University Of Colorado Denver

NIH Research Projects · FY 2024 · 2004-06

PROJECT SUMMARY/ABSTRACT Insulin produced by pancreatic β-cells is the key stimulus for glucose metabolism, and therefore it is critical that insulin secretion is adjusted to changes in energy state. Insulin secretion is acutely regulated by nutrients and hormones that change in response to feeding. If the fed state persists, as in overnutrition, adaptive control mechanisms increasingly sensitize the insulin secretory response to meet the increased insulin demand. However, long-term overnutrition can also become maladaptive and lead to β-cell failure and type 2 diabetes (T2D). How β-cells read nutrient signals and translate these signals into adaptive and maladaptive responses is poorly understood. Supported by this grant, we have shown that the nutrient-sensitive histone demethylase LSD1 mediates nutrient-induced changes to the β-cell epigenome to regulate adaptative insulin secretion. Specifically, we found that LSD1 modifies the epigenetic state of gene regulatory elements linked to β-cell nutrient response genes, thereby modulating the amplitude of the insulin secretory response. Thus, LSD1 functions as an integration hub between the β-cell’s nutrient environment, the epigenome and transcriptional output. Our preliminary studies further show that LSD1 inhibition is adaptive in a lean state, but becomes maladaptive and promotes β-cell failure in obesity. The objective of this proposal is to determine the mechanisms by which LSD1 senses nutrients and to gain insight into how metabolic cues converge on LSD1 and the epigenome to render the adaptive β-cell response maladaptive. We will employ state-of-the-art approaches, encompassing novel mouse models, human islet experiments, single cell resolved mapping of chromatin state and gene expression, and cutting-edge computational analyses. In Aim 1, we will determine how LSD1 senses nutrients and regulates chromatin state in β-cells. To investigate the nutrient sensing mechanism, we will manipulate LSD1’s metabolically regulated co-factor FAD and measure effects on LSD1- regulated processes in β-cells. Furthermore, we will dissect LSD1’s enzymatic and non-enzymatic functions in regulating the β-cell epigenome, using novel mouse models and in vitro experiments. In Aim 2, we will identify mechanisms by which LSD1 inhibition triggers maladaptive processes that accelerate β-cell decompensation in T2D. Employing genetic mouse and islet models of metabolic stress, we will manipulate LSD1 activity and study how these manipulations affect β-cell chromatin state, gene expression, and phenotypes. In Aim 3, we will examine the role of LSD1 in human β-cells. Here, we will leverage transcriptomic and chromatin maps we generated from a collection of nondiabetic, prediabetic, and T2D donor islets to determine whether the LSD1- regulated program contributes to β-cell failure in T2D. By unveiling fundamental mechanisms by which β-cells interpret nutrient signals, this proposal will prove critical for identifying strategies to prevent or reverse maladaptive processes and β-cell failure.

NIH Research Projects · FY 2025 · 2004-01

The primary cilium is a sensory organelle, found on most types of cells in vertebrates, which is required for many of the key cellular signaling pathways. Mutations in genes involved in primary cilia formation and function produce genetic disorders termed ciliopathies, affecting multiple organ systems. Renal disease is a prominent feature of many ciliopathies, as defects in the primary cilia of the polarized renal epithelial cells lead to variety of pathologies, including development of kidney cysts and loss of kidney function. Despite the importance of apical cilia in kidney function, we still understand little about the molecular machinery governing apical cilia formation in polarized renal epithelia. Our work during previous funding cycle identified Rab19 as an important coordinator between cilia formation and epithelia polarization. We have shown that Rab19 is present at the ciliated centrosome (also known as basal body) where it regulates several stages of cilia formation, including the docking of the basal body at the apical plasma membrane, as well as extension of ciliary axoneme. Despite advances in our understanding of the mechanisms mediating cilia formation in polarized epithelial cells, many questions remain. How do epithelial cells coordinate polarization and formation of apical structures such as cilia? How does Rab19 regulate apical cilia formation? Finally, how cilia formation and function is disrupted in renal disorders such as polycystic kidney disease (PKD)? To start addressing these pivotal questions we used proteomic analysis to identify TTLL12 as a Rab19- interacting protein. Importantly, we also have shown that TTLL12 is required for cilia formation in polarized renal epithelia, while having little effect on ciliation in non-polarized cells. TTLL12 belongs to a family of tubulin tyrosine ligase like (TTLL) proteins. TTLL family proteins mediate secondary modification of - and -tubulin by catalyzing tyrosination, glutamylation, or glycylation. These secondary modifications comprise so called “tubulin code”, that affects microtubule stability, polymerization and function. Interestingly, the enzymatic activity of TTLL12 remained unclear since it was shown to have little effect on tubulin tyrosination, glutamylation, and glycylation. Our recent studies suggest that TTLL12 may be tubulin-specific methylase, and that tubulin methylation may play and important role in regulating microtubule dynamics. Thus, based on our studies, we hypothesize that: Rab19-TTLL12 complex regulates apical cilia formation and function in polarized renal epithelial cells, possibly by mediating tubulin methylation. To test this hypothesis in vitro and in vivo we will first determine the role of TTLL12 in regulating microtubule dynamics and stability (aim#1). Additionally, we will determine the role of Rab19 and TTLL12 during apical cilia formation in renal polarized epithelial cells in vitro and in vivo (aim#2).

NIH Research Projects · FY 2025 · 2003-09

ABSTRACT Cellular differentiation, development and homeostasis depend on regulation of gene expression, which is largely focused on the DNA transcription initiation process. During transcription initiation, Mediator, a large multi-protein complex conserved throughout eukaryotes, conveys regulatory signals to RNA polymerase II (RNAPII), the enzyme responsible for transcription of all protein-coding genes. Mediator includes 25-30 different polypeptides (depending on the specific organism) organized into Head, Middle and Tail modules, plus a dissociable kinase module (CKM) that includes the Cdk8 kinase, the only catalytically-active Mediator subunit. Mediator conformational rearrangements that stabilize preinitiation complex (PIC) components have explained the effect of Mediator on basal transcription. However, conformational rearrangements alone are insufficient to explain the response of Mediator to transcription factors (TFs) that enables transcription activation and repression. Here we propose biochemical, functional and cryo-EM studies of mammalian Mediator (mMED) that build on our previous work and explore the significance of mMED’s antagonistic interaction with the CKM and with MED26, a metazoan-specific, dissociable mMED subunit closely linked to modulation of mMED–RNAPII interaction. The CKM and MED26 interact with Mediator around a Head-Middle module interface (the CTD-binding gap) where RNAPII interaction is initiated by binding of the carboxy- terminal domain of the largest RNAPII subunit (the CTD). CKM-bound (CKM-mMED) and MED26-bound (MED26-mMED) forms of mMED were independently identified by various research groups shortly after Mediator’s discovery and we propose to test a mMED activation mechanism based on CKM-mMED to MED26- mMED interconversion that we hypothesize controls mMED interaction with RNAPII and PIC formation. In Aim1 we will Investigate the connection between CKM – MED26 antagonism and Mediator activation. We posit that the crux of the mMED activation mechanism is control of the initial CTD-dependent mMED– RNAPII interaction by antagonistic effects of the CKM (limits RNAPII interaction) and MED26 (required for RNAPII interaction) at the CTD-binding gap. We will use in vitro and in vivo approaches including biochemical, functional and genomic analyses to understand modulation of mMED interaction with RNAPII and its effects on transcription initiation. These studies will test a proposed activation mechanism that would explain the significance of mMED subpopulations with opposite functional effects and test whether interconversion between mMED forms can explain mMED activation. In Aim 2 we will determine the structural underpinnings that enable regulation of Mediator-RNAPII interaction. We postulate that TF targeting of CKM-mMED and subsequent effects on MED26 and CTD interaction at the CTD-binding gap are enabled by mMED structural rearrangements or changes in mMED conformational dynamics. Structural analysis of well-defined intermediate steps will reveal how mMED’s structure enables activation. We will use cryo-EM to determine near-atomic resolution maps of various intermediates and use state-of-the-art image analysis approaches to understand their conformational and interaction dynamics. These studies will reveal structural factors that underpin primary (initial CTD-dependent interaction with RNAPII) and subsequent (further modulation of RNAPII interaction and PIC assembly) aspects of the mMED activation mechanism. In Aim 3 we will investigate long-range structural rearrangements in mMED that enable mMED activation by TF binding to Tail module subunits. We believe that changes in the composition, structure or conformational dynamics of the Tail module triggered by interaction with TFs can be communicated along the mMED structure to the CTD-binding gap, allowing mMED to respond to a variety of regulatory signals through the same fundamental activation mechanism. We will use cryo-EM, image analysis and biochemistry to understand how signals from TF binding to various mMED Tail subunits converge to control interaction of mMED with RNAPII. The studies described in this aim will test the generality of the proposed mMED activation mechanism and reveal how the mMED structure can integrate signals from a variety of TFs that target different mMED subunits. The studies proposed in this application are both conceptually and technically innovative. We will test a novel model for regulation of activated transcription initiation based on interconversion between “repressed” and “activated” forms of mMED by applying a multi-disciplinary approach combining state-of-the-art molecular biology, genomics, bioinformatics and cryo-EM/image analysis techniques to understand early steps of mammalian PIC assembly and transcription initiation. We will combine in vitro studies allowing examination of individual aspects of the proposed mechanism with in vivo studies to verify biological significance and functional consequences. The use of state-of-the-art cryo-EM and image analysis will allow us to build on prior accomplishments and arrive at a detailed structural understanding of the way in which the intricate mMED structure enables activated transcription regulation.

NIH Research Projects · FY 2025 · 2003-04

Project Summary The primary goal of our training program in “Cardiovascular Biomechanics and Imaging” is to continue attracting, training, and graduating highly qualified, multi- disciplinary scientists in the area of cardiovascular bioengineering at pre- and post-doctoral levels. The program was created 15 years ago in response to: 1) the dearth of training opportunities available at the intersection between cardiovascular medicine and bioengineering at the University of Colorado and indeed within the Greater Rocky Mountain region; and 2) the strong institutional commitment to build bioengineering research and training within the University of Colorado. We believe the first 15 years have produced outstanding outcomes, with all pre-doctoral (except one) and post-doctoral graduates of the program in research positions within academia or industry, and many successfully receiving research grants and trainees of their own. Further, NHLBI T32 funding has been important in leveraging a large amount of additional funding from the University of Colorado, which will be used to help support our next cycle of trainees. We believe the combination of outstanding outcomes, significant and continuing institutional support, and growing reputation continue to make a compelling argument to continue this program.

NIH Research Projects · FY 2025 · 2002-09

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 is a proposal for a Fellowship training program in pediatric endocrinology and post-doctoral psychology, designed to provide multidisciplinary training in diabetes research at the Barbara Davis Center (BDC), and Children’s Hospital Colorado (CHC) at the University of Colorado (UC). Funding is requested for two years of research training for three Pediatric Endocrinology trainees and one PhD psychologist in each year of the Program. The proposal’s rationale is: 1) There is a critical shortage of pediatric endocrinologists and psychologists trained in diabetes research 2) Diabetes is an increasingly significant healthcare problem in the pediatric population with devastating consequences from a personal and health economics viewpoint 3) Current rapid advances in diabetes investigation calls for an increased pool of physician scientists trained in clinical, biochemical, epidemiological, and behavioral research to carry out basic and translational research in pediatric type 1 (T1D) and type 2 diabetes (T2D) to bring advances in diabetes research to the clinical arena. Psychologists training in behavioral research beside physicians will add strength to the physician program and contribute diabetes research trained psychologists to the very small pool of these important specialists. The BDC and CHC have an extensive and proven record in research training of young investigators. A strong endocrinology program at CHC, in addition to a research program in T2D, will provide a well-rounded pediatric endocrinology training program. The BDC, CHC and the wider UC have well-established investigators in epidemiology, pathophysiology, immunobiology, genetics, and molecular biology of T1D and T2D. This network of experienced mentors is ideally suited to develop the next generation of young pediatric endocrinologists prepared for a career as diabetes investigators. Areas of research focus within the BDC include developmental islet cell biology, genetics, immunology, autoimmunity, prediction and prevention of T1D, as well as the epidemiology of and novel treatments for T1D and T2D. The integration of psychologists training in diabetes behavioral research will add importance and depth to the Program. These areas of research are urgently needed and have been identified as important to the national public health to address diabetes in the US. The ability to provide strong didactic courses in statistics, epidemiology, research methods, ethics and regulation of human research, medical writing, molecular biology and genetics, combined with a strong mentoring faculty will allow us to provide the training required to develop pediatric endocrinologists and psychologists into the scientists needed for the leaders of tomorrow.

NIH Research Projects · FY 2026 · 2002-09

Cholestatic liver diseases are among the most important liver disorders that occur in infants and children, leading to devastating morbidity and accounting for over 70% of liver transplants performed during childhood, thus posing a major public health burden. Although major advances in genetics of these disorders have been made over the past decade, therapeutic options are limited. Investigation of these disorders promises to advance scientific knowledge about hepatobiliary development, hepatocyte transporters, cholangiocyte biology, genetic regulatory networks, the neonatal immune response and mechanisms of injury, as well as the discovery of biomarkers of disease and testing of new diagnostic and therapeutic strategies. With the advent of next generation sequencing, genomics/epigenomics, proteomics and metabolomics, disease modeling paradigms and a pipeline of new potential therapies, the immediate translational impact of research in these disorders has never been greater. The 8 cholestatic disorders of the Childhood Liver Disease Research Network (ChiLDReN) have been studied in multi-centered research consortia at our Center for the past 22 years, during the last 15 years within ChiLDReN. Members of our Clinical Center at the University of Colorado Denver and Children’s Hospital Colorado have played major leadership roles as the Chair of the Steering and Executive Committees of ChiLDReN and Chairs of 4 protocols, among other roles. The objectives of this grant application in response to NIH RFA-DK-23-017 through our four specific aims are: to be chosen to be an active, collaborative and productive Clinical Center in the next version of ChiLDReN; to continue to enroll participants with biliary atresia and primary sclerosing cholangitis in the respective protocols and to continue to follow those already enrolled in all of the ChiLDReN ongoing protocols; complete, analyze and publish all of the ChiLDReN study protocols; to be an active participant in all new investigations, protocols and clinical trials initiated by the Network; to develop and propose new clinical and translational studies, trials and ancillary studies that will utilize our participant populations and existing data and biospecimens; to maintain leadership roles within the network and direct the Administrative Core Functions of ChiLDReN; to subcontract to Children’s Hospital Los Angeles for their ongoing work within the ChiLDReN Network; and to continue to provide PFIC/BRIC/ALGS Genotyping expertise and services. Through these activities, our Colorado ChiLDReN Clinical Center will enhance and build on the ChiLDReN goals of expanding clinical and translational research on 8 rare pediatric liver diseases focusing upon improving our understanding of these disorders and developing novel diagnostic and therapeutic strategies.

NIH Research Projects · FY 2025 · 2001-05

PROJECT ABSTRACT This application is a renewal of our aging-related Integrative Physiology of Aging T32, which is currently in its 20th year. The overall objective is to continue research training efforts on both the University of Colorado Anschutz Medical Campus (CU-AMC) and the University of Colorado-Boulder Campus (CU-B). As with the previous funding cycle, the major participating programs will be the Division of Geriatric Medicine (DGM), the Department of Integrative Physiology (IPHY) at CU-B and the VA Eastern Colorado Geriatric Research, Education, and Clinical Center (GRECC). Because we are the only T32 out of 50+ institutional training grants (T32, TL1, KL2, K12) at CU-AMC/CU-B that specifically focuses on aging physiology, we are requesting support for 5 predoctoral and 5 postdoctoral trainees each year, similar to the previous award. The specific objectives of this training program are to continue to: 1) expand the participating faculty and create an even more collaborative and effective mentoring group in support of age-related research at CU-AMC, CU-B and Eastern Colorado GRECC; 2) recruit, select, support and retain high quality predoctoral and postdoctoral (MD and PhD) trainees, with a continued emphasis on attracting underrepresented minorities (URM), women, disabled, and individuals from disadvantaged backgrounds; 3) support experiential and didactic research training opportunities in the integrative physiology of aging, including methods for enhancing reproducibility (e.g., consideration of Sex as a Biological Variable); 4) support the “T-shaped Team Training” (T3) program for trainees (and mentors); 5) monitor, support and ensure successful progress of trainees using predetermined milestones, defined in their individual Career Development Plan (CDP); and 6) enhance the mentoring skills of junior “associate mentors” through co-mentoring with a “primary mentor” and participation in the T3 program. These objectives will be carried out by offering: 1) up to 2 years of research training support (the second years dependent upon successful progress); 2) dual mentoring by outstanding primary and associate mentors; 3) a variety of relevant classroom/research experiences on both campuses and team science training programs; 4) frequent and careful review of trainee progress/success; 5) ongoing appraisal of the program goals, strength and weaknesses; and 6) sound financial and administrative oversight. Our previous success is demonstrated by our ability to: 1) fill all of our positions with outstanding trainees; 2) produce academically successful and productive trainees; 3) build a pipeline of mentors, by transitioning trainees to associate mentors and associate mentors to primary mentors; and 4) expand aging-research across campuses through ongoing collaboration among the participating programs and faculty.

NIH Research Projects · FY 2024 · 2001-02

Project Summary The proposed studies will examine mechanisms by which morbidity and mortality are greater in older burn patients. Regardless of age, most burn patients do not die from primary injuries but rather from complications, such as pneumonia and multiple organ failure (MOF). Clinical and experimental evidence reveal that healthy aged subjects are in an elevated basal inflammatory state, referred to as “inflamm-aging,” which can slow the immune system's response to invading pathogens. We and others believe that inflamm-aging is caused by translocation of bacterial products from the intestinal lumen and that continuous exposure to these products triggers the production of pro-inflammatory cytokines, including interleukin (IL)-1β and IL-6. In addition, shifts in the intestinal microbiota in the aged can promote systemic inflammation and these shifts are much more dramatic after burns. Novel preliminary data in our clinically-relevant mouse model of burn injury confirm that aged burn-injured mice have heightened circulating levels of danger-associated molecular patterns (DAMPs), a greater breach in intestinal epithelial barrier integrity, and more dramatic dysbiosis of the fecal microbiome relative to their young counterparts. Moreover, we know from our recent work on burn patients that advanced age upregulates pro-inflammatory mediators in the lungs and blood, which parallels our mouse studies. From these observations, we hypothesize that, relative to young, older subjects who sustain a burn injury have greater 1) breach in the integrity of the intestinal epithelial barrier and 2) shift in the intestinal microbiome, leading to excessive systemic inflammation and damage to critical organs, like the lung. To test the concept of the post-burn “gut-lung axis,” in Aim 1, we will examine intestinal barrier integrity in burn-injured young and aged mice over time using blood-borne and fecal biomarkers of intestinal damage and determine if restoring the intestinal barrier reduces inflammation and improves function of distant organs, including the lung. In Aim 2, we will examine levels of a subset of gut and lung antimicrobial peptides (AMPs), determine the extent of fecal microbiome dysbiosis, and investigate whether modulating AMP levels restores normal intestinal and pulmonary parameters in young and aged burned mice. Lastly, in Aim 3, we will study burn patients longitudinally to determine if the superimposed impact of burn injury with advanced age alters intestinal barrier integrity and fecal microbiota to a greater extent than seen in younger subjects with equivalent size burns. Moreover, we will see if there is a relationship between intestinal dysregulation and systemic/pulmonary inflammation by assessing blood-borne and fecal biomarkers of intestinal damage and microbial dysbiosis and determine if these biomarkers can be used to predict acute hypoxic respiratory failure and poor immune response following pulmonary infections. These studies will expand our understanding of how advanced age alters the gut in the context of burn injury and its systemic implications. Our work may also lead to the development of novel therapies for the treatment of patients with burns and other forms of trauma.

NIH Research Projects · FY 2025 · 2000-09

Project Summary/Abstract The rationale of this Paul Calabresi Award in Clinical Oncology Research (PCACOR) competitive renewal application is to continue to prepare highly qualified cancer clinical researchers, called Scholars, who will become independent leaders. These clinical research leaders will meet the ongoing need for highly trained academicians running pivotal cancer clinical trials and/or the critical clinical correlative studies that will inform ongoing translation from bench to bedside. Through this unique training program and the expertise of our mentors, the Scholars will succeed in learning to communicate and coordinate multidisciplinary teams of scientists and apply the ever-growing amount of scientific knowledge to personalize cancer treatment. Specific objectives are to: 1) Design a flexible Individual Development Program for each Scholar selected by the multidisciplinary Advisory Committee from applicants who may be oncology trained MDs, DOs, PhD clinicians with completed clinical training or PhD level basic scientists committed to an academic cancer career with heavy clinical focus; 2) Foster interdisciplinary training, communication and interaction through intentional team mentoring and regular Scholar-centered meetings, creating completed PCACOR Scholars successfully transitioned to independence; 3) Create a diverse cohort of highly trained clinical research leaders that will run novel, innovative multidisciplinary research agendas to move the translational of our current science to reduce the burden of cancer. Also, these highly trained thought leaders will become part of the ongoing mentorship cycle back to new trainees. The PCACOR design incorporates our exceptional faculty as strong mentors who will provide training and role models in the richly supported environment of the University of Colorado Cancer Center. Experienced Program Leaders are Virginia F. Borges, MD, MMSc, and Jill Slansky, PhD, both Professors with extensive relevant experience. The tailored Training Period lasts for two years, with the potential for a one- year extension and determined by the Advisory Council. The training period includes Core Requirements of a selection of Scholar-tailored didactic coursework, a Scholar designed major research project of at least one cancer clinical trial or ancillary study. The Training Period will focus on the specific individual training needs of each Scholar for which mastery of these needed skills through the mentored environment will ensure transition to independence. As an outcome metric, all Scholars submit an NIH or equivalent grant on completion of the training. The ultimate success is demonstrated by our record of accomplishment. Thirty early career faculty have trained as PCACOR Scholars. Seventeen Scholars are faculty members at University of Colorado Anschutz Medical Campus; all of which have had multiple peer-reviewed publications and additional peer-reviewed funding from NIH, DOD, ACS and/or disease-based foundations. All have designed at least one cancer clinical trial, and several are institutional leaders and/or leading national cancer clinical trials. Our PCACOR successfully produces the translational cancer clinical researchers of now and the future.

NIH Research Projects · FY 2026 · 2000-06

The Annual National MD/PhD Student Conference was established in 1986 by the University of Colorado Medical Scientist Training Program (CU-MSTP) and for the past 39 years has occurred in mid-July at a Colorado mountain resort. We obtained R13 funding in 2000 and this is the 5th competitive renewal application. The objective of this unique conference that is organized by trainees and specifically targets MD/PhD trainees is to provide a forum that brings together a broad group of ~200 dual-degree students from across the nation to learn about cutting-edge biomedical research from Keynote speakers and other MD/PhD trainees; to promote networking and life-long relationships with peers that are experiencing similar issues as they train; and to promote the progression of their career as high impact physician-scientists. The current 4th year CU MSTP students organize the conference, guided by an annually rotating Faculty Advisor and supported by the Program Director and Manager. The 2.5-day conference is multi-disciplinary, addresses the most important current biomedical problems, and includes talks by 3 to 4 nationally renowned Keynote speakers, both established (Nobel Laureates, NAS members & HHMI Investigators) and rising stars; oral student presentations and posters; timely Break Out Sessions; a Career Panel; and a Research Track Residency Recruitment Fair. Break Out Sessions include topics such as: Research Ethics, Rigor & Reproducibility, Interfacing with Industry, and Clinical Trials. A popular addition is the Research Track Residency Recruitment Fair, whereby Directors of multiple research-focused residency programs (PSTP) interact with students as they serve on career panels and attend talks and posters. A major goal of the conference is to include a broad group of attendees and speakers and welcome all participants. The R13 grant supports student Travel Awards of $1500 each, designed increase student participation. For the past 5 years, the yearly total average attendance was 223. A total of 22 Keynote speakers have participated in the past five years with 2 Nobel Laureates, 1 NCI Acting Director, 1 former NIH Director, multiple NAS members and HHMI Investigators. Publicity for the conference is accomplished via emails to the AAMC MD/PhD Directors and Administrators list-serve; our CU MSTP Conference Website; Conference Facebook, Twitter and Instagram pages; and listing the conference on the American Physician Scientist Association website. In sum, the continuous improvement of this unique conference, the consistent elite caliber of the Keynote speakers, the successful community building among students from across multiple backgrounds and institutions, the emphasis on providing opportunities for career promotion as physician-scientists, and the impactful involvement of Research Track Residency Directors justifies our request for continued R13 support.

NIH Research Projects · FY 2025 · 1999-12

Project Summary Neuroinflammation has been identified as a therapeutic target for acquired epilepsy. However, if and how mitochondria contribute to neuroinflammatory processes such as astrogliosis remains unknown. The goal of this proposal is to determine if mitochondrial reactive oxygen species (mtROS) generated within neurons can activate astrogliosis. This non-cell autonomous regulation is hypothesized to occur by posttranslational redox modification of glial fibrillary acidic protein (GFAP) resulting in long-lasting neuroinflammation and epilepsy. To test this idea, we plan to utilize a combination of tools and approaches including embryonic and inducible mouse models lacking mitochondrial superoxide dismutase in forebrain neurons, 2-photon imaging of astrocytic calcium, mitochondria-specific models, and mass spectrometry. The following specific aims will be explored. Specific Aim 1: Determine if mitochondrial oxidative stress is sufficient to increase astrogliosis in vitro. Specific Aim 2: Determine the role of neuronal mtROS in GFAP upregulation and astrogliosis in vivo. Specific Aim 3. Determine the role of mtROS in GFAP upregulation, astrogliosis, epilepsy and/or cognitive dysfunction in chemoconvulsant epilepsy models. Potential clinical application of the proposal includes identification of novel metabolism-based therapies to target astrogliosis in epilepsy.

NIH Research Projects · FY 2025 · 1999-07

PROJECT SUMMARY The Colorado Womens Reproductive Health Research Career Development Program (‘Colorado WRHR’) hereby seeks its 6th funding cycle. Our outstanding team of 27 mentors, includes 5 past WRHR primary mentors, and 14 past or current K12 Scholar mentors, the PI and Research Directors Raj Kumar, PhD and Jeanelle Sheeder, PhD. Our research theme broadly emphasizes Womens Reproductive Health at key life transitions: 1. Reproductive Aging, Cardiometabolic Fitness, and Brain Health; 2. Maternal Health and Reproductive Life Course; and 3. The Immune System and Its Links to Gynecologic Cancer. Each window of research opportunity will also include cross cutting themes incorporating Social Determinants of Health and Artificial Intelligence whenever possible. Specific Aims follow: Aim 1: Design the most effective training program for early career physician scientists in OBGYN in the nation. We will employ best practices and thoughtfully applythe recommendations from our Advisory Committees. We will provide the fundamentals that have served our scholars well in the past and will continue to incorporate feedback from program graduates and engage past Scholars in our program who are no longer at the University of Colorado through research retreats and collaborations. Aim 2. Prepare Scholars for leadership and advocacy in Women’s Reproductive Health Research. We will promote skill development in leadership and advocacy through trainings available locally at the institutional level and nationally such as the AAMC Early Career Development program. We will continue to model advocacy for Women’s Health Research through the efforts of the PI (past co-Chair of the Women First Research Coalition), RDs and mentors, and our professional organizations. These aims are designed to meet and exceed the goals the of RFA-HD-25-005 to create the next generation of leaders in our field.

NIH Research Projects · FY 2026 · 1998-12

SUMMARY The extraordinary functional range of the mature CNS requires synchronized activity of neural circuits. Formation of such circuits involves the timely specification and correct positioning of neural progenitors, and disruptions to this process are associated with neurodevelopmental disorders. Hence, a key goal of developmental neurobiology is to understand the control of neural progenitor specification in space and time in embryogenesis. One mechanism for proper spatiotemporal formation of neural progenitors involves morphogen gradients that specify distinct progenitor types at different positions. While we know that morphogens do induce different progenitor types at different positions, we do not understand how individual cells interpret morphogen signals, nor how they convert this information into distinct cell identities. An informative example of CNS morphogen action is the developing hindbrain, where retinoic acid (RA) and fibroblast growth factor (Fgf) control formation of neural compartments (rhombomeres) – each of which represents a unique progenitor population. However, we do not understand the distinct genetic programs that define individual rhombomeres, nor how they arise from the earlier hindbrain primordium. Filling these knowledge gaps is essential, but a profound barrier has been the lack of comprehensive molecular data for individual progenitors as they undergo specification in response to morphogen signals. We overcame this barrier by applying scMultiome analysis – which combines RNAseq and ATACseq of individual nuclei – to several stages of hindbrain development. At the earliest stage, we detect three populations (a.k.a., PHPDs) containing progenitors with mixed rhombomere (r) identities representing r2/r3, r4 and r5/r6 and we find that these PHPDs form in response to RA and Fgf. At later stages, our analyses – for the first time – molecularly resolve all rhombomeres and define their unique gene regulatory networks (GRNs). These advances now allow us to address several key questions: How do progenitor cells respond to morphogens? How are the mixed progenitor identities resolved into individual rhombomere identities? How are unique GRNs formed in each developing rhombomere? We will answer these questions in the context of our hypothesis that morphogens act via specific cis-regulatory elements to induce mixed identity GRNs in the PHPDs, and these are subsequently resolved via repressive genetic interactions into rhombomere-specific GRNs that specify unique progenitor types. Our project will delineate how morphogens control genetic programs for positioning and specification of neural progenitors in the hindbrain. Since morphogens control neural specification throughout the developing CNS, our findings will be broadly applicable to normal brain development, to modeling of neurodevelopmental disorders, and to the implementation of restorative or replacement strategies as clinical treatments.

NIH Research Projects · FY 2025 · 1998-04

Project Summary/Abstract Our competing renewal proposal seeks funds to continue to support attendance of Trainees and Young Investigators (YIs) at the Annual Meeting of the Perinatal Research Society (PRS). The PRS provides career development through two separate, but interrelated endeavors. One is a two-day grant writing program funded by R13HD079163 which occurs immediately preceding the Annual Meeting of the PRS. The second is the PRS YI Program funded by R13HD036244. It is the second program that we seek continued NIH R13 support. The Specific Aims of the Annual Meeting of the PRS that foster career development are based in its design and execution as follows: Aim 1) YI acknowledgement and interactions with PRS Members. PRS members are typically experienced faculty and mentors, with many leaders in the field of Reproductive Biology. Numerous activities are designed to encourage interactions between the YIs and PRS Members. Aim 2) The speaker program. Speakers present the latest research in their field. All presentations are in one room and there are no “concurrent” sessions. Aim 3) The early career speaker program. Three or four YIs give presentations at the Annual Meeting. Aim 4) Ongoing mentoring through the Associate Membership process. In recognition of the long career path to independence we realize failing to support the YI's after the end of the meeting is not the best path to success. We follow up with a Council review of all YI attendees and offer continued mentorship by Associate Membership of the society. This allows attendance and full participation in future years. We also follow up at least twice a year with email contacts to monitor career progress for workshop attendees. We have invested in the PRS Associate Member Paper Prize, which is given to the top three papers published by an associate member in the preceding year. Winners are provided complimentary registration to the next PRS meeting. Finally, as Associate Members obtain their independent grant funding, they transition to Regular Membership in the PRS. The need for this kind of opportunity for early career development is significant because of the challenging current academic and funding environment. The impact of our YI program is that it allows participants to make tangible and immediate progress in developing and expressing their research ideas and then translating that into scientific presentations, collaborations, and new mentor-mentee relationships. It is in the Main Meeting that they become integrated into the society.

NIH Research Projects · FY 2026 · 1997-04

The University of Colorado Cancer Center (UCCC) drives cancer discovery and innovation, translates these findings to the clinic, leads in patient-centered care, reduces health disparities, trains the next generation of cancer-focused scientists, and strives to optimize the impact on our catchment area – the State of Colorado. UCCC has benefited from a generous increase in funding and has made significant investments in all programs and structures. The highlights of recent progress include: 1) completion and deployment of a comprehensive strategic plan; 2) restructure of the programs; 3) recruitment of dynamic and accomplished new leaders including a Director (Schulick) and a second Deputy Director (Bradley), and recruitment of inaugural Associate Directors of Community Outreach and Engagement, as well as Informatics and Data Science; 4) establishment of two cross cutting themes of Cancer and Aging and Cancer and Metabolism; and 5) overhaul of UCCC membership criteria to better focus the activities of members and to increase emphasis on mentoring. This large infusion of resources and restructuring have better enabled multiple innovative discoveries in the laboratories to be brought to the bedside through investigator-initiated trials, FDA approval, and establishment of new standards of care. They have also enabled successful bi-directional community outreach and engagement and population-based programs within the catchment area. The physical space, organizational capabilities, transdisciplinary collaboration and coordination, cancer focus, and institutional resources have all been improved, setting UCCC on a trajectory of success well into the future. In the next funding period, UCCC will focus on the strategic aims, goals, as well as the 21 specific strategies outlined in the strategic plan. Highlights of these strategies include efforts to: 1) provide outstanding research infrastructure and state-of the art shared resources; 2) expand clinical research, clinical trials and investigator-initiated trials; 3) expand research and interventions across programs to address health outcomes, disparities, and underserved populations; 4) nurture the pipeline of prospective oncology researchers, clinicians, and administrators; strengthen alignment and collaboration within the UCCC and across all sites; and 5) grow philanthropic support. As necessary, UCCC leadership will modify the course to adjust to unexpected challenges and opportunities. Modified Specific Aims Modified

NIH Research Projects · FY 2025 · 1997-01

This is an application for a 5 year competitive renewal for the Colorado Nutrition Obesity Research Center (NORC). The MISSION of the Colorado NORC is to advance the science of nutrition and obesity by facilitating interdisciplinary, collaborative, translational research and by fostering the development of the next generation of scientists in the Rocky Mountain region. We are in our 30th year of operation, and we now support 125 NORC Members, 26 Associate trainees, and 47 Associate faculty scientists and educators. Colorado NORC Members have a healthy, nutrition and obesity-related annual funding portfolio. Building upon a multi-million dollar institutional investment in nutrition and obesity research over the past 30 years, we have secured an additional investment in institutional resources over the next 5 years to support the Colorado NORC Programs and Cores and to support the administration, building operation, programmatic development, and recruitment/retention efforts, with the Anschutz Health and Wellness Center (AHWC). This investment is important, because the AHWC is the administrative home of our center, its programs, much of one of the biomedical research cores, and a substantive portion of our Members who are performing clinical and translational research on nutrition and obesity. The CENTRAL THEME of the Colorado NORC is the prevention and treatment of obesity and its metabolic complications across the lifespan, through better nutrition, lifestyle modifications, and medical treatments. We specifically build and foster interdisciplinary research on nutrition and obesity in five areas that have emerged naturally in our Research Base: 1) Early Life Influences; 2) Biological Sex Differences; 3) Exercise and Energy Expenditure; 4) Metabolic Regulation and Dysfunction; and 5) Nutrition and Weight Management Interventions for Better Health. The Colorado NORC has strong leadership with Dr. Paul MacLean (Director), Dr. Daniel Bessesen (Associate Director), and Dr. Darleen Sandoval (Associate Director). During this next funding cycle, they will be taking substantive steps to help NORC members engage in T3/T4 research for translation. After 30 years of operation, the Colorado has never been as well-positioned to advance the science of nutrition and obesity on so many fronts and to foster the next generation of leaders in these fields.

NIH Research Projects · FY 2025 · 1991-09

PROJECT SUMMARY/ABSTRACT This T32 training grant renewal application is to support the training of 6 predoctoral and 4 postdoctoral trainees in Immunology. Training is to be performed at 3 institutions: The University of Colorado Anschutz Medical Campus (UCD-AMC), the Barbara Davis Center for Childhood Diabetes (BDC; at AMC), and National Jewish Health (NJH). All training faculty are primary or secondary members of the UC-AMC Department of Immunology & Microbiology. There are 43 faculty trainers and similar in number to our last renewal application in 2015. This cohort of faculty provide expertise in both basic, human and translational immunology. The grant will be directed by two Program Directors, Dr. Raul Torres and Dr. Ross Kedl, whose laboratories are both located at UC-AMC. This arrangement is to ensure appropriate attention is directed to both predoctoral and postdoctoral trainees. Dr. Torres is a well-known B cell biologist who has had continuous NIH funding for over 20 years and has successfully trained a number of graduate students that have gone on to secure positions as faculty, in biotech, journal editor and strong academic postdoctoral positions. He has been the PI on this T32 for the previous 5 years. Dr. Kedl has successfully trained a number of predoctoral and postdoctoral fellows that also currently occupy (tenure-track) faculty and biotech positions. He was the Associate Director of the Ph.D. training program in immunology at UC-AMC for almost 10 years and is well known for his work on T cells. Responsibility for various aspects of the Training Grant will be shared by these two PDs as described in the Multiple PD Leadership Plan. Trainees will be selected for support by the Program Steering Committee after application by trainee and mentor. Selection will be based on the merit of their previous work and their proposal for future research and appointments will typically be for 2 years. Trainees are required to take Responsible Conduct in Research every 4 years, to attend and present in our weekly “Research in Progress” forum and to complete each year an Individual Development Plan that is discussed with their mentor. Evaluation of progress of research and advice about career direction will be provided for predoctoral students by their Thesis Committee and for postdoctoral fellows by a mentoring committee consisting of at least 3 faculty members, chosen by the fellow. Predoctoral and postdoctoral trainees must meet their committees at least once every 6 months, and the committees must submit written reports describing the meetings. This Training Grant has a long and distinguished record, with almost all of its trainees going on to successful careers as scientists, ranging from academia to public service at NIH or the CDC to research in Biotech or with big Pharmaceutical companies. Particularly notable is its continued successful recruitment and training of a number of underrepresented minorities. The Program continues to be a major focus for immunological research in the Rocky Mountain region in particular and the USA in general.

NIH Research Projects · FY 2025 · 1991-07

PROJECT SUMMARY This application seeks renewal of a long-standing, innovative and highly successful postdoctoral training program in Nutrition that has a primary goal of training the next generation of physician scientists and PhD researchers committed to disease prevention and health promotion through careers in Nutrition. As suboptimal diets have surpassed tobacco smoking as the leading global mortality risk factor, the rationale for an outstanding nutrition research workforce is compelling. The training program, based in a major academic medical center, addresses 3 broad themes: 1) obesity and its co-morbidities; 2) nutrition, diet and eating behavior; and 3) lifecourse and development. Faculty mentors include a balanced mix of MD (10) and PhD (14) researchers representing 5 academic units in the university, and whose nationally recognized research spans basic (T1) to translational (T4) approaches. All faculty are affiliated with the NIH-supported Colorado Clinical and Translational Sciences Institute, which offers excellent support for translational research and numerous training and career development programs. Trainees to date typically have been physicians (1 position) seeking a career in academic medicine and nutrition-related research and PhD’s (2 positions) pursuing postdoctoral training in one of our thematic areas. The specific objectives of the training program include achieving excellence in all aspects of the scientific literature and methods relevant to trainees’ areas of research, with an emphasis on understanding scientific rigor and reproducibility and guiding principles of ethical conduct in research; mastering basic principles of Nutrition Science; preparing and submitting research grants and manuscripts; and acquiring skills for professional development and communication to undertake a successful academic career. The program provides 2-3 yr of research training, which includes ongoing rigorous work with a primary mentor, and interaction with faculty leadership and all trainees supported by this T32 plus other pre- and postdoctoral trainees through participation in weekly Nutrition Journal Club, professional development series and Nutrition seminar series. Of recent past trainees, two-thirds have obtained grant funding, and 80% have progressed to careers in academic settings. Based on increases in the quality and number of applicants, including physicians, relative to number of available slots, we propose an additional position for the next cycle, thus projecting 2 MD and 2 PhD trainees. The training environment promotes collaborative research and fosters productive interaction between physicians and PhD researchers, a model that prepares trainees to be exceptionally well positioned to emerge as future leaders in Nutrition to address the most compelling global health challenges.

NIH Research Projects · FY 2025 · 1991-07

Program Summary/Abstract The fundamental objective of our Trauma Research Center Training Program is to inspire the next generation of surgical trainees to pursue a career that includes basic science and translational investigation. At a time when surgical scientists are becoming rare, this training grant is critical to maintain the participation of clinically active surgeons in basic research. While our Center’s primary focus is Trauma-Induced Coagulopathy and Inflammation, we believe the diversity of opportunities within our Program, and general applicability of the coagulation/inflammation process to other disciplines, ensures relevance to any surgical trainee. Our secondary objective is to recruit capable surgeons to the field of academic trauma surgery in which lack of research has become a national crisis. Trauma remains the leading cause of death in the US for individuals younger than 46 years of age with bleeding the primary cause of these preventable deaths. Hence the need for basic and translational investigation in this area is essential and remains our inspiration. The design of our Program is a two-year full-time commitment to basic investigation conducted primarily in well-established surgical research laboratories within the University of Colorado Anschutz Medical Campus, supplemented by patient data generated from prospective clinical studies at our regional Level I Ernest E Moore Shock Trauma Center at Denver Health. Experimental work will focus on the mechanisms disrupting coagulation-inflammatory homeostasis, and includes investigation with proteomics, metabolomics, and genomics. The major funding for this basic research will be the 1RM1GM13968 “Resuscitation Strategies for Achieving Thrombo-inflammation Homeostasis” funded by NIGMS, UM1HL120877 TACTIC funded by NHLBI, and DOD-W81XWH – 12-2-0028 “Acute Coagulopathy of Trauma” funded by the Department of Defense. Clinical studies will be funded dominantly by the DOD W81XWH–12-2-0028 W81XWH-16-D-0024 “Linking Investigation in Trauma and Emergency Service Clinical Research Network” (LITES) as well as a number of institution and industry supported trials. The surgical research fellows will commence participation in the Program after the second or third year of general surgery residency (PGY 2 or 3) for two consecutive years. We encourage pursuit of advanced degrees, e.g. MS, MPH, and PhD, with training in the School of Graduate Medical Education. As in the past, we are requesting funding for four positions annually, sequenced so that first and second year fellows are assigned to our core laboratories.

NIH Research Projects · FY 2025 · 1986-12

Project Summary/Abstract Interleukin-38 (IL-38), a member of the IL-1 family, has not been studied until recently despite its discovery 20 years ago. It is a neglected cytokine because no receptor was identified for how IL-38 functioned. However, in 2012, we reported that recombinant human IL-38 bound to the IL-36 receptor (now IL-1R6) and suppressed the production of IL-17 and IL-22. In that study, we proposed that IL-38 acted as a receptor antagonist for IL-1R6. However, the dose-response of IL-38 did not behave as receptor antagonist but rather acted as an inhibitor of cell activities. New data suggests that IL-38 requires IL-1R6, an orphan receptor in the IL-1 Family, to suppress IL-17. Formerly termed IL-1 Receptor Associated Protein Like-1, IL-1R9 will be studied for its putative role in the suppression of innate immunity by recombinant IL-38. Using CRISPR/Cas methods, we have generated a colony of mice that are deficient in IL-38. These mice are used to determine a requirement for endogenous IL-38 in mouse models of human inflammatory diseases. In those models where disease severity worsens in IL-38 deficient mice, we will use recombinant IL-38 to treat mice for suppression of innate inflammation. In order to fully understand the role of IL-38 in innate immunity, we generated a mouse colony deficient in IL-1R9. We will study the requirement of IL-1R9 for the function of recombinant IL-38 in those mouse models where treatment with IL-38 has significantly reduced disease severity. A unique aspect of this application is that IL-1R9 in on the X-chromosome, an unusual finding in cytokine biology. Because IL-1R9 is on the X-chromosome, we can address how suppression of innate immunity is affected in males compared to females. With most autoimmune diseases having a 70% predilection for females and with each autoimmune disease there is an inflammatory contribution, we have designed studies for comparisons of homozygous IL-1R9 deficient males to homozygous IL-1R9 females. In these studies, we will also evaluate the role of IL-38 to inhibit the activation of the NLRP3 inflammasome using a specific oral NLRP3 inhibitor presently used to treat patients. In addition to AIM 1 and AIM 2 studies on recombinant IL-38 suppression of innate immunity and the putative role of IL-1R9, we will produce and test an IL-38-Fc fusion protein (AIM 3). The rationale for producing an IL-38-Fc fusion protein is to provide pre-clinical data for an IL-38 therapeutic. In AIM 4 we address the issue of IL-38 release from the cell. IL-38 circulates in healthy subjects but levels are significantly low in subjects at risk for a cardiovascular events. However, IL-38 being a B-cell product suggests that processing of the IL-38 precursor and release from the cell is not via traditional pathways. We will examine pathways for secretion that are used by other members of the IL-1 Family : inhibition of NLRP3 and inhibition of calpains. The overall goal of these studies is to advance the biology and clinical significance of IL-38 as well as to exploit the anti-inflammatory properties of IL-38 as a therapeutic.

NIH Research Projects · FY 2025 · 1986-09

Project Summary The primary goal of this T32 program is to provide outstanding research training in the pathogenesis of rheumatic and autoimmune diseases for individuals at several key levels of career development (medical student, predoctoral graduate student and postdoctoral MD and PhD fellow). The longstanding scientific opportunities in Denver in basic, translational and clinical research, as well as health services and outcomes research, provide a highly supportive environment to promote the broad goals of this training program. Newly developed programs have increased the emphasis on studying diseases more prevalent in under-represented minority populations as well as recruiting members of these groups into academic pathways. These efforts join together with longstanding innovation programs available through the NIH-funded Colorado Clinical and Translational Sciences Institute (CCTSI), with its Clinical Research Training Program and formal didactic training in clinical investigation for MD and PhD trainees, to increase opportunities and positively affect our T32 program. In addition, >20 Graduate School PhD programs provide substantial formal training as well as intellectual and laboratory resources for trainees interested in basic research. Extensive mentoring and educational programs in responsible conduct of research and scientific rigor continue to enhance the quality of the training experiences. Additionally, the ongoing support of the NIH-funded Autoimmunity Prevention Center, the Studies of the Etiology of Rheumatoid Arthritis (SERA), a rapidly expanding Colorado Center for Personalized Medicine, newly established programs in spondyloarthritis pathogenesis and prevention, and the Colorado School of Public Health further facilitate a comprehensive training environment. The training program will remain for this competing renewal under the direction of Dr. Michael Holers, Professor of Medicine and Immunology and Head of the Division of Rheumatology. Dr. Holers will work closely with an Executive Committee and Co-Director Dr. Kristi Kuhn. Thirty Participating Faculty Members will contribute. Notably, based on opportunities through development of a new Pediatric Rheumatology training program by Dr. Rob Fuhlbrigge and two years of successful research productivity by one of those fellows funded through an INCLUDE Down Syndrome T32 supplement, support is requested to expand from three to four postdoctoral fellows per year. Continued support for two medical students and two pre-doctoral graduate students per year is requested. The curriculum for the School and Department of Medicine includes research-intensive programs that provide access to highly motivated medical student short term trainees as well as individuals participating in the Medical Scientist and Physician Scientist Training Programs. The primary criteria by which the program will continue to be judged is the successful development of academic investigators who drive basic and clinical research advances necessary to improve the health of patients with rheumatic and autoimmune diseases. Leadership of this T32 hope to have the opportunity to continue support for their trainees.

NIH Research Projects · FY 2024 · 1983-07

Project Summary This application is a competitive renewal of the Division of Renal Diseases and Hypertension at the University of Colorado Anschutz Medical Campus T32 Training Program, currently in its 45th year. The program proposes to train 4 post-doctoral Trainees (MD and PhD) per year. The mission is to train the next generation of MD and PhD scientists in academic nephrology. To accomplish this objective, the training program requires the T32 Trainees to have mentored, comprehensive and specialized career development training. Our program stresses a wide range of science, including basic molecular mechanisms, translational science, genetics, epidemiology and clinical trials. Each Trainee will undergo an intense 2 to 3 year training program, including didactic courses, education seminars and career development training. Formal graduate school training is available through the Colorado Clinical and Translational Sciences Institute and Colorado School of Public Health. Each Trainee is given the choice to pursue his or her research training either in the laboratory setting or in any Training Faculty Member's clinical research group deemed integral to that Trainee's research training experience. The laboratories have modern, state-of-the-art equipment and staff that provide the best possible basic, translational and clinical research environment offered at the University of Colorado Anschutz Medical Campus. The diverse faculty is expert in a wide range of research topics, including parenchymal and cystic renal diseases, fluid and electrolyte disorders, acid-base disorders, hypertension, acute and chronic kidney disease, acute renal replacement therapies, chronic dialysis (peritoneal and hemodialysis, both at home and in-center), and all aspects of kidney transplantation. There are 31 mentors, of which 45% are women, and 8 emerging mentors from 4 schools and 5 departments. The inclusion of emerging mentors is to provide a continuum of mentorship and leadership development for the Training Program. For a sustainable workforce, the Training Program also focuses on academic persistence, resilience and wellness. Between 2009-2019, 25 Trainees completed their training and 60% remain in academic nephrology. In the past 5 years, 100% of our MD Trainees entered into academic nephrology. Our Trainees have been awarded 10 Career Development Awards during this time period. Four faculty have recently achieved the "K to R" transition over the past 5 years. Our training program is highly committed to the recruitment, training and career success of women and underrepresented minorities. Overall, 40% of Trainees over the past 10 years were women and 8% came from underrepresented minorities or disadvantaged backgrounds. All these factors, a history that dates back over 40 years and a strong track record of success enables the Division of Renal Diseases and Hypertension at the University of Colorado Anschutz Medical Campus to be an ideal site for training future leaders in nephrology.

NIH Research Projects · FY 2025 · 1982-07

Project Summary The overarching goal of our research is to develop new biomarkers of islet autoimmunity and to translate these discoveries to the prevention of human type 1 diabetes (T1D). During the last funding period, we developed a multiplexed assay to detect serum autoantibodies to islet and non-islet autoantigens with excellent sensitivity and predictive value when compared to our traditional radio-binding assays. The proposed studies will broaden the spectrum of relevant autoantigens to those that are post-translationally modified (PTM) as well as provide insights into when native versus PTM binding autoantibodies develop in the natural history of T1D. Our recent findings have strengthened the evidence for autoreactive T cells responding to (pro)insulin, including those activated by hybrid insulin peptides (HIPs) and defective ribosomal insulin products (DRiPs), within the residual pancreatic islets of T1D organ donors. However, we still lack robust T cell biomarkers that could reflect the activity of autoreactive T cells, especially at the earliest stages of islet autoimmunity or during immunomodulation to prevent progression to clinical diabetes. Assays for islet autoantibodies and autoreactive T cells that measure both natural and modified islet autoantigens, combined with a better understanding of their relationship, will enhance our knowledge of T1D pathogenesis and improve prediction of progression through the stages of T1D. In specific aim 1, we will optimize and multiplex autoantibody assays to native and PTM modified islet autoantigens to determine the temporal development of these antibodies in longitudinal samples from children prospectively followed from birth to development of islet autoimmunity and clinical diabetes. Specific aim 2 focuses on developing a non-cellular T cell biomarker of islet autoimmunity measured from whole blood DNA using the T cell receptor sequences obtained from residual islets of T1D organ donors. The successful completion of this proposal will result in scalable assays for the measurement of islet autoantibodies and autoreactive T cells, and improved understanding regarding the timing of autoantibody and T cell immune responses to native and post-translationally modified islet autoantigens during the development of T1D.

NIH Research Projects · FY 2025 · 1981-07

This is a resubmission of a renewal application for a highly successful training grant funded for 35 years from the Department of Dermatology at the University of Colorado Anschutz Medical Campus (UCAMC). This program began as an Immunodermatology training vehicle but has greatly changed in the past 13 years as the research focus in this Department has broadened. The remarkable expansion of the research program in the Department of Dermatology at UCAMC provided a perfect opportunity to restructure the scope and focus of this training grant which initially occurred in the renewal application in 2014. To better reflect the breadth of our training program, we have changed the title of our resubmission to the following: “Training in Translational Research of Skin Diseases.” Dennis Roop and David Norris will continue to serve as Co-Directors and lead an impressive multidisciplinary team of trainers and mentors, 15 of 26 of the current training faculty are new. Several new developments in the Department of Dermatology and the Gates Center for Regenerative Medicine have influenced the focus and organization of this renewal application, including the following: 1) the opening of the Gates Biomanufacturing Facility (GBF), a state-of-the-art cGMP manufacturing facility translating innovative research discoveries into safe and effective cell therapy and protein biologic products for human clinical trials; 2) establishment of the EB iPS Cell Consortium, comprised of Colorado, Stanford and Columbia universities, to fight the rare and debilitating genetic skin blistering disease Epidermolysis Bullosa (EB); and 3) the establishment of a new Ehlers-Danlos Syndrome (EDS) Center of Excellence on the UCAMC. Historically, this has been a highly successful training grant, preparing clinical scientists for academic careers, stimulating extensive research collaborations between scientists in Dermatology and other departments, and supporting a highly productive research program in the Department of Dermatology. Of the 13 trainees who have been supported by this T32 during the last funding period (2015 to present), 10 are still in training, 1 is completing a residency in Dermatology and 2 have taken positions in industry. This training grant will support 2 predoctoral and 4 postdoctoral training positions per year. Research training in preceptor’s labs will be supplemented with a substantial program of enrichment including research seminars, research in progress, lectures on relevant topics, and training in ethics. Unique training opportunities for our T32 trainees include: 1) Training in the Manufacturing of Cells and Biologics for Clinical Use; 2) Workshops on “Protection of Intellectual Property and Strategies for Commercialization”; and 3) Lectures on “Introduction to Bioinformatic Tools for Analyzing Next Generation Sequencing Data”. Recruitment of candidates for this program will be managed by a well-organized administrative structure and will include a robust plan to attract underrepresented minorities and enhance diversity of this training program.

NIH Research Projects · FY 2025 · 1979-07

PROJECT SUMMARY The primary goal of this Training Program is to develop academic leaders among Neonatologists, Maternal- Fetal Medicine (MFM) Specialists, and Non-Clinician Scientists who know how to work together collaboratively and have the capacity, based on personal meritorious accomplishments in research, to identify, understand, and solve important problems in Perinatal Biology and Medicine. This training program provides basic and clinical-translational research training, including concepts and techniques in perinatal/developmental physiology, biochemistry, and cell and molecular biology. To our knowledge, ours is the only T32 Training Program designed with this focus, structure, and goal. Trainees will be appointed after demonstrating research commitment and accomplishment during their late 1st or 2nd yr of fellowship/postdoctoral training. During their 1st year, trainees work with Faculty Mentors on their primary research projects while integrating into the broader perinatal biology community at CU-AMC through our T32 Curriculum. This gives them the multidisciplinary training required in Perinatal Biology. Formal training in Responsible Conduct of Research and Rigorous Research Design and Relevant Data Science and Quantitative Approaches are required. The 2nd year is devoted to the completion of the research projects, manuscript submission, expansion of the Fellow's research with interdisciplinary approaches, and further career development training. Seminar programs in research design, abstract and manuscript preparation, laboratory and career management, and grant writing are provided. This approach allows us to fulfill our two Specific Aims. Our primary specific aim is to provide the training and environment that ensures short- and long-term success of T32 Fellows as defined by: 1) publications, 2) appointment to research-intensive positions, 3) Career Development Awards, 4) Pilot and Project Grants, 5) research based awards and honors, and 6) Scientific Leadership Positions. Our secondary specific aim is to further strengthen the research endeavors related to Perinatal Biology and Medicine at CU-AMC as defined by 1) collaborative manuscripts with T32 Fellows, 2) co-mentorship of Trainees, 3) joint grant submissions, and 4) joint establishment of innovative programs. This aim directly benefits the T32 Fellows as it models the type of collaborative research we want them to engage in long-term. We also will place a major emphasis on the recruitment of NIH defined underrepresented minorities to our T32 Program with supplemental research and career development funds ($12,000 per URM Trainee) provided by the Section of Neonatology. Our approach equips the trainees with the capacity to move more successfully into academic careers in either basic or clinical-translational science. Over many years, graduates of this training program have achieved high academic positions nationally and internationally and have had major impact on developing and supporting research training programs. Attaining such ultimate research/academic leadership among our trainees is a major aim of this program.