University Of Colorado Denver

NIH Research Projects · FY 2026 · 2013-07

Project Summary/Abstract Stimulation of hippocampal NMDA-type glutamate receptors (NMDARs) can induce long-term potentiation (LTP) or depression (LTD), two opposing forms of synaptic plasticity that are thought to be required for higher brain functions such as learning, memory and cognition. This proposal will focus on the molecular computation that mediates the decision between these two forms of plasticity. LTP has long been known to require the Ca2+/calmodulin(CaM)-dependent protein kinase II (CaMKII) and its autophosphorylation at T286 (pT286), which generates Ca2+-independent “autonomous” CaMKII activity (~20% of the maximal Ca2+/CaM-stimulated activity). In the first funding period of this proposal, we have shown that CaMKII and pT286 are additionally required for LTD. But how can pT286 CaMKII possibly mediate both of these two opposing forms of plasticity? We have started to address this question in the second funding period: LTP additionally requires CaMKII binding to the NMDA-type glutamate receptor (NMDAR) subunit GluN2B, and two mechanisms suppressed this binding during LTD. By contrast, LTD instead requires a distinct additional autophosphorylation, the inhibitory pT305/306, which blocks Ca2+/CaM binding, limits further pT286, and also directly inhibits GluN2B binding. The three regulatory CaMKII mechanisms (pT286, pT305/306, and GluN2B binding) are all thought to require the CaMKII holoenzyme structure and all cross-regulate each other. In order to elucidate how this cross- regulation works to enable the LTP vs LTD decision by CaMKII, we will here determine the specific holoenzyme rules underlying each mechanism. In three related but independent aims, the project will determine the holoenzyme rules for CaMKII phosphorylation at T286, phosphorylation at T305/306, and GluN2B interaction. A fourth aim will test effects on neuronal functions, which are expected to differ for LTP versus LTD.

NIH Research Projects · FY 2024 · 2013-07

T32 Abstract Rapid introduction of basic science discoveries into clinical fields requires close collaboration of basic and clinician scientists. Accomplishing this in otolaryngology requires a multidisciplinary approach to better define and treat the many disorders of the head and neck. Residency programs today offer limited research training for otolaryngologists, but few become independent investigators. In contrast, the basic sciences postdoctoral training offers little exposure to the clinical setting, making translational research difficult. The goal of this application is to provide research training in otolaryngology and its related sciences. Support is requested for in-depth training for 1) residents, 2) postdoctoral fellows, 3) predoctoral students and 4) short-term medical students. All trainees will receive an interactive basic research experience with ongoing exposure to and interaction of trainees in the clinical setting through conferences and courses. The pre- and postdoctoral trainees will have a 24-month block of training. Research training for MDs will begin in medical school with students doing short-term (3-month) projects and continue through the residency program with a 2-year block midway through the clinical training. One resident will be admitted each year into this research track. Early introduction and continued research involvement throughout the residency will increase our ability to attract academically oriented faculty members into the field, with backgrounds to become independent investigators. This cross-field exposure will enhance the experience for trainees and promote clinical and basic science interactions as a faculty. A major strength of the program is drawing members of the faculty from a wide variety of departments of the School of Medicine and across the UC campuses involved in otolaryngology related research. This will enhance collaborative efforts in related fields of hearing, balance, smell, taste, speech, language and head and neck cancer. All faculty members have primary or secondary appointments within the Department of Otolaryngology, creating an ideal environment for translational research between basic and clinician scientists. Through this multidisciplinary approach to research training for different levels of trainees from a variety of fields, recruitment and retention of a research-oriented academic faculty involved in research into disorders of the ears, nose and throat will be increased.

NIH Research Projects · FY 2026 · 2013-04

PROJECT SUMMARY/ABSTRACT The Clinical Research/REproductive Scientist Training Program (CREST) has trained 88 physician scientists in the conduct of clinical research, has enabled these physicians to participate in multicenter clinical trials and secondary data analyses, and has helped further their careers and create a cadre of leaders in Obstetrics and Gynecology. Didactic training in biostatistical methodology is combined with a hands-on mentored team science project (MTSP) to guide Scholars through the data analysis and publication process. Throughout the program, mentorship and networking are emphasized to provide early career interactions with academic societies within their specialties and pathways to leadership. CREST Scholars have produced a total of 1,874 unique peer-reviewed publications since the start of the program in 2005. Specific aims of the current proposal will build upon past successes to: 1. Train the Reproductive Medicine workforce to meet the national need for clinical research. It is more critical than ever to educate trainees in clinical research and apply these principles to clinical practice. We shall continue to train up to 6 Scholars per year. 2. Promote diversity in the biomedical research workforce by recruiting and training Scholars who are underrepresented in Medicine (URiM) and equipping them for success. We shall improve our data capture and followup of URiMs recruited to CREST and apply best practice to welcome new URiM candidates to the program. 3. Provide ‘holistic’ mentoring for Scholars extending to their home institutions. We shall identify an experienced, senior faculty member who will provide one-on-one support for each Scholar’s research experience as well as longer term mentoring beyond the program’s 2-year time frame. By building upon our programmatic strengths and extending the reach of our mentoring protocols, we plan to continue our exceptional success in preparing for the future of the field. CREST has trained 7 academic Division Chiefs in Ob/Gyn, one department Chair, and two Presidents of the Society for Reproductive Endocrinology and Infertility to date, along with multiple committee members of the American Society for Reproductive Medicine.

NIH Research Projects · FY 2026 · 2012-09

PROJECT SUMMARY (ABSTRACT) The long-term goal of the proposed research is to elucidate the systems and circuits, within and across brain regions, responsible for decision making. The objective of this proposal is to determine how a single target for movement is selected among competing alternatives. An experimentally-tractable approach to achieving this objective is to interrogate neural circuitry in behaving animals engaged in deciding where to move, a particularly important form of decision making for survival that we refer to here as spatial choice. The midbrain superior colliculus (SC) integrates input from several systems representing variables critical for goal-directed behavior, topographically represents contralateral spatial targets for orienting movements, and is a critical node in the network of brain regions responsible for spatial choice. However, the mechanisms underlying how spatial choices are made is poorly understood. In a series of experiments in awake and behaving mice, we test the overall hypothesis that inhibitory SC neurons, driven by local pre-motor output, mediate spatial choice by suppressing SC populations representing competing targets. The premise for this hypothesis is supported by decades of foundational research in behaving animal models and brain slices, and by our and others’ recent work leveraging the behaving mouse model to unify these two branches of research. In particular, in the previous funding cycle we found that inhibitory SC neurons do not merely suppress local pre-motor output, but instead mediate spatial choice via long-range projections. To test our overall hypothesis, In Aim 1, we examine how activating inhibitory SC neurons modulates activity across both SCs, under baseline conditions in quiescent mice and in behaving mice performing an odor- cued spatial choice task requiring selection of a left or right reward port. We then record and perturb specific projection-based subtypes of inhibitory SC neurons during spatial choice. These experiments test the hypothesis that inhibitory SC neurons shape activity at distal sites in the SC, providing a mechanism for selecting among competing spatial goals. In Aim 2, we will determine the relationship between the activity of pre-motor SC neurons and distally-projecting inhibitory SC neurons by recording and perturbing their activity during quiescence and behavior. These experiments will test the hypothesis that pre-motor SC neurons inhibit activity in populations representing distal targets by locally activating long-range inhibitory SC neurons. If successful, the outcome of this proposal will be the elucidation of the neural circuit basis for spatial choice, a key basic function of the nervous system. In addition, our research will lay the groundwork for elucidating how SC circuitry is extrinsically modulated by other brain regions to subserve goal-directed behavior.

NIH Research Projects · FY 2026 · 2012-09

The Colorado Specialized Center of Research Excellence (SCORE) on Sex Differences and Women’s Health is focused on Bioenergetic and Cardiometabolic Consequences of the Loss of Gonadal Function. The overarching scientific goal of the SCORE is to address gaps in knowledge of the impact of gonadal aging on the regulation of energy homeostasis and metabolism by conducting mechanistically driven research across the basic-to-clinical translational spectrum. The scientific premise is that gonadal aging, through the accompanying decline in sex hormones, mediates biological changes that increase risk for chronic disease. Importantly, because gonadal failure is inevitable in women in mid-life, but rare in men until much later in life, this meets the objective of the SCORE Program to study sex differences that impact women’s health. The Colorado SCORE is focused on the increased propensity for abdominal fat accumulation and the associated cardiometabolic dysfunction that are consequent to the loss of ovarian function. The overarching educational objective is to engage new investigators in research aligned with the focus of the SCORE. The SCORE will provide mentored career development and training on how to conduct research that appropriately addresses sex as a biological variable (SABV). These overarching objectives will be accomplished through the following Aims: 1) Advance novel translational research on mechanisms that are postulated to contribute to the abdominal adiposity and cardiometabolic dysfunction that accelerate following estradiol withdrawal. All projects include a focus on the kynurenine pathway because of the emerging evidence that it regulates energy homeostasis, metabolic function, and the aging process. The Colorado SCORE investigators are experts in these areas and are exceptionally well positioned to advance this research. 2) Attract and support outstanding early career investigators with research interests related to the SCORE and engage them in CEC training activities that will successfully advance their careers. The SCORE will support one Scholar and four Pilot Project Awardees each year. 3) Provide infrastructural support for the SCORE Research Projects, Scholars, and Pilot Awardees through the LAC, including biostatistics, data management, financial oversight, and administrative support. 4) Partner with and leverage resources provided by other programs at CU-AMC (Multidisciplinary Center on Aging, Ludeman Center for Women’s Health Research, Nutrition Obesity Research Center, Clinical and Translational Sciences Institute) to elevate the visibility and multi-disciplinary reach of women’s health research and the importance of investigating SABV. Activities will target both scientific and lay communities at local, regional, and national levels. Achieving these Aims will position the SCORE as a national leader in women’s health research, and the CEC as a model program for training the next generation of scientists to integrate SABV in their research with a goal of improving women’s health.

NIH Research Projects · FY 2026 · 2012-06

Project Summary/Abstract Mucosal inflammatory diseases such as Crohn's disease and ulcerative colitis, collectively termed Inflammatory Bowel Disease (IBD) remain among the most debilitating chronic disorders of the western world. It is estimated that more than 3 million Americans suffer with IBD, with incidence rates on the rise in many populations. The precise etiology of IBD is not known but emerging evidence implicates shifts in the constellation of microbes in the intestine (dysbiosis) as a contributing factor. This proposal is focused on a precise understanding of the role of microbiota-derived molecules in promoting intestinal resiliency during ongoing inflammation. In particular, we have identified a class of microbiota-sourced molecules derived from dietary tryptophan, termed indoles, that contribute fundamentally to mucosal barrier function and wound healing. Our work in progress has focused on defining endogenous host molecules that interact with indole(s) and become potential targets for the development of new drugs to treat IBD. Ongoing unbiased work has identified neutrophil myeloperoxidase (MPO) as a candidate target for indole(s) with potential importance in promoting healthy barrier function. In this proposal, we hypothesize that this indole-MPO axis determines the extent of bystander tissue damage in active mucosal inflammation. Three synergistic specific aims are proposed to address this goal. In Aim 1, we will profile the direct impact of neutrophil MPO on epithelial tight junction proteins in modeled inflammation. Aim 2 will elucidate the specificity of indole derivatives on MPO inhibition and suppression of epithelial junctional damage. Specific Aim 3 will test the indole-MPO axis in multiple models of intestinal inflammation. Results from these experiments will provide new insights into innate regulation of mucosal barrier and an expanded physiological role for indoles produced by commensal bacteria. The studies proposed in this project are intended to elucidate a novel role for microbial metabolites in innate immune responses during mucosal inflammation using in vitro and in vivo models. This work will provide new insight to the development of improved therapeutic approaches for treating IBD.

NIH Research Projects · FY 2025 · 2011-09

Center Overview: Project Summary American Indians and Alaska Natives (AI/ANs) are at substantial risk of diabetes. They exhibit rates of diabetes often greater than other citizens; they suffer serious, debilitating complications thereof; a disproportionate share of their scarce health care resources are consumed by a small percentage of patients with diabetes. A growing body of knowledge about evidence-based policies, programs and practices promises to redress these disparities. The Center for American Indian and Alaska Native Diabetes Translation Research: 1) Provides an administrative structure that promotes diabetes-related translational research capacity. Its Pilot and Feasibility Program supports small-scale, innovative projects; 2) Offers related training, technical assistance, and consultation to investigators and key stakeholders through an Enrichment Program; 2) Sustains and expands a Research Base of funded faculty whose research either directly targets diabetes prevention and treatment or is translational in nature with clear potential for application to diabetes translational research; 3) Supports a Translational Research Core that offers resources in the cultural adaptation of interventions, health literacy, health technologies, dissemination and implementation science, cost of care/ sustainability, and qualitative research methods to advance a culturally grounded, multidisciplinary problem-oriented translational research program of major scientific and programmatic importance to Native people; 4) Serves as a National Resource Core for other investigators and programs pursuing diabetes translational research with AI/AN communities; and 5) Establishes a Community Outreach and Engagement Core that promotes the dissemination of lessons learned and ensures key stakeholder participation in their extension to this special population.

NIH Research Projects · FY 2025 · 2011-08

This R13 meetings grant application seeks partial support for three upcoming annual Alcohol and Immunology Research Interest Group (AIRIG) meetings in conjunction with either the Midwest Autumn Immunology Conference (AIC) in Chicago, IL, the Research Society on Alcoholism (RSA), the Society for Leukocyte Biology (SLB), the Shock Society or Keystone Symposia. Funding is requested to cover travel costs and housing for students, postdoctoral fellows, junior faculty, and meritorious scientists, as well as some of the costs for invited speakers, for a one-day meeting which will cover broad aspects of alcohol and immunology. The 2023 AIRIG meeting will be held at the University of Colorado Denver (UCDenver) Anschutz Medical Campus on Friday December 8, 2023.The topic will be “Alcohol, Infection & Immunity.” This full-day AIRIG meeting will include 3 plenary sessions: 1) Alcohol, infection and host immunity, 2) Alcohol, inflammation and tissue damage, and 3) Alcohol and the gut-brain axis, along with posters, short presentations selected from submitted abstracts and a lunch time mentoring/educational workshop. We will hold the 2024 AIRIG meetings at Loyola University Chicago (Loyola) and 2025 AIRIG meeting as a satellite in conjunction with the RSA meeting in New Orleans, LA. Themes of the latter two meetings will be “Alcohol, Inflammation and Tissue Injury” and “Alcohol and Multi-Organ System Responses.” The 2024 meeting will be held the day before the annual meeting of the AIC. The AIC is a regional meeting with an impressive list of invited plenary speakers and ~440 registrants, while RSA meetings are much larger, especially when held along with ISBRA. Holding the AIRIG meeting with AIC and RSA will allow greater visibility of alcohol and immunology research. Additionally, these meetings have numerous opportunities for young developing scientists to participate on many levels, including selecting a large number of oral presentations selected from submitted abstracts. In addition, these meetings have Career Development Programs, including sessions on Grant Writing. These sessions allow time for career building and collaborative opportunities, which may not be feasible or readily available at other meetings. Thus, in addition to providing a forum for the discussion of current research in the field, overall goals of the co-organizers are to expand the participation of students, postdoctoral trainees, and other meritorious scientists and to increase the critical mass of scientists working on alcohol and immunology issues. We believe that these goals will be best achieved by scheduling upcoming AIRIG meetings in conjunction with the AIC and RSA.

NIH Research Projects · FY 2026 · 2011-08

PROJECT SUMMARY/ABSTRACT The Colorado Pulmonary Alcohol Research Collaborative (CoPARC) Resource, centered at the University of Colorado Anschutz Medical Campus (CU Anschutz), provides an essential service for investigators to conduct clinical and translational research to understand the pathogenesis of alcohol misuse in pulmonary infections (particularly community-acquired pneumonia, or CAP) and their sequelae. In 2011, CoPARC launched an infrastructure to obtain and distribute biospecimens and data from otherwise healthy participants with well- characterized alcohol misuse, and healthy controls, including bronchoalveolar lavage and bronchial brushings obtained by bronchoscopy, and blood. During the last renewal, CoPARC's repertoire expanded to meet evolving investigator requests, with addition of new biospecimen types (e.g. stool, nasal epithelial brushings), new clinical data (e.g. assessment of neuromuscular weakness), and new participant cohorts (e.g. inhaled cannabis users). Expansions provided novel opportunities for studies of the gut-lung brain axis, and dual use disorders, among others. The last renewal also featured new enrollment of critically ill patients with respiratory failure and the acute respiratory distress syndrome (ARDS) that often complicates CAP in association with alcohol misuse. Biospecimens collected from critically ill patients include BAL, tracheal aspirates, serial blood samples, and stool. Infrastructure was also implemented to longitudinally assess patients who survive critical illness for up to a year post-discharge to evaluate domains of respiratory, physical, and mental health, including substance use habits. R24 Supplement funding provided an added opportunity to study critically ill patients with Coronavirus (COVID)-19. CoPARC has had consistent support from consortia (Emory University, Louisiana State University, Loyola University Chicago, University of California San Francisco, and University of Nebraska Medical Center) to enroll participants and patients in parallel who augment the size and representativeness of CU Anschutz cohorts. For the renewal, CoPARC's long-term objective is to magnify the utility and impact of existing Resource services based on evolving needs of investigators, under the guidance of a committed Steering Committee. Aims for CoPARC's renewal include: 1. Generate and diversify the Resource's biological specimen and data inventory to facilitate research targeting the impact of alcohol misuse across pulmonary and extra-pulmonary organ axes, based on dynamic and evolving investigator requests. 2. Expand access to biorepositories and data generated through a) consortia arrangements, and b) new multi- center clinical research and data networks, whose participants have well-characterized alcohol use habits, to enable clinical and translational research on a larger scale. 3. Strengthen connections in the pulmonary-alcohol research community through novel services provided by CoPARC to enhance impactful, collaborative research, and disseminate Resource content and availability.

NIH Research Projects · FY 2026 · 2011-06

The specific aim of our competitive 11-15 year renewal is to continue our short-term research education program that provides underrepresented minority undergraduate students (“diversity trainees”) with an inspirational, perspective and career transforming biomedical research experience in the Colorado Undergraduate Summer Program (“CUSP”). Overview: CUSP uses NHLBI funding to recruit and enroll 8 diversity trainees each summer from colleges nationwide. Trainees prepare for and conduct laboratory research for 9 weeks with an experienced investigator/mentor. Trainees design their own project, perform hands-on research, attend daily interactive research discussions, and present their research at our concluding school-wide graduation poster session. Trainees also participate in complementary clinical conferences, receive career advice from admissions directors, faculty, and students, shadow physicians in clinical settings, and are integrated into a wide range of supportive social and academic diverse campus communities. Innovative Aspects: CUSP (1) is a well established program that consistently attracts many top applicants from colleges nationwide, (2) uses a holistic selection process that targets students who are not committed to biomedical research careers, (3) amplifies learning by adding additional diversity and non- diversity trainees using additional non-federal matching support from participating colleges, donors, and CU, (4) is located on a new all inclusive contiguous medical campus that offers many research and learning opportunities, (5) employs a unifying inflammatory, immunologic, and oxidative stress related mechanisms focus that is relevant to NHLBI related health and disease, (6) emphasizes using experienced enthusiastic diversity and non-diversity mentors, (7) embodies a comprehensive instructional and laboratory program that engages trainees by integrating research training, medical perspectives, career counseling, campus tours, and social activities, (8) provides additional shadowing, educational, and social activities, and (9) is independently reviewed by education evaluation experts working at CU. Program Success: Independent evaluations indicate that CUSP diversity trainees (1) appreciate and embrace all aspects of CUSP, (2) become more knowledgeable and enthusiastic about research and research careers, (3) continue CUSP mentor relationships, (4) conduct research again, (5) frequently enter health professions schools, and (6) actively promote CUSP to new trainee candidates. Program Director: John E. Repine, MD will continue his leadership as an experienced physician- scientist-teacher-mentor who founded and directed CUSP for the last 9 years. Significance: CUSP motivates and prepares biomedical research uninformed diversity trainees for continuing in biomedical research and undertaking biomedical research and health care related careers.

NIH Research Projects · FY 2024 · 2011-04

Abstract This competing renewal application is to continue our studies on the role of hybrid insulin peptides (HIPs) as neo-epitopes for autoreactive CD4 T cells in autoimmune diabetes. During this very productive period we have established that HIPs formed from insulin C-peptide and other β-cell granule proteins are antigens for large numbers of autoreactive CD4 T cells in the pancreas of NOD mice, can be used in strategies to induce antigen-specific tolerance, and are also present in human islets and PBMC of patients with type 1 diabetes (T1D). Our objective in the next period of the award will be to characterize new insulin (Ins) B- chain HIPs and T cells reactive to these HIPs, in both NOD mice and human subjects. We hypothesize that antigenicity of the much studied insulin B chain peptide, B:9-23, is due to hybrid peptide formation between sequences of B:9-23 and cleavage peptide products of other granule proteins. The rationale for pursuing these studies is based on preliminary data indicating that B-chain HIPs containing B9-23 sequences are strong agonists for insulin-reactive T cell clones and that T cells reactive to these HIPs can be identified in the polyclonal T cell population of NOD mice. Our aims for this next project period are to (1) investigate the role of B-chain HIP-reactive T cells in disease pathogenesis; (2) determine whether tolerance can be induced with insulin B-chain HIPs coupled to biodegradable nanoparticles in NOD models of spontaneous disease and islet transplantation; and (3) investigate the presence of B-chain HIP-reactive CD4 T cells in human T1D and develop new human tetramers containing B-chain HIPs. The B-chain HIPs will provide further tools for understanding the autoimmune response to pancreatic β-cell antigens in general, and specifically the contribution of insulin-reactive T cells to the disease process. We predict that these studies will lead to generation of reagents not only for tracking disease-relevant T cells, but also for developing and expanding approaches to antigen-specific therapy.

NIH Research Projects · FY 2025 · 2010-08

SUMMARY. The programmed death receptor 1 (PD-1) drives immune escape in head and neck squamous cell cancer (HNSCC), whose incidence is rising due to human papillomavirus (HPV). Inhibitors of PD-1 (PD-1i) in T cells, or its ligand in cancer cells (PD-L1i), are in use in many cancers including recurrent/metastatic (R/M) HSNCC, but most patients do not respond. While the PD-L1 extracellular domain (ECD) is a ligand mediating T- cell anergy, the intracellular domain (ICD) role is less known. We found that PD-L1 over-expression increased key pro-tumorigenic properties. ICD deletion disrupted this signaling and reversed the pro-growth features. We hypothesized that PD-L1 ICD interacts with intracellular proteins, and to define the PD-L1 interactome we used a proximity-dependent labeling assay to measure interacting proteins with/without PD-1 binding. The PD-L1 interactome was consistent across cell lines, and top partners included nuclear factor of activated T cells (NFAT) subunits interleukin enhancer-binding factor 2 and 3 (ILF2/ILF3), that stabilized the signal transducer and activator of transcription 3 (STAT3) to induce pro-growth properties. In humanized mouse models of HPV+ and HPV- HNSCC, PD-1 binding induced PD-L1 signaling, and combined PD-L1i and STAT3 inhibitors (STAT3i) were required to achieve durable tumor control. In sum, PD-1 binding to PD-L1 led to an elegantly coordinated effect of simultaneous immune evasion and tumor progression, the latter driven by STAT3. In this project we will study the full spectrum of the PD-L1 interactome in HNSCC from its mechanistic basis to in vivo validation, and then patient clinical testing. First, we will identify the ICD precise domain responsible for the PD-L1 interactions with its cytoplasmic targets, by deploying HNSCC (and melanoma as control) cell lines with a set of mutated ICD constructs on a PD-L1 knockout by CRISPR/Cas9 backbone. PD-L1 interactome and signaling disruption will be assessed by changes in intrinsic and extrinsic properties. This will be followed by structural modelling and analyses of PD-L1 interactions that will enable discovering pharmacological disruptors. Secondly, we will test the role of STAT3 signaling in PD-1i response in a HNSCC humanized model with patient-matched, autologous thymic education capability. To elucidate complex immune events, in vivo testing is key, and to be relevant, mouse models need to be able to elicit a true, immune-based antitumor response. Thirdly, we will conduct a proof-of-concept trial of the STAT3i NT219 combined with the approved PD-1i pembrolizumab in PD-1i-naïve HNSCC patients. The planned immuno/pharmacodynamic and efficacy testing will explore if combined therapy increments response, and if baseline parameters including STAT3 signaling expression predict efficacy, setting the stage for practice-changing trials. The notion that the PD-1:PD-L1 axis, the most validated checkpoint inhibitor pathway in human cancer, is a bi-directional and synchronized process rather than a simple ligand- receptor step is a paradigm shift. By understanding canonical PD-L1 signaling and the coordinated role of the PD-L1:PD-1 interaction we will unlock the full potential of immune modulation as an anticancer therapy.

NIH Research Projects · FY 2025 · 2010-07

Summary The fundamental role of the immune system is to distinguish self from non-self, and healthy from unhealthy tissue. The ligands and receptors encoded by the major histocompatibility complex (MHC), the killer cell immunoglobulin-like receptor (KIR), and the natural killer complex (NKC), interact to help accomplish this role. Successful pathogen defense drives the genetic diversity of these regions but may also lower the threshold for autoimmunity. At the extremes of this spectrum is HLA-B*27, which protects against certain chronic infections but is also the major risk factor for spondyloarthritis (SpA) and acute anterior uveitis (AAU). Spondyloarthritis (SpA) refers to inflammatory musculoskeletal diseases, axial SpA (includes ankylosing spondylitis), psoriatic arthritis (PsA), colitis-associated arthritis, and reactive arthritis (ReA). Painful, inflamed joints are a common feature of SpA. Progression of axial disease leads to spinal fusion and reduced mobility. AAU, inflammation of the anterior chamber of the eye, commonly occurs in SpA patients, and can cause blindness. Although recent therapeutic advances have improved symptom management, whether they also improve long-term outcomes is controversial. The frequency of HLA-B*27 is high among these groups, especially in axial SpA, with up to 90% carrying HLA-B*27. About 2 million adults in the United States have SpA, with an estimated minimum of 44 million globally. Thus, SpA carries a significant health and economic burden, and there is a pressing need to understand more about the underlying disease mechanisms. The MHC region and KIR are associated with SpA. The receptor KIR3DL1 recognizes HLA-B*27 as a ligand, but how this relates to SpA is unknown. Given the shared associations but distinct manifestations of SpA, PsA, ReA and AAU, we hypothesize that the genetic polymorphism of KIR3DL1, in combination with the MHC, determines risk for specific SpA subtypes. Although genome-wide association studies have contributed to understanding the genetic landscape of SpA, the diversity of KIR and MHC limit the utility of these methods for uncovering more precise allelic associations. The Norman Lab is uniquely equipped to answer how the genetic diversity of the MHC, KIR, and NKC impacts SpA disease susceptibility and severity. In Aim 1, we will use our sequencing and bioinformatics approach to analyze these loci at high throughput and resolution from a total of 3,335 patients and 4,165 controls, with additional replication cohorts. Our cohort includes patients with axial SpA, PsA, colitis-associated arthritis, ReA, and AAU. We will generate per-individual haplotypes and fine-map the associations and interactions underlying SpA. In Aim 2, we will focus on the relationship between KIR3DL1 and HLA-B*27, by investigating how KIR3DL1 allotypes impact NK cell responses in axial SpA. In Aim 3 we will apply our targeted long-read sequencing methods to the extended MHC region and develop a method to reliably infer allele-specific expression of MHC, KIR, and NKC from single-cell RNA sequencing. Through these Aims, we will supply in-depth genetic analysis of MHC, KIR, and NKC in SpA, determine how KIR3DL1 allotypes impact NK cell function in disease, significantly advance the field with reference haplotypes and innovative bioinformatic methods, and provide a model for how to elucidate HLA/KIR associations in general.

NIH Research Projects · FY 2024 · 2010-07

This is a competing renewal application for a short-term research education program originally funded as a T35 in 2000 and as an R25 in 2010. Our major objective is to continue providing annual short-term research education experiences for highly motivated students from under-represented backgrounds in order to expose them to biomedical research in the area of pulmonary and cardiovascular disease. Locally known as GEMS (Graduate Experiences for Multicultural Students), over the past 10 years, >138 undergraduate (UG) and 29 health professional students (HPS); >125 supported by the R25 and the rest supported by other programs. Collectively, these students have published 93 manuscripts; >70% have earned terminal degrees, are working in science or health-related fields or are still enrolled in school. More than 70% of student participants were under-represented ethnic minorities. The program builds upon our established infrastructure and uses the significant strengths of one of the top pulmonary medicine programs in the country. We continue the tradition of addressing the pipeline by requesting 10 undergraduate and 4 health professional student slots. Here, in addition to the usual didactic and hands-on research activities, we will use the model of academic “coaches” who are not intended to supplant the mentor, but rather complement this relationship. Coaches will be past GEMS participants who are still at Anschutz Medical Campus. Coaches will maintain contact with the students throughout the year and will guide them through a successful career path. Furthermore, to ensure student success, we propose to use social science approaches and provide the students with a toolkit that will create an environment, a community of practice, where they feel safe to talk about personal, academic and professional issues and to bond through shared norms and values. We will also implement implicit bias workshops and mentoring best practices for students and mentors. We incorporate a set of targeted questions in the application that will aid in selection of students highly motivated to pursue biomedical research. We believe that these approaches will continue the GEMS tradition of excellence in training students from under-represented backgrounds while at the same time enhancing student’s academic success beyond the summer GEMS internship.

NIH Research Projects · FY 2025 · 2010-05

PROJECT ABSTRACT Vision healthcare should be based on science. Cochrane is the only international organization that has as its core mission to synthesize (via systematic reviews) and keep up to date the world’s literature for all of healthcare. Systematic reviews use a highly structured and reproducible methodology to identify, appraise, and synthesize available evidence for specific clinical questions. Cochrane Eyes and Vision (CEV) aims to prepare and promote access to systematic reviews of interventions used to prevent, diagnose, or treat eye conditions and visual impairment. Since 2002, the CEV US Project (CEV@US) has published 101 systematic review protocols, 104 systematic reviews, and 75 methodological papers and book chapters; has educated over 100,000 individuals in methods for systematic reviews; and has informed 121 clinical practice guidelines in the US and internationally. Since its start, CEV@US has been laying the groundwork for having an impact on practicing clinicians: demonstrating to all who are interested how to conduct an excellent systematic review (e.g., education, journals, methods research), how to apply this knowledge (e.g., clinical practice guidelines and decision-support applications), and forming important partnerships (e.g., with clinicians and patients/consumers). The focus of our 2022-2027 competitive renewal is impact – impact of CEV reviews on clinical practice. Aim 1: We will prepare systematic reviews that address prioritized questions in collaboration with clinicians and methodologists, publish the reviews, and regularly update them as new research emerges. We will apply state-of-art methods in producing these reviews. Aim 2: We will partner with individual (e.g., clinicians, patients) and organizational stakeholders (e.g., professional societies, academic medical centers, journals) to ensure that the knowledge underpinning clinical practice is reliable, available, and applied. Aim 3: We will build research capacity by educating health professionals. Our education is flexible and tailored, and is offered online and in-person, via professional organizations, clinical programs, and one-time workshops. Aim 4: We will conduct foundational methodological research related to systematic reviews and influence the field of evidence synthesis. We will evaluate the impact CEV@US activities using a mixed-method study. Aim 5: We will promote evidence-informed decision making by disseminating the results of our research widely.

NIH Research Projects · FY 2026 · 2009-08

PROJECT SUMMARY In many lung diseases--including asthma--excessive mucus disrupts clearance and obstructs airflow, but mucus dysfunction is not effectively treated by existing therapies. The chief macromolecules in airway mucus are MUC5AC and MUC5B. To study them causatively, we made Muc5ac and Muc5b knockout mice. We discovered that Muc5b is required for mucociliary clearance (MCC) in health, but Muc5ac is dispensable. Instead, Muc5ac causes mucus plugging and is required for airway hyperreactivity (AHR) in models of asthma. Nonetheless, despite its homeostatic requirements in mice and humans, we now know that excessive MUC5B/Muc5b can itself be detrimental in lung fibrosis (PF). These findings show significance, but they also highlight the need to find ways to prevent mucus dysfunction while also preserving defense. We postulate that this can be accomplished in part through an improved understanding of MUC5AC and MUC5B assembly mechanisms. MUC5AC/Muc5ac and MUC5B/Muc5b are very large proteins that form even larger polymers via linkages between their carboxyl (C-) and amino (N-) termini. Their pathologic properties depend on covalent disulfide bonds whose reduction reverses AHR and improves MCC, suggesting that therapeutic intervention targets could be revealed by determining precisely how mucins assemble. Mucin C- and N-terminal polymerization domains are homologous with the protein von Willebrand Factor (VWF). Cysteines required for VWF assembly are conserved in mucins. We have identified which are required for airway mucin assembly and function. Here we seek to continue determine structural requirements and cellular mechanisms for mucin polymerization and function. Emerging data suggest that a cluster of three cysteines referred to as a “cysteine triad” is crucial. One of member of the triad is the cysteine that forms an inter-molecular disulfide. This requires the acidic environment provided in the Golgi to facilitate disulfide exchanges that liberate cysteines needed for intermolecular disulfides. Low pH also protonates histidines in domains that surround the triad. The cysteine triads and pH-sensing histidines in VWF are conserved in mucins. We hypothesize that airway polymeric mucin biosynthesis is regulated by Golgi specific mechanisms that mediate mucus function and dysfunction. We will test this by 1) examining how assembly is regulated by Golgi-localized cysteine-triad dependent mechanisms, 2) determining the conserved histidine residues are required for pH-dependent N-terminal assembly, and 3) demonstrating how these affect mucin polymerization airway mucus functions. Studies here will advance the mucin biology field while laying groundwork for approaches to prevent mucus dysfunction while preserving defense. While research here focuses on asthma, results will have implications for diseases such as chronic obstructive pulmonary disease (COPD), CF, and other conditions where mucus dysfunction is prevalent.

NIH Research Projects · FY 2025 · 2009-08

Project Summary/Abstract Significant research efforts are currently underway to understand and prevent the pathogeneses of Type 1 and Type 2 diabetes, both of which are associated with the gradual loss of functional insulin-producing b cells. Progress has been made with using human pluripotent stem cell (hPSC) populations as alternative sources of islet cells; however, the efficient production of pure populations of fully functional b cells has not yet been achieved. Furthermore, the current protocols result in highly variable differentiations depending on the stem cell source, differentiation procedure and user. In addition, both endogenous and stem cell-derived islet cells display inherent phenotypic instability and often undergo spontaneous dedifferentiation and/or reprogramming in pathophysiological conditions, which results in the loss of cellular identity and phenotypic dysfunction. These challenges suggest that there is still a great need to identify the precise transcriptional and epigenetic mechanisms that promote the specification and maintenance of islet cell identities. We and others have demonstrated that development and maintenance of b cell identity requires the continuous activity of several non-redundant transcriptional programs. Although many of the transcription factors that are essential for maintaining a and b cell phenotypes have been characterized, we lack a complete understanding of how cell-specific gene programs are regulated to specify and maintain these closely related cell lineages in mice, and there are varying reports related to their respective roles in human pancreas development. This proposal will allow us to gain a more complete understanding of the basic molecular mechanisms underlying islet cell differentiation and the maintenance of islet cell identity/function; this knowledge will pave the way to improved human islet cell differentiation protocols and the identification of better treatments for islet cell dysfunctions in diabetes. These studies are facilitated by increased access to novel and improved molecular technologies. Furthermore, our studies in rodent models are providing important biological and physiological context that will be directly translated into the hPSC differentiation system. Our first aim will characterize the differential molecular mechanisms that regulate a vs b cell programs in mice. We will focus on the combined molecular activities of NKX2.2, NKX6.1, KLF4 and CHD4 in regulating endocrine cell fate decisions. We will also take an unbiased approach using combined single cell ATAC-Seq and RNA-Seq to characterize the transcriptional landscape associated with islet cell fate decisions in vivo. The second aim will characterize the role of the chromatin modifier protein CHD4 in regulating b cell development and function, and in modulating NKX2.2 and NKX6.1 regulatory activities. The third aim will begin to apply this knowledge to the directed differentiation of human islet cells from hPSCs. We propose to identify the necessary and sufficient functions of NKX2.2 and NKX6.1 in regulating a and b cell fate decisions. We will also use this platform to identify the chromatin and gene expression landscapes regulated by NKX2.2 and NKX6.1 during these lineage choices.

NIH Research Projects · FY 2026 · 2008-07

The alternative pathway of complement contributes to a wide range of glomerular diseases. Factor H is the main regulator of this innate immune cascade. Although some patients with kidney disease have underlying defects in factor H, complete deficiency of the protein is quite rare. It is not clear why kidney is so frequently the target of alternative pathway-mediated injury. One possible explanation is that endogenous proteins can function as factor H antagonists, effectively creating microenvironments locally deficient in the regulator. The factor H related proteins (FHRs), for example, antagonize binding of factor H to tissue surfaces, including the glomerular basement membrane (GBM). We have also discovered that another class of proteins - the annexins - act as factor H antagonists on kidney surfaces. Although the FHRs and annexins probably block factor H from binding other tissues in the body, the high concentration of alternative pathway proteins within the glomerular capillaries causes preferential activation at this location. Based on these findings, the primary hypothesis of this grant is that is that the GBM is uniquely susceptible to alternative pathway activation because the dysregulatory proteins (FHRs and annexins) effectively block factor H from binding this surface, yet it is simultaneously exposed to high concentrations of activating proteins. Alternative pathway activation within the glomerulus generates C3a and C5a, which ligate receptors on resident myeloid cells and create an inflammatory milieux within the kidney. Complement fragments generated in the kidney also stimulate T cells and B cells systemically. To test this hypothesis, the following specific aims will be pursued. Aim 1) Characterize the effects of dysregulatory proteins in the kidney. We will use in vitro and in vivo models to explore the molecular mechanisms that cause alternative pathway activation in the kidney. Aim 2. Examine the downstream effects of AP activation in the kidney. In this aim we will explore the downstream effects of glomerular complement activation on cells throughout the kidney. We will also examine whether complement activation in the glomeruli affects the adaptive immune response systemically. This could represent a mechanism by which inflammation in the kidney increases production of autoantibodies, creating a positive feed-back loop. Aim 3. Test novel strategies for blocking complement activation in the kidney. In this aim we will test the ability of novel engineered proteins to reverse alternative pathway dysregulation in the kidney. The project is expected to provide an overarching explanation for why the kidney - among all the organs - is so uniquely susceptible to alternative pathway-mediated injury. These studies will also test strategies for specifically blocking kidney inflammation. By targeting the underlying molecular triggers of complement activation in the kidney, drugs can be designed to inhibit these pathologic processes without blocking complement-mediated immunity systemically. This approach is expected to be both more effective and safer than currently available treatments.

NIH Research Projects · FY 2024 · 2008-01

The long term goals of this project are to understand the molecular and biophysical mechanisms for the regulation of cardiac pacemaking in sinoatrial node myocytes (SAMs) across the gamut of physiological conditions. SAMs function as cardiac pacemaker cells by firing spontaneous action potentials (APs). As in other excitable cells, the precise shape of sinoatrial APs reflects the composite activity of the unique complement of ion channels and transporters on the plasma membrane. AP waveforms are not static; they vary in response to short- and long-term changes in physiological context. In principle, differences in AP waveforms should lend insight into the changes in ionic currents that underlie cellular electrophysiological responses. However, our ability to decode the causal relationships between ionic currents and AP shape remains an elusive goal in all excitable cells. This gap in understanding is caused by a lack of information about AP waveforms and currents in different physiological contexts and by difficulties inherent to the study of interrelated systems using conventional research approaches. The present proposal addresses these general questions by focusing on the mechanisms by which aging slows cardiac pacemaking. Proposed experiments follow from work in prior funding periods and new preliminary data which show that aging slows pacemaking in part by decreasing the spontaneous AP firing rate of SAMs in association with changes in a limited subset of AP waveform parameters and reductions in the funny current (If) and voltage-gated Ca2+ currents (ICa,L and ICa,T). They also address the prior observation that age-dependent reductions in pacemaker activity and If in SAMs can be reversed by high concentrations of exogenous cAMP via a cAMP-mimetic mechanism. Proposed experiments will use new research tools developed during the current funding period (1) to define age-dependent changes in the relative contributions of currents active during different phases of the AP in SAMs, (2) to test the ability of different currents, singly and in combination, to transform the AP phenotype of young and aged SAMs, and (3) to test the hypothesis that age-dependent reduction in a novel If regulatory protein is responsible for the hyperpolarizing shift in voltage-dependence and resulting slowing of AP firing rate in SAMs and heart rate in mice. Results of these studies will experimentally define for the first time causal links between individual ionic currents and AP waveform parameters in SAMs that are responsible for cardiac pacemaking in general and will reveal how these mechanisms are changed during normal aging.

NIH Research Projects · FY 2025 · 2007-09

Project Summary This application is for the 4th cycle of funding for the Colorado Building Interdisciplinary Research Careers in Women’s Health Research (BIRCWH) K12 program, established in 2007. Over the past 15 years, we have had 21 Scholars in the program, 18 of whom have remained in academic medicine and are funded. Collectively, these Scholars have published over 550 publications since their time in the BIRCWH program (not including abstracts). For our proposed 4th cycle, we will continue to work with our Scholars and their carefully chosen primary mentors in the rich academic environment of the University of Colorado-Anschutz Medical Campus. We will help them form innovative interdisciplinary Scientific Advisory Committees (SACs). Scholars and their entire SACs as well as BIRCWH leadership will meet twice yearly and participate in monthly trainings using an evidence-based curriculum focused on developing academic and career development skills. In addition, learning from our experience in 3 prior cycles of BIRCWH and in addition to our successful ongoing methods, we propose several key program innovations: o The evidence-based curriculum now will also include focus on how to integrate sex and gender into all steps of research from study design to analysis. o Encouraging Scholars to begin working on their individual K award relatively early in the course of their tenure as a BIRCWH Scholar (i.e., 8-9 months after initial BIRCWH Scholar appointment) with strong mentor/SAC/BIRCWH leadership support. o Ongoing interaction for BIRCWH Scholars with their SACs following completion of the BIRCWH Scholar period (through the next academic steps) to extend guidance while also working to establish independence. o Addition of bioinformatics as a cutting edge, cross-cutting tool that will serve to interweave our foci. o Increased biostatistician availability to meet the complex statistical needs of the Scholars. o Addition of a BIRCWH facilitator, Dr. Laura Brown, a successful former BIRCWH Scholar, who will work with Scholars by focusing on research and career development issues including helping them work on their next grants- grants that will help them transition successfully to their post-BIRCWH careers. o A strong focus on working to increase diversity of our BIRCWH Scholars. Our overall goal is to: Support top applicants from a diverse pool of exceptionally motivated and talented individuals, and provide them with an appropriately tailored, top-quality interdisciplinary mentored research and career development experience. The excellent institutional support for the Colorado BIRCWH, with strong focus on women’s health, will also fuel the growth of this important program which aims to develop successful, independent interdisciplinary scientists in women’s health and sex differences research and thereby improve the health of women.

NIH Research Projects · FY 2025 · 2007-07

The University of Colorado Anschutz Medical Campus Computational Bioscience Program (CPBS) Program is an independent, Ph.D.-granting and postdoctoral training program based in the University of Colorado School of Medicine, with a 20-year track record of innovative and effective training of pre- and post-doctoral fellows for research careers. We are a second-generation teaching program, informed by the experience of the many biomedical informatics training models that have come before us. Our program is designed to produce graduates with depth in both computational methods and biomedicine, an intimate familiarity with the science and technology that synergizes the two, and the skills necessary to pioneer novel computational approaches to significant biomedical questions. We are aware of the difficulty of achieving both breadth and depth in a reasonable amount of time, and believe we have identified a novel approach that is capable of training productive interdisciplinary scientists in a relatively short period. The program is tightly focused on transforming already strong students and recent Ph.D.'s into mature and productive scientists. Our program is structured around a set of four categories of educational goals and objectives: knowledge, communication skills, professional behavior, and self-directed life-long learning. Our graduates demonstrate the knowledge of core concepts and principles of biomedical informatics, and have the ability to apply computation to gain insight into important biomedical problems. Their knowledge includes mastery of the fundamentals of biomedicine, clinical and translational research, statistics, and computer science, as well as proficiency in the integration of these fields. Our graduates have contributed to the discovery and dissemination of new knowledge. They demonstrate interpersonal, oral, and written skills that enable them to interact productively with scientists from both the biomedical and the computational domains, to communicate the results of their work in appropriate formats, and to teach others biomedical informatics skills; they effectively bridge the gap between biomedical and computational cultures. Our graduates demonstrate the highest standards of professional integrity and exemplary behavior, as reflected in a commitment to the ethical conduct of research, continuous professional development, and thoughtfulness regarding the broader implications of their work. Our unique approach to teaching computational bioethics has been adopted by many others around the world. Our graduates demonstrate habits and skills for self-directed and life- long learning, and recognize that biomedical informatics is a rapidly evolving discipline. Our program itself is also undergoing continuous improvement, carefully tracking our efforts and quickly responding to changes in the field and in our situation. We are justifiably proud of our outstanding track record as well as of our dynamic and adaptive approach to the training of adept, flexible, and curious scientists able to comfortably assimilate new ideas and technologies during the course of their professional careers. Based on our successful track record, we are requesting that our current slot allocation be increased to 9 predoctoral, 6 postdoctoral and 4 short term positions.

NIH Research Projects · FY 2026 · 2007-01

PROJECT SUMMARY Due to the overwhelming success of the new mRNA vaccines during the SARS-CoV2 pandemic, vaccines have entered the public consciousness to a level not seen since the days of smallpox eradication and the polio epidemic. It seems reasonable to assume that the established rules governing T cell responses to infections might be useful in developing better subunit vaccine formulations. However, there is mounting evidence that the immunological mechanisms relevant to adjuvant-elicited cellular immunity are vastly different than those observed in response to infectious challenge. Over the last 20 years, we have published extensively on a growing list of factors that highlight the mechanistic distinctions between vaccine-elicited (Tvacs) and infection-elicited (Tinf) T cell responses in mice and non-human primates. Our most recent data has revealed additional features mechanistically unique to Tvacs. Whereas Tinf split their functions between 2 different cell types and either divide or become memory, Tvacs do both within the same cell. Our data support an entirely novel model of CD8+ T cell activation whereby two transcription factors normally opposed to one another in function instead cooperate to sustain Tvacs exponential clonal expansion as well as a memory cell fate. The goals of this project will be to understand how these transcription factors cooperate instead of competing to support a vaccine-elicited T cell's transcriptional and metabolic needs.

NIH Research Projects · FY 2025 · 2005-07

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. This application seeks the fourth renewal of the Institutional Training Program (T32) in Pediatric Gastroenterology, Hepatology, and Nutrition at the University of Colorado School of Medicine (CUSOM) and Children’s Hospital Colorado (CHCO) on the CU Anschutz Medical Campus in Aurora, Colorado. Our T32 program’s overarching goal is to provide pediatric post-doctoral trainees exceptional training and mentorship to develop or refine their research skills, knowledge, and grantsmanship for successful careers in research related to gastrointestinal tract, liver, and pancreatic health and diseases in infants and children. Our program capitalizes on the robust physical and scientific environment at the Anschutz Medical Campus, featuring 32 dedicated research faculty members and a talented pool of potential applicants. We request continuation of five training positions to nurture pediatric physician scientists in two pathways: Basic Laboratory Scientists or Clinical/Translational/Health Outcomes Scientists, utilizing CUSOM's resources in basic, translational, clinical, and health outcomes-based research. Research training will focus on four overarching scientific themes: Mechanisms of Tissue Inflammation and Injury, Innate & Adaptive Immunity and Host-Microbiome Research, Clinical/Translational and Health Outcomes Research and Stem Cell Biology and Regenerative Medicine. Our T32 faculty comprises 32 scientists with over $47 million in grant funding and a strong track record of successful mentoring and training. The successes of our trainees include that as of early 2024, 75% of the 40 trainees completing the T32 training will have secured and remain in full-time academic positions. Of the 12 trainees completing the T32 during the current cycle, 5 (42%) have obtained K or K-like awards. To increase success in research careers our program’s basic research experiences are supplemented with graduate courses, seminars, and multidisciplinary meetings. Trainees in clinical-translational or health outcomes research will engage in the Master of Science in Clinical Sciences Graduate Program at CUSOM and utilize the infrastructure of the Colorado Clinical and Translational Sciences Institute. The pediatric GI research training program is structured as a 2-3 year initiative, commencing in the second year of subspecialty fellowship. Eligible candidates primarily come from the distinguished fellowship in Pediatric Gastroenterology, with potential candidates from related disciplines such as neonatology and nutrition. Fellows receive a minimum of 80% protected time for mentored research training in a structured environment. We closely monitor the quality of educational and research experiences through trainee, mentor, and Executive Committee feedback, with defined metrics and ongoing improvement processes.

NIH Research Projects · FY 2025 · 2005-04

Project Summary/Abstract (ASDs) an Individuals communication, can spectrum disorders results from the loss of expression of the Fragile X mental retardation protein (FMRP), mRNA-binding protein encoded on the X chromosome involved in suppressing protein translation. with FXS can exhibit a range of debilitating deficits in cognitive abilities, social interactions and and sensory processing. Deficits in sensory processing, often observed as hypersensitivity, contribute to other deficits associated with FXS (e.g., in social interactions). The Fragile X syndrome (FXS) is the single most common monogenetic cause of autism in humans. FXS broad objective of this proposal is to examine dysfunction in the olfactory system of mouse models for Fragile X with reduced expression of the mouse gene for FMRP, Fmr1. Surprisingly, despite the overwhelming importance of olfaction for rodents, there are few studies in this sensory modality in Fmr1 KO mice. Our goals, divided across three Aims, will be: Aim 1: To test the hypothesis that Fmr1 regulates inhibitory synaptic connections in the bulb. Aim 2: To test the hypothesis that Fmr1 regulates local network activity in the bulb by controlling inhibitory synaptic connections. Aim 3. To test the hypothesis that Fmr1 regulates olfactory behavior and bulbar oscillations in awake behaving mice by controlling inhibitory synaptic connections. Our studies will utilize a variety of approaches including patch-clamp recordings in brain slices, ultrastructural analyses, behavioral experiments, and in vivo electrophysiological recordings. Mouse models will include whole-animal Fmr1 KO mice, which is the most widely used mouse model of FXS, along with conditional KO and recovery mice in which Fmr1 expression is selectively and inducibly manipulated in GAD65-expressing GABAergic interneurons. This multidisciplinary approach will enable us to identify dysfunction in olfaction that results from altered Fmr1 expression at levels ranging from single synapses to whole-animal behavior and also link the changes observed at the different levels.

NIH Research Projects · FY 2025 · 2004-09

The complement system is a major pro-inflammatory and immunomodulatory pathway and plays a central role in the mechanisms that drive the pathogenesis of experimental murine models of human rheumatoid arthritis (RA). In these murine models, inappropriate complement activation that is directed to self-tissues drives initial cellular influx into the joint as well as synovial inflammation and bone erosions. However, despite the extensive insights into the murine disease that we have developed with support of this grant, including the activation pathways involved, how control of the system is overcome and how individual effector pathways promote tissue damage, we do not know how the complement system plays a pathogenic role in patients with RA. Recent findings in our studies of the human disease have identified a prolonged preclinical phase in RA characterized by the presence of circulating autoantibodies and mucosal inflammation that appears to drive the initial break in tolerance to citrullinated self-antigens. Following that asymptomatic phase, where complement activation is present in the mucosal site but not systemically, it is likely that complement activation and effector mechanisms are then especially important as the disease transitions to the very early phases of synovitis when circulating autoantibodies directed against citrullinated proteins initially react with antigens which develop and are displayed in the joint. To build our understanding of the human disease and translate information from models of disease to patients themselves, the major focus of this competing renewal proposal is to understand how the complement system is involved in the early synovitis in RA. We are especially well positioned to accomplish this important goal, as with other support mechanisms we are able to identify and follow subjects from the preclinical period into the very first appearance of synovitis and the diagnosis of RA. By obtaining synovial biopsies, a skill set also developed in our program in the last 3-4 years through other funding, and informative blood samples from patients in this important transition period, we will work to characterize the role of complement activation as well as its regulatory and effector mechanisms in the initial development of inflammatory arthritis and synovitis in patients. In addition, as there are distinct sub-types of RA, designated pathotypes, which have prognostic importance, we will determine what complement activation processes are associated with individual pathotypes and how that influences clinical outcomes. Finally, we will focus special attention on a major synovial cell type in human RA, which are highly inflammatory fibroblast-like synoviocytes, whose importance is increasingly understood and for which there are reported complement signatures in unbiased omics studies. We will define the mechanisms by which complement interacts with this cell type, both with regard to how complement is regulated by inflammatory cytokines as well as activated, and then defining the subsequent phenotypic changes. A major goal in these studies is to use information gained to inform the use of current and next generation complement therapeutics in this important human autoimmune disease.