University Of Alabama At Birmingham

NIH Research Projects · FY 2024 · 2024-09

Abstract The Metabolism Core Laboratory at the University of Alabama at Birmingham (UAB) proposes to purchase and install an indirect calorimeter system to support leading research programs in metabolism, nutrition, diabetes, obesity, and other chronic metabolic diseases. Indirect calorimetry, via a whole-room indirect calorimeter system, is the gold standard for measuring gas exchange to estimate energy expenditure (EE) via oxygen consumption and carbon dioxide production, providing precise and accurate measures under regulated environmental conditions. UAB has a history of innovation in studies of metabolism utilizing indirect calorimetry and, in the 1990s, was among the first to install an indirect calorimeter in an academic research facility. This original indirect calorimeter reached the end of its functionality following 20 years of service, and researchers have been required to shift measurement modalities toward mobile metabolic monitors or doubly labeled water, both of which have significant limitations relative to the 24-hour measurement capability of a whole-room indirect calorimeter. Despite these limitations, multiple NIH-funded investigators have incorporated measures of EE into study designs, resulting in high-impact scientific studies and publications while demonstrating a sustained need for the return of whole-room indirect calorimetry capabilities. The room calorimeter system proposed in this application will meet these research needs and provide a replacement for the previous system to again enable appropriate, controlled environmental conditions for both short- and long-term EE measures. Components of daily EE include resting EE, activity-related EE (both exercise/physical activity and non-exercise activity), thermic effect of feeding, respiratory exchange ratio (for nutrient substrate utilization), sleeping metabolic rate, total daily EE, and other measures as required by study designs. While some aspects of these comprehensive EE measures may be collected via a portable metabolic monitor or doubly labeled water techniques, effects on EE associated with nutrition composition, timing of feeding, energy balance, and health and disease states often require the precision and accuracy available only in the whole-room indirect calorimeter system. Combined with the ability to deliver interventions (nutrition, activity, pharmaceutical, environmental), the room calorimeter system will support the current research base and be available to a broad network of investigators connected to the UAB Nutrition Obesity Research Center, Diabetes Research Center, and O’Neal Comprehensive Cancer Center and restore indirect calorimetry capabilities to the state of Alabama and the broader region, where none currently exist.

NIH Research Projects · FY 2024 · 2024-09

Abstract Alzheimer’s disease (AD) affects millions of people worldwide, and its prevalence is expected to increase dramatically in the coming years. Although some people are resilient to AD pathology with normal cognition despite tau and amyloid deposits, understanding the factors that drive such resilience will be critical for identifying treatments for those at the highest risk of impairment. In the research proposed here, we will use cognitive neuroscience methods to develop a systems-level understanding of the mechanisms that underlie early-stage AD. We will examine tau deposition and functional network health in a sample of participants with normal cognition to understand the etiology of AD and individual differences in cognitive resilience. Recent results have shown that tau is present in different parts of the brain in different people. Thus, we hypothesize that tau deposits in a particular brain network disrupt functional network health, and patients who appear cognitively normal despite pathology have tau deposits in locations that have a minimal impact on functional networks. To test this hypothesis, we will use tau positron emission tomography (PET) to quantify the spatial distribution of tau pathology. We will also determine the role of brain network connectivity and atrophy in the health of a cohort of participants enriched for individuals at risk for AD. In Aim 1, we will use data from the publicly available Alzheimer's Disease Neuroimaging Initiative dataset to test our hypotheses by determining how tau deposits affect the health of pre-defined brain networks. We will measure functional network health based on network segregation, neural dynamics, and cortical atrophy. In Aim 2, we will determine how segregation, dynamics, atrophy, and tau deposits affect network flexibility. We will recruit 150 cognitively normal participants aged 55 or older from populations enriched in participants with blood markers indicating tau and, thus, at high risk for developing AD. They will undergo tau PET and resting-state functional magnetic resonance imaging (fMRI) as part of a University of Alabama at Birmingham Alzheimer's Disease Center study and will undergo fMRI during an auditory/visual attention task. In Aim 3, we will determine how these variables interact to affect cognitive and sensory performance and whether they predict longitudinal cognitive decline. Successful completion of this work will result in a better understanding of how tau deposits affect the normal patterns of brain activity and patterns of brain atrophy. These studies will provide new information on the mechanisms through which tau results in a system-wide decline and the conditions that lead to resilience. This knowledge may suggest treatments for patients with AD and patient-specific training or behavior modification strategies that, applied early in the course of AD, could mitigate its long-term effects.

NIH Research Projects · FY 2025 · 2024-09

Although child occupant fatalities have decreased dramatically since 1975, ~2 children die in crashes each day and ~60k more are treated each year in emergency departments (EDs) for crash injuries. The bulk of research to date examining factors that contribute to crash outcomes, including results from my K99 period, are focused on the time period before and during a crash. Very little research has examined post-crash factors and their influence on children’s long-term health. The high burden and costs of crashes among children ($530M annually [CDC 2019]) underscore the critical need to improve child occupant safety across the crash-injury continuum. Enhancing child occupant safety would prevent thousands of childhood disabilities and deaths annually, while ameliorating the financial, physical, and psychological burden of crashes experienced by children, their families, and society. This R00 is utilizing a breadth-to-depth approach that integrates knowledge obtained from population- (breadth) and individual-level (depth) analyses to improve children’s long-term health outcomes following a crash. The K99 phase of this work leveraged novel, state-level data to identify factors present pre- and during- a crash that contribute to child restraint system use and children’s injury risk. The R00 phase of this work seeks to expand these findings, specifically by conducting mixed-methods studies at two pediatric hospitals to characterize and understand factors that contribute to post-crash outcomes for children. Aim 1 of the R00 is to examine potential variations across hospitals in diagnostic, treatment, and discharge approaches for crash-involved children. To achieve this aim, we will interview physicians and other health care providers who make decisions about children’s treatment needs/options following a crash. We will also conduct a large, retrospective review of health records across study sites. Aim 2 is to characterize post-crash quality of life and health outcomes for a large cohort of crash-involved children and their families. We will use longitudinal surveys and semi-structured interviews completed by caregivers to characterize post-crash outcomes (e.g., physical and mental health, family dynamics). The knowledge gained by this project will ultimately reduce overall rates of pediatric crash injury and death by bolstering postcrash treatment strategies for children.

NIH Research Projects · FY 2025 · 2024-09

Project Summary/Abstract Polycystic kidney disease (PKD) is the leading genetic cause of renal failure, affecting an estimated 500,000 Americans. There is currently no cure for the disease, as we do not know what pathway drives disease pathogenesis. What we do know is that the driver of this disease localizes to an organelle known as the primary cilium. Our preliminary data using a novel genetic mouse model provides strong evidence that ARL13B, a regulatory GTPase that localizes to the primary cilium, drives cyst formation. This proposal aims to uncover the mechanism of ARL13B action in the cilium, by specifically dissecting its interaction with three effector proteins using a combination of biochemical and genetic approaches. We prioritized a list of 30+ candidate interacting proteins based on their links to renal cystogenesis, ARL13B interaction, and known pathways in other tissues. From this list, we are bringing the top three candidates–CDK1, ARL3, and INPP5E– forward for investigation in this proposal. These top candidates will provide knowledge how they are involved in kidney cystogenesis and provide Dr. Van Sciver with a framework in which to test other putative interactors of ARL13B. In Aim 1, we identify critical amino acid residues required for interaction between ARL13B and each effector. In Aim 2, we directly test ARL13B’s enzymatic activation of ARL3 in driving kidney cystogenesis using a point mutant ARL13B in vivo mouse model. In Aim 3, we directly test the enzymatic activity of INPP5E, which requires ARL13B for its localization to cilia, in driving kidney cystogenesis using a point mutant INPP5E mouse model. Aim 1 will be performed under the co-mentorship of Dr. Rick Kahn, a renowned expert in GTPase biochemistry, specifically in ARL and ARF family GTPases, and Dr. Tamara Caspary, an accomplished mouse geneticist. Dr. Van Sciver will receive additional training through didactic lectures in kidney physiology and purification and analysis of protein complexes. In addition, the Atlanta Network Training in KUH Scientific Research (ATLANTIS) program offers Dr. Van Sciver opportunities through three cores: research training, professional development, and networking. The detailed training plan coupled with this proposal will contribute to the success of Dr. Van Sciver’s overarching goal of becoming a tenure-track faculty member at an academic or research institution. Long-term outcomes from this proposal will define precise mutations mediating ARL13B’s interaction with each effector molecule, providing fundamental knowledge with a high probability of dissecting ciliary ARL13B’s pro-cystic mechanisms in part or in whole. In addition, this proposal will determine whether ARL13B functions via ARL3 and INPP5E and set up Dr. Van Sciver to test whether ARL13B specifically acts via CDK1 by generating a mouse in analogous experiments to Aims 2 and 3 in the future. This 5-year research and training plan will allow Dr. Van Sciver to establish an independent research program identifying mechanisms driving PKD.

NIH Research Projects · FY 2025 · 2024-09

Membrane transporters are essential for cellular life and are generally understudied compared to other membrane proteins, such as GPCR and ion channels. The long-term goal is to pursue a fundamental mechanistic understanding of membrane transporters closely related to human diseases. In this proposal, the overall objective is to extend our previous research on members of human SLC and bacterial siderophore ABC importers. The rationale is that, with detailed knowledge about the structure-function relationship of these membrane transporters, it will be possible to design small molecules explicitly targeting them to have physiological effects. To accomplish the goal, we expect to pursue the following three projects: 1) understanding the substrate selectivity and promiscuity; 2) probing the connection between transporters’ unique structural features and their function; and 3) searching for small molecule inhibitors/activators of the transporters. The approach involved in this proposal is multidisciplinary, including biochemistry, structural biology, microbiology, cell biology, molecular dynamics simulation, and various methodologies of biophysics. This proposal is significant because it is expected to reveal the basic molecular mechanisms of essential membrane transporters and provide scientific justification for further development of drugs targeting these transporters.

NIH Research Projects · FY 2026 · 2024-09

PROJECT SUMMARY Dementia such as Alzheimer's disease (ADRD) is the most expensive medical condition20 in the US and affects more than 5 million Americans21. Analyses from the ACTIVE study showed that a specific type of computerized cognitive training (CTa) reduced risk of ADRD among older adults 29% across 10 years22. [Recent follow-up analyses indicate that ACTIVE participants with signs of mild cognitive impairment (MCI) at baseline randomized to CTa were 23% less likely to be diagnosed with ADRD across 20 years23]. While these results are encouraging, MCI was not clinically diagnosed and thus evidence is inconclusive to recommend CT for ADRD prevention. Many efficacious CT techniques now exist but have distinct cognitive effects. Given that cognitive deficits are varied among those with mild cognitive impairment (MCI), who are at higher risk for ADRD, a combination of CT techniques may be most efficacious. Significant knowledge gaps remain as the best CT exercise(s) for those with MCI is unclear. We were awarded a clinical trial planning grant to design and establish the feasibility of the ACTIVE MIND trial (AG062368). We propose phase II of ACTIVE MIND, an adaptive random- ized trial to identify the most efficacious CT exercises to improve everyday function in MCI. We will further quantify the effect size of CT to reduce incident ADRD among persons with MCI. In this phase II trial, our primary objective is to determine which CT arm results in the largest functional improve- ments and has the best probability to reduce ADRD incidence. Our investigators include international experts in CT, MCI/ADRD, recruitment and retention, neuropsychological assessment, neuroimaging, biomarkers, and adaptive trial design. Our approach is to compare different CT arms to a stringent active control condition with equivalent participant expectations. Measures will include innovative indi- ces of instrumental activities of daily living (IADL), standard cognitive assessments, as well as neu- roimaging and novel blood-based biomarkers. Potential moderators of CT will be assessed to identify who benefits. This study is innovative, in the application of adaptive trial methodology to efficiently identify the most efficacious CT exercises to reduce ADRD incidence in MCI. We further explore neu- roimaging and blood-based biomarkers as potential moderators of CT outcomes. Our premise is that targeted CT improves everyday function (i.e., IADL), which subsequently delays ADRD onset. Our long term goal is to improve older adults' functional trajectories and thereby curb ADRD prevalence. The contributions will be significant, advancing our understanding of how CT may be successfully im- plemented to curb ADRD prevalence. Significance is considerable given that an intervention delaying the onset of Alzheimer's disease by only one year will result in 9.2 million fewer cases of the disease by 205024.

NIH Research Projects · FY 2024 · 2024-08

PROJECT SUMMARY & ABSTRACT The overarching objective of our T32 application “AdvanceSGM Health for Research Diversity” (hereafter referred to as Advance SGM) is to support the development of predoctoral trainees who will have exceptional research and community engagement skills in advancing sexual and gender minority (SGM) health equity in the Deep South. The rationale motivating our program is twofold: First, the need to recruit, retain, and raise-up researchers from underrepresented backgrounds in an effort to understand and address the dramatic health inequities facing SGM communities across our region. Second, the need to address SGM health holistically, not just from a disease or deficits approach. We will provide predoctoral trainees with education and experience in rigorous, reproducible, and transparent scientific approaches. We have designed our program to include the following key activities: 1) Provide advanced training in SGM research through weekly seminars focused on health equity, multi-level prevention interventions, methods, and their intersections; 2) Prepare trainees for an academic research career via application workshops focused on professional development training; 3) Engage trainees in SGM research via a shadow experience and community intensive for trainees to ‘learn by looking,’ followed by an immersive community-based participatory research project for students to ‘learn by doing’; 4) Expand trainees’ personal and professional networks through formalized mentoring and networking opportunities; 5) Evaluate the effectiveness of AdvanceSGM in increasing the diversity of scholars addressing SGM health equity across racial, ethnic, SGM, and other minoritized statuses. The planned duration of appointments is two years per cohort. We project a total of 15 trainees across three cohorts (1st cohort years 1 and 2, 2nd cohort years 2 and 3, final cohort begins in year 5 and continue their final year upon T32 renewal). We will draw on our diverse personal and professional networks to recruit applicants from underrepresented backgrounds designated by NIH (e.g., experience with homelessness, educationally and/or financially disadvantaged, raised in rural and/or Centers for Medicare & Medicaid Services-designated low- income areas) alongside other systematically excluded groups (e.g., race and ethnicity, sexual minority, gender minority). Prospective UAB doctoral students, as well as current doctoral students already enrolled in the Schools of Public Health, Nursing, or College of Arts and Sciences and who are in years 1 or 2 of their program, are eligible. Intended trainee outcomes will include traditional NIH evaluation metrics (e.g., presentations, publications, and grants submitted and awarded) in addition to other critical metrics such as the extent to which the training contributed to the development and growth of professional networks among trainees. This will be assessed through a robust mixed methods evaluation plan.

NIH Research Projects · FY 2025 · 2024-08

The societal and patient-centered impacts of osteoarthritis (OA) are profound – total costs for OA treatment in the US exceed $486 billion annually. OA management is limited to symptom management (e.g., pain, inflammation). As such, OA often progresses to end-stage at which time only surgical options are available in the form of total joint replacement (TJR). Alarmingly the prevalence of moderate-severe functional limitations 2- 5 years post-surgery remains high (>30%) post TJR. OA is a heritable trait which we recently estimated to be as high as 50% in Hispanic Veterans in the Million Veteran Program (MVP). Despite two recent seminal manuscripts on the genetics of OA, gaps remain in our understanding of the role of specific genetic variation to OA susceptibility, heterogeneity, and progression. These gaps impede development of personalized approaches essential for guiding OA risk reduction and therapeutic intervention. Previous research has demonstrated OA joint site reflects underlying etiological heterogeneity demonstrate distinct genetic region associated with OA at specific joint sites. Similarly, women are more likely to develop OA however, the MVP is comprised of predominantly male Veterans limiting power to identify genetic variants which may be sex specific. Our overall goal is to decipher the genetics of OA susceptibility, heterogeneity, and progression. Toward this goal in Aim 1, we identify new and fine-map known genetic loci associated with susceptibility to OA and total hip/ total knee arthroplasty (THA/TKA); in Aim 2, we disentangle OA heterogeneity by identifying distinct genetic variation associated with OA at specific joint sites; and in Aim 3, we identify sexually dimorphic genetic variants associated with OA in women. We will identify genetic regions associated with OA susceptibility, heterogeneity and progression enabling risk stratification and in turn advancing patient care.

NIH Research Projects · FY 2025 · 2024-08

There are healthcare gaps in health in multiple older communities, resulting in potentially preventable morbidity and mortality in older adults. Addressing gaps in health and healthcare for older adults requires innovation in aging research at the nexus of geriatric medicine, gerontology, and palliative care research. Community Based Participatory Research (CBPR) is an important and promising strategy to meet this challenge because it has been shown to, increase trust, and result in positive health outcomes. The goal of this K07 Academic Career Leadership Award is to leverage the applicant’s national leadership in partnering with communities of older adults using Community Based Participatory Research (CBPR) to create a sustainable training, mentoring, an incubator program in CBPR that will result in a path towards improved health. The overarching objective is to build a cadre of geriatrics, gerontology and palliative care researchers, clinicians, and administrators across the US with the necessary skills to harness the power and promise of CBPR in fostering impactful and sustained conduct healthcare system change for older adults. To accomplish this, the applicant proposes the following Specific Aims: (1) Build a national CBPR training program focusing on health improvement in older populations; (2) Measure program outcomes, and (3) Develop a program sustainability plan. The applicant is a senior investigator, program innovator, and leading national expert in designing and executing rigorous, federally-funded, CBPR research to improve care for older adults with serious illness and (b) Training and mentoring healthcare professionals in aging and palliative care research that is visioned, designed, executed, and disseminated in equal, bidirectional partnership with local communities. Her long-term career goal is to make a broad-reaching and sustainable impact on the approach to addressing health gaps in aging and end-of-life research for older adults by establishing a nationally-available CBPR training center for aging research. To achieve these goals, she has designed a training and skills development program that includes a focus on 5 key areas: (1) Curriculum development for adult learners; (2) Institutional Change leadership skills; (3) Mentorship of beginning investigators in healthcare; (4) Implementation science, and (5) Further expertise in aging focused research. The creation of an impactful and sustainable CBPR training program for aging research and clinical professionals promises to change the state of research and implementable solutions to address health gaps among older adults across the US. This academic-community partnership approach promises to overcome barriers in communities, catalyze development of local culturally appropriate interventions, and ultimately change approaches to gaps in aging care for varied populations of older adults across the US.

NIH Research Projects · FY 2025 · 2024-08

While promising for improving the lives of Alzheimer’s disease (AD) patients, new disease modifying drugs, like lecanemab, are expensive, require costly clinician assessment and brain imaging to determine eligibility, and are administered by intravenous infusion. These features may make new drugs prohibitively costly and potentially perceived as less worthy by patients of states like Alabama (AL), where poverty is widespread and medical resources are limited. To ensure that all Alabamians who may benefit from new AD drugs can have access to them, there is an urgent need to understand: 1) access to care challenges that contribute to the full cost of new drugs, i.e., cost that includes copayments for drugs, specialist visits and imaging, travel and productivity losses of clinic-based treatment, etc; 2) perceived value of new drugs, and 3) overall long-term costs and benefits of new drugs. The objective of this project is to address these knowledge gaps by conducting a mixed methods study and develop an economic analysis for estimating long-term costs and benefits of AD new drugs for older adults in AL. In a R61 planning phase (Year 1), we will conduct qualitative analyses to understand challenges of AD patients in 5 domains of access to care: Availability, e.g., of specialists, infusion clinics, imaging; Accessibility i.e., how easy it is to reach such medical resources; Affordability of drugs, specialist visits, imaging; Accommodation, i.e., how easy it is to use medical resources; and Acceptability, i.e., perceived benefits and side effects of new drugs, and preferences for treatment attributes including costs. These analyses will inform framing of the economic analysis, i.e., defining the analysis perspectives and costs and benefits to include under them. We will then develop a Markov model based on published models and set up data collection for its inputs. In the R33 phase (Years 2-5), we will collect model inputs using a survey of 240 patients and/or caregivers exposed to new (N = 80) and traditional AD drugs, and medical record abstraction for 120 patients who had visits with specialists (neurologists, psychiatrists, or geriatricians). Other inputs will be collected from the literature and public sources. Because access to care challenges, perceived new drugs’ value, and long term costs and benefits, may differ across patients’ groups, e.g., for African American (AA) and white patients, we will obtain base case estimates of costs and benefits, and run extensive sensitivity analyses to evaluate the robustness of results to the chosen model inputs, overall and for different AL populations. This work will occur under the guidance of a Patient/Caregiver and Provider Advisory Board (PPAB) to reflect the voice and relevant contexts of patients, caregivers, and providers in our findings. The PPAB will also help us define hypothetical interventions to enhance uptake of new drugs, for which we will examine cost-effectiveness using our economic model. Lastly, we will work with the PPAB to summarize results and develop recommendations for broadening use of new drugs in AL to disseminate to academic and non-academic audiences, including provider, patient and caregiver groups, and other stakeholders. In the supportive environment of the University of Alabama at Birmingham, with the proposed strong community engagement, the investigative team of this project will shed some light on the costs and benefits of new AD drugs, and contribute significantly to achieving broader use of these drugs among all older adults who may benefit from them, thus improving AD outcomes in a state like Alabama.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY Chorioamnionitis (chorio) is the most common cause of preterm birth. Chorio increases the risk for fetal and neonatal mortality and bronchopulmonary dysplasia (BPD). There are no effective therapies for preventing preterm birth, chorio, or BPD. I have previously shown that preterm birth due to chorio is propagated by IL-1 receptor-associated kinase 1 (IRAK1). Preliminary data from my human and mouse transcriptome & cytokine analysis demonstrate increased Th17 activation, and poor lung and cardiac function in mice exposed to chorio. The central hypothesis is that chorio induces specific alterations of the fetal immune system via stimulation of the IRAK1 pathway, resulting in lung parenchymal and vascular injury. We will test the hypotheses with the following Specific Aims: Specific Aim 1: To test the hypothesis that human infants exposed to chorio have increased Th17 cell activation and subsequent pro-inflammatory cytokine generation leading to long-term adverse lung function & pulmonary hypertension. I will conduct a prospective cohort study on 152 preterm infants (<32 weeks GA) (exposed and non-exposed to chorio). I will determine changes in IRAK1 mRNA levels and immune cell profiles in blood and tracheal aspirate, and measure lung mechanics using non-invasive oscillometry. The analysis will test the relationship of mRNA and cytokine levels and immune cell population with lung function and echocardiogram. Specific Aim 2: To test the hypothesis that newborn mice exposed to chorio have increased Th17 cell activation via the IRAK1 pathway leading to long-term immune cell dysfunction and adverse lung physiology. I will test if exposure to chorio alters pro-inflammatory cytokine concentrations and immune cell populations in pups of wild type (WT) and IRAK1 KO mice. Lung mechanics will be tested by oscillometry, and cardiac function assessed by echocardiography. Lungs will be harvested for histology, transcriptomic & characterization of immune cell profile.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY Anti-3-hydroxy-3-methylglutaryl-CoA reductase (HMGCR) immune mediated necrotizing myopathy (IMNM) is an increasingly recognized, severe subset of autoimmune myopathy that causes significant muscle weakness, morbidity, and often permanent physical disability. Randomized, placebo-controlled trial (RCT) data on anti-HMGCR IMNM treatment are highly limited, and no treatment trials specifically of anti-HMGCR IMNM have been completed to date. This lack of placebo-controlled interventional trial data is a critical barrier to improving outcomes for anti-HMGCR IMNM patients and represents an increasingly urgent research gap as the number of anti-HMGCR IMNM patients increases. Recent observational evidence strongly suggests that intravenously administered pooled human immunoglobulin (IVIG) may be particularly effective in treating anti- HMGCR IMNM. Corroboration of these data via a placebo-controlled trial is a fundamental next step towards establishing the role of IVIG as an effective treatment for anti-HMGCR IMNM. In addition, understanding of anti- HMGCR IMNM immuno-pathogenesis and its response to immunomodulatory therapy, such as IVIG, is limited. Overcoming this second knowledge gap would also facilitate identification of effective therapies. Our group’s data suggest that HMGCR-specific CD4+ T cells may play an important role in anti-HMGCR IMNM disease propagation. Our overarching goal is to conduct a pilot, exploratory clinical trial to test the central hypothesis that IVIG is an effective treatment for HMGCR myopathy in improving both key clinical outcomes and immunologic markers of disease pathogenesis. Our multidisciplinary team will innovatively combine clinical and translational approaches to generate urgently needed placebo-controlled data on the initial clinical efficacy of IVIG in anti- HMGCR IMNM and to determine the relationships of HMGCR-specific CD4+ T cells with disease activity and response to immunomodulatory therapy. This proposal is responsive to PAR-21-045 which calls for exploratory clinical trials in autoimmune and musculoskeletal conditions. There are two Aims. The goal of Aim 1 (RCT) is to demonstrate the safety, tolerability, and initial efficacy of IVIG for anti-HMGCR IMNM in a phase 2, double-blinded, placebo-controlled trial that will provide critical pilot data towards a future phase 3 trial. The goal of Aim 2 (T cell immunology) is to determine whether immunologic features of HMGCR-specific CD4+ T cells (prevalence, effector function, and epitope specificity) 1) are biomarkers of disease activity and 2) are affected by IVIG in anti-HMGCR IMNM.

NIH Research Projects · FY 2024 · 2024-08

PROJECT SUMMARY New disease modifying drugs, like lecanemab, if used equitably across populations disproportionately affected by Alzheimer’s disease (AD), like African Americans (AAs), could potentially reduce AD outcome disparities. However, these drugs are expensive, require costly clinician assessment and brain imaging to determine eligibility, and are administered by intravenous infusion: these features can be challenging for patients of economically disadvantaged states like Alabama. To understand how to deliver new drugs equitably in this state, there is an urgent need to understand: 1) access to care challenges that contribute to the full cost of new drugs, i.e., cost that includes copayments for drugs, specialist visits and imaging, travel and productivity losses of clinic-based treatment, etc; 2) perceived value of new drugs, and 3) overall costs and benefits of new drugs, for AA and white AD patients. The objective of this project is to address these knowledge gaps by conducting a mixed methods study and develop an economic analysis for estimating long-term costs and benefits of AD new drugs for AA and white older adults in AL. In a R61 planning phase (Year 1), we will conduct qualitative analyses to understand challenges of AD patients in 5 domains of access to care: Availability, e.g., of specialists, infusion clinics, imaging; Accessibility i.e., how easy it is to reach such medical resources; Affordability of drugs, specialist visits, imaging; Accommodation, i.e., how easy it is to use medical resources; and Acceptability, i.e., perceived benefits and side effects of new drugs, and preferences for treatment attributes including costs. These analyses will inform framing of the economic analysis, i.e., defining the analysis perspectives and costs and benefits to include under them. We will then develop a Markov model based on published models and set up data collection for its inputs. In the R33 phase (Years 2-5), we will collect model inputs using a survey of 240 patients and/or caregivers exposed to new (N = 80) and traditional AD drugs, and collecting medical records for 120 patients who had visits with specialists (neurologists, psychiatrists, or geriatricians). Other inputs will be collected from the literature and public sources. We will obtain base case estimates of costs and benefits for AA and white AL populations, and run extensive sensitivity analyses to evaluate the robustness of results and the health equity impact of new drugs. This work will occur under the guidance of a Patient&Caregiver and Provider Advisory Board (PPAB) to reflect the voice and relevant contexts of patients, caregivers, and providers in our findings. The PPAB will also help us define hypothetical interventions to enhance equity in new drug use, for which we will use the economic model to examine their cost-effectiveness. Lastly, we will work with the PPAB to summarize results and develop recommendations for equitable delivery of new drugs in AL to disseminate to academic and non-academic audiences, including provider, patient and caregiver groups, and other stakeholders. In the supportive environment of the University of Alabama at Birmingham, with the proposed strong community engagement, the investigative team of this project will shed some light on the costs and benefits of new AD drugs for AA and white populations, and inform the equitable distribution of these drugs to positively impact diverse populations suffering from AD in a disadvantaged US state.

NIH Research Projects · FY 2025 · 2024-08

Project Summary/Abstract Complex diseases are driven by intricate interactions among various cell types present at the disease sites. Recent advancements in single-cell RNA sequencing (scRNA-seq) and spatial transcriptomics have enabled cell-type-specific characterization of molecular phenotypes at the RNA-level. However, systematic profiling of the protein-level dysregulation for different cell types remains a technical challenge. Addressing this unmet need is critical for developing and optimizing therapeutic strategies, as proteins constitute the majority of clinical biomarkers and druggable targets. In recent years, advanced mass spectrometry (MS)-based proteomics has been applied to primary bulk disease samples to detect genome-wide protein expression and modifications. Moreover, by simultaneously profiling the disease samples via next generation sequencing (NGS), a new multiomics field, “proteogenomics”, has enabled deeper understanding of biological regulations at different molecular levels. Our previous studies have demonstrated the values proteogenomics data in understanding the functional consequences of genetic abnormalities and elucidating signaling pathway cascades. However, due to the moderate sensitivity of MS and the high requirement of NGS read depth, proteogenomics data are mostly generated from bulk disease samples and lack firsthand information about constituent cell types. In this project, our goal is to develop innovative computational approaches to infer the cell-type-specific protein phenotypes from MS-based proteogeomics data. First, for canonical proteins annotated by public databases, we will develop a data deconvolution pipeline to infer their cell-type-specific expression and active modifications. Second, we will create a proteogenomics pipeline to identify noncanonical peptides and proteins and map them to their cell type of origin. Of note, our computational approaches will address the unique challenges in MS proteomic data quantification and leverage the integrative nature of the proteogenomics data analysis. We will create two data portals, CellPathDb and cskAtlas, to host our methods and inferred results from current proteogenomic datasets. These data portals will enable interactive queries of cell-type-specific protein phenotypes for experimental biologists and clinical researchers.

NIH Research Projects · FY 2026 · 2024-08

SUMMARY Dental caries is a polymicrobial disease that affects much of the human population worldwide, where the equilibrium of a cooperative eco-organization among commensal microbes shifts towards a dysbiotic framework with an overrepresentation of pathogenic microorganisms, especially by Streptococcus mutans (SM). The progression of this disease begins with bacterial attachment and a complex cascade of events that include the production of soluble (a-1,6-linked) and insoluble (a-1,3-linked) glucans by glucosyltransferases (Gtfs) from the GH70 hydrolase family. This class of enzymes utilizes dietary sucrose to produce various isomeric glucan polymers through two important steps: (i) first is the sucrase (invertase) activity, where cleavage of sucrose results in glucose and fructose; (ii) subsequently, the polymerization of glucose molecules forms extended a- glucans. SM’s three different Gtfs, GtfB, GtfC, and GtfD, have been biologically well characterized, and their synthesis of glucans is crucial for biofilm formation, bacterial colonization, and virulence. However, despite more than 30 years of research and numerous studies, there remains a fundamental knowledge gap on their enzymatic mechanism; specifically, how do they produce the soluble and insoluble glucans via distinct functional domains? The goal of this application is to determine the specific molecular mechanism(s) that drive SM’s Gtfs to produce various glucans, particularly how they synthesize soluble and insoluble glucans. Our structural studies show that the sucrase activity pocket is very tight and cannot accommodate an a-retaining double displacement reaction like the GH13 enzymes. Therefore, polymerization must take place at another distinct site in the nearby vicinity. We hypothesize that ‘The GH70 glucosyltransferases of SM adopt a novel enzymatic mechanism to produce the soluble a-1,6-linked and insoluble a-1,3-linked glucans.’ We will address our hypothesis through three specific aims (SAs) by (a) determining the structures of Glucosyltransferases (GtfB & GtfD) of SM (SA1), (b) elucidating the catalytic mechanism of SM’s glucosyltransferases (SA2), and (c) characterizing the role of (i) the sucrase site, (ii) the polymerization site, (iii) the glucan binding domains and (iv) their inhibitors on biofilms, colonization, and virulence potential through in vitro and in vivo models of dental caries (SA3). The results from this study will (1) determine the novel structures adopted by these Gtf enzymes, (2) assign specific roles to domains/regions, (3) identify key residues involved in the dual-step enzymatic action, (4) specify how each GtfB and GtfD polymerize glucans, (5) reveal the importance of the sucrase site, the polymerization site, and the glucan binding domains on disease outcomes, and (6) develop inhibitors that selectively target each Gtf’s enzymatic activity. These investigations would provide a mechanistic foundation for the 800+ GH70 hydrolases, as multiple biotechnology interests exist to engineer these glucansucrases to dispense varied sizes of dextran for chromatography, food preservation, and pharmaceuticals. The long-term objective of this study is to explore innovative strategies for specifically addressing the cariogenic dysbiosis mediated by SM’s Gtfs.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY/ABSTRACT The overall goal of this K23 proposal is to provide Olivia Van Gerwen, MD, MPH with the necessary mentorship and career development to become an independent investigator whose work will contribute to improving the sexual health of key populations. The focus of her research is identifying sexual health priorities of patients and the providers who care for them, particularly pertaining to vaginal health. Symptoms such as irritation discharge, odor, and dyspareunia are seen in the setting of atrophy and dysbiosis and not only do they cause significant discomfort to those experiencing them, but these changes can increase the risk of HIV and sexually transmitted infection acquisition. Despite their significance, vaginal symptoms and infections are poorly characterized. There is a critical knowledge gap in how healthcare providers should assess these symptoms (both through history-taking and clinical specimen collection) in a patient centered manner. In this study, we will perform mixed methods research to inform the development of clinical tools for use by healthcare providers to collect both vaginal history data and clinical specimen data. This research will allow providers to better assess and manage vaginal symptoms, ultimately optimizing the sexual health of patients and decreasing the impacts of HIV and sexually transmitted infections. Aim 1: Develop a survey instrument measuring the vaginal symptoms experienced by patients for use in clinical settings. Aim 2: Examine the psychometric properties of the items to finalize the instrument. Aim 3: Characterize the acceptability and feasibility of vaginal specimen self-collection and quantify the frequency of bacterial vaginosis among a subset of patients. With guidance from my mentorship and advisory team, I have created a training plan that will allow me to gain expertise in mixed methods research, community engagement, and sexual health outcomes research through didactic coursework, hands-on training, and expert mentorship. Upon completion of the research aims in this proposal, I will develop an R01 proposal which will utilize the tools and data generated to develop a multi-level intervention focused on improving the patient-provider experience in sexual health settings.

NIH Research Projects · FY 2025 · 2024-08

Title: Pathophysiological mechanisms in the brain’s endogenous glucagon-like peptide 1 system mediated by obesogenic diets Pharmacotherapies that target the glucagon-like peptide 1 receptor (GLP-1R) system are commonly prescribed for the treatment of type II diabetes and, more recently, for weight loss. Although successful at reducing appetite and bodyweight, there are several limitations of GLP-1R agonists that limit their widespread use. Moreover, the role of the endogenous GLP-1 and GLP-1R system, particularly in the brain, and its role in obesity pathogenesis is unclear. In this application, our multidisciplinary team will address this primary gap in knowledge by dissecting the function and activity states of GLP-1-producing neurons in the nucleus of the solitary tract (NTS) and their outputs to GLP-1R-expressing neurons in the paraventricular nucleus of the hypothalamus (PVH) in a model of diet-induced obesity. In Aim 1, we will use convergent electrophysiological, anatomical, and optical methods to measure the wiring and activity states of GcgNTS neurons, the primary source of GLP-1 in the brain, after exposure to obesogenic high fat diet. Further, we will use custom tools and novel transgenic mice to dissect neurotransmission between GcgNTS neurons and one of their primary outputs in the PVH, Glp1rPVH neurons, ex vivo and in vivo. In Aim 2, we will determine the causal role of GcgNTS neurons and outputs to the PVH in progressive and interventional models of diet-induced obesity using intersectional viral tools. These exciting experiments will yield important insight into the pathophysiological role of the brain’s endogenous GLP-1/GLP- 1R system and establish if targeting GcgNTS neurons is a therapeutically tractable strategy for new obesity treatments.

NIH Research Projects · FY 2025 · 2024-08

The FUture Surgeons and Scientists Investigating ONcology (FUSSION) YES program at the University of Alabama at Birmingham (UAB) is designed to increase interest in cancer-related research among college students to learn about biomedical sciences, with the goal of creating a pipeline of future physician-scientists with cancer-related research programs. To achieve this goal, FUSSION will provide a 2-year mentored research experience augmented by workshops on cancer-related topics, scientific communication, and career and academic development. The NCI Youth Enjoy Science program provides an ideal avenue to achieve the following aims: SPECIFIC AIM 1: Provide cancer-related research experiences for undergraduate students with mentoring from successful physician-scientists. To achieve this aim, students will participate in a 2-year experience on cancer-related issues, mentored by scientists and physician-scientists, including two consecutive summers of full-time, research with dedicated scientists. SPECIFIC AIM 2: Establish a curriculum that provides a foundational understanding of a variety of cancers, research-related knowledge, responsible conduct of research, professional and career development skills, academic development, and dissemination of research results. Students will receive instruction on foundational and clinical cancer topics, along with training in scientific communication, career development, and academic preparation to enter graduate and medical school. SPECIFIC AIM 3: Expand awareness of current cancer research and the importance of the FUSSION program by integrating outreach opportunities into students’ research dissemination strategies. Students will disseminate their research to the UAB campus and wider communities: at a departmental research event, a campus symposium that family and friends can attend, and to the community through an outreach panel discussion. Participants will share the knowledge and experiences gained and the impact it has had on their understanding of cancer, with an emphasis on the importance of research in addressing cancer prevention, treatment, and outcomes. The FUSSION project is unique in its focus on physician-scientists. To our knowledge, no other YES program has a specific focus on physicians as a part of cancer research. Given UAB’s exceptional success in research and training, and the Department of Surgery’s experience developing similar research and training programs, UAB is well positioned to implement such a program. There is a great need for more physicians who are also researchers, to translate scientific discoveries to the bedside. Increasing the number of physician-scientists will directly impact patient diagnosis, treatment, and outcomes to mitigate health inequalities.

NIH Research Projects · FY 2024 · 2024-08

PROJECT SUMMARY Influenza A virus (IAV) is commonly complicated by secondary infections with Streptococcus pneumoniae (Spn) and this increases the morbidity and mortality of IAV, particularly during IAV pandemics. Highly pathogenic avian influenza (HPAI) currently poses a risk as the etiological agent for an influenza pandemic. However, little is known about how this pathogen may increase susceptibility to secondary bacterial infections. The airway epithelium serves as the first line of defense against bacterial pathogens in the lungs and it is severely damaged during IAV infection. Despite this, the role of the airway epithelium in increased susceptibility to secondary bacterial infections after IAV is not well defined. IAV induces a robust endoplasmic reticulum (ER) stress response in the airway, contributing to a disruption in host protein production during IAV infection. Interestingly, induced ER stress in the airway epithelium leads to a reduction in cystic fibrosis transmembrane conductance regulator (CFTR) function. Prior work from our lab demonstrates that IAV causes CFTR dysfunction, disrupting the airway surface liquid and subsequently increasing the burden of Spn in primary differentiated human bronchial epithelial cells (HBECs). Our preliminary data indicates that correction of ER stress during IAV infection reduces the burden of Spn in the airway epithelium without interfering with IAV replication. The broad objective of this proposal is to determine how ER stress during IAV infection is involved in both a loss of CFTR and an increase in susceptibility to secondary Spn infections. Aim 1 will test the hypothesis that ER stress during IAV disrupts host protein production, damaging the airway epithelium by causing a loss of CFTR and a reduction in secreted host proteins. To specifically study IAV infection in the airway epithelium, we will use an HBEC culture system. To identify the role of ER stress in a loss of CFTR after IAV, we will infect cells with IAV and treat them with pharmacological agents to manipulate the ER stress response and then quantify CFTR abundance and function by Western blot, Ussing chamber analysis and micro-optical coherence tomography. Additionally, we will evaluate the effect of ER stress during IAV on the secretion of host proteins by proteomic analysis of the airway surface liquid. Aim 2 will test the hypothesis that ER stress during IAV increases susceptibility to Spn and disrupts the host response to bacterial infection. To evaluate the role of ER stress in increased susceptibility to Spn, we will coinfect HBECs with IAV and Spn and treat them with pharmacological agents to manipulate the ER stress response. We will quantify Spn in both the airway surface liquid and the airway epithelium. To determine the effect of IAV infection on the ER stress response to Spn, we will coinfect HBECs with IAV and Spn and measure ER stress markers by Western blot. We will evaluate various strains of Spn to identify strain-dependent effects on the induction of ER stress. Additionally, will use both H1N1 and HPAI strains of IAV throughout this proposal to identify strain- dependent differences that may contribute to the increased pathogenicity of HPAI.

NIH Research Projects · FY 2024 · 2024-08

Project Summary Chronic rhinosinusitis (CRS) is a common inflammatory disease that affects a large portion of the U.S. population, resulting in poor quality of life for those affected and utilizing billions of dollars of health care resources. Unfortunately, CRS presents more like a heterogeneous syndrome than a distinct diagnostic entity, resulting in variability in both phenotype, symptoms, and inflammatory signatures. Consequently, CRS pathophysiology and associated mechanisms of disease remain poorly understood. Our group recently identified a unique inflammatory signature specific to elderly CRS patients, which is characterized by profound elevation of IL-1 and other pro-inflammatory cytokines. The central hypothesis of this proposal is that CRS in aged patients is associated with an age-dependent, IL-1-associated mechanism that derives from dysfunctional innate immunity and activation of the inflammasome. We will test this hypothesis by analyzing a large prospectively enrolled cohort of CRS patients. Specific Aim 1 will determine whether aging is associated with altered Toll-like receptor expression or function, with resultant ‘priming’ of the innate immune system. Aim 2 will determine whether aging in CRS patients results in increased inflammasome activation. We will subsequently analyze the ability of common microbial ligands and endogenous age-related inflammatory stimuli to activate inflammasome-associated cytokine production and release. Finally, in Specific Aim 3 we will determine whether there are functional associations between the sinonasal microbiome and/or individual pathogens with aging, innate immune function, and the IL-1-driven pro-inflammatory signature. This proposal seeks to characterize a previously unrecognized inflammatory subtype of CRS that affects aged individuals, a vulnerable population with limited medical and surgical options. Findings from this study will provide further insight into the mechanism of CRS and reveal previously unidentified associations between innate immune function, the sinus microbiota, and different types of chronic mucosal inflammation in the sinonasal cavity.

NIH Research Projects · FY 2025 · 2024-08

The University of Alabama at Birmingham (UAB) Undiagnosed Diseases Program (UAB UDP) was established as a fee-for-service clinical program in 2013 with support by UAB Medicine. The program was accepted as an affiliate site of the UDN in 2022, fully IRB-approved in April 2023, and in Sep. 2023, was added as an X01 site. The UAB UDP seeks to continue the affiliation, working within the UDN to provide services to groups in the Southeast and beyond who historically have not benefited from modern diagnostic investigations. The UAB UDP provides diagnostic evaluations for children and adults with chronic, undiagnosed diseases. Patients are referred by physicians using HIPAA-compliant secure communications systems or are referred from the UDN through the UDN Gateway. A UDP nurse practitioner contacts accepted patients for informed consent and works with the patient and the referring physician to collect and summarize medical records and obtain laboratory and imaging studies. A short summary is then produced which is entered into the PhenoTips system and data are shared with the UDN Gateway. After physician review of the summary an in person or telemedicine appointment is made (billed to insurance) with the physician and any relevant specialist consultants. Laboratory tests and imaging studies are ordered as clinically indicated. Progress in evaluations is reviewed by UAB UDP staff at monthly meetings, and a monthly conference is held with UDP staff and consultants to discuss ongoing unsolved cases. Genetic testing, including genome sequencing, is ordered if clinically indicated. Patients who live in Alabama can be offered free-of-charge whole genome sequencing at HudsonAlpha Institute for Biotechnology under a state-supported program, the Alabama Genomic Health Initiative (AGHI). HudsonAlpha is also able to do long-read sequencing in instances where a genetic diagnosis has not been achieved with short-read sequencing. Several innovations are proposed which include increased use of telemedicine and a “genome-first” diagnostic approach, the inclusion of basic scientists in a set of diagnostic teams, collaboration with a federally qualified health center to broaden patient access, and incorporation of a set of bioinformatic and AI-powered tools to improve diagnostic success rates and reduce turnaround time.

NIH Research Projects · FY 2026 · 2024-08

Metabolic dysfunction-associated steatotic liver disease (MASLD) and steatohepatitis (MASH) (aka NAFLD and NASH) affects 1 billion people, yet no approved therapy is available. Interestingly, our new antidiabetic small molecule thioredoxin-interacting protein (Txnip) inhibitor, TIX100, effectively protected obese and diabetic mice against steatosis and showed anti-inflammatory, anti-fibrotic effects in a non-diabetic, non-obese, model of MASH. Of note, Txnip is known to promote oxidative stress, apoptosis and inflammasome activation and to be elevated in livers of humans and mice with NAFLD/NASH. Thus, our overall hypothesis is that TIX100 protects against MASLD/MASH by improving glucose and lipid homeostasis and inhibiting inflammation and fibrosis. We propose 3 Specific Aims: #1: Determine the therapeutic effects of TIX100 in the context of diet-induced obesity and non-obese MASLD and MASH models. #2: Define the cellular TIX100 effects controlling MASH-associated inflammation and fibrosis and #3: Elucidate the molecular mechanisms underlying the beneficial effects of TIX100 in MASLD/MASH. To this end, we will assess liver histology, markers for lipogenesis, inflammation and fibrosis, hepatic and circulating lipids, and liver dysfunction, as well as any potential changes in whole body metabolism. This will be done in the context of MASLD, induced by high-fat diet obesity and of MASH and fibrosis, induced by a validated choline-deficient L-amino acid-defined high-fat diet. Involvement of carbohydrate response-element-binding protein, a key transcription factor of Txnip and lipogenic genes will also be determined by liver chromatin-immunoprecipitation studies. In addition, we will elucidate the specific TIX100 effects on hepatic stellate cells (HSC) and fibrogenesis, Kupffer cells and inflammation and cell autonomous effects using isolated primary hepatocytes. Using liver microRNA sequencing, we further discovered miR-34a as the top microRNA downregulated by TIX100. Intriguingly, miR-34a has been reported to promote lipogenesis and inflammation and to be upregulated in livers of humans and mice with MASLD/MASH. Now, our preliminary studies reveal that TIX100 treatment reduces the elevated miR-34a levels of MASLD/MASH and we will therefore continue to define the effects of TIX100 on miR-34a signaling in our liver, hepatocyte and HSC samples. To validate Txnip as a TIX100 target in the treatment of MASLD/MASH, we will take advantage of our whole body and liver cell-specific Txnip-deficient mouse models and expose them to DIO and CDAA-induced MASLD/MASH with and without TIX100 treatment. This will allow us to tease apart any Txnip-mediated (and potential Txnip- independent) effects and further define the mechanism(s) of TIX100 action in the treatment of MASLD/MASH. Thus, the results of these studies will provide mechanistic insight and critical preclinical data for the development of novel therapeutic approaches for MASLD/MASH at the intersection of metabolism, inflammation, and fibrosis. Since TIX100 has already undergone extensive preclinical safety toxicology and pharmacokinetic studies in preparation for first-in-man trials, the proposed studies also promise a high translational impact.

NIH Research Projects · FY 2025 · 2024-08

Project Summary The purpose of this NIH F31 application is to obtain support for the PI, Ashley Adamson, to carry out mentored research and career development activities such that the PI obtains the necessary experience and training to become a rigorous independent researcher. The project goal is to leverage the PI’s current skillset with a rigorous training program to position her to study the key proteins, pathways, and mechanisms behind environmentally caused neurodegenerative diseases, namely Parkinson’s disease dementia. The primary objective of this research proposal is to investigate the role of cell cycle regulatory proteins in facilitating cognitive impairments caused by trichloroethylene exposure in the context of Parkinson’s disease dementia. Work from the lab of Dr. Briana De Miranda, the PI’s sponsor, has shown that exposure to trichloroethylene inhalation induces nigrostriatal dopaminergic neurodegeneration, motor deficits, and alpha-synuclein accumulation in mice. Furthermore, the PI showed that trichloroethylene exposure induces cognitive deficits congruent with those commonly observed in Parkinson’s disease. However, it is currently unknown by what mechanisms trichloroethylene facilitates these deficits. This project aims to increase our understanding of the mechanisms by which cognitive impairment is induced by trichloroethylene exposure by evaluating the roles of aberrant CDK5 activation in pathogenic protein accumulation (Aim 1) and increased cellular senescence in neural stem cell populations (Aim 2). The long-term objective of the PI’s research is to identify the role for environmental exposures in influencing the significant heterogeneity of cognitive symptomology in Parkinson’s disease, and to identify disease modifying therapeutic strategies to stop the progression of the disease. The proposed training plan for Ashley Adamson is sponsored by Dr. Briana De Miranda and co-sponsored by Dr. Erik Roberson. The overall goal of the training plan is to provide Ashley with the conceptual and technical skills necessary to establish a strong foundation for a career in academic research. The training plan has been developed with activities to foster Ashley’s training in 3 crucial areas: 1) conceptual and technical research, including studies in neurodegeneration, cognition, and neurotoxicology; 2) statistical rigor, reproducibility, and transparency; including preparation in ethical research practices; and 3) career development, including training in mentorship, teaching, and scientific communication. This proposal will uniquely prepare the PI to conduct rigorous hypothesis-driven research in neurotoxicology and neurodegenerative disease while also curating the skills necessary for a becoming a competent scientist, teacher, and mentor in academic science.

NSF Awards · FY 2024 · 2024-08

Highly accurate copying of DNA into RNA is essential for cell growth and proliferation across all domains of life. This project will fill a fundamental gap in our understanding of the chemical reactions involved in copying the information contained within DNA into RNA with a low occurrence of errors. The project will produce a detailed description of how the enzymes involved extend the RNA chain and perform error correction. The results produced will aid the broader scientific community in understanding why higher organisms have evolved three specialized enzymes to copy specific classes of genes when lower organisms use a single enzyme to copy all genes. The project will train students and produce Ph.D.-level experts who will be sought by both the public and private sector for their ability to apply rigorous biophysical approaches. Undergraduate students will also be trained in these approaches. This exposure will allow the students to be more competitive for future careers in STEM. High-fidelity (low probability of error) gene expression is essential for organisms in all domains of life. Archaea and Bacteria employ a single RNA polymerase, RNAP, to transcribe all genes in the prokaryotic cell. In contrast, eukaryotes have at least three RNA polymerases (Pols). Pol I synthesizes ribosomal (r) RNA, Pol II synthesizes messenger RNA and most regulatory RNA, and Pol III synthesizes the 5 S rRNA and transfer RNA. It has been hypothesized that Pol II evolved from the archaeal RNAP and Pol I and III evolved from Pol II via sequence divergence of core subunits and incorporation of transcription factors as bona fide subunits. These observations suggest that the eukaryotic polymerases have evolved divergent mechanisms of transcription and transcription fidelity making them uniquely suited to transcribe their target genes. The project will interrogate the role of subunits within these multi-subunit RNA polymerases that are related to error correction. The overall goal of this work is to employ transient state kinetic approaches to inform the evolutionary divergence by quantitatively defining the kinetic mechanisms of transcription and transcription fidelity for the three Pols. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-08

Exposures to chlorine (Cl2) or bromine gas (Br2) may occur during industrial accidents and in the military arena. Acute lung injury (ALI), followed by chronic lung injury (CLI) characterized by airway reactivity, fibrosis and remodeling are significant concerns following halogen gas exposure; however, the mechanisms underlying this transition to CLI after exposure to Cl2 or Br2 remain to be determined. An important tenet in CLI mechanisms is protease dependent degradation and remodeling of the lung with neutrophil elastase (NE) considered a key mediator. However, α1-anti-trypsin (α-1AT), an endogenous inhibitor of NE is typically present at higher levels than NE raising the question how can NE mediate CLI in vivo. We recently discovered a novel mechanism where NE is also bound to the surface of small exosome-sized extracellular vesicles (EVs) released by PMNs during their activation and degranulation. Importantly, when associated with EV’s, NE is resistant to inhibition by α-1AT explaining how NE can induce pathogenic effects in the face of abundant α-1AT. We showed that NE+EVs are proteolytically active in vitro and in vivo; administration of NE+EV’s to naïve mouse airways led to reactive airway disease and a progressive destruction of the airways in an NE activity dependent manner. Thus, NE+EVs are novel mediators of CLI; a role for these proteolytic EVs in halogen gas toxicity has not been investigated. We hypothesize that PMN-derived NE+EVs mediate chronic lung injury after halogen gas exposure. Preliminary data supporting this hypothesis include demonstrating that NE+EVs are elevated in the airways post Cl2 gas exposure, and importantly cause progressive lung injury when administered to naïve mice. We propose two aims: Specific Aim 1: Determine the role of PMN-derived EVs in mediating CLI after halogen gas exposure. Proposed studies will determine the temporal formation of EVs and changes in NE activity following exposure of male and female mice, or rats, to either Cl2 or Br2 gas and relate these to the development of CLI assessed by reactive airways, fibrosis and airway remodeling (morphometry). In addition, we will isolate EVs from lungs after Cl2 or Br2 gas exposure, and then transfer them to naïve mice to evaluate their potential to cause CLI. Finally, PMN depletion experiments will be conducted to confirm that formation of pathogenic EVs after halogen gas exposure are derived from PMNs. Specific Aim 2: Determine the role of NE activity associated with EVs in mediating CLI after halogen gas exposure. We will determine the role of NE by comparing Cl2 or Br2 toxicity in WT or littermate NE-/- mice and the lung injury causing potential of EVs from these mice will be compared. Finally, to validate and identify EV associated NE activity as a novel target for countermeasure development, we will develop and test a strategy that specifically inhibits NE activity on EVs and then determine the ability of the latter to cause CLI after transfer to naïve mice. Collectively, the proposed studies will delineate a new and common mechanism for Cl2 and Br2 toxicity mediated by PMN-derived EVs and identify EV-associated NE activity as novel target for future countermeasure development.