Wayne State University

NIH Research Projects · FY 2025 · 2025-08

Project Summary/Abstract The goal of this proposal is to bring a Thermo Fisher Scientific Orbitrap Astral mass spectrometer with a High-Field Asymmetric Waveform Ion Mobility Spectrometry (FAIMS Pro Duo) devise, an Ardia Instrument Control and Data Management system plus a nano-LC to the Proteomics Core at Wayne State University. The Proteomics Core supports an extensive user base including 7 Major Users and 19 Minor Users who have contributed to the proposal and who have NIH-funded projects that will be advanced by having access to the Orbitrap Astral. The Proteomics Core at Wayne State also supports the Karmanos Cancer Institute and the P30 funded Cancer Center as well as the NIEHS funded CURES P30 Center and CLEAR P42 Superfund Center. This Orbitrap Astral has unique capabilities for deep profiling of complex proteomes using Data Independent Analysis (DIA) acquisition. Advances in the Orbi Astral that make it unique for DIA are the 200 Hz scan speed with 80,000 Resolution and superior sensitivity. These features allow the Astral to acquire DIA data using 2 Da windows, something no other MS system can achieve. The new mass analyzer in the Orbitrap Astral make it an exceptional instrument for DIA proteomics. Combined with the experienced mass spectrometrists in the Core, the Orbitrap Astral will be a major asset to research programs of NIH-funded investigators at WSU and in the SE Michigan region. As investigator needs have evolved they have developed an increased need for higher resolution, greater sensitivity and higher mass accuracy proteomic mass spectrometry. The Orbitrap Astral is designed to address todays needs in proteomic analysis. The instrument excels at deep sequencing and quantitative proteomics as well as accurate characterization of post translational modifications. The current mass spectrometers in the Core lack fast duty cycles and the advanced ion optics that make detection of low abundance species routine on the Astral. The ability to unambiguously localize post translational modifications (PTM) in peptides is a critical feature in any proteomic MS system and the Astral is outstanding in this due to the high degree of coverage within the identified proteins. Wayne State University is strongly committed to this proposal as evidenced by a combined $350,000 in new support for the instruments management, operation, maintenance, informatics and usage. The technical expertise of Proteomics Core personnel as well as the physical and administrative infrastructures are all in place and ready to immediately put this new mass spectrometer to work on NIH-funded biomedical research projects. This instrument will provide transformative technologies for advancing dozens of NIH- funded, ongoing projects, as well as catalyzing new research directions for investigators at Wayne State University.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Over 2 billion people suffer from vision impairment worldwide, largely due to chronic, age-associated retinal degenerative diseases. This presents a significant health and cost burden on our society. In addition, given that humans possess limited regenerative ability, there are currently no cures for these diseases. In contrast, zebrafish maintain a remarkable capacity to regenerate the retina following intense forms of acute damage, such as phototoxic ablation of photoreceptors, neurotoxicity by pharmacological agents, or even physical “pokes” using a wide-gauge needle. In 2008, resident Müller glia (MG) were identified as the stem cell source that were triggered to re-enter the cell cycle following these acute damage events. Since then, zebrafish MG have become a rich source to identify several genetic pathways involved in retinal regeneration. Despite these advances, most human retinal degenerative diseases are not acute, but instead manifest under slower, chronic stress conditions. The current work builds on two newly-established chronic models of photoreceptor degeneration: our chronic low light model and P23H mutant zebrafish that express a mutated Rhodopsin protein that is associated with retinitis pigmentosa in humans. Both models show a slow decay of rod photoreceptor outer segments and loss of ~50% rod photoreceptor nuclei. Interestingly, in neither model do we observe MG reactive gliosis or MG-mediated regeneration in this model. This differs substantially from our previously established acute light damage model, in which photoreceptors are rapidly destroyed and MG undergo two stages of reactive gliosis surrounding their cell-cycle re-entry. Here, we will compare the transcriptional, cellular, and functional changes that occur during chronic and acute photoreceptor damage, assessing 8 timepoints along a 28-day protocol. These data will provide the template for addressing photoreceptor degeneration and neuroprotection mechanisms (Aim 1), and MG reactivity to neuronal damage (Aim 2). This proposal aims to address a critical need to better understand genetic and glial support mechanisms in the context of chronic vs acute damage. Using state-of-the art bioinformatic approaches, cell sorting, pharmaceutical treatments, morpholino-mediated gene knockdown, and visually-evoked optokinetic responses, this proposal aims to directly test the relationship between acute and chronic photoreceptor stress, coupling cellular and morphological changes with transcriptomic and functional genetic studies.

NIH Research Projects · FY 2025 · 2025-08

Among women with endometrial cancer, certain high-risk populations have up to twice the mortality rate compared to others. This is due to a combination of social and biological factors, including socioeconomic differences, histologic subtype, stage at diagnosis, and access to quality care. Some high-risk populations have higher rates of aggressive histologic (e.g., non-endometrioid) and molecular (e.g., P53 abnormal) subtypes of endometrial cancer compared to others but tend to have worse survival across histologic subtypes. Recent studies have demonstrated that a substantial proportion of the excess risk of death experienced by high-risk populations similar to those in Detroit, Michigan cannot be explained by known social and biological factors, underscoring an urgent need for additional research. Chronic stress, through both environment and sociodemographic stressors, is thought to be associated with cancer development, progression, and survival. We hypothesize that chronic stress may modify the relationship between molecular subtypes of endometrial cancer and survival. We have assembled a team of population scientists, bioinformatics researchers, basic scientists, and clinicians to investigate this hypothesis among a cohort of women with endometrial cancer in Metropolitan Detroit. Our preliminary data demonstrated that among women with endometrial cancer, area-based disadvantage measures, which have been shown to correlate with chronic stress, are associated with both overall- and endometrial cancer-specific survival. For this project, we will leverage a comprehensive dataset from the population-based Detroit Research on Cancer Survivors (Detroit ROCS) cohort. This dataset includes annual survey data, geocoded addresses, and longitudinal clinical and vital statistics data for 320 women with endometrial cancer living in Metropolitan Detroit at diagnosis. Additionally, it includes whole exome sequencing of paired normal and tumor biospecimens from 132 women from the Detroit ROCS cohort. Aim 1 will evaluate differences among clinically important molecular characteristics of tumors and associations with survival. In Aim 2, we will investigate associations between chronic stress, molecular characteristics, and survival among the cohort. Aim 3 will involve data collection of a pilot cohort of women with aggressive endometrial cancer from the gynecologic oncology clinics at Karmanos Cancer Institute in Detroit, Michigan, to inform feasibility of the development of a prospective collection of stress measures, molecular characteristics, and endometrial cancer-relevant outcomes in support of a future R01 submission. Collectively, this project aims to generate preliminary data on socio-genomic associations associated with endometrial cancer outcomes, furthering the field of research in improving outcomes for those with gynecologic cancers.

NIH Research Projects · FY 2025 · 2025-08

Project summary/Abstract: The role of the corneal epithelium is to: 1) act as a barrier against physical trauma, pathogens, and chemicals, 2) maintain a constant level of stromal hydration, and 3) serve as an optical interface focusing light onto the retina with optimal quality. A healthy corneal epithelium requires coordinated actions of multiple dynamic cellular processes and signaling pathways. To date, studies of extra- and pericellular proteases in eye and vision research have mostly centered on their harmful effects in corneal wound healing and infection. Based on our preliminary data, we hypothesize that the cell-surface anchored serine protease matriptase, in contrast, is essential for normal ocular surface function and promotes recovery from injury to the corneal epithelium. We generated matriptase hypomorphic mice and observed an abnormal irregular corneal surface cell pattern and increased inflammation. Our preliminary data using human corneal epithelial cells suggest that matriptase silencing causes decreased barrier function accompanied by impaired tight junction (TJ) integrity and aberrant proteolytic processing of the epithelial cell adhesion molecule (EpCAM). Under injury conditions, we identified the pro-form of hepatocyte growth factor (pro-HGF) as a candidate substrate for matriptase with key functions in corneal wound healing. Matriptase efficiently cleaves pro-HGF and converts it into the active form needed for activation of its cell-surface tyrosine kinase receptor c-Met and for subsequent stimulation of cell migration. Our hypothesis is that matriptase promotes corneal epithelial barrier function via regulation of tight junction formation and promotes tissue repair by activation of the HGF/c-Met signaling pathway. Aim 1: Determine the role of matriptase in corneal epithelial maintenance and function. Novel matriptase knockout mouse models as well as 3D cellular models of the human cornea will be utilized. We will use a balanced combination of hypothesis-driven targeted experimentation and unbiased approaches to comprehensively characterize the role of matriptase in corneal homeostasis and to identify critical pathways required for proper regeneration and differentiation, TJ formation, EpCAM processing, and barrier function. Aim 2: Identify functions for matriptase in corneal epithelial tissue repair. Matriptase-deletion mouse models and 3D epithelium/stroma cell models will be used to determine the impact of matriptase on corneal repair in experimental wound repair. Comparative RNA-Seq and Mass Spectrometry analysis will be performed to identify matriptase substrates and matriptase-mediated pathways in homeostasis and injury response. The impact of stimulation with Internalin B321 ( InlB321 ), an agonist of the HGF receptor c-Met, on cell migration/tissue repair in vitro and in vivo will be tested as a potential therapeutic strategy to treat corneal epithelial injury. Significance: Implementation of the approaches described in this proposal will provide a mechanistic understanding of the role of matriptase in corneal epithelial homeostasis and injury and identify actionable targets (factors/pathways) for therapeutic options to promote corneal epithelial repair leading to restoration of function.

NIH Research Projects · FY 2025 · 2025-08

Tarca, A.L., 2022 The fetal mortality rate in the USA is 5.73 per 1,000 births, and there are significant disparities among racial and ethnic groups. While factors such as complications of the placenta, umbilical cord, congenital anomalies, and maternal conditions may be involved, one-third of fetal deaths are classified as unexplained. Our preliminary data suggest that novel placenta-specific proteins offer more discriminating power for fetal death cases compared to current maternal plasma biomarkers. However, the question of whether detection of such protein dysregulation in samples collected earlier in gestation from asymptomatic patients will improve prediction of fetal death compared to known biomarkers remains to be answered. The studies proposed in this R21 application are important because reliable identification of women at risk for fetal death is needed to enable use of preventive strategies such as pravastatin, a drug that prevents recurrent fetal death and placenta and maternal vascular diseases. Therefore, we will profile most promising maternal plasma protein candidates in samples collected weeks before fetal death diagnosis and in gestational age- matched controls. Samples were identified in the Wayne State University Perinatal Biobank. Using machine learning and predictive analytics pipelines as we have previously described, we will generate a placenta-specific plasma protein signature that is predictive of fetal death in asymptomatic patients. Multi-protein machine learning models, such as elastic net and random forest, will be developed and tested using cross-validation to ensure generalizability. Furthermore, we will assess the specificity of the fetal death proteomic model in an independent existing longitudinal dataset that includes data from normal pregnancies and cases of preeclampsia and small-for-gestational-age neonates. Finally, we will explore the molecular subclassification of fetal death cases using clustering algorithms based on plasma protein dysregulation prior to fetal death. The goal of such analysis is to generate additional hypotheses and tackle the heterogeneity of the fetal death phenotype, which has historically hindered the identification and validation of biomarkers for this syndrome.

NIH Research Projects · FY 2025 · 2025-08

Project Summary/Abstract This request is for funds to purchase a Canopy Biosciences CellScape instrument to fill a critical void in highly multiplexed spatial biology capabilities, becoming the only instrument of its kind on the Wayne State University (WSU) campus. The purchase of this instrument will directly support scientific advancement of NIH-funded research in cancer, metabolic disorders, and infectious diseases of the eye and allow our Users to perform deep analysis of various pre-clinical and patient sample types for better understanding of disease development and identifying new treatment options. The CellScape instrument will be housed within the Microscopy, Imaging, and Cytometry Resources (MICR) Core, a centralized instrumentation core facility of WSU and the Karmanos Cancer Institute (KCI). The MICR Core staff has extensive experience in all aspects of flow cytometry, confocal microscopy, and small animal imaging and currently serves over 75 PIs and 125 individual Users, both internal and external to WSU. While the MICR Core is well-equipped with several flow cytometry analyzers/sorters and light microscopy systems, the CellScape instrument will allow us to bridge these two technological arms of our core and introduce a new breadth and depth to the research capabilities of our facility. 16 PIs have been identified for the proposed instrument, 6 Major Users and 10 Minor Users. These 16 PIs have over $20M in NIH-funded direct costs from at least 5 different institutes. The new instrument will be maintained by the MICR Core which has extensive infrastructure and expertise in place to oversee User training, assay development and troubleshooting, instrument operation, data analysis, and maintenance.

NIH Research Projects · FY 2026 · 2025-07

Project Summary/Abstract Objective/Hypothesis: Pseudomonas aeruginosa is an opportunistic bacterium known to induce microbial keratitis and is associated with rapid corneal destruction and blindness, especially in individuals who wear contact lenses or are immunocompromised. Microbial keratitis triggers a robust inflammatory response driven by the influx of innate immune cells. During infection, neutrophils (PMNs) and macrophages (Mϕ) are recruited in the acute phase of inflammation to effectively remove the pathogen. However, their persistence and chronicity can lead to extensive tissue destruction, including corneal opacity, stromal fibrosis, and ulceration, if not properly treated. Eosinophils are another type of innate immune cell known to secrete cytokines, chemokines, and growth factors. Although their contribution to microbial keratitis has not yet been explored, an emerging body of literature suggests that eosinophils not only possess antibacterial properties but contribute to tissue remodeling and inflammation resolution. The overarching objective of this proposal is to characterize the role of eosinophils in modulating the corneal response to Pseudomonas aeruginosa-induced infection. Specific Aims: Experiments proposed in Aim 1 will characterize the spatiotemporal distribution of eosinophils in normal and infected corneas. Aim 2 will define the phenotypic profiles of eosinophils during infection. Aim 3 will comparatively analyze proteomic profiles during corneal infection. Study Design: To investigate the role of eosinophils during corneal inflammation and infection, we will use established mouse models of P. aeruginosa-induced keratitis with two disease outcomes: susceptible C57BL/6 (B6) mice that exhibit corneal perforation and resistant BALB/c mice that heal over time. We will largely utilize a combination of microscopy, immunostaining, flow cytometry, proteomics, and eosinophil depletion to comprehensively achieve the three stated aims. Impact: The proposed work is expected to elucidate the currently undefined role of the eosinophil within the corneal microenvironment, filling a current gap in knowledge regarding eosinophil biology in the eye. Therapeutically, the eosinophil may serve as an effective cellular target to help improve disease outcomes.

NIH Research Projects · FY 2026 · 2025-06

HYPERTENSION is considered the leading cause of “loss of health” worldwide, involving the kidney’s inability to excrete excess salt. During high salt intake the kidneys excrete the extra salt load by increasing the blood pressure, phenomenon known as the pressure-natriuresis response. On the other hand, salt-sensitive hypertension is a sustained increase in blood pressure caused by an acute salt intake, which is prevalent in 50% of African-Americans and in 30% of Caucasians. Enhanced salt retention by the thick ascending limb of Henle’s loop (TAL) involving the Na+/K+/2Cl- cotransporter (NKCC2) has been described in patients and genetic animal models of salt-sensitive hypertension. However, the molecular mechanism for this defect is not fully understood. Ubiquitination is a post-translational modification that regulates expression of channels and transporters. Recently a novel E3 ubiquitin ligase adaptor F-Box leucine-rich domain 13 (FBXL13) was identified as novel locus for blood pressure regulation in humans. We found that FBXL13 recognizes and interacts with NKCC2. We found that the high salt-induced increase in NKCC2 ubiquitination is blunted in FBXL13-KO mice. Global FBXL13-KO mice show exacerbated total NKCC2 expression. However, the global FBXL13-KO mice show high levels of ubiquitinated NKCC2, indicating that other E3-ubiquitin ligases or adaptors mediates the ubiquitination of NKCC2. Moreover, global FBXL13-KO mice are not salt sensitive, nor they develop hypertension. Therefore, this R03 project aims to discover other E3-ubiquitin ligases that mediates NKCC2 ubiquitination and play a role on NaCl reabsorption and blood pressure regulation under normal or high salt diet. In Aim I, we hypothesize that high salt diet stimulates the 48-linked poli-ubiquitination of NKCC2 via multiple E3-ubiquitin ligases. This proposal is significant to human health because in most hypertensive patients and in animal models of hypertension, the natriuretic effect of nitric oxide is decreased, yet the mechanisms involved in the development of hypertension are poorly understood. Although we already have drugs (loop diuretics) that effectively block NKCC2 activity, they are not used to treat hypertension due to their side effects and offside targets combined with the lose potency over time. This proposal will explore the mechanism and signaling cascade by which high-salt diet stimulates NKCC2 ubiquitination and will characterize a new E3-ubiquitin ligases that may play a critical role in salt-sensitive hypertension. This application will focus on specific interactions between E3-ubiquitin ligases and adaptors with NKCC2. Moreover, this proposal will increase the knowledge on post translational mechanism that regulates blood pressure. The proposal also will help us to understand a post-translational mechanism that we know little about and how it regulates NKCC2 expression, which would be crucial for the development of new strategies for the treatment of hypertension in special salt-sensitive hypertension, which could lead to the development of novel and specific loop diuretics.

NIH Research Projects · FY 2026 · 2025-06

Project Summary The prevalence of metabolic disorders (e.g., obesity), continues to increase and imposes significant costs on the US health care system. In the US, ~42% of adults (20+ years of age) are classified as obese Exposure to environmental contaminants has been implicated as a causal factor, and some of these contaminants, termed metabolic disrupting chemicals (MDCs), increase weight and lipid accumulation and/or perturb energy balance and/or metabolic homeostasis, contributing to metabolic dysfunction including obesity. Polyethoxylated surfactants (alkylphenols and alcohols) are emerging contaminants with ubiquitous human exposures and robust effects on metabolic health. Used widely in hard surface cleaners, detergents, paints, pesticides, and other products, polyethoxylated surfactants persist in the environment and contribute to chronic human exposure. We have reported an obesogenic effect in zebrafish following developmental exposure to nonylphenol polyethoxylates at environmentally relevant concentrations. In contrast, cetyl alcohol polyethoxylates increased overall adiposity without any apparent impacts on body weights. Given the growing use of these polyethoxylates in consumer products and the lack of comprehensive toxicity assessments on them, it is crucial to determine how polyethoxylates differentially act to disrupt metabolic health. Our central hypothesis is that these polyethoxylates preferentially promote an unhealthy visceral adipose expansion through thyroid hormone receptor antagonism, in part through skewing the commitment of mesenchymal stem cells to the adipose lineage at the expense of osteoblasts. Aim 1 will comprehensively assess the role of thyroid receptor disruption in the observed effects through a combination of in vitro experiments (co-exposures, siRNA knock-down, and chromatin immunoprecipitation with sequencing) and in vivo experiments (co- exposures and genetic knock-down approaches) to rigorously assess the role of thyroid hormone in the observed polyethoxylate-induced metabolic health effects. Aim 2 will directly interrogate the nature of polyethoxylate-induced adiposity via comprehensive assessment of adipose tissue in developmentally exposed zebrafish (histology and adipocyte morphometrics, immune cell infiltration, and metabolomics to measure inflammatory mediators). Paired in vitro experiments will assess impacts on mesenchymal stem cell commitment and differentiation through adipogenesis and osteogenesis assays and single cell transcriptomics to evaluate how polyethoxylates disrupt cell fate and differentiation decisions. This research will move the field forward by supporting a more integrated and physiologically relevant model of contaminant-induced metabolic disease. Given the wide environmental prevalence of polyethoxylates, clear human exposure pathways, and metabolic health impacts in a model with the potential for clear translation to human health, there is a critical need to better characterize effects and their mechanisms. We expect results of our proposed studies will drive additional research into these chemicals, most notably the AEOs that have received limited attention.

NIH Research Projects · FY 2025 · 2025-06

The long-term objective of this R61/R33 is to evaluate cannabidiol as a “cognitive enhancer” to increase efficacy of cognitive behavioral therapy (CBT) for generalized anxiety disorder (GAD). Emotion regulation difficulties, particularly a diminished ability to manage negative emotions, are central to the development of GAD and other anxiety disorders, which are often linked to irregularities in prefrontal brain regions. However, despite the general effectiveness of CBT, nearly half of all patients do not show significant improvement. Successful response to CBT is evidenced by improved emotion regulation and enhanced activity of the dorsomedial prefrontal cortex (dmPFC) during cognitive reappraisal, which is a proxy of the interventions practiced in CBT (e.g., cognitive restructuring). Our prior work suggests that administration of an exogenous cannabinoid facilitates emotion regulation and enhances dmPFC activation in posttraumatic stress disorder— an anxiety-related condition sharing similarities in prefrontal engagement during emotion regulation. Further, cannabidiol, a well-tolerated non-intoxicating component of cannabis, has been linked to significant reductions in anxiety. Acute cannabidiol administration also modulates dmPFC activity during emotion processing. Therefore, both cannabidiol and CBT have converging effects on dmPFC activity, indicating that a combined therapy may have synergistic benefits. The objective of this project is to test the novel hypothesis that the combination of CBD+cannabidiol will result in greater reductions in GAD symptom severity compared to CBT alone, and this effect will be associated with increased dmPFC activation during an emotion regulation task. In the R61 phase, individuals with GAD will be randomly assigned to one of three arms to identify potential dose- dependent effects of cannabidiol (FDA-approved EPIDIOLEX®)-assisted CBT: (1) 5-week Brief CBT plus moderate-dose cannabidiol (5 mg/kg BID); (2) Brief CBT plus low-dose cannabidiol (2.5 mg/kg BID); or (3) Brief CBT plus placebo. The “go” criterion for progression from R61 to R33 is target engagement, defined as a large effect size (d≥0.8) in within-subject change in dmPFC response during the emotion regulation task from pre- to post-CBT following CBT+cannabidiol. In the R33 phase, we will combine the minimum effective dose from the R61 phase with a standard 8-week Brief CBT protocol to (1) replicate target engagement and (2) test whether cannabidiol augments the reduction in symptom severity and the maintenance of treatment gains between once-weekly sessions and post-CBT. Individuals with GAD will be randomized to either a (1) CBT+cannabidiol or (2) CBT+placebo arm. Like the R61 phase, participants will complete the emotion regulation fMRI task pre- and post-CBT to determine whether CBT+cannabidiol leads to greater increases in dmPFC activation vs. CBT+placebo. We will also assess treatment response (reduced GAD symptom severity) at each CBT session and at 3 months post-treatment to explore long-term effects. Together the R61 and R33 phases will provide the most directly translational and critical test of cannabidiol-assisted CBT for GAD.

NIH Research Projects · FY 2026 · 2025-05

The ability to image the presence or absence of oxygen is of paramount importance to the study of biochemistry, medicine, and multiple diseases. Hypoxia is linked to a variety of diseases including kidney disease; hepatic and neurological toxicities; and the progression, proliferation, and therapy resistance of many cancers, making hypoxia an important diagnostic and therapeutic target. This proposal describes plans to study a new class of phosphonate-containing complexes of EuII that are compatible with systemic delivery, where the lack of systemic delivery is the largest obstacles to the widespread use of EuII for imaging hypoxia in multiple diseases. Specifically, we propose to address the two greatest challenges preventing systemic delivery: persistence time and relaxivity. With the advent of hypoxia-responsive contrast agents for magnetic resonance imaging (MRI), including the recent development and characterization by our team of a novel EuII-based agent that persists in blood, the ability to systemically deliver EuII for imaging hypoxia is expected to become a reality for the stud of multiple diseases. Our overarching goal is to develop hypoxia-sensing probes for MRI to target unmet needs in diagnostic medicine relevant to a range of diseases. We will build on our discovery from the previous funding period of the first EuII-based molecule compatible with systemic delivery by increasing the persistence and relaxivity of EuII through control of the phosphonate arms that are at the center of kinetic resistance to O2 and through conjugation to macromolecules. Our hypothesis is that new phosphonate complexes of EuII based on our lead complex will enable systemic delivery of EuII by increasing the persistence time and relaxivity of EuII in vivo. Aim 1 studies the properties of new derivatives of our initial discovery that contain electron-withdrawing groups. Aim 2 studies dendrimeric conjugates of our original phosphonate complex. Aim 3 defines the toxicity and biodistribution of Eu-containing phosphonate complexes in mice. The expected outcome of this proposal is an understanding of the design criteria of phosphonate complexes of EuII that enable systemic delivery and make the complexes compatible with bioconjugation to enable study of a wide-range of hypoxia-related diseases. We envision that our results will serve as the basis for future translation for monitoring new and existing therapies for hypoxia-related diseases.

NIH Research Projects · FY 2026 · 2025-05

Project Summary/Abstract Objective/Hypothesis. Diabetes is expected to reach epidemic levels by 2030. Despite the prevalence of diabetic retinopathy, many adult diabetic patients develop visually disruptive corneal complications, including impaired wound healing and corneal neuropathy. Unfortunately, treatment for diabetes-induced damage to the cornea remains severely limited to strict glycemic control. Our objective is to investigate a promising peptide-based combination therapy that effectively targets hyperglycemia-responsive circuits in the cornea. By doing so, we hypothesize that Tβ4/VIP combination treatment can prevent high glucose-induced damage to corneal structure and function. In addition, we examine one such responsive circuit – the DDR-TRP axis – a novel pathway that has yet to be explored in the cornea. Specific Aims. This proposal intends to: 1) establish the therapeutic role of Tβ4 and VIP as a combination treatment against diabetes-induced corneal complications; and 2) elucidate the DDR-TRP axis under hyperglycemic conditions. Study Design. Our experimental approach is designed to include both in vitro and in vivo cell-specific analyses. In vitro assessments are focused on corneal epithelial cells and stromal fibroblasts complemented by extensive, comprehensive in vivo assessments that include morphological changes to the epithelium, stroma, epithelial-stromal interface, endothelium, changes in corneal nerves and tear film, AS-OCT, endothelial cell density measurements, fibroblast differentiation, and phenotypic profiling to distinguish between keratocytes, fibroblasts, myofibroblasts, immature/mature dendritic cells, monocytes/macrophages, and neutrophils. In vivo studies include the use of two (2) type 1 diabetic mouse models (STZ-induced and spontaneous Akita) to account for any potential effects due to STZ exposure and will assess changes in corneal structure and function, including corneal integrity and barrier function, sensitivity and innervation, inflammation, and wound healing; all of which not only focus on improved disease outcome but mechanistically examine the role of the DDR-TRP axis and how Tβ4/VIP therapeutically regulates this axis. Impact. Overall, the studies proposed seek to delineate a newly identified mechanism of diabetes-induced damage while providing key preclinical evidence for the development of a novel therapeutic approach with robust translational relevance and potential for significant impact on diabetic corneas.

NIH Research Projects · FY 2026 · 2025-05

Abstract / Project Summary The opioid epidemic has impacted the lives of millions of individuals, including women that use opioids, such as fentanyl, during pregnancy putting the fetus at risk. A shocking NBC news story in late 2023 reported on a clinical case study that indicates a novel syndrome in babies exposed to fentanyl during pregnancy may have been found. However, polysubstance use in humans often complicates research, which may be the case here; therefore, an urgent need exists to determine the effects of gestational exposure to fentanyl (alone) on offspring well-being. In response to this need, we propose a set of studies using a rodent gestational exposure model to measure the effects of fentanyl during pregnancy on offspring at birth through early development. Our central hypothesis is that gestational fentanyl exposure causes malformation of the body and brain and abnormal behavior in offspring. We will test this hypothesis by determining the impact of gestational fentanyl exposure on morbidity and mortality, developmental milestones, external morphology, blood cholesterol, brain morphogen signaling, and neonatal opioid withdrawal and related behavioral measures in offspring. Furthermore, we will determine the impact of gestational fentanyl exposure on brain volume, morphology, and neurochemistry of offspring using magnetic resonance imaging and spectroscopy in vivo. The results of these studies will directly impact the clinical situation and our understanding of the direct role of gestational exposure to fentanyl in the reported cases indicating that a newly discovered fetal fentanyl syndrome may exist.

NIH Research Projects · FY 2025 · 2025-04

Individuals from financially limited communities have limited access to evidence-based interventions (EBIs) for mental health yet also experience greater psychiatric and medical comorbidities and poor treatment outcomes. Alarmingly, rates of major depressive disorder (MDD) and substance use disorder (SUD) have risen significantly for adults living in economic hardship over the past two decades, resulting in worsening clinical and functional outcomes. There is a pressing need to disseminate EBIs for MDD-SUD that are acceptable, scalable and sustainable in economically limited settings, including Certified Community Behavioral Health Clinics (CCBHCs) which seek to expand access to services in economically strained areas. Peer recovery specialists (PRSs), individuals in recovery from SUD, have promise for increasing access to EBIs and are widely employed in CCBHCs; yet, few studies have evaluated the clinical effectiveness, implementation, and cost-effectiveness of PRS-delivered EBIs, even as PRS models are rapidly scaling in the US. Preliminary data: PIs Felton and Magidson have led two open-label trials demonstrating the feasibility, acceptability, and preliminary effectiveness of a PRS-delivered model (Peer Activate) to improve MDD and SUD outcomes among economically-limited individuals. The team has shown that behavioral activation (BA), an EBI based on reinforcement theory and focused on scheduling of valued, adaptive behaviors is particularly suitable for PRS delivery and promising to address gaps in EBI access. Preliminary studies established the feasibility and acceptability of this approach; next steps are to conduct a large-scale randomized trial to evaluate clinical and cost effectiveness, test potential modifiable mechanisms of treatment effectiveness, and evaluate longer-term implementation. Methods: We propose to evaluate Peer Activate in a fully-powered randomized hybrid Type 1 effectiveness-implementation trial (n=250) in Detroit, MI. Leveraging a well-established partnership with a community-based, PRS-led CCBHC, we will compare Peer Activate to enhanced treatment as usual (ETAU; non-specific, PRS-delivered supportive care) on depressive symptoms (primary) and substance use (Aim 1), and evaluate potential mechanisms of this approach (environmental reward; Aim 2). Guided by the EPIS framework and Proctor’s model, we will evaluate longer-term implementation outcomes (Aim 3), including cost effectiveness. A community advisory board will guide all aspects of the trial to promote the potential for sustainability. Impact: Our proposal is consistent with the goals of PAR-21-130, NIMH strategic objectives to develop innovative service delivery models for economically limited individuals with mental health and SUD comorbidity, and NOT-MH-22-170 promotion of partnerships between academic institutions and CCBHCs. Findings have the potential to increase availability of sustainable EBIs for mental health and SUD comorbidity in hard-hit areas.

NIH Research Projects · FY 2025 · 2025-04

Project Summary Infertility affects approximately 16% of couples in the U.S., posing a significant reproductive health challenge. Spermatogenesis, the process of sperm development in adult males, involves intricate epigenetic mechanisms in the testis and maturation through interactions with extracellular vesicles in the epididymis called epididymosomes. These processes enable sperm to adapt to their environment through epigenetic changes. Our previous research linked preconception phthalate exposure to alterations in sperm methylome profiles, RNA cargo of seminal plasma extracellular vesicles, and embryonic development. However, the impact of other endocrine disrupting compounds (EDCs), such as Per- and polyfluoroalkyl substances (PFAS), on male reproductive health remains poorly understood. Emerging evidence suggests that PFAS exposure may affect fertility by influencing testosterone levels and sperm quality. However, the specific mechanisms by which PFAS influence spermatogenesis, sperm epigenetics, and epididymosomes remain largely unexplored. This proposal aims to investigate these mechanisms and explore potential connections between PFAS exposure and adverse effects on male reproductive health. We will conduct single-cell RNA sequencing (scRNA-seq) of the testis and study the small non-coding RNA (ncRNA) cargo of epididymosomes to better understand how PFAS impacts male reproductive health. Leveraging our established mouse PFAS exposure model, we have generated compelling preliminary data, identifying over 2,800 sperm differentially methylated regions (DMRs) associated with behavior, developmental pathways, and lipid metabolism. Thus, the central hypothesis of this proposal is that PFAS exposure negatively affects overall male reproductive health. Our proposed aims will focus on scRNA-seq analysis of testicular somatic and germ cells to decipher the influence of PFAS exposure on spermatogenesis trajectory and transcriptomes. Additionally, we will characterize epididymosomes and evaluate the effect of PFAS exposure on their sRNA cargo. The expected outcomes of this research will enhance our understanding of how PFAS mixtures impact spermatogenesis and the loading of sRNA cargo in epididymosomes. These insights will advance translational research, leading to the development of novel approaches for treating and preventing adverse reproductive health outcomes. Furthermore, this project will provide advanced training in bioinformatics, toxicology, and single-cell techniques, contributing to progress in environmental and reproductive health research.

NSF Awards · FY 2025 · 2025-03

The transition to a wider range of energy solutions is important for addressing global challenges, including mitigating carbon dioxide (CO2) emissions, integrating renewable energy, and electrifying transportation systems. Although lithium-ion batteries dominate the market, the scarcity, high cost, and supply chain vulnerabilities of lithium hinder their scalability for large-scale applications. This CAREER project focuses on nonaqueous magnesium (Mg)-CO2 batteries, an alternative next-generation energy storage technology that combines the natural abundance, low cost, and high capacity of Mg anodes with the ability to harness CO2 consumption for generating electricity. The primary bottlenecks in Mg-CO2 batteries involve designing efficient electrocatalysts and gaining an understanding of the interfacial reaction mechanisms. This research will address these obstacles and provide insights to guide the development of sustainable, high-capacity rechargeable batteries. The educational outreach efforts aim to inspire middle and high school students to pursue STEM careers and engage in cutting-edge scientific research in advanced battery technologies. To achieve this, summer workshops at the PI's lab will introduce students to the domain of energy storage and computational chemistry through immersive virtual reality (VR) demonstrations of battery chemistry, molecular simulations, and computer programming. Graduate and undergraduate students will obtain hands-on experience in advanced materials research that will prepare them as a skilled workforce to drive innovation in sustainable energy technologies. This project aims to develop predictive computational tools for designing electrocatalysts, elucidating electrolyte decomposition chemistries, and investigating interfacial reaction mechanisms in nonaqueous Mg-CO2 batteries. Key challenges, such as sluggish reaction kinetics and the formation of blocking interfaces, currently impede the practical realization of these batteries. To address the limitations, the research integrates multi-scale computational techniques—including density functional theory (DFT), machine learning (ML), and eReaxFF reactive molecular dynamics (MD) simulations—to explore electrocatalyst design strategies and electrode-electrolyte interfacial chemistries at the atomistic level. Specifically, this project will (i) predict structure-activity relationships and establish design principles for single-atom catalysts supported on nitride MXenes and transition metal oxides, (ii) investigate electrolyte decomposition pathways and solid electrolyte interphase formation on Mg anodes, and (iii) study reaction kinetics and mechanistic details of cathode-electrolyte interfaces. The computational results will be validated through experimental collaborations. The knowledge gained from this project will provide foundational guidelines to accelerate the development of sustainable, high-capacity metal-CO2 batteries for renewable energy and transportation electrification. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2025 · 2025-03

The rapid advancement of embedded systems, driven by system-on-a-chip (SoC) technologies, is accelerating the deployment of autonomous machines in fields like robotics, drones, and autonomous vehicles. These innovations demand more than correct operation; autonomous machines, particularly those powered by deep neural networks (DNNs) in autonomous vehicles, must meet strict timing requirements that necessitate rigorous real-time safety certifications. Such certifications rely on advanced analytical methods combining worst-case execution time analysis with schedulability analysis to ensure operational safety and reliability. However, significant challenges persist in integrating worst-case execution time and schedulability analysis, especially in evaluating the timing accuracy of systems that use computing accelerators for autonomous driving. The project’s novelties are its development of an integrated architecture that leverages hardware-software co-design to address these complex issues, with the aim of significantly enhancing the safety and reliability of autonomous driving systems and other autonomous machines. The project's broader significance and importance lie in its commitment to integrating this research into educational and outreach activities to encourage diversity in STEM fields and broaden participation in computing and engineering. To achieve these research goals, the project is organized around three core objectives: (1) developing an accelerator-enhanced SoC that enables real-time scheduling decisions directly at the hardware level, supporting efficient task management by the runtime scheduler; (2) designing an accelerator-aware real-time scheduling framework that accurately characterizes and assesses the complex execution behaviors and timing requirements of DNN tasks within the operating system layer; and (3) implementing an outlier management strategy to derive reliable worst-case execution time values for DNN tasks on accelerators at the application layer. Together, these objectives advance system software to reliably manage DNN-intensive applications on customized heterogeneous SoCs and establish precise, temporally accurate resource allocation schemes. This project sets the foundation for certifiable, safety-critical autonomous machines that deliver assured real-time performance and reliability. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2026 · 2025-02

Project Abstract The ability to image oxygen in vivo has important diagnostic, prognostic, and therapeutic implications because hypoxia is correlated with aggressive disease states and resistance to many types of treatments. EuII/III-based systems for magnetic resonance imaging (MRI) are an attractive approach for imaging hypoxia due to their potent oxygen-sensing capabilities. EuII produces bright T1-weighted images, but oxidation of EuII by O2 produces EuIII that does not positively enhance images. Our lab has demonstrated that EuII-containing contrast agents can be used for imaging hypoxia in tumors in vivo and that EuII/III-based contrast agents are compatible with multiple imaging modalities including 1H- and 19F-MRI, chemical exchange saturation transfer, and photoacoustic imaging. Although these results are promising, the short half-life of EuII in vivo is the critical barrier to imaging hypoxia in a wide range of environments using the EuII/III redox couple because the rapid oxidation of EuII at physiologically normal levels of pO2 limits the administration of the EuII-containing contrast agents to injection directly at sites of interest and prevents ratiometric imaging using multiple modalities. Our overarching goal is to increase persistence of EuII long enough to enable systemic delivery and ratiometric imaging of hypoxic regions in a wide range of diseases. We propose to address the critical limitation of the persistence time of EuII in oxygenated environments by modulating factors that kinetically hinder oxygen approach in the outersphere. We previously showed that a perfluorocarbon-soluble EuII-containing complex, EuII1, dispersed in a perfluorocarbon nanoemulsion has increased persistence times in vitro and in vivo as seen by 19F MRI. Our working hypothesis is that altering factors of gas diffusion will afford control over persistence of EuII1 in oxygenated environments. Aim 1 studies the solubility of O2 in nanoemulsions consisting of different perfluorocarbons with varying abilities to dissolve O2. Aim 2 investigates the influence of Henry’s and Graham’s laws of gas diffusion on the persistence of EuII using different gasses to perfuse into perfluorocarbon nanoemulsion to act as a counter pressure against O2 diffusion. In Aim 3, the effect of the size of nanoparticles in the perfluorocarbon nanoemulsion is studied with respect to the persistence of EuII. The expected outcomes of this proposal are (1) a better understanding of factors that influence the kinetic outersphere approach of O2 to EuII dispersed inside perfluorocarbon nanoemulsions and (2) persistence times of EuII1 in vivo that are long enough to enable systemic injection and ratiometric imaging.

NIH Research Projects · FY 2026 · 2025-02

Project Summary Opioid Use Disorder (OUD) poses serious consequences including overdose or death from excessive and prolonged use of prescribed or illicit opioids. The opioid epidemic has also resulted in a striking, nationwide increase in infectious diseases including HIV, hepatitis, and invasive bacterial and fungal infections, which worsen long-term health outcomes. The consensus is that opioids are immunosuppressive and thereby increase the vulnerability to infections. However, the effect of acute or repeated opioid exposure on immunity remains elusive. In the eye, substance use is an important risk factor for developing endogenous endophthalmitis, a blinding infectious disease primarily caused by fungal (Candida sp.) and bacterial (S. aureus) pathogens. Several clinical and epidemiological studies have reported an increasing number of opioid use-associated endophthalmitis cases. Our preliminary data showed that cultured human retinal pigment epithelium (RPE) cells, human monocytes, and mouse retina express the mu-opioid receptor (MOR) which is activated in response to morphine exposure. Acute morphine exposure increased, whereas repeated morphine exposure decreased, inflammatory response in cultured cells. Similarly, repeated morphine administration in B6 mice reduced cytokine levels in the plasma but elevated them in the retinal tissue. Additionally, systemic Candida auris infection in morphine-dependent mice increased fungal burden in the eye, brain, and kidneys. Based on these findings, we hypothesize that repeated opioid exposure (e.g., morphine and fentanyl) impairs both systemic and ocular innate immunity resulting in increased incidence and severity of infectious endophthalmitis. The overall goal of this proposal is to evaluate the effect of opioids on innate immunity and its impact on the pathogenesis of endophthalmitis. Here, we will elucidate mechanisms underlying impaired innate responses of retinal and myeloid cells under opioid exposure (Aim 1) and determine whether OUD increases the susceptibility and severity of endophthalmitis in mouse models (Aim 2). The knowledge gained from this study could have a significant impact on ocular infections among substance abusers and could contribute to the advancement of therapeutic interventions in the field, offering new hope for preserving vision and improving outcomes in patients affected by these conditions.

NIH Research Projects · FY 2024 · 2025-02

Early childhood adversity is associated with significant risk for engaging in problematic substance use across the lifespan. Adolescents living in poorer neighborhoods are both more likely to experience adverse childhood events and less likely to have access to evidence-based preventative substance use interventions. Thus, there is a critical need to identify interventions that can effectively prevent the escalation of substance use among these youth and be feasibly disseminated in these communities. Delay discounting has emerged in the literature as a critical etiological marker of substance use risk and a potential pathway from early life disadvantage to problematic alcohol and drug use. Elevated delay discounting is characterized by an exaggerated tendency to select rewards that are immediately available. Higher (more problematic) rates of delay discounting during adolescence are associated with more severe and frequent substance use and are predictive of steeper escalations in alcohol, marijuana and tobacco use. Working memory, the capacity to store and manipulate information utilized in decision making, is closely linked to delay discounting and a plausible neurocognitive mechanism linking early adverse environments to problematic discounting tendencies and subsequent substance use. Guided by an experimental therapeutics framework, recent findings from our own research team and others suggests the efficacy of utilizing a computer-based working memory training program to improve discounting among mid-life adults. Computer-based training programs may be particularly suitable to implement in poorer areas for at-risk adolescents. Specifically, adolescence is a critical period for the development of both working memory and delay discounting, and interventions targeting working memory during this life stage have evidenced favorable clinical results. Further, computerized interventions require limited staff and space resources, making them both feasible and scalable in communities with limited funding. Building on promising findings from the investigator team’s formative research, this application proposes to pilot a computer-based working memory intervention among adolescents exposed to early life adversity in Detroit, Michigan. Primary aims include establishing the preliminary feasibility, acceptability and appropriateness of delivering a computerized working memory intervention in a community setting serving adolescents as well as conducting a small-scale Stage I randomized control trial to examine the effectiveness of this intervention in reducing rates of delay discounting. The subsequent impact of this intervention on changes in adolescent substance use over a three-month follow-up period will also be examined as an exploratory aim. The proposed research will provide valuable insight into the utility of targeting a specific pathway from early adversity to later substance use during adolescence and will establish a model for disseminating interventions in youth-serving community agencies. Results of this study will also set the stage for future large-scale (R01) prevention research.

NIH Research Projects · FY 2026 · 2025-01

Abstract High-throughput experimentation has revolutionized organic synthesis and pharmaceutical lead identification by enabling the design, execution, and analysis of previously unimaginable quantities of experiments. This paradigm shift over the past two decades has transformed the landscape of chemical research by accelerating nearly every aspect of synthesis and analysis. These advances have been realized, in part, through engineering developments such as automated liquid handlers. However, these engineering innovations have also led to certain chemical limitations. Many reagents and catalysts, despite being highly useful in conventional synthesis, are excluded from modern HT approaches because they are not compatible with automated liquid handlers used in standard HT workflows. The omission of these important compounds has resulted in significant limitations in current HT approaches. This proposal outlines efforts to address some of these gaps. Specifically, it outlines efforts to adapt heterogeneous materials and macromolecules to modern HTE approaches to better utilize their unique properties in organic synthesis. In the first goal, we aim to develop colloidal suspensions of commonly insoluble inorganic reagents, allowing them to be processed as homogenous liquid solutions. These reagents will undergo benchmarking in important cross-coupling reactions, including cross-electrophile couplings and in engineering challenges. In the second goal, we focus on the development of material and macromolecular analogs of commonly used, yet cytotoxic reagents, facilitating direct to biology synthesis and assays. Our research emphasizes macromolecular analogs of organic super bases and metal scavengers that are particularly relevant to high-throughput catalysis. Lastly, in the third goal, we plan to synthesize and evaluate easily accessible site-isolated catalysts for transformations historically constrained by bimolecular decomposition. This includes electrophilic carbon-hydrogen bond functionalization and mild fluoro-dediazotization reactions. The completion of these studies will yield innovative tools for modern synthesis and pharmaceutical development. By bridging the gap between heterogeneous reagents/catalysts and common automated reaction platforms, this research has strong potential to expand the impact of high-throughput experimentation in organic chemistry thereby accelerating the synthesis and identification of molecules important to human health.

NIH Research Projects · FY 2026 · 2024-12

Project Summary/Abstract The long-term goal of our research program is to explore how mitophagy promotes tissue health. Mitochondria generate cellular energy, and a natural byproduct of executing this essential function is susceptibility to damage. A wide assortment of human diseases, including metabolic and neurodegenerative diseases, shares the pathological feature of improper accumulation of damaged mitochondria. Mitophagy, a type of mitochondrial quality control (MQC), degrades damaged mitochondria to prevent their inappropriate accumulation; and therefore, represents a critical mechanism required for the maintenance of tissue health. Most of our understanding of mitophagy comes from research performed in cell lines in culture, and thus, little is known about how the diverse mechanisms of mitophagy promote tissue health in intact organisms. Our lab seeks to address this knowledge gap by leveraging the genetic advantages offered by the model organism Drosophila melanogaster. We previously discovered that disrupting the ataxia-associated gene Vps13D results in severe perturbations in mitophagy such that mitochondria fated for mitophagy become stalled prior to degradation. These data have enabled us to elucidate molecular mediators of in vivo mitophagy in the fly, allowing us to generate unprecedented genetic tools and preliminary hypotheses that will facilitate a better understanding of the utilization of mitophagy in physiological contexts to support tissue function. In the next five years, we propose to generate genetic tools to effectively manipulate all known forms of mitophagy in fruit flies through engineering of the genes required to execute mitophagy: mitophagy adapter and receptor proteins. With these tools we will test the hypothesis that the two mechanistic forms of mitophagy degrade mitochondria with functionally distinctive objectives. We will test this hypothesis by individually investigating the physiological roles of the two forms of mitophagy in flies: ubiquitin-dependent mitophagy (in Research Direction #1) and receptor-dependent mitophagy (in Research Direction #2). In the third research direction, we will probe the relationship between these distinct forms of mitophagy to better understand how their interplay maintains tissue health and homeostasis. Overall, the information generated from this proposal will provide foundational knowledge on a critical cell biological process closely associated with human disease. In the future, this knowledge can be used to facilitate the development of mitophagy-targeting therapeutics that have the potential to treat diseases and conditions associated with mitophagy perturbations such as Parkinson’s Disease, diabetes, and ischemia-reperfusion injuries.

NSF Awards · FY 2024 · 2024-12

This grant provides partial travel support to U.S.-based early career researchers and students to attend a workshop that will be held together with the International Congress on Sustainability Science and Engineering (ICOSSE) in Auckland, New Zealand, Feb. 20-22, 2025. The purpose of the workshop is to exchange emerging ideas about ways and means of advancing research and enhancing education on industrial and environmental sustainability using AI and digital technologies. The workshop will help the U.S. workshop attendees to gain comprehensive understanding of the research and education opportunities in the challenging areas, define research and educational directions, and develop ideas for advancing their research and educational activities in the next step. As it will be jointly held with the ICOSSE’25 that will cover various topics on sustainability, the workshop will further benefit the U.S. workshop participants to learn more broadly the frontier research and education in engineering sustainability. The workshop will provide an international platform for the U.S. based scholars to engage with global leaders, innovators, and visionaries in multi-disciplinary research from around the world, especially from the U.S. and Asian and Pacific countries; to identify research directions for shaping the future of sustainability engineering using AI and digital technologies; and to develop international collaboration opportunities. Junior faculty members, early-career researchers, graduate and undergraduate students will be strongly encouraged to attend the workshop. A publication based on the workshop will be generated. The workshop will help develop broad international collaboration in this very challenging area. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT: Since 2015-26 epidemic in Brazil, Zika virus (ZIKV) infection has been linked to cause microcephaly in newborn infants and World Health Organization declared it a global public health emergency. One third of these infants were also reported to have ocular anomalies, yet the ocular pathology of ZIKV remain poorly understood, with no available treatments. Being RNA virus, ZIKV can evolve rapidly, underscoring the need for continued research. My long-term goal is to investigate how flaviviruses affect the blood-retinal barrier (BRB) and cause ocular issues. During my postdoctoral training, I conducted high-throughput transcriptomics on ZIKV-infected RPE cells and identified potential role of Ang/Tie2, S1P, and AMPK pathways involved in barrier integrity. However, the role of these pathways in ZIKV-induced BRB disruption leading to ocular anomalies remains unexplored. My hypothesis is that ZIKV infection dysregulates these pathways, compromising barrier integrity, and enabling viral entry into the eye, resulting in ocular complications. Therefore, therapeutic strategies aimed at restoring BRB integrity may alleviate ZIKV-induced ocular pathology. To test this hypothesis, I propose three aims: 1) Investigate the role of Ang/Tie-2 and S1P signaling in ZIKV-induced BRB permeability; 2) Examine the impact of AMPK activation on BRB integrity during ZIKV infection; and 3) Study the effects of RPE-specific knockout of AMPK, Ang/Tie2, and S1P pathway signaling on ZIKV-induced chorioretinal atrophy in mice. During the mentored phase, I will establish a congenital ZIKV-infection model in immunocompetent mice with guidance from Dr. Kumar (mentor) and Dr. Mor (collaborator), both experts in host-pathogen interactions and ZIKV-pregnancy models. I will also learn to culture iPSC lines to generate retinal organoids and perform gene knockouts using the CRISPR/Cas approach, guided by Dr. Arumugaswami (co-mentor), an expert in stem cell and retinal organoid culture. The mentored phase will involve regular meetings with mentors, co-mentor, and advisory committee members, attendance at scientific conferences, and ongoing career development. In the independent phase, I will establish my own laboratory and complete the proposed aims by utilizing an RPE- specific knockout ZIKV-congenital infection model. Successful completion of these aims will elucidate (i) the roles of host Ang/Tie2 and S1P pathways, and AMPK in BRB regulation; and (ii) the feasibility of mitigating ZIKV-mediated ocular tissue damage via restoring barrier integrity. These findings will provide crucial insights into the pathobiology of ocular ZIKV infection. I am well-positioned for this research and my career development, thanks to Dr. Kumar's state-of-the-art facility at Wayne State University and collaborations with renowned scientists, Dr. Arumugaswami, Dr. Mor, Dr. Yu, Dr. Shukla, and Dr. Ljubimov, which will contribute significantly to my research efforts and facilitate the establishment of my independent laboratory at an academic institution in the US.

NIH Research Projects · FY 2025 · 2024-09

Project Summary Endophthalmitis is a devastating complication caused by a wide range of microorganisms which can result in irreversible vision loss if not diagnosed and treated promptly. The current diagnostic approach heavily relies on microbial culture to determine the etiology of endophthalmitis. However, only ~40% of suspected endophthalmitis cases are culture-positive, leaving many potentially infectious culture-negative cases. Therefore, the discovery of potential biomarkers is urgently needed for both the diagnosis of endophthalmitis and in guiding appropriate antimicrobial therapy. Here, we propose to use metabolomics, an emerging high-throughput technology that can identify, quantify, and characterize hundreds to thousands of low molecular weight biochemicals (metabolites), using targeted or untargeted analytical approaches. Since metabolism is directly or indirectly linked to every aspect of cell function, metabolomics is believed to reflect the phenotype of what is occurring in the body. Recently, we performed untargeted metabolomics in an experimental model of endophthalmitis and discovered distinct metabolic profiles of uninfected and bacterial-infected mouse eyes. Moreover, metabolomics analysis led to discovering a therapeutic target, i.e., itaconate/Irg-1 signaling, to ameliorate experimental bacterial endophthalmitis. Our pilot vitreous metabolomics analysis showed distinct metabolic profiles in patients with uveitis, endophthalmitis, and other inflammatory retinal diseases. Based on preliminary data and our prior publications, we hypothesize that “intraocular infections induce biochemical changes in the vitreous leading to distinct metabolic signatures with translational potential for therapeutic targeting and/or diagnosis of infectious endophthalmitis”. This will be tested by performing untargeted metabolomics in vitreous of culture-positive bacterial and fungal endophthalmitis (Aim-1). We will further devise a metabolic profile to aid in predicting endophthalmitis in culture-negative chohort. This will be accomplished this by developing an artificial intelligence (AI) model that can predict endophthalmitis' etiology based on vitreous metabolic profiles (Aim-2). The experiment will also involve validation using an independent cohort of culture-positive vitreous. The proposed study will be the first to utilize metabolomics profiling to identify the metabolic sigantures in the vitreous of endophthalmitis patients. The resulting profiles and/or biomarkers will be readily translatable for future clinical studies, ultimately guiding precision medicine for diagnosing and treating ocular infections.