Douglas E. Wright - US grants

DESCRIPTION: A significant subpopulations of nociceptive sensory neurons (IB4/Ret neurons) has been recently found to switch their neurotrophins requirements from nerve growth factor (NGF) to glial cell line-derived neurotrophic factor (GDNF). IB4/Ret neurons are quite sensitive to trophic deprivation caused by injury. GDNF administration appears to reverse the deficits in these neurons. This proposal will test the hypotheses that IB4/Ret neurons are vulnerable to diabetic neuropathy and they respond differently to injury than other populations in diabetes. This information may reveal that these cells are sensitive to therapeutic actions of GDNF, and not NGF. The specific aims are to determine the sensitivity of IB4/Ret neurons to diabetes by measuring changes in selective markers expressed by IB4/Ret neuron in streptozotocin (STZ)-induced mice. The study will characterize differences between NGF-responsive (trkA) and IB4/Ret neurons in their capacity to express genes that are required for regeneration but are suppressed in diabetes. Changes in gene expression for GAP-43 and Ta1 a-tubulin will be compared between TrkA and IB4/Ret populations in axotomized, STZ-induced mice. The ability of GDNF or NGF to enhance expression of these genes in IB4/Ret neurons will be tested by supplying trophins to axotomized, STZ-induced mice. The long-term goal of this project is to analyze the responses of sensory neurons to experimental diabetes and couple these results with new information about the trophic needs of affected neurons to design treatments with better specificity for DN. Results from this study will provide specific evidence tha sensory neurons have varied susceptibilities to diabetes related to their functions and neurotrophic responsiveness, and suggest GDNF as a candidate to be used in treatments for diabetic neuropathy.

The role of nerve growth factor-related neurotrophins as survival factors during development is now well established. The neurotrophins also have great potential to regulate neurons in adulthood and thus may serve as potent therapeutic agents. Because neurotrophin-3 (NT-3) supports proprioceptive neurons, it is anticipated that NT-3 will be effective in the treatment of disorders that involve large myelinated sensory fibers. The long-term objective is to characterize the cellular and molecular actions of NT-3 on postnatal primary sensory neurons to understand how NT-3 may improve sensory impairment. The proposed studies will focus on two paradigms that compromise proprioceptive neurons: peripheral nerve injury and neuronal degeneration in disease. Three animal models, NT-3 transgenic, NT-3 null mutant, and hereditary mutant mice that undergo degeneration of NT-3-dependent proprioceptive neurons will be studied. With this approach, the trophic support of neurons following perturbation can be characterized in vivo, and NT-3 transgenes can be introduced into other mouse models to test the therapeutic effects of- NT-3. The first Specific Aim will characterize the interactions between injured proprioceptive neurons and their 5 trophic support at different stages postnatally. This Aim will then test if NT-3 stimulates proprioceptive axons to regenerate, branch, and reinnervate targets in muscle. The second Aim will examine the role of NT-3 in the degeneration of proprioceptive neurons in dystonia musculorum (dst) mice, a mouse model for sensory ataxia. This Aim will then test the therapeutic potential of NT-3 by breeding a muscle-expressed NT-3 transgene into dst mice. Finally, this Aim will explore how the disruption of newly-identified genes in dst mice cause the selective vulnerability of proprioceptive neurons. Results from these studies will define new biological actions of NT-3 and reveal how manipulation of the trophic support of neurons may be useful to treat pathologic neural conditions.

[unreadable] DESCRIPTION (provided by applicant): Diabetic neuropathy (DN) is a serious neural complication that develops in many diabetic patients. Small unmyelinated sensory fibers are commonly affected, leading to abnormal cutaneous sensation and pain. It is believed that interrelated mechanisms contribute to DN and insufficient neurotrophic support has recently been added to the list of possible deficits. Nociceptive neurons affected in small-fiber DN respond either to nerve growth factor (NGF) or glial cell line-derived neurotrophic factor (GDNF). Whereas studies have noted deficits in NGF support to sensory neurons in DN, little is known about the role of GDNF. The long-term goal of this project is in an animal model of diabetes, examine the biology of GDNF-related ligands and receptors and correlate deficits with physiological and behavioral deficits that are caused by diabetes. Our purpose is to uncover mechanisms underlying the development of DN, thus providing information that will aid in developing novel treatments for DN. Our previous studies demonstrate that the central processes of GDNF-responsive neurons are sensitive to diabetes and GDNF administration can reverse deficits in GDNF-responsive spinal terminals. This proposal will test the hypotheses that GDNF support to primary sensory neurons is impaired in diabetes and anatomical/physiological deficits in GDNF-responsive neurons can lead to impaired responses to cutaneous stimuli. The first specific aim will characterize deficits in GDNF/GDNF-receptor synthesis and transport in STZ-induced diabetic mice, and test whether GDNF treatment can improve ligand/receptor abnormalities. The second aim will characterize deficits in GDNF-responsive fibers in the spinal cord and skin, and then test the ability of GDNF to stimulate sensory axon growth and reinnervation. The final aim will characterize deficits in GDNF-responsive neurons by performing electrophysiological recordings of single, identified neurons in an in vitro skin-nerve preparation. Physiological deficits will be correlated with abnormalities in the response of diabetic mice to noxious mechanical, chemical and thermal stimuli. The capacity of GDNF to modify neuronal physiology and behavioral responses to cutaneous stimuli will also be tested. In sum, this grant proposes to use molecular, anatomical, physiological and behavioral approaches to understand the biology of GDNF in DN and to test whether GDNF has therapeutic actions on cutaneous neurons affected in diabetic animals. Results from this study will 1) provide evidence that impaired GDNF support contributes to the development of DN and 2) establish GDNF as a candidate to be used in treatments to improve cutaneous function in DN. [unreadable] [unreadable]

? DESCRIPTION (provided by applicant): Diabetic neuropathy (DN) is one of the most prevalent and debilitating complications of diabetes. However, DN remains largely an untreatable condition and current strategies only address pain control. A number of cellular mechanisms have been identified that lead to DN, but no explanation exists as to why certain patients develop pain, or alternatively develop insensate sensory loss. This paucity of information has hampered clinicians in developing prophylactic treatment strategies for patients that adequately prevent painful or insensate neuropathy. Our long-range goal is to understand how changes in peripheral axons can lead to varied sensory symptoms in diabetes. In previous funding years, we have identified rodent models that better reflect the varied symptoms of human DN and used these models to understand how changes in axonal subtypes within the epidermis may be related to specific symptoms of DN. Epidermal innervation is derived from two distinct axonal populations that are differentiated by different neurochemical markers and neurotrophic responsiveness. These two sensory axon subtypes are referred to as peptidergic or nonpeptidergic axons. We believe that key modifications occur in these epidermal axon subtypes early in the progression of DN that can lead to pain, or alternatively, loss of sensation. The central hypothesis is that the ratio and damage to two axonal subtypes innervating the skin is critical in specifying sensory dysfunction in diabetes. The aims of the grant include 1) testing whether changes in epidermal axons can drive pain or loss of sensation in diabetes, and testing whether inflammation and NGF responsible for changing epidermal axon phenotypes, 2) whether exercise intervention prevent or reverse changes in epidermal axon phenotypes, and 3) whether skin biopsy analyses be used to predict and follow epidermal changes associated with small fiber DN in humans. These proposed experiments are unique as they present a simple, but testable hypothesis that may explain how the divergent sensory complications can develop in diabetes. Importantly, our approaches are easily translatable to human studies and may be very relevant to other forms of neuropathy. The analyses of skin biopsies as performed here may provide new predictive power to identify patients at risk for developing neuropathy, and may also be used as a better indicator of neuropathy improvement in future clinical trials of diabetic neuropathy.

DESCRIPTION (provided by applicant): The University of Kansas Medical Center (KUMC) presents an application for continued support for the Kansas IDeA Network for Biomedical Research Excellence (K-INBRE). The K-INBRE links KUMC (Lead Institution) with the two major doctoral-degree-granting institutions in Kansas (University of Kansas-Lawrence, KU-L; Kansas State Univ., KSU) as Graduate Partner Institutions (GPIs), and with seven Undergraduate Partner Institutions (UPls). UPls include six Kansas undergraduate campuses (Emporia State Univ., Ft. Hays State Univ., Haskell Indian Nations Univ., Pittsburg State Univ., Washburn Univ., Wichita State Univ.) and Langston Univ. (Langston, OK). Haskell Indian Nations Univ. and Langston Univ. increase diversity in the network as the first is devoted to education and training of native Americans and the second enrolls primarily black undergraduates. The long-range objective of the Kansas program is to strengthen the state's research capacity in Cell and Developmental Biology by building on the successes of the current K-INBRE. The structure and operational principles of the K-INBRE, which focus on training for biomedical research, networking and intercampus communication and the presence of a sophisticated bioinformatics program, were established during the previous years. These goals remain similar as the KINBRE has had a significant impact on biomedical research in the State of Kansas, but novel programs are tailored to fit new emerging areas associated with translational research. Programs conducted by the KINBRE have had measurable success in reaching their stated goals. The Specific Aims proposed for the next phase of the K-INBRE are to (1) maintain and improve the current multi-disciplinary research network in Cell and Developmental Biology in the State of Kansas, strengthening both communication channels and research infrastructure, (2) enhance science and technology knowledge and integration in Kansas by offering sophisticated bioinformatics technology and education, (3) stimulate basic and translational research in the State of Kansas via mentored, interdisciplinary research opportunities. Within these Aims, new features that improve the K-INBRE include broadening funding for research careers together with improvements in oversight and the mentoring process, promoting an integrated systems biology approach within our bioinformatics network, and incorporating training for translational research into the K-INBRE goals so as to smooth the progress of scientific discoveries into the clinical arena.

Functions of the Administrative Core. The overall goal of the Administrative Core is to provide efficient service to the K-INBRE Network, assuming most of the administrative responsibilities of this complex, multi-campus initiative so as to improve the ability of Kansas researchers to compete successfully for NIH research grants. The strength of the K-INBRE Administrative Core is the longstanding continuity and success of the experienced administrative personnel who have been effective in all administrative operations over its long history. The major functions of the Administrative Core remain the same as in previous years, and have adjusted to meet the increasing compliance requirements for NIH. Achievements for the previous years are outlined in the Progress section. In brief this Core: ensures continuity of the INBRE program by preparing accurate and timely progress reports and renewal applications builds good working relationships with faculty, students and administration on all ten campuses; manages the budget, administers and disseminates funds for all K-INBRE programs; plans, coordinates and organizes all aspects of the Annual Symposium, external evaluations and reviews, Kansas hosted Regional IDeA meetings, as well as meetings of the EAC, Executive Committee, Network Steering and Incentives & Awards Committees; provides updates and information on programs, awards, important dates and the symposium to the Communication Core for posting on the webpage/Facebook, as well as providing important key research advancements to NIH officials; provides oversight and facilitates outside reviews of grant applications; processes vendor payments and arrange travel/reimbursements to national and regional meetings for K-INBRE participants; collaborates with statewide and local funding agencies, as well as statewide bioscience entities to promote state and community engagement in biomedical research; ensures compliance with federal regulations (public access policy, acknowledgements in publications/presentations, plagiarism education).

In response to the call for proposals for Alterations and Remodeling support, applications from six KINBRE network universities submitted applications for Alterations and Renovations. These applications were reviewed by the Incentives & Awards Committee, and four applications (ESU, HINU, PSU, and WU) were selected to be submitted with this competitive renewal application. The selected applications were picked on the basis of need, university commitment, and their relevance to the scope of the proposed research at the requesting institution. In total, the K-INBRE is requesting $250,000 to assist these undergraduate universities the opportunity to upgrade their facilities, which in turn, will increase their capacity to conduct research. Working closely with the campus coordinators and administration, the K-INBRE was able to leverage substantial support from two of the four universities. This allowed the K-INBRE to stretch its resources in order to impact the greatest number of faculty and students in the state of Kansas. The letters of support are included within the individual institutional commitment letters that are located after the Overall Network and Management Plan.

? DESCRIPTION (provided by applicant): The Institutional Development Award (IDeA) Program within the National Institute of General Medical Sciences (NIGMS) promotes the development, coordination and sharing of research resources that increase the competitiveness of investigators in 23 IDeA-eligible states and Puerto Rico. The IDeA Program includes the Centers of Biomedical Research Excellence (COBRE), the IDeA Networks of Biomedical Research Excellence (INBRE), and the IDeA Infrastructure for Clinical and Translational Research (IDeA-CTR). Each is designed to enhance research infrastructure and faculty development in IDeA-eligible states. Each program has common needs and faces similar challenges, and the current proposal will enhance the success of these programs by providing a unique forum for the exchange of ideas and solutions to common problems. In partnership with the IDeA community, NIGMS IDeA officials and HelmsBriscoe, this renewal application requests support for multi-year conferences in response to NIH Support for Conferences and Scientific Meetings. The goal is to organize and convene the National IDeA Symposium of Biomedical Research Excellence (NISBRE). These NISBRE conferences provide unique technical workshops, working groups, and forums to promote best practices across IDeA states that enhance mentoring and career development. The first aim will plan and organize a biennial scientific symposium to showcase research and enhance communications among IDeA programs. A second aim will be to provide educational workshops to expand technological repertoire and career skills that improve competitiveness. A third aim will provide training of scientific skills and career development for student/trainees. A final aim will provide opportunities for concurrent parallel programs/organizations that enhance communication and sharing of best practices among IDeA programs. The proposed NISBRE will continue to incorporate innovative planning to meet the immerging interests and needs of IDeA investigators and to maximize interactions between senior scientists, junior faculty and postdoctoral fellows. We expect that at the conclusion of this five-year award we will have enhanced the cohesion within the IDeA community of scientists and positively impact the future funding opportunities for investigators at institutions within the IDeA Program.

PROJECT ABSTRACT- OVERALL The Kansas IDeA Network of Biomedical Research Excellence (K-INBRE) is a statewide network of graduate and undergraduate universities in Kansas and Oklahoma that work cooperatively to improve biomedical research in Kansas. Kansas formed such a network in 2001 (Kansas BRIN) and has successfully continued as the Kansas INBRE. The University of Kansas Medical Center (KUMC) is the Lead Institution and works closely with 2 other Graduate Partner Institutions (GPI) (University of Kansas-Lawrence, KU-L; Kansas State University, KSU;) and 7 institutions focused on undergraduate student training [Undergraduate Partner Institution or UPI: Emporia State University (ESU), Fort Hays State University (FHSU), Haskell Indian Nations University (HINU), Langston University (LU, Langston, OK), Pittsburg State University (PSU), Washburn University (WU), and Wichita State University, WSU]. The K-INBRE is focused on 1) ethnic diversity and geographic considerations in our large but sparsely populated state, 2) the power of mentored, team science, and 3) a skilled Kansas workforce and integration with existing educational and training programs. The long-range objectives are to promote multidisciplinary research networks with a focus on Cell and Developmental Biology; increase the research base and capacity through research support; provide research opportunities for trainees; serve as a pipeline for students to continue in health research careers; and enhance science and technology knowledge of Kansas workforce. To achieve the objectives of the K-INBRE, the proposal contains key elements as 4 Specific Aims and 3 Cores. Aim 1 seeks to improve the multidisciplinary research network in Kansas. The K-INBRE Administrative Core provides oversight of all K-INBRE functions and includes the Incentives & Awards Committee and the Undergraduate Research Office. The Communications Core focuses on network communications, scholarship and internal evaluations. Aim 2 enhances science and technology knowledge through sophisticated bioinformatics technology and education. The K-INBRE Bioinformatics Core continues to focus on enhanced genomics bioinformatics, bioinformatics education, outreach and education. Aim 3 stimulates basic, translational and entrepreneurial research in Kansas via mentored, interdisciplinary research opportunities. The Mentoring Core oversees 2 mentoring programs and the Developmental Research Project Program. The K-INBRE operates as a very successful program and continues to improve and expand our efforts in Cell and Developmental Biology research in Kansas. New innovations include an expansion of our pipeline to high school students and integration with existing educational/training programs in Kansas, as well as strong collaborations with other IDeA (INBRE and COBRE) programs in Kansas and in the Central IDeA Region.

PROJECT ABSTRACT Renovated, functional research space is imperative for continued growth and excellence in IDeA State universities. Here, the K-INBRE puts forth applications from 3 K-INBRE network universities requesting Alterations and Renovations at their respective universities. These applications were selected on the basis of nee, and their relevance to the scope of the proposed research at the requesting institution. In total, the K-INBRE is requesting $250,000 to assist 3 applications: Emporia State University (ESU), Fort Hays State University (FHSU), and Wichita State University (WSU). This application will allow these K-INBRE partner institutions to upgrade their facilities and increase their capacity to conduct research. ESU requests funds to renovate two spaces in Bruekelman Science Hall (SH55 and SH145) to create two modern, functional laboratories and a shared, common equipment area. ESU institutional support for this project includes up to $50,000 to cover any remaining balance, items not included in the cost estimate, and unanticipated cost overruns. ESU requests $33,468 from the K-INBRE, which is $4,474 less than the quote below. The $50,000 institutional commitment from ESU will be used to cover the difference. FHSU requests funds to renovate space within the 1st and 4th floor of Albertson Hall (AH) on the FHSU campus. On the first floor, Albertson Hall 166 Complex will be renovated into laboratory space and have an autoclave installed. On the fourth floor, AH 405 will be also be renovated into research space. A total of $100,000 is requested by FHSU for these renovations. WSU requests funds to replace the HVAC system for their Animal Care Facility (ACF) located in Hubbard Hall (HH). Alterations will include changes all of the ductwork and 8 Variable Air Volume (VAV) supply air boxes within the ACF itself and replacement of the ACF system air handling unit in the HH penthouse. The total project cost is estimated at $180,958 ($116,532 from K-INBRE and institutional support of $64,426 is committed from WSU. Overall, these alterations and renovations will have significant impact on these undergraduate institutions within the K-INBRE network in Kansas.

PROJECT SUMMARY / ABSTRACT The Institutional Development Award (IDeA) Program within the National Institute of General Medical Sciences (NIGMS) promotes the development, coordination and sharing of research resources that increase the competitiveness of investigators in 23 IDeA-eligible states and Puerto Rico. The IDeA Program includes the Centers of Biomedical Research Excellence (COBRE), the IDeA Networks of Biomedical Research Excellence (INBRE), and the IDeA Infrastructure for Clinical and Translational Research (IDeA-CTR). Each is designed to enhance research infrastructure and faculty development in IDeA-eligible states. Each program has common needs and faces similar challenges, and the current proposal will enhance the success of these programs by providing a unique forum for the exchange of ideas and solutions to common problems. In partnership with the IDeA community, NIGMS IDeA officials and HelmsBriscoe, this renewal application requests support for multi-year conferences in response to NIH Support for Conferences and Scientific Meetings. The goal is to organize and convene the National IDeA Symposium of Biomedical Research Excellence (NISBRE). These NISBRE conferences provide unique technical workshops, working groups, and forums to promote best practices across IDeA states that enhance mentoring and career development. The first aim will plan and organize a biennial scientific symposium to showcase research and enhance communications among IDeA programs. A second aim will be to provide educational workshops to expand technological repertoire and career skills that improve competitiveness. A third aim will provide training of scientific skills and career development for student/trainees. A final aim will provide opportunities for concurrent parallel programs/organizations that enhance communication and sharing of best practices among IDeA programs. The proposed NISBRE will continue to incorporate innovative planning to meet the immerging interests and needs of IDeA investigators and to maximize interactions between senior scientists, junior faculty and postdoctoral fellows. A core business workshop led by the DRIVEN Administrative Committee (Celdara Medical: Dr. Jake Reder and Dr. Julie Coleman; Simbex: Dr. Rick Greenwald and Jonathan Beckwith) will be implemented with the goal of providing basic business knowledge to assist Research Core Participants in maintaining their footprint. We expect that at the conclusion of this five-year award we will have enhanced the cohesion within the IDeA community of scientists and positively impact the future funding opportunities for investigators at institutions within the IDeA Program.

PROJECT ABSTRACT The Kansas INBRE (K-INBRE) network proposes an Administrative Core to oversee operations of the statewide K-INBRE program. The long-range objective for the Administrative Core is to provide leadership, oversight, financial and reporting duties, and oversee the evaluation components of the K-INBRE program. In addition, this Administrative Core houses 2 key components of the K-INBRE program that include oversight and decision-making of the funding selections (Incentives and Awards Committee) and oversight, tracking, and scholarship associated with undergraduate research opportunities (Undergraduate Research Office). Our purposes are to provide professional oversight to maintain this long-standing and highly functional statewide program, and to expand and integrate with existing educational and training programs operating in Kansas. The Administrative Core will function through the following 3 Aims: 1) provide administrative oversight to the overall K-INBRE program, 2) provide oversight and assistance to the two working committees in the administrative core and 3) expansion of the K-INBRE pipelines into existing development programs in Kansas, including high schools. A strength of the K-INBRE Administrative Core is the longstanding continuity and success of the experienced leadership and administrative personnel. The major functions of the Administrative Core continue to adjust to meet all compliance requirements for NIH. The Incentives & Awards Committee oversees decisions on funding and supervision of all awards within the K-INBRE. The Undergraduate Research Office supports biomedical research experiences for undergraduates on K-INBRE campuses. This Office will oversee programs directed by the 10 Campus Coordinators to attract and retain promising students into biomedically- related science majors and to involve these students in cutting edge research; and assist in the implementation of student focused symposia and workshops, and supervise the award of funds to support in unique undergraduate training opportunities. A new innovation for the current iteration of the K-INBRE is an expansion of our workforce training pipeline into the level of high school students, as well as new integration with current and existing educational and training programs in the state of Kansas. These new features will augment the success of these Kansas programs and provide access to our expansive number of K-INBRE faculty participants and laboratories.

Abstract: Emerging infectious diseases account for at least 12% of all human pathogens. Increased globalization among other factors led the World Health Organization to predict that novel infectious agents will continue to appear at an unprecedented rate. To protect society against these pathogens, it is essential to know all of the potential mechanisms by which infectious agents can cause disease. Viral infections are known to cause 15- 20% of cancers. For example, persistent genus ? human papillomavirus (?-HPV) infections cause non- melanoma skin cancers. ?-HPV?s role in these malignancies is through a novel mechanism that could be shared with emerging pathogens. Specifically, ?-HPV infections act as co-factor that along with UV, blocks DNA repair and reduces host genome fidelity. The resulting mutations can drive tumorigenesis without continued exposure to UV or ?-HPV. In addition to abundant supportive epidemiological, animal model, and cell culture evidence from other labs, we have established the ability of a ?-HPV gene (?-HPV E6) to attenuate the expression of four cellular DNA repair factors. ?-HPV E6?s inhibition of repair stem primarily from the viral protein?s degradation of a cellular transcription factor, p300. However, ?-HPV E6 simultaneously disrupts DNA repair through p300- independent mechanisms. This leaves uncertainty as to the extent that p300 disruption alone hinders DNA repair. This results in a critical need to define the role of p300 in DNA repair in a cleaner, complementary system. This proposal uses a small molecule p300 inhibitor (CCS1477) to address this need and advance towards our long-term goal of understanding the genotoxic/oncogenic potential of viral proteins that destabilize p300. Overall objective in this proposal is to define the extent that p300 inhibition by CCS1477 (i) hinders DDR signaling, (ii) increases the cytotoxicity associated with DNA damage, and (iii) exacerbates the frequency of mutations associated with DNA damage. It is an even split between two independent research groups (Chung and Wallace labs) and represents a burgeoning collaboration. Our research teams will use a combination of cutting edge techniques as well as traditional molecular biology and biochemical approaches to test our central hypothesis is that inhibition of p300 activity by CCS1477 will impede activation of three essential DDR kinases (ATM, ATR, and DNA-PKcs) reducing the cellular response to DNA damage. In the process of testing our hypothesis, we will provide excellent training opportunities for undergraduate researchers at our home institutions (Kansas State University and Pittsburg State University) and improve the overall research environment in the state of Kansas.