in the process of developing a strategy to integrate research into the educational experience of all medical students as a required educational activity. While the school of medicine has always had ad hoc programs for medical students to participate in research, the program proposed in this application will provide a structured program that involves research but includes an additional seminars series on career development, including how to incorporate scholarship, as well as responsible conduct of research. The Program Directors will be assisted in the management of the program by an Executive Committee and an Internal Advisory Committee. Six students per year will be accepted into the DRPMS. There will be specific strategies employed to increase the number of trainees from diverse and disadvantaged backgrounds. Selection of the trainees will be done by the Executive Committee. The research progress will be followed by the mentor(s) and the PDs. After completion of the program trainees will be invited back to become peer mentors or to continue on with research outside of the program. Intermediate and longitudinal follow- up of the trainees will be performed as part of the program's evaluation.