University Of Alabama At Birmingham

NIH Research Projects · FY 2026 · 2022-04

Project Summary G-protein coupled receptor (GPCR) kinase-2 (GRK2) is a key regulator of GPCR recycling and desensitization that is upregulated in several cardiac pathologies, including hypertrophy and heart failure (HF). Cardiac ischemia-reperfusion injury induces ERK-mediated phosphorylation of GRK2 at S670, which results in the translocation of GRK2 from the cytosol to mitochondria. We discovered that in the adult cardiomyocyte mitochondrial GRK2 regulates glucose-mediated oxidative phosphorylation by inhibiting pyruvate dehydrogenase, the rate limiting enzyme of glucose oxidation. Although the physiological impact of mitochondrial GRK2 was reported, it remains largely unknown how mitochondrial GRK2 regulates cardiac mitochondrial metabolism. This proposal focuses on deciphering mechanistically how GRK2 phosphorylation and mitochondrial translocation regulate cardiac glucose metabolism. Additionally, we propose that mitochondrial GRK2 participates in key metabolic signaling post-myocardial infarction (MI) by acting as an amplifier of pyroptosis- a novel lytic cell death mechanism. Thus, we hypothesize that GRK2 phosphorylation at S670 is paramount for cardiomyocyte responses in consequence of altering metabolic availability and cardiac injury. Using a novel GRK-S670A and two Gasdermin E mouse models, we propose to carry out two specific aims: 1.) Determine how mitoGRK2 regulates cardiac mitochondrial metabolism; 2.) Assess whether phosphorylation of GRK2 at S670 modulates cardiac pyroptotic signaling. Overall, our work will shed light on the role of GRK2 phosphorylation at S670 in cardiomyocytes and how this post-translational modification regulates metabolic signaling and chronic-injury responses. The overarching goal of this research is to exploit novel mechanistic signaling for the development and identification of new pharmaceutical drugs for HF treatment.

NIH Research Projects · FY 2026 · 2022-03

1 Project Summary: “Role of Rab10 in Alzheimer’s disease, K99/R00” 2 Candidate: My long-term career goals center on the establishment of a leading research laboratory in 3 academia focused on defining key molecular mechanisms underlying neurodegeneration and Alzheimer’s 4 disease (AD). Specifically, my recent research interests have narrowed to ask how key regulators of endocytic 5 trafficking pathways, particularly Rab GTPases and their modifiers, contribute to neuroinflammation associated 6 with AD. I am a biochemist with training in biophysics and cell biology, with the included proposal for additional 7 training in mouse models of disease. These training experiences will facilitate future investigations in molecular 8 mechanisms underlying endocytic dysfunction that manifest in disease relevant phenotypes. 9 Training: Here I propose a series of training that include a translational in vivo experiment in mouse models 10 coupled with formal coursework to accelerate my trajectory towards independence in the R00 phase. These 11 training experiences will be facilitated through mentorship from Dr. Andrew West, in addition to a Transition 12 Advisory committee at Duke University composed of Dr. Patrick Sullivan, an expert in mouse AD models, Dr. 13 Carol Colton, an expert in microglia function in AD models, and Dr. Robert Lefkowitz, a Nobel-laureate expert 14 in biochemistry and cell biology. Formal course work that is planned will further enhance training on in vivo 15 study approaches, grant writing, mentoring skills, lab management and responsible conduct in research. 16 Research: Recently, a single nucleotide polymorphism in the Rab10 3’UTR rs142787485 has been linked with 17 strong resiliency to AD susceptibility. Transcriptomic analysis reveals Rab10 mRNA as higher in AD patient 18 brains compared to healthy controls. My latest post-doctoral work disclosed that the Rab10 GTPase, highly 19 expressed in phagocytic cells, operates in a specific compartment of the endolysosomal pathway to control 20 fluid phase macropinocytosis. Preliminary data included herein suggests knockdown of Rab10 decreases the 21 internalization of aggregated tau in mouse primary microglia. Recent findings suggest internalized tau 22 aggregates break endolysosomes in microglia that stimulate inflammasome activation and the secretion of 23 damaging cytokines. Inflammasome activation may accelerate tau pathology, potentially linking Rab10 with 24 neuroinflammatory pathways important in AD. I plan to explore this novel disease-associated cycle in K99 and 25 R00 work. In the K99 phase, I would test the hypothesis that suppression of Rab10 in the P301S Tau/APOE4 26 mouse model ameliorates neuroinflammation and neuropathology. In the R00 phase, I plan to investigate how 27 Rab10 might regulate aggregated tau uptake, processing, and responses in microglia, as well as key 28 regulators of Rab10 activity that influence different endocytic trafficking pathways and immunological 29 responses. These proposed experiments would start to elucidate the potential roles of Rab10 in AD, and 30 identify novel pathways that might be exploited in the future for therapeutic gain. 31 32 33

NIH Research Projects · FY 2026 · 2022-03

Abstract O-linked ß-N-acetylglucosamine (O-GlcNAc) is a sugar attachment to the side chain hydroxyl of a serine or threonine residue on proteins. O-GlcNAcylation controls key signaling and biological processes such as signal transduction, transcription, cell cycle progression, and metabolism. Perturbations in O-GlcNAc homeostasis have been linked with diabetes, cancer, and neurodegenerative diseases. Increased glucose levels channel flux through the Hexoseamine Biosynthetic Pathway (HBP), culminating in increased O-GlcNAc levels. The activity of HBP and consequently cellular O-GlcNAc-ylation are elevated in several cancer types, including breast cancer. We recently reported that inhibiting HBP activity significantly decreased the invasive phenotype of breast tumor cells. We surmised that abundance of glucose, a readily-metabolizable carbohydrate, will drive flux through HBP, resulting in enrichment of a portfolio of proteins that are modified by O-GlcNAc-ylation. Using unbiased proteomics analysis, we identified that elevated glucose culture conditions enrich for O-GlcNAc- modified GLI proteins, transcription factors of the Hedgehog (Hh) pathway. Importantly, we identified that in elevated glucose conditions, O-GlcNAc-modification of GLI exacerbates Hh/GLI activity; and inhibiting HBP mitigated this effect. We hypothesize that HBP-directed O-GlcNAc-ylation fundamentally programs invasive and chemoresistant attributes in tumor cells through activating Hh/GLI signaling. In Aim 1 we will determine the molecular underpinnings of HBP-directed O-GlcNAc-ylation of GLI. We will determine the causes and consequences of GLI O-GlcNAc-ylation. We will first identify engagement of the HBP in O-GlcNAc-modification of GLI proteins. Next, we will undertake investigations to identify establish the mechanistic basis of how HBP signaling engages O-GlcNAc-modified GLI to program invasive and chemoresistant attributes in tumor cells. In Aim 2 we will evaluate the impact of an elevated O-GlcNAc landscape on molecular and cellular attributes of the mammary tumor and the associated immune microclimate using two distinct and complementary syngeneic mouse models of mammary cancer. To enrich the relevance, we will also evaluate human TNBC and PDX model systems. We will test if inhibiting GLI activity, in the context of elevated O-GlcNAc, uncouples the influence of O- GlcNAc-ylation on invasive and chemoresistant attributes of mammary tumor cells. Relevance: Our proposed studies are structured to systematically investigate how O-GlcNAc-driven metabolic reprogramming in cancer cells connects at the molecular level to aberrantly activate Hh/GLI signaling. The cumulative outcomes will create mechanistic understanding of how O-GlcNAc-ylation programs tumor invasion, progression and response to anti-neoplastics.

NIH Research Projects · FY 2026 · 2022-03

PROJECT SUMMARY This K23 award will allow Ronnie M. Gravett, MD, to train under multidisciplinary, expert mentorship to become an independent patient-oriented outcomes researcher dedicated to improving the HIV outcomes affecting persons in the Deep South. HIV disproportionately affects the Deep South, a region in the United States with the highest burden of new HIV diagnoses. Pre-exposure prophylaxis (PrEP) effectively prevents HIV acquisition, but there is a significant lag in PrEP uptake and persistence in the Deep South when compared to other regions of the US. Barriers prevent persons at risk for HIV from adequately accessing and using this effective strategy. The high burden of HIV and inadequate PrEP uptake are NIH priorities that need novel strategies to address, such as empowering and effective PrEP promotional strategies to increase PrEP uptake. Yet, there is a critical gap in understanding HIV prevention messaging that is informed by the greater context of the known barriers to PrEP uptake in the Deep South. To address this gap, this project will examine preferences for PrEP promotional messaging in order to create authentic and informative PrEP messages through crowdsourcing, an innovative strategy wherein a group solves a common problem and the solution is given back to the group. Integrating constructs of Andersen’s Behavioral Model of healthcare utilization and community engagement into a human-centered design model will serve as the framework for this project. This project will address the gap through the following specific aims: 1) explore preferences for PrEP messaging content in the Deep South through qualitative interviews, 2) using a discrete choice experiment (DCE), define the preferred attributes for PrEP messaging content to inform the format for the crowdsourcing open call, and 3) compare preferences for crowdsourced services to standard services to inform a digital intervention designed to increase PrEP uptake in the Deep South. Crowdsourced content created during this project will be compared to health authority content to show that community-derived content is preferred. The training plan will combine formal coursework, intensive seminars, and expert mentorship focused on the following topics: 1) quantitative, qualitative, and mixed methodologies, 2) design and conduct of discrete choice experiments, and 3) crowdsourcing to create authentic PrEP promotional messaging. This K23 award will support the development of effective content to improve HIV prevention strategies while also providing mentorship from renowned content and methodological experts, structured, multidisciplinary training in a collaborative environment, and the avenue for Dr. Gravett to develop into an independent physician investigator and leading expert in HIV prevention in the Deep South.

NIH Research Projects · FY 2026 · 2022-03

PROJECT SUMMARY Coordination of Innate and Adaptive Immunity in Intestinal Barrier Defense. Intestinal barrier defense requires the actions of both innate and adaptive immune cells on the mucosal epithelium. Tissue-resident and mobile innate and adaptive immune cells each contribute to barrier defense in the intestines, but how these cell populations are coordinated under conditions of pathogen threat are not fully understood. Interleukin-22 (IL-22) is a cytokine of the IL-10 family produced by type 3 immune cells, such as group 3 innate lymphoid cells (ILC3s) and cells of the Th17 pathway that acts on epithelial cells of barrier tissues to prevent invasion of extracellular pathogens. How IL-22 acts to coordinate intestinal barrier function remains undefined. Like many immune cytokines that participate in host defense, IL-22 is upregulated in chronic immune-mediated diseases, and it appears to play a protective role in inflammatory bowel disease (IBD), presumably by restraining epithelial damage caused by dysregulated T cell responses to constituents of the intestinal microbiome. However, pro- proliferative actions of IL-22 have also been implicated in malignant transformation of colonic epithelial cells that leads to colorectal cancer (CRC). We and others have shown that during infectious colitis modeled by the enteropathogen, C. rodentium, there are two phases of IL-22 production that can be distinguished: an early phase dominated by IL-22+ innate immune cells, which is followed by a late phase dominated by IL-22+ T cells. While both innate and adaptive immune cells produce IL-22 during infection, the respective contributions to barrier protection are unknown, as are details of the mechanisms by which IL-22 acts. In preliminary studies that have employed novel IL-22 reporter/conditional knockout (cKO) mice with which to track and/or delete specific subsets of IL-22-producing immune cells, we have found that the locations and functions of IL-22–producing cells are distinct during C. rodentium infection. Innate immune cells, dominated by ILC3s, are primarily located in and restricted to isolated lymphoid follicles, and their release of IL-22 activated by IL-23 acts long-range to activate surface colonic epithelial cells at initial sites of bacterial colonization. Remarkably, however, ILC3s fail to protect the intestinal crypts, which are invaded by bacteria in mice with IL-22 deficiency targeted to T cells. Thus, IL-22–producing T cells are indispensable for protection of the intestinal crypts via their activation of crypt- lining epithelium. Moreover, we have discovered new heterogeneity within colonic absorptive enterocytes and find that IL-22-producing T cells differentially activate these populations for increased shedding and production of neutrophil-recruiting chemokines. In this proposal we will define mechanisms by which innate and adaptive immune cells are specialized for distinct IL-22–dependent actions on different subsets of colonic epithelial cells, focusing on novel functions of IL-22+ T cells to protect colonic crypts from bacterial invasion. These studies hold promise to reveal new insights into specialization of immune cell subsets in intestinal host defense and mechanisms that control intestinal inflammation in IBD and CRC.

NIH Research Projects · FY 2025 · 2022-02

Advanced Glycan End Products (AGEs) are toxic and highly reactive dicarbonyl molecules produced by most life on earth from routine metabolic processes. As such, conserved and dedicated detoxifying systems have emerged for dicarbonyl removal. Owing to their importance, these removal systems are required to maintain longevity, thereby emphasizing the importance of dicarbonyl detoxification in maintaining health. One of the most prominent dicarbonyl species is glyoxal, which is predominantly produced as a byproduct of glycolysis. Glyoxal acts by mounting specific attacks on certain amino acids, namely arginines, cysteines, histidines and lysines in key proteins, thereby adversely altering protein function. In humans, these modifications can result in many diseased states, including: cancer, diabetes, nervous system disorders, heart disease, hypertension, atherosclerosis and aging. Unfortunately, although dicarbonyl stress-related toxicity is now regarded as important as oxidative stress, knowledge about how cells are able to detect and respond to glyoxal buildup is, by comparison, severely lacking. Our lab has discovered a novel class of Antibiotic Monooxygenase (ABM) domains that we hypothesize sense and respond to glyoxal and related dicarbonyls from bacteria to humans. This project proposes to elucidate the mechanism by which one of these ABM domains, we named Glyoxal-ABM Domain 1 (GAD1) responds to glyoxal in the bacterial pathogen Pseudomonas aeruginosa. We have thus far shown that GAD1 from P. aeruginosa, which is co- transcribed with the glyoxal detoxification enzyme GloA2, binds heme directly and is also covalently modified by glyoxal on a conserved arginine residue (Arg49). We hypothesize that GAD1 and its many homologs are specifically modified on conserved residues, which, in turn, signals to switch cellular metabolic flux away from glycolysis other pathways unable to produce the glyoxal toxin. Our studies here will Aim to (1) map GAD1 regulation, (2) determine its cellular distribution and its interactome and (3) solve the structures of its apo and holo forms, and in complex with interacting partners in P. aeruginosa. Studying GAD1 in P. aeruginosa is expected to reveal novel pathways that have potential as new antimicrobial targets, and at the same time advance our basic understanding of glyoxal toxicity sensing in humans and other multicellular organisms.

NIH Research Projects · FY 2026 · 2022-02

Project Summary Cystic fibrosis (CF) is one the most common life-shortening single gene defective disorders with over 70,000 cases worldwide. Mortality of CF patients has significantly decreased over the last decades and we are facing new treatment challenges for an aging CF population. While CF transmembrane conductance regulator (CFTR) modulator therapy is effective for many CF manifestations, airway inflammation as a hallmark of CF lung disease is not consistently impacted leading to impaired mucus clearance and susceptibility to chronic airway infections. Therefore, it is of great interest to identify novel therapeutic approaches that improve CF-associated chronic inflammation, mucociliary dysfunction and recurrent infections to be ultimately applicable for all CF patients. We have recently published that Fibroblast Growth Factor (FGF) signaling, a well characterized pro-inflammatory and aging signaling pathway, is activated in CF lung disease. We hypothesize that FGF receptor (FGFR) inhibition will attenuate CF-associated inflammation and impaired mucociliary clearance. We will test our hypothesis by employing a multidisciplinary approach combining cell culture with animal models including a chronic airway infection model. Aim 1 will establish the effects of FGFR inhibition in vitro on airway inflammation and mucociliary clearance in the human CF lung using primary human bronchial epithelial cells, cultured at the air liquid interface from different CF genotypes. The in vivo Aim 2 will recapitulate these findings in established CF rat models and assess FGFR inhibition and its impact on CF-associated inflammation, muco-obstructive disease and lung function. Aim 3 will further establish FGFR modulation in the chronically infected CF lung in vivo. The overall aim of this proposal is to establish the role of FGFR modulation in CF associated lung complications as an amenable therapeutic target to ultimately improve functional outcomes, quality of life, and long-term survival in the growing population of aging individuals with CF.

NIH Research Projects · FY 2026 · 2022-02

SUMMARY The purpose of this Mentored Patient-Oriented Research Career Development Award (K23) is to provide Colm Travers, MD, with the mentorship, training, and research experience needed to become an independent clinician scientist and leader in neonatal pulmonary outcomes research. His long-term career goal is to reduce the increasing burden of respiratory morbidity among the 360,000 survivors of preterm birth per year in the US through novel observational and interventional studies and large scale multi-center clinical trials of interventions that reduce lung injury. To achieve these goals and transition to independence, Dr. Travers and his mentors developed a comprehensive research and career development plan that includes mentorship from an exceptional team of scientists with proven track records of mentorship; intensive didactic training including completion of a Master of Science in Public Health in Outcomes Research; and a research plan that is purposefully designed to provide experiential learning in advanced research methods to study pulmonary outcomes among preterm infants. Survivors of preterm birth are at increased risk for wheeze, asthma, and abnormal pulmonary function tests (PFTs) during childhood. Studies of lung function in preterm infants have been limited by the difficulties with completing infant PFTs, restricting the ability to measure illness severity, assess response to treatments, and understand developmental lung health trajectories. Dr. Travers recently completed two prospective cohort studies using an innovative non-invasive oscillometry technique to quantify pulmonary mechanics in spontaneously breathing term and preterm infants. His early work observed that differences in lung mechanics between term and preterm infants can be reproducibly measured and persist until discharge. In the research plan outlined in this K23 proposal, Dr. Travers will expand upon this novel work, and his specific research aims are directly aligned with his training plan as follows: (1) To determine the extent to which differences in lung mechanics between healthy term infants and preterm infants with and without lung disease after birth persist until discharge from hospital or 40 weeks’ postmenstrual age, (2) To quantify changes in lung mechanics among preterm infants receiving medications for lung disease, and (3) To determine the extent to which abnormalities in pulmonary mechanics detected before discharge persist and predict longer-term pulmonary outcomes at 24 months. A non-invasive method to measure lung mechanics and predict longer-term pulmonary outcomes before discharge from hospital can fundamentally change our understanding of the effect of early postnatal lung development on lung function in preterm infants. In addition, a non-invasive bedside device that objectively measures response to treatments would be both a vital research and clinical tool. Building upon the skills and insights acquired through the proposed training and research plan, Dr. Travers will apply for an R01 to determine the effects of early interventions and therapies on lung mechanics and pulmonary health in early childhood.

NIH Research Projects · FY 2025 · 2022-02

With extensive application of treatment strategies, the acute mortality of patients having myocardial infarct (MI) has significantly reduced but these patients turn to ischemic cardiomyopathy (IC). Importantly, the incidence of IC is much higher in diabetic patients, a major risk factor for cardiovascular disease. Despite the major advances in cardiac interventions, diabetic IC (DIC) morbidity and mortality continue to rise. It is increasingly recognized that integrative approaches, rather than purely focusing on cardiomyocytes, must be undertaken for DIC prevention/treatment. Supplementation of adiponectin (APN), a protein identified as an adipokine, protects the acute ischemic heart in animal model. However, several clinical studies demonstrate that elevated APN levels are strongly associated with poor prognosis of chronic HF. We and others previously report that APN function is markedly attenuated in diabetic animals and patients. Our most recent study demonstrates that diabetes injures heart not only on cardiomyocytes, the impaired coronary endothelial cells (EC) in diabetes leading to microcirculation dysfunction dramatically enhancing cardiac dysfunction. Our recently published work and preliminary experiments further demonstrate that 1) in human diabetic coronary EC, APN receptor 1 knockout (AdipoR1, prototypic APN receptor in EC) mRNA expression is unchanged, but AdipoR1 protein expression is significantly reduced; 2) coronary vascular dysfunction (CVDy) is markedly exaggerated in diabetic animals as well as in AdipoR1KO mice; in type 2 diabetic mice, AdipoR1 expression is significantly reduced in the coronary EC. 3) proteomics reveal that AdipoR1 phosphorylation is the most significant post-translational modification in diabetic coronary EC; 4) GRK5 (not GRK2), the prototypical GRK family member in EC, is markedly upregulated in diabetic coronary EC; 5) GRK5OE in coronary EC has no effect upon AdipoR1 mRNA expression. However, GRK5OE largely reproduces the pathologic phenotypes in human coronary EC concerning APN/AdipoR1 signaling; 6) conversely, GRK5KO restores APN angiogenic effect in diabetic coronary EC. Based upon these exciting preliminary results, we will test a novel hypothesis that diabetic GRK5 upregulation and resultant AdipoR1 phosphorylation plays causative roles in diabetic CVDy and contributes to the deleterious consequences of DIC. GRK5-AdipoR1 system may be a novel therapeutic target against diabetic CVDy, ultimately ameliorating the DIC. This hypothesis will be rigorously tested by addressing the following 3 scientific questions: 1) what is the molecular mechanism that causes AdipoR1 desensitization and blocks AdipoR1 function in diabetes? (Aim 1); 2) How is APN’s coronary vascular protective action impaired when AdipoR1 is phosphorylated? (Aim 2), and 3) which intervention is most effective in restoring coronary vascular function in the diabetic heart against DIC? (Aim 3). Successful completion of proposed studies will greatly advance our knowledge in understanding the basis of diabetic cardiac injury through integrating regulation of coronary microcirculatory function, and identify novel therapeutic targets against DIC.

NIH Research Projects · FY 2026 · 2022-02

ABSTRACT — Combination chemoradiation is utilized to treat multiple gastrointestinal (GI) cancers including rectal cancer. Rectal cancer affects 40,000 people per year in the US. Approximately 85% of patients have an incomplete or poor response to treatment increasing their risk of recurrence. We have found that poor responders harbor sub-clones that are more resistant to treatment, and that the enzyme ST6Gal-1 is enriched in these sub- clones. ST6Gal-1 is a Golgi glycosyltransferase that adds the negatively-charged sugar, sialic acid (SA), to specific proteins destined for the cell surface. SA can have profound effects on the structure and function of proteins. ST6Gal-1 is one of the most pervasively upregulated glycosyltransferases in cancer cells. ST6Gal-1 has been shown to specifically promote tumor cell survival and resistance via sialylation. In addition, ST6Gal-1 has been found in extracellular vesicles (ECVs) made by cancer cells. ECVs are particles with a lipid membrane that contains RNA and protein cargo; thus, they are potential mediators of transferable resistance between cancer sub-clones. The role of ST6Gal-1 and ECVs in resistance to chemoradiation has not been investigated. The overall objective of this application is to ascertain the role of ST6Gal-1 in innate and transferable resistance to chemoradiotherapy in rectal cancer. Based on our preliminary data, we hypothesize that ST6Gal-1 mediates resistance to chemoradiation in individual sub-clones in rectal cancer, that this resistance is transferred to other sub-clones via ECVs spreading resistance, and that this resistance is regulated by ST6Gal-1 cleavage by BACE1. We have found that ST6Gal-1 is increased in rectal cancer models after treatment with chemoradiation. We will investigate our hypothesis with 3 aims: AIM 1 — Determine the role of ST6Gal-1 in chemoradiation resistance in human rectal cancer. We hypothesize that ST6Gal-1 causes treatment resistance after chemoradiation by inhibiting apoptosis. We will employ cell sorting, sequencing, and shRNA approaches. We will also conduct studies to investigate its function in patient samples. AIM 2 — Determine if ECVs carrying ST6Gal-1 transfer resistance to chemoradiation between sub-clones in rectal cancer. We hypothesize that ECVs act as vectors that impart resistance to chemoradiotherapy from sub-clone to sub-clone by trafficking ST6Gal-1, and thus, glycoprotein sialylation, in rectal cancer causing decreased apoptosis in the recipient sub-clones. AIM 3 — Determine if BACE1 promotes chemoradiosensitivity in rectal cancer due, in part, to ST6Gal-1 cleavage. We show that BACE1 mRNA is increased in tumors from patients who completely respond to chemoradiotherapy. BACE1 is known to cleave ST6Gal-1, and we found through inhibitor studies that BACE1 appear to regulate SA due to cleavage of ST6Gal-1 by BACE1. This research will evaluate a previously unknown mechanism of resistance to chemoradiotherapy in rectal cancer, with future potential for development of novel therapeutics that could target multiple resistant sub-clones across multiple GI adenocarcinomas, where the standard of care is pre-operative chemoradiation treatment.

NIH Research Projects · FY 2026 · 2022-02

Candidate: I am an Assistant Professor in UAB’s Department of Psychiatry with a background in molecular methods used in preclinical models and the role of intracellular pH dysregulation in neurodevelopmental illnesses. Additionally, I have expertise in diagnosis and treatment of psychopathology having received my MD- PhD from LSU Health Shreveport and completed a Psyhciatry Residency at Brown University. Career Goals and Development: I hope to gain expertise in assessing schizophrenia (SZ)-associated molecular disruptions in postmortem brain, in generating SZ patient-derived induced pluripotent stem cells (iPSCs) and differentiating them into disease relevant cell types, and in measuring cellular trafficking and luminal pH through the use of fluorescently-tagged protein constructs. By acquiring these skills and completing the studies laid out in this proposal, I will be well positioned and competitive for independent funding. Research Project: Deficits in protein post-translational modifications (PTM) and trafficking are reported in schizophrenia (SZ) brain, but the underlying cause is unknown. The function of organelles involved in PTM and trafficking is greatly impacted by pH disruptions, and Na+/H+ Exchangers (NHE) 6-9 are major regulators of organelle pH. In cancer cells, hypoxia causes altered energy metabolism and redistribution of NHE6 from endosomes to the plasma membrane. Similar metabolic alterations are reported in SZ brain suggesting that NHE6-9 intracellular distribution may also be affected, which could contribute to disrupted protein PTM and trafficking. So far, I have found that NHE7/8 expression is decreased in SZ dorsolateral prefrontal cortex (DLPFC) while NHE6/9 is unchanged. Still, NHE6/9 show increased expression in a tissue fraction enriched for synapses suggesting altered distribution of these proteins. Here, I propose to more extensively determine the expression and distribution of NHE6-9 first in SZ postmortem DLPFC and then in excitatory cortical neurons and astrocytes differentiated from patient-derived iPSCs. I will also determine how the introduction and removal of an acute stressor (hypoxia) affects the distribution of these proteins in these cells. Finally, I will transfect cells with fluorescently-tagged protein constructs to measure NHE6-9 and neurotransmitter receptor trafficking as well as organelle pH in live cells. These studies could help identify novel treatment targets for SZ and lead to high throughput assays to identify drugs that reverse SZ-associated molecular disruptions. Mentorship: The primary mentorship team for this proposal consists of Dr. James Meador-Woodruff, a world renowned expert in molecular disruptions in schizophrenia brain and analysis of postmortem brain tissue, Dr. Marek Napierala, an expert on molecular mechanisms of repeat expansion disorders and of modeling these illnesses using iPSCs differentiated into a variety of cell types including cortical neurons, and Dr. Vladimir Parpura, an expert in glial biology and visualization of vesicular trafficking in live cells through the use of fluorescently-tagged protein constructs.

NIH Research Projects · FY 2025 · 2022-02

Project Summary In eukaryotic organisms, transcribed RNA is processed from precursor messenger RNA (pre-mRNA) into mature RNA in a process known as splicing. During this RNA processing mechanism, the non-coding regions of pre-mRNA are removed, and the flanking regions are joined by a large molecular machine known as the spliceosome. Spliceosomes do not exist pre-assembled into splicing active conformations. Instead, splice sites (SS) are specifically chosen through the stepwise assembly of five small nuclear ribonuclear protein complexes consisting of a small nuclear RNA and a large number of associated proteins. These spliceosome assemblies are charged with correctly identifying and juxtaposing splice sites that are not explicitly sequence encoded in the pre-mRNA. Adding to the complexity of splice site selection, >90-95% of human pre-mRNAs are alternatively spliced by varying the configuration of which regions are joined and which are removed from multi-exon containing genes. Splicing errors associated with alternative usage of splice sites are implicated in a large number of human diseases such as Hutchinson-Gilford progeria syndrome (alternative 5'SS), dilated cardiomyopathies (alternative 3'SS), Myelodysplastic syndromes (altered 3'SS preference) and early-onset Parkinson Disease (cryptic splice site usage). Despite decades of research to characterize splicing mechanisms, the mechanisms that control splice site usage are incompletely understood. To fill this knowledge gap, the long- term goal of the candidate is to characterize the mechanisms that control splice site selection and the splicing factors involved. In this project, I propose to investigate protein-driven RNA rearrangements during splicing catalysis using single-molecule fluorescence microscopy methods through three specific aims. In aim 1, I will implement a single molecule Förster resonance energy transfer (smFRET) approach to characterize a conserved spliceosome rearrangement driven by the Prp22 helicase that leads to displacement of ligated mRNA from a conserved region in the spliceosome catalytic core, U5 snRNA loop 1. A Prp22 variant will be used to stall spliceosomes onto a surface immobilized pre-mRNA just after exon ligation but prior to release from the spliceosome. Prp22-driven displacement of the ligated mRNA will subsequently be monitored using fluorescent reporters installed on U5 snRNA loop 1 and the RNA substrate, respectively. Specific Aims 2 and 3 propose the investigation of a human-specific protein, FAM32A, hypothesized to stabilize the interaction between the 5' exon and U5 loop 1 in order to facilitate ligation to the 3' SS. Together, this work will answer questions about conserved and metazoan-specific mechanisms involved in the late stages of pre-mRNA splicing catalysis. This project will advance the applicant's career goal of running an independent laboratory at an academic institution in a way that combines her graduate training in mechanistic enzymology with her ongoing postdoctoral training in RNA molecular biology and biophysics to characterize the mechanisms and assembly of complex macromolecular machines whose proper functions are vital to human health.

NIH Research Projects · FY 2026 · 2022-01

SUMMARY. Vaccination is one of the most important public health achievements in history. However, we are still unable to induce protective immunity against important human pathogens, such as influenza. Thus, infectious diseases remain a major cause of disability and death. An essential component of a “successful” vaccine is the ability to generate long-lived plasma cells (LLPCs) AND memory B cells, which produce protective antibodies (Ab) and provide long-term prophylactic immunity. Importantly, the development of LLPCs and memory B cells occurs in the germinal center (GC). Thus, it is essential to understand the mechanisms that control the GC reaction. However, despite significant advances in the field, our understanding of the mechanisms that control the GC responses is still limited. One of the critical gaps in our knowledge is how “GC fate decisions” are regulated, particularly how GC B cells “choose” between staying in the GC to differentiate into highly mutated LLPCs or becoming memory B cells and leave the GCs. The lack of precise knowledge of the mechanisms that fine-tune the output of the GC is one of the main limitations when designing new vaccination strategies to overcome individual pathogen adaptions. In this regard, previous studies demonstrate that preexisting influenza-specific memory B cells in the lungs provide critical protection after reinfection. However, the factors that control the generation of lung memory B cell responses remain elusive. We believe this knowledge will be essential for designing more efficient vaccination strategies against respiratory viruses, such as influenza or SARS-CoV2. Importantly, CD4+ T follicular helper (Tfh) cells play a fundamental role in promoting GC B cell responses. In fact, in the absence of Tfh cells, GC responses and Ab-mediated protection are impaired. Thus, it is generally believed that an “enhanced” Tfh cell response after vaccination will significantly improve the efficacy of vaccines. Unfortunately, we still do not know what functional properties define a “high-quality” Tfh cell response. Our preliminary data demonstrate that, as the immune response progresses, the influenza-specific Tfh cell response “evolves.” As a consequence, different subsets of Tfh cells are present at different times after influenza infection. Based on our data, we hypothesize that GC B cells interacting with different “flavors” of Tfh cells at different times after infection receive qualitatively different signals, which temporarily fine-tunes the output of the GC and the generation of lung memory B cells. The long-term goals of this application are 1) To determine the role played by distinct subsets of Tfh cells in controlling the memory/LLPC differentiation balance. 2) To define the mechanisms that regulate the generation of “high quality” Tfh cells with the ability to promote enhanced B cell-mediated protection against respiratory viruses. 3) To determine the molecular and transcriptional mechanisms that control the generation of pulmonary memory B cells and the memory/LLPC differentiation balance in the GCs. We believe this knowledge will be essential for designing new vaccination strategies tailored against respiratory viruses.

NIH Research Projects · FY 2025 · 2022-01

PROJECT SUMMARY/ABSTRACT Impaired skeletal muscle regeneration and associated pathological tissue remodeling (loss of muscle, gain of fibrotic and adipose tissues) following injury underlie functional and metabolic decline—hallmarks of aging. Regeneration is dependent on a well-orchestrated myogenic program that includes the activation and expansion of skeletal muscle stem cells/progenitor cells (MPCs) and terminal differentiation of MPCs into mature multinucleated muscle cells. We previously demonstrated that MPCs rely on extracellular availability of the nutritionally, non-essential amino acids L-serine (Ser) and glycine (Gly). Decreased availability of Ser/Gly impairs MPC expansion, induces intramuscular adipocytes following injury, and induces toxic deoxysphingolipid accumulation in the muscle. Further, we demonstrated that endogenous Ser/Gly levels decline with age. The metabolic product (i.e. requirement) of Ser/Gly for MPC expansion and the efficacy of dietary Ser/Gly for muscle regeneration and the cell (MPC)-extrinsic environment need to be resolved. We propose to use isotope tracing of Ser and Gly to define the metabolic requirement of Ser/Gly for MPC population expansion. Further, using models that we have demonstrated reduce (depleted diet) or enhance (supplemented diet) endogenous Ser/Gly levels, we will quantify the effects of Ser/Gly availability on age- and injury-related muscle regeneration and the cell (MPC)-extrinsic muscle environment. Based on preliminary data, we hypothesize that MPCs require glutathione synthesis, from extracellular Ser and Gly, to mitigate oxidative stress. Additionally, we hypothesize that diet-induced reduction of endogenous Ser/Gly exacerbates age-related (i) impairments in muscle regeneration and (ii) toxic non-canonical sphingolipids in the cell-extrinsic muscle environment. Further, we expect Ser/Gly supplementation will counter these effects. To capture the efficacy of dietary Ser/Gly to modulate age-related impairments in muscle regeneration and remodeling we will use novel sphingolipidome profiling and transcriptomics. Successful completion of this project will transform the fundamental understanding of the metabolic essentiality of Ser and Gly for skeletal muscle regeneration and the relationship of this loss to age- related muscle deterioration. The results will enable testable scientifically grounded therapies to improve the regenerative capacity in populations that have impaired muscle regeneration, such as older adults.

NIH Research Projects · FY 2026 · 2022-01

ABSTRACT Despite notable success in EGFR-driven lung cancer, precision therapeutics have failed in EGFR-driven gliomas, the most common and deadly primary brain tumors. Reasons for failure of EGFR therapies in this clinical context include the lack of preclinical models that faithfully recapitulate the biology of EGFR-driven gliomas, including intra-tumor heterogeneity, drugs specifically designed to target invasive brain tumor cells located behind and intact blood-brain barrier (BBB), and adaptive drug resistance. Here we will develop and molecularly credential novel, EGFR-driven human glioma models for use in preclinical development of EGFR tyrosine kinase inhibitor (TKI)-based therapies. The foundation of the proposal comes from the Furnari Lab, who developed a novel platform (iGBM) for engineering glioma models using CRISPR genome editing and has established intra-tumor genetic heterogeneity as a symbiotic driver of tumorigenesis. The Miller Lab has extensive experience in small molecule experimental therapeutics using genetically engineered gliomas model and next-generation sequencing. He also used a novel chemical proteomics method, multiplex inhibitor beads coupled with mass spectrometry, to assess the glioma kinome en masse and showed that dynamic kinome reprogramming contributes to targeted drug resistance in glioma models. He is now at the University of Alabama at Birmingham, where local collaborators have extensive experience with biologically faithful human patient-derived xenograft (PDX) models. The O’Rourke Lab is a pioneer in development of sophisticated glioblastoma organoid (GBO) models that faithfully recapitulate the biology of molecularly and cellularly heterogeneous human tumors. In this Multi-PI project, we will combine our expertise to address the following Aims: (1) To develop novel genetically engineered human models driven by the most common EGFR extracellular domain mutations. We will then biologically and molecularly credential these models against genetically-matched PDX and GBO using genomics, epigenomics, transcriptomics, and kinome proteomics, and therapeutically challenge them using a panel of EGFR TKI, including one designed to specifically target invasive glioma cells behind the intact BBB. (2) To credential heterogeneous EGFR mutant iGBM models via biological, molecular, and EGFR TKI therapeutic profiling. We will thus develop human models with defined driver mutations that will be useful adjuncts to PDX/GBO for preclinical drug development. Models will be used to develop future rational combination therapies that combat drug resistance and enhance EGFR TKI efficacy. This work will therefore help realize the unmet need of precision therapeutics in neuro-oncology.

NIH Research Projects · FY 2026 · 2022-01

PROJECT SUMMARY Non-dipping type of nocturnal blood pressure (BP) is common even among treated hypertensives and is associated with a nearly two-fold higher risk of adverse cardiovascular events. Obesity impacts nearly 40% of American adults, and obese individuals have a high prevalence of hypertension and non-dipping type of nocturnal BP. The natriuretic peptides (NPs) are hormones produced by the heart that regulate BP rhythm by causing dilatation of vessels, sodium excretion, and inhibition of the renin-angiotensin-aldosterone system (RAAS). We have demonstrated that there exists a diurnal rhythm (24-hour rhythm) of NPs, which tracks closely with the BP rhythm and is in an antiphase relationship with the rhythm of the RAAS hormones. Obese individuals have lower NP levels throughout the day and a misaligned NP-RAAS-BP rhythm. LCZ696 is an FDA approved inhibitor of neprilysin (an NP degrading enzyme) that augments the NP levels and also suppresses the RAAS. Hence, the confluence of putative NP deficiency and the NP- RAAS-BP rhythm misalignment in obese may contribute to the high-risk nocturnal non-dipping BP profile seen commonly among obese individuals. Chronophamacology (controlling the time of medication administration) to synchronize the NP-RAAS-BP axis inherent biological rhythms may help control the nocturnal BP dipping pattern in obese individuals. We hypothesize that nighttime administration of NP- RAAS-BP axis therapy in obese hypertensive individuals will resynchronize their physiological rhythm patterns and help to improve their nocturnal BP profile. We plan to conduct a patient-oriented physiological, clinical trial to assess the effect of timing of administration of NP-RAAS-BP axis therapy (to target the rhythm misalignment) and the type of NP-RAAS-BP axis therapy (to target NP deficiency) on restoring the nocturnal BP dipping in obese hypertensives with non-dipping. We will conduct a 2x2 factorial design trial, wherein individuals will be randomized to one of the four arms; 1) daytime dosing of LCZ696; 2) nighttime dosing of LCZ696; 3) daytime dosing of valsartan; or 4) nighttime dosing of valsartan. We will study the effect of timing of NP-RAAS-BP axis medication inhibiting therapy (factor 1: morning vs. evening dose) on the nighttime BP profile in obese hypertensive patients with nondipping nocturnal BP. We will also assess the effect of the type of NP-RAAS-BP axis therapy (factor 2: LCZ696 vs. valsartan) on the nocturnal BP profile in obese hypertensives with nondipping nocturnal BP. We will examine the impact of timing and type of NP-RAAS-BP axis therapy on the NP levels, RAAS levels, and their diurnal rhythms. This study will assess an innovative physiologically-driven precision medicine approach of using chronopharmacology for resynchronizing the NP-RAAS-BP rhythm axis and restoring the normal BP rhythm in obese hypertensives with non-dipping BP.

NIH Research Projects · FY 2025 · 2022-01

Abstract The overall goal of this K23 proposal is to provide Caroline Presley, MD, MPH the essential mentorship, career development, and research experience necessary to become an independent investigator whose patient- oriented research will contribute to improving emotional distress, self-management, and outcomes in vulnerable people with type 2 diabetes. Diabetes distress — the emotional burden of living with and managing diabetes on a daily basis — affects 36% of people with type 2 diabetes and is closely linked with poor adherence to diabetes self-management behaviors, as well as suboptimal glycemic control, the presence of cardiovascular disease risk factors, and diabetes complications. Low-income individuals are disproportionately affected by both diabetes and diabetes distress. Currently, interventions are lacking to simultaneously help adults with type 2 diabetes to improve their diabetes self-management and glycemic control, as well as to reduce diabetes distress. In this four-year K23 project, Dr. Presley plans to study Mindfulness-Based Diabetes Education — an intervention specifically adapted for adults with type 2 diabetes and elevated diabetes distress that targets stress reduction and improvement of diabetes self-management behaviors and outcomes. In safety-net healthcare clinics, she will conduct a pilot randomized controlled trial (RCT) comparing this intervention to standard Diabetes Self- Management Education (DSME) in low-income adults with suboptimally controlled type 2 diabetes and moderate-severe diabetes distress to assess feasibility and acceptability. Additionally, Dr. Presley will use mixed methods to evaluate key contextual factors related to intervention delivery and implementation obtaining input from provider and patient stakeholders. Assessing context is critical in designing for implementation and sustainability. These results will inform a future fully-powered efficacy study of the intervention in safety-net healthcare systems. We hypothesize that Mindfulness-Based Diabetes Education will be acceptable and feasible — tested through these specific aims. Aim 1: To conduct a pilot RCT of Mindfulness-Based Diabetes Education versus standard DSME to determine feasibility and acceptability among adults with suboptimally controlled type 2 diabetes and moderate-severe diabetes distress. Aim 2: To characterize the contextual factors relevant to delivery and implementation of Mindfulness-Based Diabetes Education using PRISM. Along with this mentored research experience, the career development plan has been created for Dr. Presley to gain skills and experience in 1) behavioral interventions and trials, 2) clinical application of mindfulness-based interventions, 3) mixed methods research and implementation science through expert mentorship, advanced trainings, coursework, and conferences, as well as opportunities for continued professional development. Upon completion of the project, Dr. Presley will be poised to conduct a fully-powered efficacy study of the Mindfulness-Based Diabetes Education intervention supported by an R-series grant.

NIH Research Projects · FY 2026 · 2022-01

The overarching goal of this project is to determine the role of angiotensin converting enzyme 2 (ACE2) in the diabetic gut, how it impacts hyperglycemia and glycemic variability, and thus contributes to the pathogenesis of diabetic retinopathy (DR). The protective arm of the renin angiotensin system (RAS) consists of ACE2, which converts angiotensin II (Ang-II) to angiotensin 1-7 (Ang-1-7). Ang-1-7 opposes the effects of Ang-II by virtue of its actions on the MAS receptor. While the systemic (endocrine) RAS works with local (tissue) RAS such as that in the eye and gut to achieve homeostasis in health, in diabetes loss of key components of the protective RAS can lead to widespread pathology. The literature and our preliminary data support that diabetes results in loss of expression of ACE2 in the gut, bone marrow, and retina. Glycemic variability is implicated in DR pathogenesis. Intestinal ACE2 can regulate glucose homeostasis by modulating tryptophan absorption and incretin release and by generating Ang 1- 7 from luminal Ang II. Ang 1-7 by binding to Mas receptor can block glucose transport in the gut similar to what has been described in the pancreas. Based on this, we hypothesis: In T2D, loss of enterocyte ACE2 decreases: i) tryptophan uptake and incretin secretion leading to hyperglycemia; ii) MAS receptor activation increasing gut glucose absorption; and iii) gut barrier integrity resulting in leakage of gut microbial peptides into the circulation. All three mechanisms increase retinal permeability and activating immune cells promoting DR pathology. Aim 1 will test if dysregulation of ACE2 in the gut epithelium results in i) interruption of tryptophan transport by B0AT1 decreasing incretin secretion and ii) reduced MAS receptor activation leading to increased glucose absorption in the gut. Aim 2 will test if in db/db mice, loss of intestinal ACE2 will result in increasing circulating levels of gut microbial peptides that will activate TLRs on retinal endothelial cells and lead to increased retinal leukostasis and blood retinal barrier dysfunction. Aim 3 will examine if nutraceuticals or probiotics can restore the balance of the intestinal RAS (ACE2/Ang-1-7/MAS) in db/db mice to prevent development of DR. Impact: We propose a novel mechanism for deterioration of glucose homeostasis and increased glucose variability in diabetes- the loss of function of the ACE2:B0AT1 oligomer form of ACE2 (unique to the intestinal epithelium) and reduced levels of intestinal Ang 1-7 resulting in less intestinal MAS receptor activation. The dysregulated intestinal RAS can lead to serious retinal pathology promoting DR.

NIH Research Projects · FY 2026 · 2022-01

Functional peripheral and central vagal neural circuits of interoception inhibiting pain Interoception is the sense of the physiological condition of the body, and is critical for maintaining homeostasis and regulating cognitive and emotional processes. The neural processing of interoception can be regulated by electrical stimulation of the vagus nerve, which led to an FDA-approved therapy for seizure and depression. Interestingly, vagal stimulation also modulates intractable chronic pain in patients. However, the road to improving chronic pain management through the regulation of interoceptive inputs is blocked by our ignorance of the neurobiological mechanisms whereby vagal activity modulates chronic pain, posing a significant hurdle. To overcome this hurdle, we will focus on the neural mechanisms of vagal modulation in a mouse model of inflammation in temporomandibular joint (TMJ), a surrogate model of temporomandibular disorders (TMD). TMD is a prevalent form of chronic pain that often occurs as a comorbidity with other chronic pain conditions, such as migraine and fibromyalgia. Since chronic pain involves a wide range of neural processes ranging from peripheral nociception to affective and cognitive processing in the brain, it is possible that interoceptive inputs regulate pain pathways through multiple peripheral and central mechanisms. Vagal stimulation was suggested to inhibit transmission of pain signals at spinal cord through the regulation of descending pain modulatory pathways. Considering the high comorbidity of chronic pain and affective disorders, such as anxiety or depression, vagal inputs likely modulate pain through regulation of brain regions involved in emotional regulation. Furthermore, pain is driven by nociceptive processing by pain-sensing nerves at peripheral tissues, but vagal regulation of nociception in the periphery has not been reported. Here, our objective is to determine functional neural mechanisms by which interoception inhibits pain. Our central hypothesis is that vagal interoceptive circuits intersect with peripheral and central nociceptive pathways to inhibit pain from TMJ. We will test this hypothesis in the following specific aims.

NIH Research Projects · FY 2026 · 2022-01

Project Summary/Abstract: Exacerbations of chronic obstructive pulmonary disease (ECOPD) are a key driver of morbidity, mortality, and health care costs. A subset of COPD patients experience frequent ECOPD, and have a particularly poor prognosis. ECOPD are often caused by infections with encapsulated bacteria such as Streptococcus pneumoniae (pneumococcus), and there is growing evidence that frequent exacerbators have impaired adaptive immune responses. Prior studies have demonstrated associations between ECOPD and low IgG and IgG subclass levels, as well as downregulation of genes associated with adaptive immune pathways. Impaired pneumococcal antibody function (PAF) and specific IgG2 variants (allotypes) are associated with increased risk of encapsulated bacterial infections in primary immunodeficiencies, however these factors have not been studied in COPD. The multiplexed opsonophagocytosis assay (MOPA) measures PAF via killing of pneumococci by serum antibodies in vitro, and is the primary method for measuring immune responses to pneumococcal vaccines in adults. Preliminary studies indicate that PAF can also be used to evaluate immune function in COPD, and that lower PAF is associated with frequent exacerbations over the previous year. The central hypothesis for this proposal is that PAF and IgG2 allotype can predict a COPD subgroup with increased ECOPD risk. To investigate this hypothesis, PAF and IgG2 allotype will be measured in blood samples previously collected from the multicenter SPIROMICS cohort. Aim 1 of this proposal will determine whether low baseline PAF predicts risk of future ECOPD. The objective of Aim 2 is to determine whether low PAF predicts a chronic bronchitis COPD phenotype with neutrophilia and airway-dominant imaging. Reference levels for PAF will be established using results from a non-COPD cohort, then used to identify a PAF-deficient COPD subgroup. We will determine whether PAF deficiency is associated the above phenotypes. Aim 3 will investigate whether the IgG2 allotype associated with low PAF is more common among frequent exacerbators, versus non-exacerbating COPD and non-COPD controls. The results of this study could identify novel COPD subgroups and risk factors for exacerbations. Findings from this study may also promote the development of individualized therapeutic approaches tailored to those with low antibody function. The proposed research and career development plans are made possible through the mentorship of Dr. Moon Nahm, an international expert in pneumococcal immune responses, and Dr. Mark Dransfield, a leader in clinical and translational COPD research. The proposal also includes training in laboratory techniques, biostatistics, clinical and translational research, microbiology, and immunology, in order to foster an independent research career with a focus on immune dysfunction in COPD.

NIH Research Projects · FY 2025 · 2022-01

Preeclampsia (preE), defined as the onset of hypertension paired with proteinuria and/or other organ complication after 20 weeks of gestation, is a common pregnancy complication affecting 2-8% of pregnancies worldwide. The condition is even more common among mothers with chronic mild hypertension at pregnancy onset (affecting >25%). Importantly, preE confers a significantly increased risk of maternal and fetal morbidity including cardiovascular complications, preterm delivery, low birth weight and even death. Furthermore, recent studies demonstrate that a history of a preE is associated with an increased risk of cardiovascular disease for the mother later in life. The pathophysiology of preE is not completely understood but abnormal placentation, vascular dysfunction and oxidative stress is thought to cause maternal endothelial dysfunction resulting in the onset of clinical symptoms. To date the only definitive diagnosis is through blood pressure and urine protein measurement in the second or third trimester. However, the pathology is suspected to start in the first trimester and earlier identification can allow for better treatment and outcomes. The epigenome is recognized as an important driver of the gene expression changes necessary to support pregnancy. Numerous epigenomic studies of placental tissue have identified differentially methylated regions (DMRs, a type of epigenetic modification) associated with preE in genes and pathways suspected to underlie disease. A handful of studies have identified changes in the maternal epigenome from blood which could be identifiable earlier in pregnancy. Overall, additional research is needed to determine if methylation sites in maternal blood cells are useful to understand preE risk. This study will leverage the rich resource of the Chronic Hypertension And Pregnancy (CHAP) study designed to determine the efficacy and safety of antihypertensive treatment during pregnancy. Our ancillary study will use a nested case-control design (650 cases and 650 controls) to discover CpGs and DMRs for preE using existing data and blood samples. Findings will be replicated among participants from the Magee Obstetric Maternal & Infant Biobank database (N~650). PreE CpGs and DMRs (validated through replication) will be further tested for association with maternal cardiovascular outcomes in CHAP as well as in parous women from observational cohorts with existing metylation data from the National Heart Lung and Blood Institute’s Transomics for Precision Medicine (TOPMed) Program. The proposed research seeks to better understand the pathophysiology of preE and identify potential new biomarkers to facilitate early detection, management, and treatment of this serious pregnancy condition.

NIH Research Projects · FY 2026 · 2021-12

PROJECT SUMMARY/ABSTRACT Acute myeloid leukemia (AML) with MLL (KMT2A) gene rearrangement (MLL-r) is an aggressive disease with uncontrolled proliferation of myeloid progenitor cells and a failure of proper cell differentiation. Despite conventional chemotherapy, the overall survival of MLL-r AML remains poor and therapeutic options are limited, highlighting an unmet need to understand MLL-r AML pathogenesis and discover new genetic vulnerabilities. MEF2 transcriptional factors (MEF2A, 2B, 2C, and 2D) play important functions in the development of muscle, neuronal, and lymphoid lineages. Despite the known role of MEF2C as a direct MLL-r target essential for leukemogenesis, it remains unknown whether additional MEF2 family members are deregulated or involved in MLL-r leukemia. In this study, we identified that MEF2D gains aberrant super-enhancers and is highly upregulated in MLL-r AML. We demonstrate that MEF2D is selectively required for MLL-r AML, and depletion of MEF2D results in profound leukemia differentiation through transcriptional repression of CEBPE. We further show the MEF2D-CEBPE axis is critically involved in the anti-leukemia effects of DOT1L and Menin inhibitors. Furthermore, we discovered a novel interdependency of MEF2 paralogs in MLL-r AML. These preliminary data have provided us scientific rationale and enthusiasm for our central hypothesis that MEF2D, a novel transcriptional dependency highly expressed in MLL-r AML, maintains leukemia through inhibition of a CEBPE- centered myeloid differentiation program. This hypothesis is supported by extensive preliminary data and will be further tested by two specific aims: (1) establish the oncogenic function of MEF2D in MLL-r leukemogenesis and therapeutic response, and (2) investigate the mechanisms of MEF2D-mediated oncogenic regulation in MLL-r AML. In Specific Aim 1, we will determine the role of MEF2D in our pre-established genetically defined AML mouse models in vitro and in vivo; we will also evaluate the role of MEF2D-CEBPE axis in Menin inhibitor- mediated anti-leukemia effects. In Specific Aim 2, we will determine the molecular mechanisms by which MEF2D represses CEBPE, define MEF2D target genes using unbiased genome approaches, and evaluate the role of MEF2D-MEF2C interaction in MLL-r AML. The long-term goal of this project is to understand the MEF2 regulatory network in MLL-r AML, and to develop novel therapeutic approaches targeting oncogenic MEF2 factors for leukemia therapy. The main objective of this proposal is to establish the oncogenic function of MEF2D and determine how it regulates leukemia cell self-renewal and differentiation. Results from this proposal will provide significant new knowledge on the critical role of MEF2D in AML, reveal a new mechanism for suppression of normal hematopoietic differentiation in leukemia, and serve as the basis for targeting MEF2-related pathways as a potential therapeutic strategy for AML patients.

NIH Research Projects · FY 2025 · 2021-12

Abstract The overall goal of our project is to validate a diagnostic tool for dementia with Lewy bodies (DLB), and Parkinson’s disease dementia (PDD), two common neurodegenerative diseases affecting 1.4 million people in the U.S. Currently, definitive diagnosis of DLB and PDD often requires the postmortem detection of disease- associated alpha-synuclein (αSynD) aggregates in the brain. Clinically, DLB and PDD can be easily misdiagnosed with other dementias and parkinsonisms such as Alzheimer’s disease (AD) and tauopathies. An unmet medical need is to identify biomarkers for early and differential diagnosis of DLB and PDD in more easily accessible tissues. We have taken advantage of the emerging technology known as the real-time quaking induced conversion (RT-QuIC) assay to develop a robust platform for ultrasensitive detection of αSynD in peripheral tissues. In preliminary studies, we are able to detect prion-like seeding activity of αSynD in the skin of DLB and PD patients with 100% specificity and sensitivity. In addition, we were remarkably successful in detecting αSynD seeding activity in multiple peripheral tissues including skin, sigmoid colon, and submandibular glands in autopsied specimens. We hypothesize that RT-QuIC of peripheral αSynD is a highly sensitive and robust diagnostic biomarker for premortem diagnoses of DLB and PDD. To test this hypothesis, we propose to pursue the following four Aims: (1) Establish peripheral αSynD as a biomarker for postmortem diagnosis of DLB and PDD using RT-QuIC assay; (2) Assess skin αSynD as a biomarker for premortem diagnosis of DLB and PDD; (3) Determine peripheral αSynD and tau as a biomarker for differentiating DLB and PDD from other dementias and parkinsonisms such as AD and tauopathies; (4) Explore gut αSynD as a biomarker for premortem diagnosis of DLB and PDD using colon biopsy. Successful implementation of this proposal will establish RT- QuIC assay utilization for early diagnosis of DLB and PDD using readily available peripheral specimens.

NIH Research Projects · FY 2026 · 2021-12

ABSTRACT The reasons for individual variability in the physiologic response to dietary patterns are not well understood but hamper efforts to provide optimum diets to our population. There is an urgent need to understand the complex interaction of demographic, genetic, metabolic, behavioral, psychosocial, and environmental factors that affect the responses to dietary patterns in order to prevent and treat nutrition-related chronic diseases. The field of “precision nutrition” holds great promise for elucidating these interactions to eventually predict the optimal diet for an individual or groups of individuals. The overall objective of this application is to demonstrate that the University of Alabama at Birmingham (UAB) is uniquely positioned to join the consortium as a Nutrition for Precision Health Clinical Center (RFA-RM-21-005). The study team will collect a wide range of physiological and metabolic data from individuals in response to free-living (module 1) and controlled diets (modules 2 & 3), that will be used in analyses to determine potential predictors of response to diet. Sophisticated data methods (artificial intelligence, machine learning, mathematical modelling) will then be employed by the study group to identify the comprehensive phenotypes needed for individualizing diet prescriptions. We aim to accomplish the following three specific aims: Specific Aim 1 (module 1): Conduct an observational study of 2000 free-living individuals consuming their usual diet for 14 days. The physiologic responses to a standardized test meal challenge will be assessed while they are consuming their usual diet. Specific Aim 2 (module 2): Conduct a free-living controlled feeding study in 400 subjects fed three isocaloric diets varying in macronutrient composition at maintenance energy requirements. Diets are designed to elicit a wide range of responses among participants. The physiologic responses to standardized test meals and diet-specific meals will be measured at the end of each 14-day diet period. We will also collect measures of 24-hr glucose, 24-hr blood pressure, 24-hr physical activity, cardiorespiratory fitness, and sleep during each diet period. Specific Aim 3 (module 3): Conduct a domiciled controlled feeding study in 150 subjects of three isocaloric diets (same diets as in aim #1) fed at maintenance energy requirements. In addition to module 2 outcomes, assessments including room calorimetry, doubly labelled water, cardiorespiratory fitness, and muscle and fat biopsies will be completed in module 3 participants while they are domiciled in cottages at the Lakeshore Foundation Campus near UAB. Achieving these aims will create a database that allows sophisticated data analysis (e.g., AI, machine learning) to develop algorithms to match people to optimum diets. UAB, with access to >16,000 All of Us participants in Birmingham, outstanding facilities for conducting diet interventions, and an outstanding research team, can be a valued member of the Nutrition for Precision Health Consortium.

NIH Research Projects · FY 2025 · 2021-11

Each puff of an e-cigarette generates micromolar amounts dihydroxyacetone (DHA) from the combustion of propylene glycol and glycerol. Up to 40-55% of e-liquid content is converted to DHA in each puff from an e- cigarette, making DHA a high-volume component found in all e-cigarette vapors, which the vaper inhales with each puff of the e-cigarette. DHA is approved for external use as a sunless tanning agent, but serious concerns have been raised about inhalation exposures through spray tanning and now e-cigarette use. We have shown that DHA is genotoxic, cytotoxic, and induces mitochondrial dysfunction in skin and kidney cells, but the effects of inhalation exposures to DHA are currently unknown. The long-term goal of the proposal is the identification and validation of markers for cellular and metabolic stress induced by DHA exposure that can be examined in tissues from vapers to understand the consequences of repeated inhalation exposures to DHA. The objective of this proposal is to address the gap in existing studies, which have only focused on skin models, by examining the exposure effects of DHA at both acute and chronic doses in pulmonary and cardiovascular cells. Our central hypothesis is that exposure to DHA alters metabolic pathways, promotes oxidative stress, disrupts Ca2+ homeostasis, and leads to mitochondrial dysfunction. The rationale for this work is that DHA exposures to the lung and cardiovascular system allow direct absorption of DHA into cells. DHA-induced changes in metabolism and mitochondrial function would compromise overall cellular function, leading to disease. Three specific aims will test the central hypothesis: 1) DHA incorporation into metabolic pathways alters glycolysis and induces glycosylation protein damage; 2) DHA exposure alters NAD(P)H pools inducing oxidative stress, and 3) DHA exposure alters cytosolic Ca2+ levels and disrupts mitochondrial function. The first aim will test the sub-hypothesis that DHA alters metabolic pathways by tracing DHA metabolism using isotopologues of DHA and identifying metabolite disequilibrium. The second aim will test the sub-hypothesis that an excess of DHA changes cofactor pools and induces oxidative stress. The third aim will test the sub-hypothesis that DHA alters Ca2+ signaling to induce mitochondrial dysfunction, in addition to causing metabolic stress and oxidation-reduction imbalance. The study is innovative because it extends beyond the genotoxic and cytotoxic characterization of DHA to measure DHA’s ability to reprogram pulmonary and cardiovascular cells metabolically. The research is significant because e-cigarette users are chronically exposed to DHA, which will directly impact pulmonary and cardiovascular cell homeostasis and cause severe declines in cellular function or even induce cell death. This work will establish essential markers for DHA exposure to allow future epidemiological work to associate DHA exposure to disease.