Melanie Simpson

Overview North Carolina State University, North Carolina A&T State University, and the University of North Carolina at Charlotte, three institutions with long-standing collaborations, join as an Alliance to develop the Department-Led Model to Promote Doctoral to Faculty Diversity (DFD) project. The DFD project engages faculty, department heads, and directors of graduate programs in transforming the culture and practices in their doctoral programs to prepare diverse dissertation students for faculty careers in their disciplines. This proposed project seeks to advance knowledge about a ����������������deep change��������������� model focused on faculty. Faculty have direct influence on doctoral students������������������ progress, level of preparation and motivation for faculty careers, as well as holding responsibility for setting the requirements of the doctoral program. Companion social science research will advance knowledge about how mentoring models can adequately prepare mentors in cross-cultural contexts to engage in effective mentoring of URM doctoral students. Recognizing that changing faculty attitudes, practices and formal components of doctoral programs requires ����������������deep change,��������������� the DFD project implements five key strategies needed for institutional change: (1) department leadership support and commitment, (2) collaborative leadership involving faculty fellows, department head, graduate coordinator and the DFD leadership team, (3) a robust design that continuously uses information gained from surveys and interviews of graduate students and faculty, (4) development of faculty awareness, understanding and skills for mentoring dissertation students across cultures and developing a truly inclusive doctoral program, and (5) a comprehensive plan for sharing the goals, methods and outcomes with the department heads, graduate directors and department faculty. Departmental DFD Teams consisting of a faculty fellow, the department head, and the graduate director form the core of the DFD program. The faculty fellows commit to two-year terms developing their understanding and skills related to cross-cultural mentoring, dissertation advising, facilitating discussions about charged topics related to diversity, and change agency. These teams will lead their department faculty in examining the pathways through their doctoral programs, obstacles to success for underrepresented minority (URM) students, and best practices for promoting URM progression to faculty careers. DFD Teams will then lead the department faculty in developing a formal departmental plan designed to promote success of URM doctoral students and address barriers that may exist. Dr. Rebecca Brent will conduct formative evaluation and the Research Institute for Studies in Education will conduct summative evaluation. Social science research will employ an embedded case study method design using primarily qualitative interview methods and a descriptive survey. Research questions will address (1) roles department heads, directors of graduate programs, and faculty advocates play in promoting or hindering success of doctoral mentoring for diverse STEM students, (2) how mentoring models can prepare faculty to provide effective dissertation and career mentoring to students of a different race or gender, and (3) how institutions can build capacity to support and sustain effective mentoring of URM doctoral students. Intellectual Merit. The DFD project adapts methods for promoting institutional change to making change in dissertation advisors������������������ awareness, understanding and practices. Longitudinal data on the attitudes, experiences, and skills of faculty and students will be combined with in-depth descriptions of how elements of the DFD project translate to the mentoring experience from the perspectives of doctoral students and faculty dissertation advisors. This project will advance knowledge by testing a flexible model of promoting deep change at three very different institutions. Critical Inquiry serves as a framework for the social science research because it is well suited for questioning

Dr Meilleur will work with others in the use of neutron technology in biology. The faculty member will lead a vigorous research program in the determination of protein structure by neutron analysis. The faculty member will also develop and improve the technology of structure analysis by neutron scattering.

Prostate epithelial cells partly control the potency and availability of androgens by inactivating and excreting them through a process termed glucuronidation. Function of the glucuronidation pathway in prostate cancer is a significant component of the response to androgen deprivation therapy (ADT). The inevitable recurrence of prostate cancer on ADT (castration resistant prostate cancer, CRPC) has few treatment options and ultimately results in mortality. UDP-glucose dehydrogenase (UGDH) is required for glucuronidation and its elevated expression predicts ADT recurrence. Because UGDH is the sole enzyme responsible for synthesis of the UDP-glucuronate precursor for glucuronidation, UGDH is a promising target in the battle against CRPC. Though UDP-glucuronate has three fates in the prostate cell, we have shown that the manipulation of UGDH expression can selectively control activity of the glucuronidation pathway and resultant growth potential of tumor cells. In this collaborative proposal, we will advance work we have done to determine how activity of UGDH in the cell is controlled by the AGC kinase RSK3, and capitalize on a novel peptide inhibitor of UGDH we identified. Hypothesis: Selective inhibition of UGDH will delay, prevent, or reverse emergence of castration resistant hallmarks in prostate tumor cells, thereby optimizing response of prostate cancer to androgen deprivation therapy. Aim 1: Determine the efficacy of RSK3-UGDH combined inhibition on cellular androgen response and prostate tumor growth kinetics in vitro, using isogenic AD and CR cells and PDX models. Aim 2: Use a high throughput approach to identify next generation peptides and small molecule lead compounds for selective inhibition of UGDH.

UDP-glucose dehydrogenase (UGDH) provides an essential precursor for elimination of excess androgen hormones from the prostate by glucuronidation. Its expression is elevated in cancerous prostate glands within biopsied tissue, relative to normal glands and to normal appearing adjacent glands, in which expression is depressed. Using tissue microarrays, which consisted of pathologist-graded biopsy specimens collected from patients prior to beginning androgen deprivation therapy, we evaluated whether UGDH levels could predict response to therapy. The patients had been followed for five years and evaluated for response versus recurrence. Each microarray slide consisted of 100 biopsies. One slide contained specimens from patients who responded. The other slide contained those of patients who experienced recurrence. We immunostained the slides and quantified fluorescence intensity of cancerous and normal appearing adjacent glands in each specimen. All cancerous glands overexpressed UGDH relative to normal glands. Importantly, a significant difference between cancerous and normal adjacent glands was found to correlate with response to androgen deprivation therapy. These preliminary data suggest UGDH will be useful as a predictive biomarker for success of androgen deprivation therapy. Additional work by our lab demonstrated that UGDH expression can be manipulated to control intracellular androgen levels, suggesting it may also be a good therapeutic target to sustain strong androgen response during therapy, thereby sensitizing prostate tumor cells to drugs such as abiraterone or other anti-androgens. In this proposal, we will test this hypothesis by implanting androgen sensitive and castration resistant prostate tumor cells with manipulated UGDH expression into intact and castrated mice. We will evaluate tumor growth kinetics, as well as a complete endpoint profile of glucuronidation pathway proteins and metabolites to determine effects of UGDH modulation on hormone dependent tumor growth. Successful completion of these studies will functionally implicate UGDH in prostate cancer progression and highlight it as a new drug target.

Matrix remodeling and stromal-epithelial cellular communication contribute to aggressive progression of prostate cancer. Hyaluronan (HA), is a glycosaminoglycan polymer synthesized and turned over when appropriate for cell proliferation and motility. Normally, its levels are tightly controlled by HA synthase enzymes (HAS), hyaluronidases, and specific HA receptors such as the HA receptor for endocytosis (HARE). HA isnegligible in normal adult prostate, but abundant in prostate tumors and bone metastases. Quantification of tumor cell associated HA and its turnover enzyme, Hyal1, predicts invasive progression and biochemical recurrence after resection. This proposal is focused on determining how the HA synthesis and turnover enzymes work together to influence matrix morphology and cell communication. The hypothesis is that surface HA borne by tumor cells increases metastatic efficiency by facilitating arrest in HARE-expressing vasculature and/or entry of the tumor cells into lymph and marrow tissue. In addition, excess tumor-borne HA may accelerate tumor cell endocytosis and/or endocytic recycling if Hyal1 is present, activating lymphatic remodeling. Rate of endocytic recycling determines the surface density of growth factor and adhesion receptors and thereby impacts tumor cell motility and metastatic survival. In aim 1, tumor cells selected for inducible HA synthesis or HA turnover will be used to test respective roles of these enzymes in clinically relevant mouse models of prostatic growth and bone metastasis. Pharmacological agents and in vivo knockdown will be used to examine how HARE functions in host target tissues to regulate prostate tumor cell colonization. These strategies will also determine how HA signaling and turnover may be therapeutically targeted to delay or prevent prostate cancer progression. Aim 2 will examine metastasis mechanisms by comparing the effects of tumor versus stromal components of HA metabolism and HA signaling on lymphatic vessel morphology, as well as lymph node and bone metastasis. The molecular format for delivery and propagation of the HA signals that trigger morphological changes to support metastasis will be characterized. Aim 3 will pursue the novel observation that elevated Hyal1, which is both a secreted and a lysosomal enzyme, increases the rate of endocytic recycling in the prostate tumor cells stably selected for its expression. The working hypothesis is that Hyal1 impacts several specific signaling pathways concurrently by modulating the rate of vesicular trafficking, thus contributing to tumor growth by maintaining surface presentation of important receptors and by re-externalizing biologically potent digestion products of HA that serve as signals. Its autocrine effects, as well as its impact on prostate stromal cells, will be tested.

UDP-glucose dehydrogenase (UGDH) is a unique, essential enzyme with the central role of providing UDPglucuronate,a rate-limiting precursor for plasma membrane hyaluronan synthesis, Golgi proteoglycan production, and ER-localized modification of hormones for elimination. Our laboratory has shown that insufficiency of UGDH contributes to loss of control of intracellular steroid levels, and dysregulated tumor cell growth rate in prostate cancer. It is not known how the cytosolic UDP-glucuronate is partitioned to itsrespective fates in the high levels needed for specifically timed product formation, nor how UGDH activity is controlled to limit competition with other pathways. Our hypothesis for this proposal is that hexameric and dimeric units of UGDH sense metabolic status of the cell and respond with increased or decreased enzymatic activity. Information for molecular sensing is conveyed partly through differential protein-protein interactions that occur upon exposure of the dimer-dimer interface. We will test this with two aims. Aim 1: Determine the functional outcome of specific UGDH interactions with components of the androgen elimination pathway. We will directly measure interactions of UGDH with hyaluronan synthase, the Golgi UDP-xylose transporter, and the ER UDP-glucuronate transporter, as the three proteins that mediate the demands for UDP-glucuronate flux. We will quantify UDP-glucuronate, steroid-glucuronide, notch glycosylation and hyaluronan production, which will respectively report overall UGDH activity, and functional distribution to the ER for androgen elimination, the Golgi for proteoglycan secretion, or the plasma membrane for hyaluronan synthesis. Aim 2: Characterize and validate the UGDH interactome using an unbiased approach. We will use mass spectrometry to identify proteins that differentially co-fractionate by size exclusion chromatography with our wellcharacterized hexameric and dimeric point mutants. As a complementary approach, we will identify proteins that cross-link with hexameric versus dimeric UGDH point mutants that incorporate a photo-activatable crosslinker as a non-natural amino acid. Validated interactions and/or post-translational modifications will be used to design strategies for selective partitioning of UDP-glucuronate to favor hormone elimination in future preclinical studies of prostate cancer.