University Of Alabama At Birmingham

NIH Research Projects · FY 2026 · 2023-01

Abstract Chronic kidney disease (CKD) is a worldwide public health problem affecting ~850 million people including 37 million Americans. Advanced CKD results in a number of adverse clinical outcomes leading to high rates of mortality, morbidity, and healthcare expenditure. Recognizing these staggering challenges, the Department of Health and Human Services initiated the “Advancing American Kidney Health (AAKH)” through Executive Order in July 2019. Derangements in iron metabolism are a hallmark of advanced CKD, however, the mechanistic underpinnings of such perturbations and their clinical impact on the course of CKD are not fully understood. The overall goal of this project is to fill the current gaps in knowledge and address this unmet need for the development of novel therapeutic interventions by targeting iron metabolism to slow progression of CKD and delay the need for kidney replacement therapy, two of the major goals of the AAKH initiative. By conducting preliminary studies, we have discovered that macrophage ferritin heavy chain (FtH) diminishes development and progression of CKD. Guided by our findings we propose the unifying hypothesis that myeloid FtH orchestrates iron distribution and regulates macrophage plasticity under injurious/inflammatory conditions. This premise is substantiated by (i) significant upregulation of Spic, a lineage-defining transcription factor that selectively controls development of iron recycling macrophages and (ii) marked elevation and aggregation of synuclein-alpha (Snca), a common pro-inflammatory factor in neurodegenerative conditions in in two models of CKD with substantially higher levels observed in kidneys of mice with targeted deletion of FtH in myeloid compartment. To confirm our hypothesis, we will execute the following specific aims: Aim 1: To test the hypothesis that myeloid FtH regulates monocyte/MΦ differentiation towards iron recycling phenotype through controlling transcription factor Spic. Aim 2: To test the hypothesis that myeloid FtH establishes disease tolerance to CKD via suppression of Snca. Successful completion of the proposed aims will have a significant impact on our understanding of the individual and collective effects of myeloid FtH, Spic and Snca expression in iron handling by the kidney and their detailed connotative roles in the pathogenesis of CKD, thereby paving the way for a new therapeutic approach in this disease by targeting iron metabolism in CKD.

NIH Research Projects · FY 2026 · 2023-01

Project Summary: Chronic obstructive pulmonary disease (COPD) is the fourth leading cause of death in the United States and is associated with substantial respiratory morbidity. COPD is characterized by spirometric airflow obstruction due to structural changes in lung parenchyma (emphysema) and airways. However, there exists a marked discordance between spirometry diagnosis and presence of emphysema on CT. Emphysema on inspiratory CT is defined by low-density areas <-950 Hounsfield Units (HU). By anatomically matching inspiratory and expiratory CT scans through image registration, we derived a CT measure of lung elasticity termed the Jacobian determinant of lung deformation (J) which is a point-by-point measure of lung expansion and contraction during respiration. We hypothesize that the CT-based lung mechanics will enable identification of regions that appear normal per traditional CT density criteria but are mechanically compromised during respiration. We will test the “Silent Zones” hypothesis by evaluating 10,300 current and former smokers enrolled in the Genetic Epidemiology of COPD (COPDGene) cohort with the following specific aims. In Aim 1, we will quantify Silent Zones by matching inspiratory and expiratory CT scans and to determine their associations with lung function, respiratory quality of life and functional capacity. In Aim 2, we will use 6,284 subjects who completed a second COPDGene visit after 5-years to quantify the percentage of Silent Zones progressed into emphysematous areas and also to determine the prognostic utility of Silent Zones by testing their association with FEV1 decline and mortality. In Aim 3, we will develop a deep convolutional neural network to identify Silent Zones directly from inspiratory CT scans, thus avoiding the computationally intensive image matching process. I will utilize this proposal to acquire advanced training in biostatistics, lung physiology, deep learning, parallel computing for large medical cohorts. The opportunities created by this Career Development Award will provide me with a clearly delineated path to acquire expertise and develop a research niche in the field of COPD. The aims of this research proposal and career development plan are possible through the active mentorship of Dr. Surya Bhatt, a leading expert in lung imaging research and the Director of UAB Lung Imaging Lab and Dr. Arie Nakhmani, an expert in computer vision, image registration, and machine learning methodologies. The proposed study will provide me with the skill set to achieve my long-term goal of an independent career in translational research focusing on medical imaging and machine learning applications for COPD.

NIH Research Projects · FY 2026 · 2023-01

Social anhedonia—a reduced tendency to enjoy interpersonal relationships and/or reduced interest in social interactions—poses a significant public health challenge. It is considered one of the most pervasive and debilitating features of severe mental illness, including psychosis. Social anhedonia substantially influences social functioning in psychosis, but currently there are no available treatments that target this debilitating hedonic deficit. Our limited understanding of the underlying mechanisms of social anhedonia presents a major obstacle for developing and evaluating interventions that target social anhedonia. This application aims to tackle this lack of knowledge by probing a process that is hypothesized to be central to social anhedonia: disrupted social reward learning. We developed our hypothesis from two complementary lines of work: a theoretical model of social anhedonia in psychosis and a growing body of work from affective neuroscience and behavioral neuroscience. To examine this hypothesized relationship between social reward learning and social anhedonia, this two-site study will recruit a sample enriched for social anhedonia (i.e., individuals who are within two years of their first psychotic episode) and employ a perturbation-based neuroimaging approach. The specificity of the relationship between reduced sensitivity to social reward and social anhedonia will be examined in two ways. First, we will employ two social reward learning tasks, each with both social and nonsocial reward conditions. Second, we will perturb social reward learning using Motivational Interviewing as a probe that is designed to increase sensitivity specifically to social reward. Participants will be randomized 1:1 to MI or a time- and format-matched control probe. At pre- and post-probe, participants will perform two social reward learning tasks in the scanner. We will examine the relationship between sensitivity to social reward and reduced subjective experience of social pleasure at both the behavioral and neural levels. The findings of this project will allow direct inferences about underlying mechanisms beyond demonstrating only the cross-sectional correlations between social reward sensitivity and social anhedonia. Thus, the findings of this project will provide valuable insights into the mechanistic pathways of social anhedonia and could provide novel neurobehavioral phenotypes that can serve as targets and biomarkers for developing novel treatments. Further, social anhedonia is not limited to severe mental illness, as some individuals with other mental disorders and also without mental illness report social anhedonia. The findings of this project will be valuable for our future efforts to determine whether the same underlying mechanism is related to social anhedonia across conditions and populations or not.

NIH Research Projects · FY 2025 · 2023-01

PROJECT SUMMARY Glioblastoma (GBM) is a lethal brain cancer treated by surgery, radiotherapy, and chemotherapy. While fractionated radiation therapy (60Gy delivered over 4-6 weeks) is efficacious in killing many cancer cells, a subset gains radioresistance and survive, leading to tumor recurrence. Understanding the mechanisms underlying adaptive radioresistance may lead to new therapeutic approaches to overcome radioresistance. To decipher radioresistance mechanisms, we need sophisticated models that reflect the genetic heterogeneity of GBM, including the stem cell population, which is considered to be particularly prone to acquiring radioresistance. Till now, only limited radioresistance models have been developed and the role of glioma stem cells (GSCs) has not been properly examined. To address this gap in knowledge, I developed novel preclinical models of mouse and human GSCs that were progressively adapted to repeated irradiation. Using these models, I identified a novel resistance mechanism driven by IGF1-induced N-cadherin signaling validating my approach. To identify additional radioresistance genes, I performed a genome-wide CRISPR library screening in radioresistant human GSCs and found new candidate genes, including Syndecan 1 (SDC1). I showed that SDC1 expression is increased in 4 different radioresistant GSC lines and correlates with malignancy and poor outcome of patients with malignant glioma (TCGA database). Knockout of SDC1 in radioresistant GSCs restored radio-sensitivity, decreased IGF1R expression and suppressed IGF1R signaling. As prior literature has shown that SDC1 and IGF1R interact, these findings suggest that SDC1 and IGF1R might be components of the same radioresistance pathway. Based on this rationale, I hypothesize that increased SDC1 expression induces adaptive GBM radioresistance by activating IGF1R signaling; and that blocking SDC1-induced IGF1R activation will antagonize radioresistance and increase survival. I will test my hypothesis through the following aims: 1) examine how SDC1 confers GSC radioresistance, 2) determine how SDC1 activates IGF1R signaling, and 3) evaluate the efficacy of targeting SDC1-induced radioresistance in mouse models. My project is innovative because the role of SDC1- IGF1R-mediated signaling in GSC radioresistance has never been examined. This project will lead to the development of future my research and will become a foundation for my future career. My career plan is to systematically identify the drivers and related signaling mechanisms underlying radioresistance in GSCs. I have a dual background as a neurosurgeon and basic neuro-oncology researcher and have further secured the support of advisors and professional collaborators (see letters of support). Successful completion of this project will lead to the development of better therapies for the treatment of therapy-resistant GBM and propel my transition to an independent investigator in neuro-oncology.

NIH Research Projects · FY 2025 · 2022-12

PROJECT SUMMARY Our overarching goal is to utilize biology-based mathematical models and advanced molecular imaging to dramatically decrease systemic toxicities while either maintaining or accelerating tumor control in preclinical models of breast cancer. Advances in systemic therapies have improved long-term survival in patients with locally-advanced breast cancer, however there has been a concomitant increase in the associated their long-term side effects, including cognitive deficits and cardiac problems. We have developed practical, biology- based mathematical models capable of systematically investigating the timing, order, dosing, and sequencing of combination therapies to identify therapeutic regimens that can potentially maximize response while minimizing toxicity. Preliminary results (both experimental and mathematical) reveal that alternating the order and dosing of combination chemotherapy (doxorubicin) and targeted therapy (Herceptin) can significantly and synergistically enhance response while reducing the chemotherapy dose by 50%. Furthermore, using optimal control theory, we have identified therapeutic regimens suggesting we can achieve tumor control 1.6x faster without increasing the amount of chemotherapy. We propose to develop the mathematical formalism that allows for systematically determining, on a patient specific basis, therapeutic regimens that maximize tumor response and minimize side effects. We then select the most promising options and test them experimentally against established treatment regimens and test for superior outcomes and toxicity. We also seek to develop quantitative imaging technologies capable of characterizing the temporal alterations in brain and cardiac function—organs known to be adversely affected by chemotherapies. We plan to achieve this goal with the following Specific Aims. Aim 1 will validate mathematical predictions for maintaining tumor control with minimal chemotherapy dose by employing optimal control theory to identify and biologically validate (with immunohistochemistry and overall tumor burden measurements) the three most promising combination treatment strategies. Aim 2 will implement advanced molecular imaging to quantify toxicity changes in critical organs during therapy by employing cardiac imaging of membrane potential (18F-TTP+-PET) and brain imaging of microglia activation (TSPO, measured with 18F-DPA- 714-PET) to determine longitudinal differences between long-term effects in animals treated with the standard and the optimized regimens. Completion of these aims will deliver a practical, experimental-computational approach for identifying optimal treatment strategies in pre-clinical mouse models, and appropriate for prospective testing in phase 1 clinical trials. As toxicity is the main dose-limiting factor in cancer treatments, developing methods to control it will dramatically effect patient health.

NIH Research Projects · FY 2026 · 2022-12

Project Summary Efforts to amplify the body’s immune system against cancer has faced a barrier due to the body’s own immunosuppressive tumor-promoting mechanisms, as commonly present for many cancers. We are dissecting these shared mechanisms by focusing on specific immunosuppressive cells in the tumor with the goal of developing effective therapeutic approaches by targeting these cells to treat cancer. In this proposal, we investigate the potential tumor-promoting role of a specific regulatory T (Treg) cell subset, called follicular regulatory T (TFR) cells. TFR cells are known to regulate follicular helper T (TFH) cells, B-cells and germinal center (GC) antibody responses, while the intensity of TFH cells, B-cells and tertiary lymphoid structures (TLS) in many cancers predict improved clinical outcomes and responses to cancer immunotherapy. However, the contribution of TFR cells and humoral antibody responses to the regulation of anti-tumor immunity and tumor progression remains to be largely unexplored. Our recent study of TFR cells has revealed that these cells accumulate in murine and human melanoma. The functional stability and suppressive activity of TFR cells in the tumor require the expression of the transcription factor Blimp1. Deletion of Blimp1 in Treg cells not only results in impaired suppressive activity, but also leads to the expansion of TIL TFH cells and GC B-cells, and enhanced tumoral IgE deposition secondary to TFR dysregulation. Further analysis revealed that higher tumoral TFR signatures along with PRDM1 expression indicated increased malignancy and risk of metastasis in various cancers. Increased IgE was associated with the activation and polarization of tumor-associated macrophages (TAMs) via the IgE high affinity receptor FcεRIα, potentially remodeling the tumor microenvironment (TME). These unexpected findings lead us to hypothesize that intratumoral TFR cells negatively impact the TME and promote tumor progression by repressing IgE-mediated anti-tumor immunity, and disrupting intratumoral TFR suppressive activity improves tumor control. Using genetically-modified mice and various tumor models, we will define the functional capacity and mechanistic action of TFR cells in the regulation of tumor progression, define the contribution of IgE to anti-tumor immunity and the TME remodeling. Finally, we will validate the TFR-IgE/FCERIA axis in human tumors. Completion of this study will reveal the previously unappreciated cell type, TFR cells, as a cancer prognostic biomarker and the IgE response as a predictor to the TIL TFR cell function. Insights gained from this project will facilitate the identification of new therapeutic targets and predictive markers to therapeutic outcome, and the development of effective approaches to treat a broad spectrum of cancers.

NIH Research Projects · FY 2026 · 2022-11

Human cytomegalovirus (HCMV) is responsible for significant mortality and morbidity in immunocompromised patients and results in neurodevelopmental abnormalities in infants and children infected in-utero. Currently there is no licensed vaccine and antiviral therapies have dose limiting toxicities. Thus, further understanding of the replication of this virus, including mechanisms of virion envelopment, could identify new strategies for vaccine and antiviral drug development. In this proposal, we will define interactions between an essential virion glycoprotein complex, gM/gN, and membrane associated outer tegument proteins that lead to envelopment of the virus in a specialized cellular compartment, the assembly compartment. Recombinant viruses with specific mutations in envelope and outer tegument proteins that result in shared phenotypes of defective envelopment will be used to define of viral glycoprotein/ tegument protein interactions required for virion assembly. In addition to increasing current understanding of herpesvirus assembly, results from these studies could potentially identify novel targets for development of therapeutics to limit replication of this virus.

NIH Research Projects · FY 2026 · 2022-11

A molecular understanding of the events responsible for Streptococcus pneumoniae (Spn) organ invasion during bacteremia remains elusive although its consequences are devastating. We have closely examined the interaction between capsule and vascular endothelial cells (VEC) and developed the hypothesis that capsule dynamics are complex and impact all stages of Spn translocation across VEC. This includes attachment, receptor mediated endocytosis (RME), resistance to intracellular killing, and trafficking to the basolateral surface for organ invasion. Testing of our hypothesis will reveal the intracellular mechanics of Spn trafficking that drive organ invasion. Our results will impact risk assessment strategies for serotype-based vaccine design. AIM 1. Determine how serotype impacts platelet-activating factor receptor (PAFr) and laminin-receptor initiated adhesion/uptake of Spn. Our preliminary results indicate capsule has serotype-variable antagonistic effects on the bacterium's interaction with host cells that affects RME. We will use a comprehensive panel of isogenic capsule switch mutants of low and high invasive disease capability, specifically focusing on serotypes belonging to the same serogroup, i.e., having single and defined molecular differences in their capsule structure, to determine how specific biochemical features, such as acetylation, affect interactions with both peripheral and cerebral VEC. We will quantify serotype dependence of surface exposure of the Spn adhesins phosphorylcholine (PC) and choline binding protein A (CbpA), its effect on adhesion to their ligands PAFr and LR, respectively, on receptor-initiated signaling responsible for uptake, and in summate, on invasion rate. AIM 2. Determine how serotype influences the path taken by intracellular Spn for translocation vs recycling vs removal. The degradation of cargo taken up by RME in non-phagocytic cells is mediated by the novel LC3-associated endocytic recycling pathway `LANDO'. Thus, LANDO is likely a critical pathway co-opted by the pneumococcus to cross VEC. Using low and highly invasive capsule mutant swaps, we will determine the ability of PAFr and LR to initiate LANDO. We will subsequently determine the impact of capsule and serotype on Spn trafficking across wild type vs LANDO deficient VEC in vitro. We will characterize targeting of Spn into heart and brain of pafr-/-, LANDO deficient, and LR-blocked mice, thereby assessing how capsule modulates vital endocytic processes at the crossroads of bacterial translocation vs death in the lysosome. AIM 3. Determine how capsule shedding impacts bacterial fate within VEC. Whereas pneumococci in the bloodstream are encapsulated when taken up by VEC, our results suggest capsule is shed within the endosome following RME and provides protection from low pH and oxidative stress which are deployed to kill the bacterium. We will determine the status of capsule (shed or unshed) and the importance of shedding on Spn as they cross the VEC. We will determine if serotype impacts the kinetics of capsule shedding by Spn and how capsule type and shedding influences endosome acidification and lysosome fusion.

NIH Research Projects · FY 2025 · 2022-09

Group differences have been seen in experimental pain tests and hormone levels are often estimated based on biological factors, such as age. To date, it is unknown how circulating hormone levels affect acute pain sensitivity and, given the rise in hormonal supplement usage (i.e., for low testosterone or menopause) and errors in age-based estimations, these data may be critical to assessing potential risk for chronic pain. Further, while we have shown that immune cells are critical to the persistence of chronic pain in animals, it is unclear how circulating hormone levels affect (a) these cell populations in healthy adults, (b) acute pain sensitivity in humans and (c) other psychological factors that may contribute to pain sensitivity. The current project is focused on examining the role that circulating hormones may play in various aspects that contribute to the susceptibility to chronic pain. Specific aims of the study include: 1. To determine the impact of circulating hormone level on pain sensitivity. 2. To quantify differences in immune cell populations and activity. 3. To examine social and psychological factors that contribute to pain sensitivity and the impact of hormones. We will recruit healthy individuals between the ages of 18 and 65 years of age. We will use quantitative sensory testing to assess sensitivity to cold, pressure, and heat via standardized protocols. Blood samples will be taken for assessment of circulating hormone levels (testosterone, estradiol, cortisol), immune cell populations and stimulated cytokine release, as well as salivary samples to measure stress reactivity. Questionnaires will measure pain state, quality of life, self-reported health, sleep, depression, and social support and activity/sleep monitors will provide objective measures of sleep. Together, this will be the first exploration of the impact of circulating hormones on factors that may predispose individuals to chronic pain, carried out in a broad population of healthy adults.

NIH Research Projects · FY 2025 · 2022-09

Autosomal dominant polycystic kidney disease (ADPKD), is caused by mutations in PKD1 or PKD2 genes. Disease severity is highly variable, even among families with the same PKD gene mutation. This variability has been attributed, in part, to environmental factors. Among them, a high protein diet is one of the most recognized ADPKD progression-accelerating factor. Protein composition is also a key element that can affect the rate of cyst growth. For example, soy protein compared to casein (animal-based protein) diet slows kidney cyst in rodent PKD models. Our preclinical data shows that wheat gluten (WG) diet compared to casein protein diet decreased the number of kidney macrophages (MФ), expression of Ccl2 (but not Csf1 or Ccl5) and slowed the rate of cyst growth in Pkd1-knockout mice. Therefore, animal-based protein load may have cystogenesis-promoting effects through augmentation of the cystogenesis-promoting Ccl2 pathway. Previous studies show that loss of Pkd1 increases Ccl2 in the renal tubules facilitating the recruitment of MФ, promoting kidney cyst growth and that these effects were attenuated in Ccl2-Pkd1 double knockout mice. We found that animal protein load impairs kidney mitochondrial function in Pkd1-knockout mice. When these mice were fed a WG diet enriched with top 3 amino acids abundant in casein compared to WG diet, there was increased number of kidney MФ and cyst growth compared to counterparts fed an isocaloric WG diet. This suggest that specific AAs in the casein diet promotes cyst growth. Furthermore, treatment with lysine (most abundant AA in casein compared to WG diet), but not aspartate (second abundant in casein) or glutamate (most abundant in WG), suppressed markers of mitochondrial function, impaired glucose metabolism, increased gluconeogenesis marker phosphoenolpyruvate (Pepck1) and increased tubular injury in Pkd1 mouse kidney cells. Oral lysine supplementation (1wk) in Pkd1 knockout mouse increased kidney cyst growth and levels of Pepck1 compared to counterparts given a saline. These data are well aligned with a major role kidneys play in AA metabolism through the mitochondria, an organelle that is functionally impaired by PKD mutations. Our overarching hypothesis is that specific AA(s) abundant in an animal protein-based diet exacerbate mitochondrial dysfunction, increase chemokine expression, MФ recruitment, and accelerate cystogenesis. Aim1 will test the hypothesis that high casein-protein diet impairs mitochondrial function, activates chemokine Ccl2 expression, increases kidney MФ accumulation, and promotes cyst growth; and deletion of Ccl2 in Pkd1-knockout mice have opposite effects. Aim2 will test the hypothesis that specific AA (lysine) abundant in an animal-based protein diet compared to a plant-based diet, leads to mitochondrial dysfunction, increased immune response and accelerated cyst growth. The clinical significance of this grant proposal is that patients with PKD could benefit from a plant-based protein diet and recent plant-based meat development has made dietary protein modification a feasible intervention that is safe and can have immediate impact for patients with ADPKD that require long-term treatment.

NIH Research Projects · FY 2025 · 2022-09

Knee osteoarthritis (OA) is the most prevalent form of arthritis and a significant cause of lost productivity in the U.S. As the population ages and obesity rates increase, the rates of knee OA climb as well, impacting quality of life (QOL) for millions. Current national efforts to reduce analgesic utilization highlight the critical need for safe and effective alternatives for pain relief. Low-carbohydrate diets (LCDs) reduce inflammation and pain independent of weight loss, indicating that diet interventions offer a non-pharmacological complementary treatment. However, differences between individuals exist in metabolism that are rarely addressed in diet interventions. Thus, it is important to assess the potential of different diets in a broad population of chronic pain sufferers to determine the potential of diets to reduce knee OA pain. Here, we will recruit adults (n=200) with knee OA to complete our two-phase protocol. Phase 1 will involve a 1-week diet run-up that will allow for quantification of pain measures, psychosocial variables (socioeconomic status, nutritional knowledge, proximity to grocery stores, food insecurity), and diet quality to provide a baseline for comparison. Phase 2 will be a 6-week diet intervention (LCD or USDA diet) in which both groups will be provided with all meals at the direction of study personnel and input from participants. Evoked pain tasks, measures of pain severity, catastrophizing, and interference will be assessed every 3 weeks in addition to QOL measures, mood, and depression. Physiological variables will be assessed through blood draws (inflammatory profile) and dual-energy X-ray absorptiometry scans (DXA; body composition, visceral fat) at the end of Phases 1 and 2. This will be the first study to examine the efficacy of these diets to reduce knee OA pain and interactions with biopsychosocial variables. Changes in all pain measures following Phase 2 will be assessed with respect to published measures of clinically-meaningful differences in pain, as well as for statistical significance. The central hypothesis is that the LCD will improve pain and QOL in participants with knee OA more than the USDA diet, but that both will be beneficial.

NIH Research Projects · FY 2022 · 2022-09

While genetic mouse models have been the backbone of translational biomedical research for the past four decades, mice often fail to model human disease due to species-specific differences in the cell biology of the affected organs, as well as the evolutionary divergence of their genome. The ferret has proven an excellent species for modeling infectious disease (including SARS-CoV2) and human diseases of the brain, pancreas, and lung when mice have failed. The University of Iowa (UI) is the only location in the world capable of genetically engineering ferrets with multiple precise genetic alterations previously only achievable in mice. This technology has enabled the construction of genetic ferret models capable of fate mapping stem cell compartments (i.e., lineage tracing), modeling human diseases including those with humanized genetic loci, and conditional genetics using Cre recombinase. A federally-funded National Ferret Research and Resource Center was formed eight years ago, which provides services for the creation of new genetic ferret models, performing research studies in ferrets, and the distribution of tissues and cells from existing genetic lines. This national resource has provided services to more than 129 academic investigators,108 of which are outside the UI, and currently has 8 contracts with biotechnology companies seeking to develop therapies for genetic and acquired diseases using ferret models. These services have exceeded the capacity of facilities for both performing research in ferrets and housing ferrets on the UI campus. This proposal seeks to design and construct a research facility that will house the National Ferret Research and Resource Institute (NFRRI) on the UI campus. The proposed facility has been designed to maximize synergy and minimize overlap with existing facilities at UI and will be adjacent and connected to existing ferret expansion housing (referred to as the BSRF facility). The NFRRI will contain state- of-the-art equipment for genetically engineering ferret zygotes and performing research in ferrets. Procedural space will include a USDA-compliant sterile surgical suite and separate microinjection room for model creation. In addition to wet lab research space, specialized procedural suites will accommodate nebulization, pulmonary function testing, and specialized imaging equipment. The NFRRI will primarily provide services to academic investigators outside UI and for-profit biotechnology companies, but will also provide ferret models for research that will occur on the main UI campus in existing research facilities. Furthermore, the chosen location for the facility has adjacent land for future expansion of research space and ferret housing with a long-term strategic plan that recruits next-generation leaders of the NFRRI and capitalizes on existing relationships with for-profit biotechnology companies seeking to develop therapies for human diseases in preclinical ferret models. The short- and long-term strategic goals of the NFRRI will be facilitated through financial partnerships with the University of Iowa (cost sharing of the NFRRI building), Cystic Fibrosis Foundation, and Marshall Farms (the largest breeder of ferrets for research).

NIH Research Projects · FY 2024 · 2022-09

PROJECT SUMMARY Almost all maternal and perinatal deaths worldwide are in low-income countries (LICs). Delays in accessing and receiving appropriate quality pregnancy care, particularly during labor and delivery, is a major reason for these outcomes. Reducing adverse pregnancy outcomes is a current and high global priority. The University of Alabama at Birmingham's (UAB) maternal and fetal medicine and neonatology groups are the primary referral services for high-risk pregnancies and newborns statewide in underserved Alabama. Through UAB's highly valued, busy and pioneering toll-free 24/7 Medical Information Service via Telephone (MISTTM), timely, and often life-saving point-of-care consultation and guidance is available to rural providers caring for pregnant women and newborns. Although such provider support services are now part of standard-of-care in the United States, their implementation and assessment of that implementation are often unavailable in LICs with the worst maternal and perinatal outcomes. The rapid rise in mobile phone ownership in LICs provides a tremendous opportunity to adapt, implement, and evaluate innovative, potentially life-saving interventions such as MISTTM. The mobile platform can help mitigate structural barriers and enhance healthcare access needs of the patient by providing clinical support for remotely-located providers facing medical emergencies -- while also enhancing local care systems in resource-constrained settings. Thus, the purpose of this study is to adapt and evaluate the implementation of mobile-phone based MIST (mMIST) to improve maternal and newborn outcomes in a LIC. The proposed mMIST intervention will be adapted from our ongoing experience at UAB and implemented in Cameroon, which has the 9th highest maternal mortality ratio and one of the highest rates of perinatal mortality in the world. We propose three Specific Aims: 1) Adapt and develop a 24/7 mHealth support system for peripheral providers who provide healthcare to pregnant women and newborns; 2) Test mMIST's feasibility and acceptability in one health district in northwest Cameroon, and 3) Evaluate the effectiveness - using a stepped wedge cluster randomized trial design - of full-scale implementation of mMIST in reducing maternal and perinatal deaths, and serious maternal and newborn morbidities in 19 districts (clusters) of northwest Cameroon with 65,000 deliveries per year. A participatory approach involving stakeholders and a representative task force will inform the development and delivery of the intervention. Three frameworks will inform this study, WHO Model for Quality of Care, ADAPT-ITT for intervention adaptation, and the implementation science Exploration, Preparation, Implementation, Sustainment (EPIS) model. Additional features include plans to ensure sustainability by engaging the government and mobile service providers, and attention to m-health capacity building within this rigorous research study. If demonstrated effective, scaling-up the mMIST intervention will improve maternal and newborn outcomes across similar LIC settings.

NIH Research Projects · FY 2025 · 2022-09

Summary: Chlorine and bromine (Cl2 and Br2) are highly reactive and extremely toxic halogen gases that cause debilitating cardiopulmonary injury and death. Research from our previously funded ‘Identification of Therapeutic Lead Compounds U01’ provided published and preliminary data that identified calpain inhibitor (CI) as highly efficacious and safe antidote for Br2 inhalation-induced cardiopulmonary injury and mortality. These studies were carried out in our unique halogen exposure facility and the discovery of the mitigating agent was made possible by the development of the Cl2/Br2 exposure-induced rat model of acute cardiopulmonary damage and mortality. Using the whole body Br2 exposure rat model we demonstrated that the therapeutic action of CIs is caused by reduction in Br2 induced calpain activity, reduction of proteolysis of the myocardium and preservation of cardiac function leading to decreased mortality. Calpains (calcium dependent proteases) are activated by Br2/Br2 reactant-induced loss of cardiac sarcoendoplasmic reticulum Ca2+ ATPase, SERCA activity and subsequent catastrophic cytosolic Ca2+ overload. Intraperitoneal administration of a CI (that was selected from several commercially available CIs based on an ex-vivo high-throuput evaluation of cardiac calpain inhibition) 1h after Br2 exposure significantly mitigated acute increase in cardiac calpain activity, decreased Br2-induced mortality. Administration of CI significantly improved the clinical scores, heart rate and oxygen saturation. Br2-induced pulmonary edema and cardiac hypertrophy was also reduced. Several cardiovascular parameters such as blood pressure, ventricular pressure, cardiac output and other diastolic and systolic heart functions were improved in Br2-exposed animals after CI treatment. We have also initiated the studies on its various structural analogs. These structures were characterized for their improved solubility, bioavailability and stability. Preliminary data for their activity and cellular toxicity was also evaluated. Thus, as mentioned in the CounterACT FOA we provided validation of molecular targets for therapeutic development, proof of in vitro activity of the lead compound, preliminary in vivo proof-of-concept efficacy data, and preliminary adsorption, distribution, metabolism, excretion, and toxicity (ADME/Tox) evaluations. Therefore, this lead compound ‘calpain inhibitor’ is now ready for optimization and we will also evaluate ADME/safety profile of the CI and its most effective analog. Optimization of CI/analog dose and delivery in a large animal model of halogen-induced cardiopulmonary injury will also be performed. These studies will allow the lead compound to move forward and help design the pivotal studies needed for regulatory FDA approval of CI under the animal rule.

NIH Research Projects · FY 2024 · 2022-09

Abstract This supplementary award aims to provide extensive research training in neuroimmunology for a gap-year researcher. The proposed project explores the complex landscape of age-associated neutrophil alterations in the brain after cortical brain injuries and clarifies their unique contributions to neuroinflammation and cognitive effects. This investigation focuses on understanding the role of aging-associated neutrophil subpopulations post-injury within the thalamus, leveraging single-cell RNA sequencing (scRNA-seq) data from our parental R01 award. We aim to identify specific age-associated indicators within the brain using refined analysis techniques and advanced clustering algorithms. Validation of the presence and activation status of these markers in the injured brain will be pursued through immunohistochemistry and flow cytometry, providing crucial insights into the nuanced age-specific neutrophil responses. We aim to elucidate the intricate interplay between age-specific neutrophil subpopulations, their activation dynamics, and the evolving neuroinflammatory and cognitive consequences following traumatic brain injury. We will also examine how neuroinflammation and cognitive recovery after injuries are affected by age-associated neutrophil suppression. We will use methods established in our parental R01, particularly antibody-based cell depletion, to assess the impact of age- associated neutrophil infiltration on neuroinflammatory markers, microglial activation, and cognitive recovery after injuries. These studies will be conducted in parallel to hands-on experiments and data analysis training, which will enhance the researcher’s skills and prepare the researcher for graduate studies.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT Traumatic brain injury is a risk factor for cognitive impairment and dementia, such as Alzheimer’s disease (AD) and frontotemporal dementia (FTD), particularly in the aged populations. Nevertheless, the mechanisms by which aging exacerbates cognitive deficits after brain injury are not fully understood. Human brain imaging studies reported the signs of microglial activation in the thalamus that correlate with cognitive deficits. Our preliminary studies using a local microglia depletion in mice have discovered that thalamic microglia activation is required for cognitive deficits after brain injury. In the middle-aged mice, cognitive deficits after brain injury were exacerbated and accompanied by dysregulated responses of microglia and accumulation of AT8-positive phosphorylated tau proteins (p-tau). Recent studies have reported that one of the aging-associated molecular signatures in the mouse brain is an increased interleukin-33 (IL-33) expression in oligodendrocytes. Indeed, IL- 33 expression was increased in thalamic oligodendrocytes. Notably, blocking of IL-33 in the aging thalamus ameliorated aging-associated exacerbation of cognitive deficits. These findings suggest that aging-associated changes in the thalamic environment and microglial responses contribute to p-tau accumulation and exacerbated cognitive deficits in aged mice after cortical injuries. Thus, in the proposed study, we will test our hypothesis that aging-associated oligodendrocyte-derived IL-33 exacerbates cognitive impairment after cortical injury by driving microglial dysfunction and tau pathology in the thalamus. In Aim 1, we will further evaluate aging-associated changes in thalamic pathology and cognitive impairment after cortical injuries and determine the effects of thalamic microglial depletion and neuronal tau deletion on exacerbated cognitive impairment. In Aim 2, we will investigate the requirement of oligodendrocyte-microglial IL-33 signaling in aging-associated microglial dysfunction, p-tau accumulation, and worsening cognitive impairment after cortical injuries. In Aim 3, we will examine the mechanisms by which aging-associated IL-33 signaling alters microglial responses after cortical injuries. Together, this study will determine the mechanisms by which oligodendrocyte-microglia IL-33 signaling induces microglial dysfunction, p-tau accumulation, and cognitive impairment relevant for AD/ADRD.

NIH Research Projects · FY 2025 · 2022-09

PROJECT ABSTRACT/SUMMARY. The proposed R01 is built upon the premise that: 1) Cognitive impairment (CI) persists even in the context of HIV viral suppression; 2) women living with HIV (WLWH) experience disproportionate burden of CI and have higher rates of, and stronger associations with, many of the underlying biopsychosocial risk factors for CI, as well as unique sex-specific risk factors; 3) immune activation persists in the context of viral suppression; 4) immune activation (e.g., Interleukin[IL]-6, sCD163) is a consistent risk factor of CI in HIV; 5) neurobiological pathways affecting cognitive functions are activated by HIV directly and indirectly through immune activation; 6) psychosocial factors such as stigma/discrimination, depression, and substance use impair cognitive functions and are associated with heightened immune activation, likely both directly and indirectly through reduced ART levels. By leveraging the strong infrastructure of the Women's Interagency HIV Study (WIHS), including existing behavioral data and specimen, the current longitudinal study will fill significant gaps in the field by examining the role of the neurobiological pathways whereby chronic immune activation leads to CI phenotypes in a large phenotypically well-defined group of WLWH. In a highly cost-effective approach, we will use existing biospecimen to assess biomarkers of chronic immune activation and neurobiological pathways and link them to existing neuropsychological assessments among N=500 WLWH aged 26 and older (Median age = 50) at 3 time points, to assess the following aims: Aim 1. To examine neurobiological pathways mediating the link between chronic immune activation and CI phenotypes in WLWH. Aim 2. To identify psychosocial factors affecting immune activation and neurobiological pathways. Aim 3. To examine the role of ART hair levels in mediating the effect of psychosocial factors in Aim 2 on immune activation and neurobiological pathway biomarkers. Impact. The results of this study will have several important clinical implications for the prevention and treatment of CI in WLWH, including behavioral and pharmacological therapies. Understanding the unique impact of biomarkers on cognitive outcomes will provide guidance on clinical intervention targets (e.g., immune therapy, dopamine reuptake inhibitors). Understanding the role of psychosocial factors in modulating immune activation and neurobiological pathways and the mediating role of ART adherence level, will inform behavioral intervention strategies (e.g., resilience, social support). Psychosocial factors may be particularly ideal intervention targets to curb the inflammatory cascade leading to CI.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT MicroRNAs have been studied for over two decades and found to impact extensively on various cellular functions like differentiation, proliferation and oncogenesis through regulation of gene expression using the Argonaute (Ago) containing RNA-induced silencing complex (RISC). In the cell, however, microRNAs co-exist with a nearly equal abundance of non-canonical short RNAs (ncsRNAs) that were not believed to enter the RISC. This has begun to change with our discovery that some members of the ncsRNAs, the tRNA derived fragments (tRFs) enter into RISC and silence gene expression, and others do not. The 18-26 base long tRF-3a molecules are derived from tRNAs by processes very different from the biogenesis of microRNAs, and yet repress gene expression by incorporation into Ago-RISC (RISC). In Aim 1 we will focus on specific tRF sub-classes, tRF-3b and tRF-1, that do not enter into Ago-RISC, to identify the surveillance pathways that keep short RNAs from dysregulating gene expression through RISC. We will study a methyltransferase that inactivates tRF-3b molecules by modifications on the RNA, a modification that is also regulated by demethylases that are inactivated by Isocitrate Dehydrogenase (IDH) mutations, seen in many cancers. We will also focus on an RNAse that degrades tRF-1 molecules to prevent them from entering into RISC and silencing gene expression. The results will reveal how the surveillance mechanisms work and how pathogenic or therapeutic alteration of the surveillance mechanisms will alter gene expression and improve RNA mediated therapy. In Aim 2 we will turn to ncsRNAs, exemplified by three tRF-3a molecules, that enter into RISC, silence gene expression and alter phenotypes of cancers and cancer cell-lines. We will test whether even in these cell line the tRF-3a molecules regulate gene expression by hijacking microRNA specific mechanisms and thus alter cellular phenotypes. We will also determine whether the ncsRNAs help or hinder microRNAs from doing their function. The field of short RNA mediated post-transcriptional gene regulation will be altered fundamentally by the recognition that microRNAs work in a complex milieu of other short RNAs that compete with or assist microRNAs, and that the cell has evolved mechanisms to protect the integrity of microRNA-mediated gene regulation.

NIH Research Projects · FY 2025 · 2022-09

Despite the availability of effective HIV prevention tools such as pre-exposure prophylaxis (PrEP), significant differences persist among women in the South where the rate of HIV infection is fourteen-times higher for Black women compared to White women. PrEP utilization remains low among women in the South with estimates of <7% eligible women receiving PrEP in Alabama. Moreover, algorithms to determine PrEP eligibility demonstrate poor predictive power for women. The proposal takes advantage of a dynamic and innovative collaboration of experienced HIV-prevention investigators from the University of Alabama-Birmingham, University of North Carolina-Chapel Hill, Centers for AIDS Research, the Alabama Department of Public Health (ADPH), and Social Scientific Systems. This novel research proposal utilizes a population-based approach to establish a geographically representative cohort of women (‘Camellia Cohort’) at significant risk for future HIV acquisition, based on recent gonorrhea or syphilis infection, across the state of Alabama (AL), to better understand factors associated with risk of STI and HIV diagnosis and predictors for PrEP use. The aims of this proposal are to: 1) refine the HealthMpowerment (HMP) digital platform, to include key elements that optimally engage and retain a cohort of women at-risk for HIV in AL; this will be achieved via stakeholder engaged research with women on PrEP and PrEP care team members using nominal group techniques; 2) recruit and retain a geographically-representative and rurally-enhanced cohort of 800 women ages 18-44, with recent STI positive testing and HIV-uninfected at enrollment, to participate in the Camellia digital cohort utilizing a geographically-varied sampling framework using public and commercially available annual county-level HIV and STI testing data and 3) evaluate predictors, mediators and moderators for STI/HIV incidence and PrEP use and adherence in the Camellia Cohort by collecting demographic, behavioral, and community level assessments via the HMP digital platform and HIV/STI testing through ADPH’s home testing program at regular intervals; we will purposefully sample women with high HIV vulnerability and/or PrEP use within the cohort for in-depth interviews (n = 30) to further explore factors contributing to PrEP use to guide future intervention development. This research will lay the groundwork for a larger research program testing the implementation of a further adapted HMP digital platform designed to improve HIV/STI testing, PrEP utilization and persistence among women in the South, focusing on areas with higher HIV incidence.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT Sickle cell disease (SCD) is an inherited red blood cell disorder that predominantly affects individuals of African descent. The physiologic hallmarks of SCD include chronic red blood cell hemolysis, oxidative stress, inflammation, vascular damage and end-organ damage. These physiologic sequelae lead to unpredictable vaso occlusive complications, such as pain and acute chest syndrome (ACS). Human breastmilk contains antioxidant, anti-inflammatory and immune-modulating properties proven to be protective against chronic inflammatory conditions, such as asthma, obesity and cardiovascular disease. The lasting impacts of human breastmilk have been demonstrated in adolescent and adult studies, wherein individuals that were breastfed as infants had lower levels of biological markers of inflammation in their bloodstream. Interestingly, breastmilk has even been demonstrated to be protective against the erosion of telomeres – which are the protective caps of DNA and established markers of oxidative stress and inflammation. The American Academy of Pediatrics and World Health Organization recommend exclusive breastfeeding (EBF) for the first 6 months of life, followed by 1 to 2 years of breastfeeding with nutritious supplementation. However, despite these recommendations and the well- established protective benefits of EBF, the gap between Healthy People 2020 breastfeeding goals and current rates of EBF is widest amongst African American women compared to all other racial and ethnic groups. Furthermore, the protective effects of EBF have never been explored in SCD. Several studies have noted a lack of breastfeeding role models and support networks as barriers to EBF among African American women. One community-based organization located in Birmingham (Alabama) — the Chocolate Milk Mommies —, specifically addresses these barriers through online and in-person education and support groups, certified lactation consultant services, free breast pump rentals for low-income mothers and a home visitation and peer- mentoring program. Chocolate Milk Mommies specifically targets African American mothers, given the lower rates of breastfeeding in this population. This group achieved a 6-month EBF rate of 83% in 2019, far exceeding state and national trends. Given the pervasive oxidative stress and inflammation in SCD, and the consequent risk for pain and poor lung health, it is essential to test the protective benefits of EBF and establish effective EBF- promoting interventions among this high risk population. The aims for this project are to: 1) test the association between EBF exposure and SCD complications, 2) test the association between EBF exposures and biological markers of oxidative stress and inflammation, and 3) pilot the feasibility to recruit and retain 20 mothers of children with SCD to a community-based EBF program (the Chocolate Milk Mommies) for ≥6 months, which will inform a future larger randomized controlled trial. The long-term goal for this research is to establish the protective benefits of breastfeeding in SCD and improve overall health outcomes through the promotion of EBF.

NIH Research Projects · FY 2025 · 2022-09

Latently infected CD4+ T cells are considered to be the most important HIV reservoir preventing the implementation of an HIV cure. However, myeloid cells have been shown to be infected by HIV/SIV and to establish latency in animal models1-4. Therefore, elimination of the T cell reservoir alone is not likely to result in complete virus eradication. One of our long-term goals is to aid the development of effective HIV cure strategies by gaining a better understanding of the type and location of the cells that harbor replication competent HIV under ART suppression. HIV-associated neurological disorders or HAND affect up to 50% of people living with HIV (PLWH)5-7 suggesting that HIV-infected cells may persist in the brain of ART-suppressed individuals. HIV-DNA has been readily detected in the CSF of aviremic ART-suppressed PLWH and its presence is associated with poorer neurocognitive performance8. Analyses of brain tissue obtained postmortem from PLWH indicate that macrophages and microglia are major targets for HIV infection in the brain9-11 and demonstrate the presence of HIV-DNA+ cells in brain tissue from aviremic individuals10-12. Microglia are the predominant population of myeloid cells in the brain and in contrast to macrophages, are long lived and undergo cell division13,14. For these reasons, microglia are thought to represent a key cellular reservoir of HIV in the brain11. While there is a significant body of knowledge about the mechanisms of HIV latency and persistence in resting CD4+ T cells, there is significantly less known about HIV persistence in myeloid cells and therefore, a need to establish their possible role as a source of HIV reactivation after ART discontinuation. Specifically, in the brain the contribution of microglia is relatively unknown due in part to the difficulties in sampling cells in this compartment in PLWH. Our hypothesis is that HIV maintains a persistent reservoir in the brain under suppressive ART and our objective is to utilize an innovative humanized mouse model reconstituted with human brain microglia to increase the knowledge and understanding about how microglia contribute to HIV persistence and viral rebound by analyzing 1) HIV suppression by ART in human microglia in the brain, 2) the viral reservoir present in infected microglia, and 3) the development of HIV latency and rebound in human microglia in the brain after analytical therapy interruption. The new knowledge gained from the proposed experiments will contribute to a better understanding of HIV persistence in the brain, reactivation, and aid the development of novel HIV Cure approaches that target the CNS.

NIH Research Projects · FY 2024 · 2022-09

PROJECT SUMMARY Pancreatic neuroendocrine tumors (pNETs) are the second most common malignancy of the pancreas, with an overall survival of 3.6 years and successful surgery the only treatment offering potential for cure. However, around 40-95% of pNETs are metastatic at the time of initial diagnosis, with local recurrence within the resection bed as the norm. Furthermore, patients with liver metastases from pNETs often have debilitating symptoms such as uncontrollable diarrhea, flushing, skin rashes, and heart failure. There are few systemic therapies that have proven to be clinically useful, and those that have still bear widely variable response rates and have poor side effect profiles. The goal of this proposal is to determine the role of Notch2 in the proliferation, metastasis, and hormone-secreting phenotype of NETs, and evaluate Notch2 as a predictor of patient outcomes. Preliminary data from our lab suggests that overexpression of Notch2 results in an increase of the proliferative rate of NET cells, while decreasing their hormone secretion. Our data also suggest that Notch2 is upregulated in metastatic pNETs compared to primary tumors. Seminal data from one of the largest genomic studies to date on pNETs has also identified Notch2 as a key master-regulator of pNET metastasis, representing a key convergence of dependencies required for disease progression and the establishment of metastasis. Therefore, it is my hypothesis that Notch2 functions in an oncogenic role in NETs and drives tumor progression, resulting in a more aggressive phenotype and portending worse patient prognosis. To evaluate this hypothesis, we will conduct phenotypic characterization (proliferation, hormone production, migration) on Notch2-overexpressing pNET cell lines that have been transiently and stably transfected. We will similarly evaluate pNET cell lines with Notch2 knockdown via siRNA, as well as stable knockout pNET cell lines generated using CRISPR-Cas9. We will then conduct similar studies in vivo using a liver metastasis mouse model whereby Notch2-overexpressing and Notch2 pNET cell lines are injected into athymic mice. Using this model, we will evaluate tumor growth and metastasis via microCT, in addition to hormone secretion over a 16-week period. We will then conduct endpoint analysis of tumors and mouse organs. Lastly, we will immunohistochemically analyze the expression of Notch2 using pNET tissue microarrays derived from human patients that have undergone surgical resection at the University of Alabama at Birmingham and evaluate the relationships between the expression of Notch2, Notch2 pathway components, and various patient outcome measures. This study will yield valuable information on the effects of Notch2 in pNETs that can help to guide future targeted therapeutic efforts and inform an understanding of pNET biology.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY: Alzheimer’s disease (AD) is the most common cause of dementia and the third leading cause of death in older adults. In addition to cognitive impairment, AD is associated with neuropathology, impaired metabolic function and gut microbiome dysbiosis. However, the relationships between gut health (including the gut microbiome), metabolism and cognitive decline remains largely unknown, despite strong evidence that the gut-brain-axis is an important intermediary in neurodegenerative disease. Moreover, normal aging also influences both gut microbiome composition and peripheral metabolic health, demonstrating the importance of including geroscience as a factor. Gut dysbiosis can result in impaired insulin resistance as well as obesity, both of which increase the risk of developing AD. Therefore, the long term goal of this project is to elucidate how altered gut microbiome composition can influence cognitive outcomes in an aged rat model of AD to identify potential targets for therapeutic intervention. The overall objective is to investigate whether the gut is able to exert its influence over cognition through metabolic intermediates, as the gut microbiome directly influences metabolite production and energy homeostasis. The rationale for this project is that very few, if any, strategies directly targeting brain function have been able to prevent or delay cognitive impairment with AD to date. This line of investigation is innovative in that it suggests a peripheral target, rather than targeting the brain directly, for the alleviation of cognitive symptoms by utilizing the strong, reciprocal link between gut and brain function. Our preliminary data indicates the TgF344-AD rat model mimics the cognitive decline observed with AD, as well as the peripheral metabolic impairments and gut dysbiosis associated AD. The proposed experiments will build on these findings and test the central hypothesis that AD-associated gut dysbiosis negatively influences metabolic function, thereby impairing cognitive function through an impaired ability to properly utilize and respond to energy metabolism. Data from humans with AD, as well as our preliminary data, suggest AD-associated gut dysbiosis negatively influences butyrate production. Depletion of butyrate, the most abundant metabolite produced by the gut microbiome, is associated with many metabolic diseases including diabetes and obesity, both of which are commonly associated with AD. Supplementation of butyrate in rodent models can improve insulin sensitivity, upregulate BDNF and thereby neurogenesis and promote a lean phenotype. Butyrate improves brain health and metabolism, gut health through supporting the gastrointestinal lining quality and influences microbiome composition. These synergistic effects make systemic butyrate a therapeutic strategy, or one component of an effective strategy, with high potential to ameliorate many AD-related impairments in function. Collectively, these experiments will determine if the gut microbiome influences cognitive outcomes in aged AD-model rats through a metabolic intermediary and whether this can be ameliorated through probiotic supplementation.

NIH Research Projects · FY 2026 · 2022-09

PROJECT SUMMARY/ABSTRACT Kidney stones (KS) are painful mineral deposits that affect 1 in 11 individuals in the United States. The economic burden from KS is over $5 billion dollars annually and is predicted to rise due to increasing prevalence. Approximately 80% of KS are comprised of calcium oxalate (CaOx). Several individuals with CaOx KS develop recurrent KS. The reasons for recurrence are not well defined. Intake of meals containing high amounts of oxalate are associated with increased KS risk. Oxalate-rich diets may induce CaOx crystal formation in the urine and nephron, which can stimulate reactive oxygen species signaling and monocyte recruitment into the renal interstitium. If crystals are not properly cleared by macrophages, this could result in the propagation of KS. Interleukin-10 (IL-10) is a key anti-inflammatory cytokine important for regulating monocyte and macrophage function. We previously reported patients with CaOx KS have reduced circulating monocyte cellular bioenergetics and increased inflammation. We further determined that CaOx crystals can cause similar responses in monocytes in vitro. We recently reported that intake of a single dietary oxalate load stimulates nanocrystalluria and alters circulating monocyte cellular bioenergetics in a small cohort of healthy subjects. The goals of this proposal are to examine the effects of low and high oxalate diets on nanocrystalluria and immunity using human translational studies and experimental models. The central hypothesis of this proposal is oxalate suppresses IL-10 signaling leading to reduced cellular bioenergetics, redox homeostasis, and mitochondrial quality control in macrophages. We further propose this contributes to impaired macrophage clearance of CaOx crystals from the kidney which may play a role in KS formation. Aim 1 will test the hypothesis that oxalate enriched diets stimulate nanocrystalluria and reduce monocyte cellular bioenergetics in healthy subjects and patients with CaOx KS. Aim 2 will test the hypothesis that oxalate reduces IL-10 signaling, cellular bioenergetics, and mitochondrial quality control in macrophages. This project involves an interdisciplinary team and multifaceted approaches to assess the effect of oxalate on nanocrystalluria and IL-10-mediated monocyte/macrophage immune responses, which may play a role in CaOx KS formation. The results obtained will generate new insights into the cellular mechanisms driving KS formation and should unveil novel strategies for KS prevention.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT Mechanisms that govern cardiomyocyte proliferation and remuscularization following ventricular injury Program Overall The overall goal for the treatment of myocardial infarction is to replace the scar tissue caused by ischemic injury with functional cardiac muscle. Since adult mammalian cardiomyocytes (CMs) are non-proliferative, and the engraftment rate for cardiac cell therapy is extremely low, most of the remuscularizing initiatives following infarction have been unsuccessful. However, recent preliminary studies from our laboratories using neonatal pigs have shown that when myocardial infarction (MI) is induced on postnatal day 1 (P1), CMs re-enter the cell cycle, proliferate, and completely restore cardiac function with little scarring. Furthermore, we have found that these neonatal hearts with the P1 injury, have a very active and prolonged CM proliferative machinery, and consequently a second LAD ligation injury at P28, which resulted in a large infarct (TTC) at Day 2-7 post LAD ligation, produced no visible infarct 4 weeks following injury. This was a remarkable result as it demonstrated, for the first time, that a heart of large mammal could remuscularize infarcted heart tissue by CM proliferation. The studies comprising this Program Project Grant (PPG) application will examine mechanisms whereby CMs reenter the cell cycle and new strategies to remuscularize injured hearts. Project 1 will identify the CM cell-cycle regulators that are activated by MI in one-day-old pigs and construct human cardiac muscle patches (hCMP) of unprecedented clinically relevant dimensions from layers of proliferating hiPSC-CMs with activated cell cycle regulators, and other cardiac cells; subsequent experiments will determine whether the identified factors and hCMPs can remuscularize the hearts of adult pigs after MI. Project 2 will use genetic strategies, viral vectors, and modified RNAs to investigate whether members of the Sonic Hedgehog signaling pathway including Gli1 and Sox4, which have already been shown to induce proliferation in cultured CMs, will promote CM proliferation in the injured hearts of adult mice and pigs. In addition, studies will examine the capacity of the master regulator, Etv2, to promote neovascularization and promote repair of the injured hearts of adult mice and pigs. Project 3 will be an extension of previousobservations that mammalian cell-cycle arrest is at least partially induced by the increase in oxygen metabolismthat occurs after birth, and that severe systemic hypoxia upregulates proline metabolism and induces CM proliferation in adult mice; the proposed studies will examine whether proline metabolism regulates CM survivaland proliferation during chronic hypoxia. Collectively, these three projects, the associated cores and the expertise of the investigators will accelerate and amplify the studiesto address the central objective of this P01 proposal: To remuscularize the injured ventricle from “within,” by promoting endogenous CM proliferation, and from “outside,” bytransplanting functionally mature hCMPs that are primed for in-vivo CM proliferation.