Vanderbilt University Medical Center

NIH Research Projects · FY 2024 · 2003-08

A fundamental question in organ formation is how the constituent tissues achieve their correct sizes and cytoarchitectures. Defects in a single tissue can affect the formation of an entire organ and a classic example of this is the eye, where disruptions in retinal histogenesis can cause microphthalmia, a severe ocular anomaly characterized by small, poorly formed eyes and congenital blindness. Mutations in the Visual System Homeobox 2 (Vsx2) gene cause microphthalmia. A definitive marker of retinal specification, Vsx2 functions in retinal progenitor cells to define tissue identity. Concurrently, Vsx2 is required for progenitor proliferation and several aspects of the neurogenic program including the timing of neurogenesis onset (neurogenic timing), and the fate specification of bipolar cells. Two gaps in our understanding are the mechanistic interconnectedness of the progenitor properties regulated by Vsx2 and whether progenitors change in how they utilize Vsx2 over the course of histogenesis. To address these gaps, we generated two new Vsx2 alleles in mice, one with a knock-in reporter/knock-out configuration and the other for conditional gene inactivation. In the first two aims, we will characterize the retinal phenotypes of these mutant alleles and determine how they compare to phenotypes caused by a natural null allele and two missense alleles that correspond to disease-causing mutations in humans. Conditional gene inactivation will be done with tamoxifen-inducible Cre/lox recombination to determine the temporal windows of Vsx2 utilization in progenitors and test the hypothesis that Vsx2’s control of retinal identity, neurogenic timing, and proliferation are separable. We also predict that additional roles in the balanced production of cells in each retinal cell class (neurogenic output) will be unmasked by temporal inactivation after the start of neurogenesis. In the third aim, we test the hypothesis that retinal identity control shifts from a Vsx2-dependent to independent state that is epigenetically defined for some of the earlier targets. In the fourth aim, we will incorporate an ex vivo culture paradigm to test candidate genes identified in the previous aims for their functional significance in promoting or interfering with retinal development. Completion of these studies will provide new insights into how Vsx2 orchestrates retinal progenitor properties and how retinal progenitors drive retinal histogenesis, an essential component of eye organogenesis.

NIH Research Projects · FY 2025 · 2002-12

Systemic hypertension is a leading worldwide cause of morbidity and mortality. Salt-sensitivity is seen in over half of affected individuals with systemic hypertension as well in significant numbers of normotensive individuals at risk for subsequent development of hypertension. The etiology of “essential” hypertension is multifactorial and has been described as a “mosaic”, but numerous studies indicate an important role for both the adaptive and the innate immune systems in mediation of salt-sensitivity by affecting both kidney and vasculature in hypertension development and/or exacerbation. The studies proposed in this application will investigate novel interactions of the myeloid cyclooxygenase (COX)/prostaglandin system with the adaptive immune system and mechanisms by which this interaction can mediate regulation of kidney salt handling and hemodynamic responses to dietary salt. Myeloid cells of the innate immune system, especially macrophages and dendritic cells, are known to be a rich source of prostaglandins. Prostaglandins, and specifically PGE2, can act as immune modulators and affect both macrophage/dendritic cell and T cell function. Cyclooxygenase is the rate limiting step in the synthesis of prostaglandins from arachidonic acid, and cyclooxygenase 2 (COX-2) is the predominant inducible isoform in myeloid cells. We previously found that PGE2 is the predominant prostaglandin produced by COX-2 in kidney myeloid cells (macrophages and dendritic cells), and myeloid COX-2 deficiency leads to a proinflammatory (“M1”) phenotype while COX-2-mediated activation of the PGE2 receptor, PTGER2/EP4, promotes a non- inflammatory, alternatively activated (“M2”) phenotype. Experimental and epidemiologic evidence link inhibition or abnormalities in the cyclooxygenase/prostaglandin system to the pathogenesis or exacerbation of hypertension. Our previous studies demonstrated that COX-2-derived prostaglandins from myeloid cells of the innate immune system are important mediators of kidney salt and water balance. Wild type mice transplanted with COX-2-/- bone marrow exacerbated increases in high salt-induced blood pressure. However, our previous studies did not elucidate the mechanisms mediating these blood pressure alterations. We now find that mice with selective myeloid deletion of COX-2 have increased hypertension and T cell kidney infiltration in response to a high salt diet, but myeloid COX-2-/- mice crossed with mice with the Rag-1 deletion, which lack mature lymphocytes, fail to increase blood pressure. These results indicate a potential role for myeloid COX-2 activity to decrease T cell activation in response to a high salt diet. In the present studies, Aim I will investigate alterations in T cell phenotype in response to myeloid deletion of COX-2 alterations in T cell-mediated increases in kidney sodium reabsorption and/or directly activates kidney sodium transporters. Aim II will determine the role of myeloid COX-2 activity to activate the PD-L1/PD-1 system to decrease T cell activation and the role of myeloid COX-2 deletion to increase myeloid cell isolevuloglandin production and thereby activate T cells.

NIH Research Projects · FY 2025 · 2002-09

ABSTRACT The Vanderbilt-Meharry BIRCWH Program seeks to increase the pool of well-prepared investigators dedicated to expanding knowledge about women's health via advances in sex and gender biology. Our scholars conduct research across five thematic areas of exceptional institutional strength emphasizing five methodologic approaches to actualize personalized prevention, diagnostics, and therapeutics for girls and women. Leveraging a tradition of research excellence, scholars have access to many ongoing cohorts taking a lifespan approach, existing interdisciplinary units in all of our thematic areas, and large-scale clinical and population- level data linked to DNA and biospecimens, including prominent consortia. This environment creates synergy for robust mechanistic and therapeutic research to deliver discoveries both inside and outside the lab. Our 36 former and current scholars conduct research as varied as immunologic and genetic aspects of lupus, gender differences in outcomes of intensive care unit care, sex differences in resilience to Alzheimer's, and role of sex hormones in T cell differentiation and cytokine expression in asthma onset and severity. Scholars are 81% women and 28% identify with minority race/ethnicity, sexual orientation, or disadvantaged status historically underrepresented in research. Nearly all remain in research. During training, scholars average 8 publications a year, and alumni have been awarded 107 extramural grants worth over $50 million total. BIRCWH Scholars are grounded in fundamentals of women's health and sex differences research, prepared to lead collaborative teams, trained to effectively deploy innovative interdisciplinary approaches to attack and solve problems, and committed to pursuing research that optimizes the health of all women. Scholars are selected by competitive review of applications from early career faculty. Training is tailored to the individual scholar guided by structured interdisciplinary mentorship and is overseen by a trio of Co-PIs who are all former BIRCWH Scholars. Program resources are further extended by myriad institutional resources that ensure our researchers thrive. Scholars form a mentoring panel, participate in weekly BIRCWH work-in-progress sessions and seminars, receive formal evaluation twice a year, attend twice-monthly career development seminar series with other K-awardees, and are regularly exposed to case studies on responsible conduct of rigorous and reproducible research. They have access to: 1) an array of cores; 2) biostatistics consults; 3) manuscript preparation groups; 4) technical editing of completed products; 5) studios with experts to vet scientific ideas, aims, and research designs; 6) intramural pilot funding; 7) grant writing support including grant workshops, a library of funded grants, and internal study sections; and 8) new expert consultations to design fully individualized training plans termed Pathways. We deploy evaluation tools to continuously enhance our program with oversight by an advisory committee and site visitors. Combined, this ensures we carefully support excellence and build diversity in the next generation of women's health researchers.

NIH Research Projects · FY 2025 · 2002-06

PROJECT SUMMARY - OVERALL The long-term vision of the Vanderbilt Digestive Diseases Research Center (VDDRC) is to continue to inspire interest in the study of digestive diseases and perform paradigm-shifting science that translates to benefit for the patients and communities we serve. The collaborative VDDRC research base is multidisciplinary (88 faculty in 10 Departments; 58 Full members, 30 Associate members), and highly productive as evidenced by 1) 444 peer- reviewed publications (57% collaborative) including 89 in journals with impact factors >10, and 2) robust funding support which increased from $24.7M to $27M (31% NIDDK) since the last competing renewal. The overarching theme that galvanizes members is the study of microbial and host constituents that impact digestive disease pathobiology within the context of inflammation and the environment, and investigative interests fall into three interactive areas: 1) Gastrointestinal Infections and Injury; 2) Progenitor Cells, Development, Regeneration, and Pre-malignant Lesions; and 3) Obesity, Metabolism, and Nutrition. The VDDRC supports four cutting-edge Cores: 1) Cell Imaging, 2) Flow Cytometry, 3) Translational Analysis, and 4) Mass Spectrometry/Proteomics. These cores are highly utilized and well integrated into the Center and provide members with the latest advances in technology. Based directly on investigator demand, we augmented VDDRC Core offerings by adding, among other services: lightsheet microscopy and cryo-EM; spectral high- throughput flow cytometry; inductively coupled MS; as well as digital histology, multiplex IF/IHC, and state-of- the-art probe development for molecular small animal imaging within a strategically reorganized Translational Analysis Core. The VDDRC has now expanded member access to institutional Core services through an innovative Complementary Awards Program supported by new institutional funds ($125K), and extended the impact of the institutionally-supported ($125K) VDDRC Academy of Investigators by using new NIDDK and Academy funds to launch a Diversity Initiative targeting underrepresented minority researchers. A strong Pilot and Feasibility Program generated a 32:1 return on investment for NIH funds over the last 10 years and 96% of awardees remain active in GI research. The Pilot Program receives institutional support of $100K/year and we have now established new partnerships with Vanderbilt Centers to co-fund additional content-appropriate Projects. A responsive Administrative Core oversees financial management and operations and contains a Biostatistical Component and an Enrichment Program, which sponsors seminars and retreats to promote scientific collaboration and the development of new initiatives. The VDDRC also broadened its impact as 1) member publications involved collaborators at 504 institutions worldwide, and 2) the Center is now engrafted within a newly minted NIDDK-funded Eastern Regional DDRCC Alliance to promote the professional development of junior faculty. These accomplishments exemplify the ability of the VDDRC to function as a highly effective catalyst that both coalesces and promotes high-impact collaborative GI research.

NIH Research Projects · FY 2025 · 2001-09

PROJECT SUMMARY The Vanderbilt Clinical Oncology Research Career Development Program (VCORCDP) provides junior faculty conducting patient-oriented research in cancer with the tools and experience to design and implement hypothesis-driven research and effectively lead cancer research projects. This program is a cornerstone of the Vanderbilt Ingram Cancer Center (VICC), and its activities are embedded across the entire VICC enterprise, training leading investigators from every specialty related to cancer. This program ensures that we develop a strong and diverse pipeline of emerging investigators who have the skills necessary to drive impactful programs, and who have been accelerated in their trajectory by multidisciplinary and community-engaged focused learning. The primary components of VCORCDP are mentored clinical research in oncology coupled with tailored didactic training that may lead to a Master of Science in Clinical Investigation or other approved individualized educational pathway and group skill development, with exposure to the breadth of cancer research. Our mentors are a diverse group of 39 committed, experienced faculty. Scholars conduct research across the cancer care continuum in one of three domains: 1) Clinical Trials and Drug Development, 2) Translational Cancer Biology, and 3) Cancer Health Outcomes and Epidemiology. Through regular meetings with the Program Directors, peer scholars, and mentorship committees, as well as preparation of Individual Development Plans and structured Progress Reports, VCORCDP scholars tailor their career development to be able to address the most relevant and meaningful issues in their field. The program is co-led by two physician scientists, a medical oncologist who conducts translational research and a pediatric oncologist who conducts population science and clinical research. An Internal Advisory Committee representing the diversity of VCORCDP research evaluates the progress of individual scholars and the program’s success. Metrics include diversity of program participants, program completion, retention in research-focused careers, funding success, publication, and impact. An External Advisory Committee, composed of a cancer epidemiologist, a community representative, and four oncology physician-scientists, provides input into the scientific direction of program, ensures it is meeting national workforce needs and cancer priorities, and monitors progress, accomplishments, and trajectory to further programmatic advancement. Fifty-six scholars have entered the VCORCDP since its inception, 57% male and 43% female. As of June 2021, seven scholars (12%) represent URiM groups. Our 56 scholars have published over 1,800 manuscripts over their careers to date. Of the 50 scholars who have graduated, 45 (90%) continue in research-intensive careers, and 38 of the 45 (84%) have peer-reviewed grant support, including clinical trials. The VCORCDP is poised to continue to successfully develop investigators to lead patient-oriented cancer research and advance cancer care.

NIH Research Projects · FY 2024 · 2001-08

Project Summary Helicobacter pylori is the strongest risk factor for gastric cancer and interactions between this chronic pathogen and innate immune cells dysregulate signaling pathways that influence oncogenesis. One H. pylori oncogenic determinant is the cag type IV secretion system (TFSS) which translocates pro-inflammatory effectors, such as CagA, into epithelial cells. In studies supported by R01 DK 58587, we demonstrated for the first time that the cag TFSS can also translocate microbial DNA, which activates TLR9. In addition to inducing inflammation, however, activation of certain host pattern-recognition receptors can suppress inflammatory responses, conferring tolerance to chronic pathogens. In the last funding period, we used a genetic deficiency model to demonstrate that TLR9 suppresses the inflammatory response to H. pylori. These data coalesce with our recent efforts to more broadly define consequences of microbial DNA translocation, and exciting data from our laboratory now demonstrate that H. pylori can suppress activation of the DNA sensor/adaptor STING. Chronic pathogens such as oncogenic DNA viruses utilize multiple mechanisms to abrogate STING signaling and in other models of inflammation-induced disease with pre-malignant potential (e.g., chronic pancreatitis), inhibition of STING worsens disease via promoting Th17 polarization. Pertinent to this model, our provocative new data using mouse, gerbil, and human samples indicate that 1) STING signaling is absent in H. pylori-infected gastric tissue within the context of increased expression of the Th17 cytokine IL- 17A, a key driver in cancer initiation and progression, 2) genetic deficiency of IL-17A significantly reduces the severity of H. pylori-induced inflammation, and 3) H. pylori up-regulates Th17 differentiation and stabilization factors, but not STING targets in gastroid:macrophage co-culture systems. We have also identified a focused subset of H. pylori proteins that harbor homology to viral effectors that inhibit STING signaling. Finally, we have gastric tissue from a unique longitudinal cohort in Colombia from persons who either progressed to irreversible premalignant gastric lesions or remained stable, which will provide critical clinical validation of our mechanistic studies. Our hypothesis is that active suppression of STING signaling contributes to the augmentation in carcinogenic risk conferred by H. pylori by promoting persistence and deploying immune responses (Th17) with carcinogenic potential. We will test this hypothesis via the following Aims: 1. Identify, define, and validate microbial effectors that regulate STING suppression by H. pylori 2. Define mechanisms through which STING suppression promotes H. pylori-induced injury 3. Perform targeted interventions to activate STING within the context of H. pylori infection

NIH Research Projects · FY 2025 · 1998-09

This is the fifth competing renewal of the Cancer Center Support Grant (CCSG) at Vanderbilt-Ingram Cancer Center (VICC). VICC is a matrix center within Vanderbilt University Medical Center and Vanderbilt University that integrates the cancer-related expertise and resources of the Schools of Medicine, Nursing, Arts and Sciences and Engineering, the Peabody School of Education and the fully integrated Veterans Administration Medical Center. The major clinical facilities and the majority of research facilities are located on one campus, which promotes informal interactions, sharing of resources and productive collaborations. Established in 1993, VICC functions as an organizational unit with a supra-departmental status. VICC-specific responsibilities and aims are 1) to conduct, support and enhance state-of-the-art, multidisciplinary basic, clinical and population-based research; 2) to coordinate and integrate cancer-related activities across Vanderbilt and to collaborate with our local, regional, national and global partners; 3) to train and develop the next generation of cancer investigators, cancer leaders and the continuum of cancer care providers; and 4) to assess and prioritize community needs and to leverage partnerships to address those needs through cancer research, care and control activities. The research, training, and community outreach and engagement objectives are accomplished through CCSG and institutionally supported organizational capabilities, planning and evaluation, and eight research programs. The VICC Research Programs are Signal Transduction and Cell Proliferation, Genome Maintenance, Host-Tumor Interactions, Gastrointestinal Cancer, Translational Research and Interventional Oncology, Breast Cancer, Cancer Epidemiology, and Cancer Health Outcomes and Control (new). Eleven shared resources are proposed, all previously supported. Remarkable VICC growth and scientific discovery over the past project period has led to 22 new multi-investigator grants for a total of 71, and 8 new training grants for a total of 26. In addition, two NCI SPOREs were successfully renewed. With these and many other NCI grants, peer-reviewed funding increased 35%. Significant accomplishments have been made in precision oncology, cancer epidemiology, health outcomes and control, genomics, cancer drug discovery, and early detection and prevention research. Increased VICC space and facilities, along with philanthropic and institutional funds, supported the recruitment of 64 new faculty, who join a dedicated team carrying out the VICC mission: to alleviate cancer death and suffering through pioneering research; innovative patient-centered care; and evidence-based prevention, education and community initiatives. VICC senior leadership provides effective oversight and catalyzes innovative and paradigm-shifting science that translates to benefit for our patients and the community we serve in our catchment and reduces the burden of cancer nationally and globally.

NIH Research Projects · FY 2026 · 1997-09

Project Summary Helicobacter pylori is the strongest known risk factor for gastric cancer and interactions between this chronic pathogen and immune cells dysregulate gastric epithelial signaling pathways that influence carcinogenesis. One H. pylori oncogenic determinant is the cag type IV secretion system (TFSS) which translocates pro- inflammatory effectors, such as CagA, into epithelial cells. In studies supported by R01 CA 77955, we demonstrated that the cag TFSS can also translocate peptidoglycan, which is sensed by the pattern recognition receptor (PRR), NOD1. However, negative regulation of certain PRRs by chronic pathogens can increase pro-inflammatory responses and disease, and we have also demonstrated that prolonged H. pylori infection activates a NOD1-dependent negative feedback loop in gastric epithelial cells, leading to increased NF-kB activation. These data have driven our recent efforts to more broadly define the consequences of NOD1 suppression using human samples and rodent models; these findings demonstrate that 1) NOD1 expression is reduced in H. pylori-infected gastric cancer tissue compared with uninvolved tissue, 2) genetic deficiency of Nod1 significantly increases H. pylori-induced carcinogenesis and expression of the Th9 cytokine IL-9, a key effector of epithelial damage and inflammation, 3) H. pylori up-regulates expression of the IL-9 receptor in ex vivo epithelial gastroids, and 4) treatment of H. pylori-infected gastroids with IL-9 leads to enhanced cell survival within the context of Nod1 deficiency. We have established innovative models (primary gastroid:macrophage:T cell co-culture systems) that more closely recapitulate the infected gastric niche to demonstrate that H. pylori upregulates IL-9 production in a cag TFSS-dependent manner. Finally, we have gastric tissue and sera from a unique longitudinal human cohort in Colombia that includes persons who either progressed to irreversible premalignant gastric lesions or remained stable, which will provide critical clinical validation of our mechanistic studies. Our hypothesis is that suppression of NOD1 signaling contributes to the augmentation in cancer risk conferred by H. pylori by deploying immune responses (Th9) with carcinogenic potential. We will test this hypothesis via these Specific Aims: 1. Utilize novel rodent models and ex vivo systems to define IL-9-dependent host responses linked to H. pylori- induced disease progression 2. Validate and extend mechanistic studies focused on H. pylori, IL-9, and NOD1 using samples and H. pylori strains isolated from persons who did or did not progress towards gastric cancer 3. Use targeted approaches to inhibit IL-9 immune responses within the context of H. pylori infection

NIH Research Projects · FY 2025 · 1997-08

For the past three decades, we have sought to elucidate mechanisms of vesicle trafficking mediated by Rab small GTPase that regulate apical membrane trafficking of key intestinal epithelial transporters that regulate polarized enterocyte function. These investigations led to the recognition of MYO5B and MYO5A as regulators of trafficking through its interaction with multiple Rab proteins. The interaction with Rab10 is based on the presence of a minor MYO5B splice variant coding for the 30 amino acid Exon D. Our determination that MHV M glycoprotein also interacted with MYO5B Exon D, led us to investigate a possible role for MYO5B+D in regulating M protein trafficking. We have successfully completed our major aims to characterize the regulation of M protein trafficking by MYO5B and Rab10, but we have not been able to demonstrate any obligate requirement for this interaction for MHV assembly or egress. We therefore feel that it is necessary to refocus our efforts on our main interests in regulation of gastrointestinal epithelial cells. Our overall goal in Year 5 of the grant is to establish new approaches to modifying Rab11a and Rab8a dependent trafficking pathways that are responsible for discrete aspects of apical enterocyte trafficking. We will therefore pursue two new initiatives this year: First, we will seek to identify specific small molecule inhibitors of the interaction of Rab11a and Rab8a with MYO5B. We will utilize the yeast 2-hybrid assay screening system developed in the first 4 years of the grant to perform high throughput screens of drug libraries. Candidate hits will be tested for the concentration dependence of their action against Rab11a interaction with MYO5B and high potency inhibitors will then be tested for their ability both to inhibit the interaction of endogenous Rab8a or Rab11a with GFP-MYO5B tail. Second, we will seek to identify Rab11a and Rab8a-specific trafficking pathways for apical transporters and enzymes in intestinal epithelial cells. We have created two novel mouse alleles, LSL-Green Lantern-MYO5B tail(YE/QR) and LSL-Scarlet-MYO5B tail(QL/YC) using internal departmental funds. We hypothesize that blockade of Rab11a dependent versus Rab8a dependent trafficking will lead to specific deficits in trafficking and alterations in epithelial physiology. To that end, we will cross each transgenic allele onto the Villin-CreERT2 driver allele for induction of expression in the intestine. We will also assess the sequestration of Rab8a and Rab11a as well as apically directed transporters (including NHE3, SGLT1 and CFTR) with the dominant negative trafficking inhibitors. These studies will allow us for the first time to assess the importance of Rab11a versus Rab8a dependent recycling pathways on the physiology of intestinal enterocytes in vivo.

NIH Research Projects · FY 2025 · 1997-04

PROJECT SUMMARY The Vanderbilt Vision Research Center (VVRC) promotes transformative vision research, spanning the eye and its diseases to visual cognition, memory, attention, and integration. We request continued support for 7 well-coordinated service modules coordinated through the administrative module to provide services and capabilities that otherwise would be unavailable due to expense or need for specialized equipment, technical infrastructure, or computational, informatic and data management resources. Animal Models, Histology and Pathology, Instrumentation, and Computation and Data Management represent cores intrinsic to VVRC facilities, while Sequencing and Informatics, Cell Imaging and Analysis, and Mass Spectrometry and Proteomics subsidize use of the world-class institutional cores for which Vanderbilt is known. Animal Models (1) provides essential services for use of nonhuman primates or other large mammals and supports animal imaging capabilities through the Vanderbilt University Institute of Imaging Science. Histology and Pathology (2) provides preparation, embedding, sectioning, and staining of all tissues derived from visual structures. Instrumentation (3) provides customized apparatus and expertise in digital interfaces for equipment. Computation and Data Management (4) provides server maintenance, programming for data analysis and machine interfacing, system administration, and webpage-based applications and platforms for data management. Sequencing and Informatics (5) subsidizes use of VANTAGE, or Vanderbilt Technologies for Advanced Genomics for high-throughput DNA and RNA services, bioinformatic support, data analysis, and biospecimen storage. Cell Imaging and Analysis (6) offers high-resolution confocal and laser-scanning microscopy, electron microscopy, and other imaging modalities with high-performance image processing and analysis through the Vanderbilt Cell Imaging Shared Resource (CISR). Mass Spectrometry and Proteomics (7) supports use of the Mass Spectrometry Research Center (MSRC), which provides high-throughput analysis of protein modifications, differential expression and spatial imaging, protein-protein interactions, and biomarkers of disease. Finally, the administrative module ensures coordinated and stable operation of the VVRC research and training missions. In the period since the beginning of the current grant cycle (7/2019 – 9/2023), our 52 members holding 18 active NEI R01 awards published 657 papers making made fundamental contributions to basic and clinical visual science, with 414 from 41 faculty utilizing at least one service module and 201 utilizing two or more. Each service module was utilized by no fewer than eight investigators, and 19 investigators utilized at least three modules. This Core grant has increased collaborations within and between basic and clinical vision researchers across the Vanderbilt campus and with other institutions. This Core grant has enhanced recruitment of world-class vision scientists who continue extensive NEI-sponsored research at Vanderbilt.

NIH Research Projects · FY 2025 · 1996-12

Center Overview: Project Summary/Abstract The Vanderbilt Diabetes Research Center (VDRC), in its 48th continuous year of operation as a NIH- sponsored Diabetes Center, seeks to continue its efforts to facilitate the discovery, application, and translation of scientific knowledge to improve the care of patients with diabetes. The VDRC is an interdisciplinary program involving 142 participating faculty distributed among 15 departments in two schools and three colleges at Vanderbilt and neighboring Meharry Medical College. Because of the VDRC and the environment it creates, VDRC investigators have made important scientific contributions related to diabetes, obesity, and metabolism. The VDRC consists of: 1) Administrative Component that coordinates the scientific, organizational, enrichment, training, and outreach activities; For example, the VDRC established the National Diabetes Research Center Virtual Seminar Series early in the COVID crisis and this is now part of efforts by 15 Diabetes Research Centers to promote communication and interaction. 2) Biomedical Research Component that recruits and selects VDRC investigators and supervises the research cores that facilitate and enhance their research; 3) Pilot and Feasibility Program that facilitates the development of new investigators into independent scientists and encourages scientists in other fields to enter the field of diabetes research; and 4) National Enrichment Program in which the VDRC serves as the coordinating and organizing center for the NIDDK Medical Student Research Program which has allowed more than 1000 medical students from more than 140 US medical and osteopathic schools to conduct diabetes-related research at one of 16 NIH-supported Diabetes Research Centers. As part of its efforts to promote the next generation of scientists, the VDRC also enhances the efforts of 6 diabetes-related training grants, four of which are supported/funded by NIDDK The NIH support for the VDRC is greatly amplified by: 1) Vanderbilt’s sustained commitment to provide research space and additional financial resources; 2) a diverse, comprehensive array of research core services at Vanderbilt, which allows NIH funds to target unique, diabetes-related research cores; and 3) collaborative efforts with other NIH-funded research Centers at Vanderbilt. The VDRC is evolving and dynamic, including additions to its investigator base, expansion of VDRC research areas, expanded focus on clinical and translational research, realignment and evolution of core support to provide unique, indispensable core services, and service as a national resource for the diabetes research community.

NIH Research Projects · FY 2025 · 1996-09

PROJECT SUMMARY The volume-regulated anion channel (VRAC) is expressed ubiquitously in vertebrate cells where it mediates the efflux of Cl- and organic solutes required for cell volume regulation, an essential physiological process. VRACs are activated and inactivated by cell swelling and shrinkage, respectively. They also detect changes in intracellular ionic strength, which modifies their sensitivity to cell volume changes. VRACs and the genes that encode them are implicated in multiple diseases including diabetes, obesity, cancer and immunity. Whole genome RNA interference screening led to the demonstration in 2014 that VRACs are encoded by five members of the Lrrc8 gene family, Lrrc8a–e. VRAC/LRRC8 channels are hexaheteromers and require co-assembly of the essential subunit LRRC8A with one or more other LRRC8 proteins. Subunit assembly order and stoichiometry are unknown. Cryo-electron microscopy (EM) structures of homomeric LRRC8A and LRRC8D channels were recently determined. However, LRRC8A and LRRC8D homomers do not exist in Nature. Furthermore, LRRC8A homomeric channels have non-native functional properties and LRRC8D channel properties are undefinable because they are not trafficked to the plasma membrane. Existing cryo-EM structures thus have limitations for understanding VRAC/LRRC8 structure-function relationships. Directly translating LRRC8A and LRRC8D cryo-EM structural information into functional understanding is further constrained by the unknown and likely variable stoichiometry and assembly of hexaheteromeric VRAC/LRRC8 channels. Our laboratory, funded by DK51610, has studied VRAC extensively and was the first to demonstrate many of the channel's unique functional properties. Most recently, we described novel LRRC8 chimeric channel constructs that allow detailed molecular study of homomeric channels with physiologically relevant functional properties and defined stoichiometry and assembly. Our chimera studies uniquely demonstrated that 1) the LRRC8A intracellular loop, IL1, has unique structural features, 2) it is required for cell volume sensing, 3) the LRRC8 C-terminus is required for sensing changes in intracellular ionic strength and 4) both the LRRC8A IL1 and C-terminus are required for correct cellular processing of VRAC/LRRC8 channels. The overarching goal of this R01 renewal application is to utilize these novel LRRC8 chimeras to better elucidate VRAC/LRRC8 channel structure-function relationships. We will characterize the roles of the LRRC8A C-terminus in VRAC/LRRC8 channel regulation and will test the hypothesis that the LRRC8 IL1 determines VRAC/LRRC8 channel pore properties and regulates channel gating. We will also determine the cryo-EM structure of a unique LRRC8 chimera in multiple physiologically relevant conformations. Our studies will provide novel insights into the regulation and function of VRAC/LRRC8 channels and will provide a higher confidence foundation for detailed mutagenesis-based structure-function analyses.

NIH Research Projects · FY 2026 · 1996-05

PROJECT SUMMARY (ABSTRACT) Helicobacter pylori is a Gram-negative bacterium that colonizes the human stomach. Although most H. pylori- infected persons remain asymptomatic, potentially serious sequelae of infection include gastric adenocarcinoma, duodenal or gastric ulceration, and gastric lymphoma. Gastric cancer is a leading cause of cancer-related death worldwide, and H. pylori has been classified as a type I carcinogen by the World Health Organization. One of the major secreted proteins of H. pylori is a pore-forming toxin known as VacA. There is a high level of genetic variation among vacA alleles from unrelated H. pylori strains, and the encoded VacA proteins exhibit marked differences in their ability to cause alterations in human cells. A large body of literature indicates that H. pylori strains containing certain forms of vacA (termed s1, i1, or m1) are associated with a higher risk of gastric cancer or peptic ulcer disease than are strains containing other forms of vacA (termed s2, i2, or m2). Thus, VacA is recognized as an important H. pylori virulence factor. Most studies of VacA activity have focused on its ability to cause cellular vacuolation (expansion of endosomal compartments) or mitochondrial alterations. There are important gaps in knowledge regarding host cell responses to VacA, benefits that VacA confers to H. pylori, and mechanistic roles of VacA in H. pylori colonization of the stomach and gastric disease. Our recent experiments indicate that VacA treatment of gastric epithelial cells causes marked alterations in cellular cholesterol homeostasis. VacA-induced alterations in cellular cholesterol homeostasis are likely to be relevant in multiple contexts: (i) a cholesterol precursor altered by VacA has activities relevant for cell viability and cell proliferation; (ii) dysregulated cholesterol synthesis contributes to malignant transformation and gastric cancer risk; (iii) treatment of hypercholesterolemia with statins (cholesterol synthesis inhibitors) is linked to reduced gastric cancer risk in humans; and (iv) H. pylori is a cholesterol auxotroph that can acquire cholesterol from host cells. We hypothesize that VacA intoxication stimulates efflux of cholesterol from gastric epithelial cells, leading to activation of the cholesterol biosynthesis pathway, and that VacA-induced alterations in cholesterol homeostasis are relevant for H. pylori colonization of the stomach and the pathogenesis of H. pylori-induced gastric disease. The specific aims are (i) to define VacA effects on cholesterol homeostasis in gastric epithelial cells, (ii) to define effects of VacA on cholesterol localization and accessibility at the interface between H. pylori and host cells, and (iii) to define the impact of VacA and cholesterol on H. pylori-host interactions in vivo.

NIH Research Projects · FY 2025 · 1995-04

Modified Project Summary/Abstract Section This application requests continued support for the “Vanderbilt Infection Pathogenesis and Epidemiology Research” (VIPER) Training Program. The objective is to prepare postdoctoral fellows for careers in infectious diseases basic science, translational research, and epidemiology in infectious diseases. Training is in basic laboratory research to study disease pathogenesis using molecular and cellular methods, translational science, and patient-oriented epidemiology and outcomes research. Our field has a critical labor force shortage in physician-scientists in these interrelated areas that this T32 addresses. There is broad expertise among our program faculty covering immunology and inflammation, virology (HIV and other viruses), bacteriology, personalized medicine, vaccines, clinical trials, and epidemiology and outcomes research (HIV, TB, hospital epidemiology, and global health). VIPER brings early-stage, emerging investigators together, rather than training them in silos, enhancing the potential for basic scientists to appreciate and understand “real world” problems and barriers to implementation that might inform their own experimental designs, hypotheses, and long-term goals. Similarly, we help empower translational scientists to better appreciate both clinical (“bedside”) needs, revealed by epidemiological research, and candidate, bench-based solutions, which they can then couple through translational research to advance human health. Enhancing interactions among these three “phenotypes” of investigators we also position epidemiologists to collect and analyze data that are most informative to their translational and basic science colleagues. Almost every “intractable” problem in infectious diseases will benefit from this holistic approach, which our T32 is aiming to foster. We will catalyze team science and collaborative problem solving through integrated conferences, networking events, shared classes/coursework, and regular interactions. Participating faculty are in the Infectious Disease Divisions of Medicine and Pediatrics, the Division of Epidemiology, and the Department of Pathology, Microbiology and Immunology. This program has a strong track record of interactive training of post-MD fellows from both Medicine and Pediatrics, and also includes post-PhD trainees. There is extensive collaboration among mentors. Long-term success is maximized by a customized research career plan for each trainee with a dedicated mentor, and under the supervision of an expert advisory committee and the Program Directors. VIPER provides support for Masters of Science in Clinical Investigation and Masters in Public Health Degrees. Physicians complete a year of clinical training before starting VIPER training, and a minimum of two years of research training is required for all fellows.

NIH Research Projects · FY 2024 · 1993-09

With the rapid accumulation of knowledge resulting from the development of new genetic and proteomic screens and methodologies, this is an exciting period in biomedical science, and there are now tools and techniques available to elucidate the underlying molecular etiologies of diseases and use this knowledge to provide new and more effective treatments or cures. The increasing incidence of kidney disease in the U.S. and worldwide represents a major challenge to our health systems. There is a growing urgency to train physician scientists and Ph.D. scientists who can increase our understanding of disease pathogenesis and develop, implement and assess new therapeutic modalities to combat renal disease. However, during the past decade, there has been a disturbing decrease in interest in Nephrology as a profession among potential trainees and especially a decreasing interest in committing to a career in kidney research. Therefore, it is crucial for research-rich nephrology programs to seek out, train and mentor the next generation of kidney research scientists. The goal of the Vanderbilt Nephrology Training Program is to provide promising M.D. and/or Ph.D. postdoctoral fellows with the knowledge and the tools to become the leaders in both basic and translational research efforts to generate new understanding in the pathogenesis of kidney diseases and to develop and implement new treatments and cures for our patients with kidney disease. The faculty who participate in the Vanderbilt Nephrology Training Program represent a group of talented and accomplished researchers with a broad range of scientific expertise and research interests. This group consists of 29 investigators whose primary appointment is in either Adult or Pediatric Nephrology and 15 other clinical and basic scientists who have ongoing interests in kidney disease or associated scientific questions. The training faculty are based in 8 departments: Medicine, Pediatrics, Pathology, Preventive Medicine, Bioinformatics, Biochemistry, Cell and Developmental Biology and Anesthesia. Research interests of the primary faculty include mechanisms of chronic glomerular and tubulointerstitial injury, acute kidney injury, cell- matrix and cell-cell interactions, renal development, epithelial cell biology, eicosanoid biology, epidemiology of progressive kidney disease, development of clinical biomarkers, health services research and clinical trials. Our training program is structured to provide physician/scientist (M.D. or M.D./Ph.D.) and selected basic scientist (Ph.D) trainees with a focused and productive research experience that will serve as the foundation for an independent investigative career directed toward understanding and treatment of renal disease. Project Summary/Abstract Page 6

NIH Research Projects · FY 2025 · 1992-07

Abstract The long-term objective of the Vanderbilt Training in Gastroenterology T32 Program is to identify, recruit, train, and mentor a diverse workforce of compassionate and dedicated investigators by providing a rigorous scientific foundation in 1) basic research, to reveal new disease mechanisms and identify novel therapeutic targets; or 2) clinical/translational research, with the opportunity for formal studies in clinical science, epidemiology, or informatics. Richard Peek serves as Director and is assisted by an Associate Director, Keith Wilson, as well as a Steering Committee and Internal and External Advisory Committees composed of senior faculty deeply invested in training young investigators and cultivating diversity. The design of this multidisciplinary program is to support 5 postdoctoral fellows/year (1-2 years in duration) who show exceptional aptitude for pursuing research careers. Trainees are selected from a curated pool of fellows accepted into the Gastroenterology training programs, physician-scientist applicants from other clinical training programs, and applicants that apply directly to preceptor laboratories. A customized mentoring team is constructed for each trainee consisting of a Mentor with nationally recognized expertise and a Research Advisory Committee to provide additional guidance, mentoring, and feedback. Each trainee becomes part of a translational research group that includes basic and clinical investigators (40 Preceptors/10 Departments), enabling an in-depth understanding of specific diseases from bench-to-bedside. The trainee experience is enriched by an extensive program of seminars and coursework tailored to meet individual needs and via the Academy of Investigators, which provides comprehensive career development and a Diversity Program focused on underrepresented minority (URM) investigators. This T32 is also engrafted into a continuum that provides a wealth of funding opportunities only available to fellows and junior faculty to facilitate an uninterrupted research focus resulting from funding stability. In the last 15 years, 51 trainee scientists have been or will be supported by this Program (33 MD or MD/PhD, 16 PhD, 2 DO); 26 (51%) have been female, and 10 (20%) have been from URM groups. Further, the number of female trainees and URM trainees increased substantially in the current funding period (female: n=12, 60%; URM: n=5, 25%) compared to the previous funding period (female: n=5, 36%; URM: n=1, 7%). At present, 41/51 (80%) of these trainees are pursuing research-intensive or research-related careers or are still in training, 81% of Program graduates have received research funding and, since 2014, the average number of publications by all trainees is 16.4/year. Collectively, the unique environment that supports digestive diseases research at Vanderbilt, catalyzed by rich collaborative interactions between basic and clinical researchers, a wide range of supporting Cores and Centers, robust institutional support, and exposure to state-of-the-art clinical care, provides an outstanding opportunity to train successful investigators using a team-science approach, whose discoveries can be rapidly translated into improved patient care.

NIH Research Projects · FY 2025 · 1986-09

The collagen IVα345 (Col-IVα345) scaffold, the major constituent of the glomerular basement membrane (GBM), is a critical component of the kidney glomerular filtration barrier. In chronic kidney disease, affecting hundreds of millions of people worldwide, over two thousand genetic variants occur in the COL4A3, COL4A4, and COL4A5 genes that encode the α3, α4, and α5 chains of the Col-IVα345 scaffold. The chains are the autoantigens in Goodpasture’s (GP) autoimmune disease, mutated in Alport syndrome, and dysfunctional in diabetic nephropathy (DN). The rationale development of therapy hinges on gaining new understandings of pathogenic mechanisms. GP disease, a renal pulmonary disorder, continues to serve as the vanguard for unlocking these medical mysteries. Among our discoveries are: a) the crystal structure of the NC1 hexamer of the α345 collagen IV scaffold, b) discovery of the primordial chloride pressure and its constraint against neoepitope formation in Goodpasture’s disease3, c) discovery of functionality of the α345 NC1 hexamer in the assembly of the GBM as an ultrafilter of proteins, d) development of the collagen triple-helix mapping tool, e) capacity of NC1 domain to home in the assembly of basement membranes, and f) detrimental effect of high glucose on the BM modifying enzymes. These published discoveries, together with substantial amount of unpublished pilot data, provide the framework for four specific aims. Our overarching hypothesis is: Col-IVα345 tethers macromolecules forming supramolecular complexes, perturbation of which, in genetic and acquired disorders, cause glomerulopathies. The achievement of the aims will yield new insights into the etiology of GP, the structure and assembly of collagen IV scaffolds and binding partners, and the molecular mechanisms of underlying DN. This knowledge is of fundamental importance for the rationale design of therapies that target the primary cause disease.

NIH Research Projects · FY 2026 · 1980-05

Project Summary This grant application from the Vanderbilt Student Research and Training Program (SRTP) requests continued support for medical student research training in diabetes, obesity, digestive diseases, and kidney diseases. The SRTP has provided research training for over 1200 students from 130 medical schools. Each year, the SRTP supports 32 medical students to conduct research with experts in diabetes, digestive disease (GI), and kidney disease (20 students in diabetes research, 6 in GI, and 6 in kidney). The SRTP also partners with the national NIDDK-Medical Student Research Program (DK-MSRP), which enables an additional 80-90 students to conduct research at one of 16 NIDDK-supported Diabetes Research Centers. The goal of the SRTP is to train physician-scientists and provide exposure to mentors who will inspire this career trajectory. A major focus of the SRTP is outreach to students with limited exposure to research environments due to institutional or curricular constraints that may limit their opportunities to conduct research. We also provide exposure to key clinical topics in diabetes, GI diseases, and kidney diseases to guide students’ clinical and research careers toward a focus on these important diseases. The SRTP is affiliated with two Vanderbilt research centers: one a NIDDK-supported center grant in diabetes (P30; DK020593) and the other a NIDDK-supported center grant in digestive disease (P30; DK058404) that provide preceptors for the student’s research experience. The SRTP has three main components: 1) A two-to-three-month intensive mentored research experience designed to “light the fire” for discovery; 2) A twice-weekly enrichment program focused on educating about the potential career pathways of physician-scientists and key clinical and research topics in diabetes, GI, and kidney disease; 3) A national research symposium: This is held at the end of the summer jointly with students from the DK-MSRP. The students present their work, hear keynote talks from visiting professors on career development, network with faculty and near-peer mentors, and learn about the next steps for research options in residency and fellowship. We propose aims and innovations that will help SRTP adapt to a rapidly changing world of research, clinical care, and medical education. These aims center around: 1) building better flexibility with changing medical school curricula, 2) vertical integration with other physician-scientist training programs and near-peer mentoring, and 3) continuing to develop robust post-program support to guide career paths. With continued support from the NIH/NIDDK, we propose a comprehensive and exciting experience early in students’ medical careers that will help them make career-defining decisions.

NIH Research Projects · FY 2025 · 1977-07

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. The discipline of Clinical Pharmacology seeks to apply an understanding of the fundamental mechanisms of drug action to improve the therapy of human diseases. The Division of Clinical Pharmacology at Vanderbilt offers an outstanding research-based, postgraduate fellowship program committed to training future leaders in the discipline. The mentoring faculty includes 16 members of the Division of Clinical Pharmacology, along with 17 members of other divisions or departments. Collaborations among investigators focusing on common research themes are well-established in the Division; this includes a strong focus on pharmacogenetics Collaborative efforts exist in pediatrics, cardiology, rheumatology, bioinformatics, genetics, and nephrology among others. The primary activity of trainees is research training in a mentored setting on questions directly relevant to drug action in man. Research can vary from bench-based translational work to clinical studies. The duration of training is 2-3 years. In the ~4 ½ year period since the last competitive renewal, 17 new trainees were appointed to the training grant: Of the 17 fellows supported, 11 had an MD or equivalent degree (65%), 3 dual MD/ PhD (18%), 1 Pharm D (6%), and only 2 with a PhD (12%). 4/12 fellows that graduated from the program have received K-awards from NIH (33%). Research training under the direction of individual faculty mentors is supplemented by didactic course work and seminars. Required courses of trainees include research ethics, responsible conduct of research, biostatistics and study design, drug regulation and development, and pharmacokinetics and pharmacodynamics. In addition, attendance at Clinical Pharmacology Grand Rounds and a weekly Fellows Lecture Series is required. This curriculum supplements the trainees' research experience and provides a broad knowledge base that will allow for fellows to develop into successful leaders in Clinical Pharmacology. There is a strong emphasis on teaching the principles of reproducible research. The excellence of the training program has resulted in substantially more applicants than positions. Growth of clinical pharmacology at Vanderbilt offers unique opportunities to further enhance the training program. Such growth includes the Oates Institute of Experimental Therapeutics that focuses on pharmacogenetics and personalized medicine; the Vanderbilt Center for Bone Biology; and the Vanderbilt Center for Arrhythmia Research and Therapeutics (VanCART) - all components of the Division of Clinical Pharmacology. The overall mission of the Vanderbilt Clinical Pharmacology Fellowship Program is to train investigators who will ultimately assume leadership positions in the discipline; it has been highly successful in this mission. Of trainees supported by the award in the past 15 years, approximately 88% are in academic medicine, industry, or government. Clinical Pharmacology at Vanderbilt is a vibrant and dynamic enterprise supported by a training program that is continually evolving to better meet the needs of its trainees.

NIH Research Projects · FY 2025 · 1975-07

This proposal seeks continued support for an Institutional NRSA that provides post-doctoral research training to 4 M.D. and/or Ph.D. trainees in endocrinology, diabetes, and metabolism. Also requested are 6 short-term summer medical student training positions. This multi- disciplinary program involves 32 faculty preceptors in five clinical and basic science departments of Vanderbilt University School of Medicine. It is an integral component of the training activities of the Vanderbilt Diabetes Research and Training Center. Its goal is to provide trainees with the knowledge and skills required for independent translational or clinical research careers. It was previously designed to train both M.D. and Ph.D. scientists in equal numbers. However, due to the stated NIH goal of increasing clinical research, we will focus our recruitment efforts on M.D.'s, M.D./Ph.D.'s and Ph.D.'s with more clinical research interests. This training may be in pre-clinical, translational, or clinical research areas, with emphasis on providing a basic science experience where appropriate. Trainees will be selected from the nationwide pool of applicants that traditionally seek training at Vanderbilt. M.D. or M.D./Ph.D. trainees usually have three to four years of relevant residency training before entering the program. Ph.D. trainees will be considered for their first post-doctoral experience. Trainees are selected on the basis of their potential as future researchers and educators. The training program utilizes a preceptor-based approach, in which the trainee develops a research project under the guidance of a faculty mentor and mentoring committee. Several criteria are used to match preceptor and trainee, including trainee interests, quality of the project proposed, amount of direct supervision that the preceptor can provide, and adequacy of the research funding and facilities available. Typical research topics include hormone action in humans as it relates to diabetes, obesity, and metabolic regulation. Research training on this T32 is at most two years, with the goal of extra training years provided by individual fellowship grants. Training is supplemented by conferences, seminars, coursework, and career guidance efforts. Trainee progress is evaluated by the oversight committee twice-yearly and the program is evaluated yearly by the same committee. In the 46 years of NIH support, the program has supported 132 post-doctoral trainees. Of those who have completed training, over 80% have embarked upon careers in academia, industry or government.