Wayne State University

NIH Research Projects · FY 2024 · 2023-08

Abstract Childhood trauma is highly prevalent in the United States, and can greatly increase the risk for developing anxiety disorders in youth. Experiences of trauma promote heightened emotional responses to potential threats, which in turn may impact cognitive ability. The imbalance between emotional and cognitive processes may contribute to hallmark symptoms of anxiety disorders, such as excessive fear and impaired functioning. It is critical to investigate the biochemical mechanisms underlying cognitive neural engagement to identify novel therapeutic targets and develop better targeted pharmacotherapies aimed at maintaining cognitive control ability in vulnerable populations like trauma-exposed youth. Functional MRI studies demonstrate that the dorsal anterior cingulate cortex (dACC) is a critical component of maintaining cognitive control, and that visual cues of negative affect may interfere with these functions. However, the key barrier to advancing this mechanistic understanding is that the vast majority of current investigations utilize functional MRI, which relies on the hemodynamic response function, and is an imprecise indicator of neural engagement. A more precise measure of neural engagement can be obtained using in vivo ¹H functional MR spectroscopy (¹H fMRS), which is a novel tool sensitive to temporal changes in glutamate under contrasting task-conditions. The objective in this proposal is to investigate the impact of childhood trauma on dACC neural engagement related to cognitive control with and without negative affect, and its association with pediatric anxiety symptoms. We will enroll 60 adolescents (ages 11-15, 50% female) from an urban setting with high rates of trauma exposure (30 trauma-exposed, 30 control). Participants undergo ¹H fMRS scanning while completing a cognitive control task specifically designed to allow direct characterization of the dACC neural engagement during cognitive control following negative affect interference. The task includes two modes that target different components of cognition: sustained attention and response inhibition. Both modes will be tested in the context of threatening faces (negative affect condition), as well as neutral shapes (no affect condition). This proposal hypothesizes that childhood trauma will potentiate negative affect interference with dACC neural engagement necessary for cognitive control functioning – demonstrated by reduced dACC glutamate modulation during the negative affect conditions of the task. This innovative approach will facilitate investigation into the neural processes which maintain cognitive control in the presence of threat in trauma-exposed adolescents. This training project will provide PI France with training in conceptual (neurobiology of trauma and anxiety) and methodological approaches (¹H fMRS and psychological assessments). It will also prepare the PI for a successful F32 submission and future academic research career.

NIH Research Projects · FY 2025 · 2023-08

ABSTRACT Crohn’s Disease (CD) is an autoimmune disease which leads to chronic inflammation and scarring of the digestive tract. The available treatments involve immunosuppressants and surgery which are costly, with many individuals still experiencing a decreased quality of life. The cause of CD is unknown, but is believed to occur due to both genetic and environmental factors, including diet. The gut microbiome is altered in individuals with CD, leading to dysregulation of the host and microbial transcriptional and metabolic landscape. Furthermore, many genetic risk variants associated with CD are in host genes known to function in host response to the microbiome. Thus, characterizing host-microbiome interactions in CD is imperative for understanding the pathogenesis of CD and identifying potential new therapeutic routes. In this proposed research, I aim to characterize the interactions between host genetic variation and the gut microbiome that regulate host gene expression in CD. I will use two complementary approaches. I will use a microbiome/intestinal organoids co-culturing approach to identify host gene expression changes in response to CD-derived microbiomes, and host genetic variants that modulate these changes using allele-specific expression analysis. I will also utilize data available through the Human Microbiome Project Inflammatory Bowel Disease Multi’omics Database to perform eQTL mapping, and integrate all of my findings with GWAS and TWAS to validate the significance of GxM for CD risk. These studies will provide insight into host-microbiome interactions that modify genetic risk for Crohn’s Disease. Furthermore, I will identify specific microbes that could become potential therapeutic targets for CD.

NIH Research Projects · FY 2025 · 2023-08

Androgen receptor (AR) plays a critical role in prostate cancer (PCa) in all stages. While a majority of patients with PCa initially respond to androgen deprivation and/or anti-androgen therapies, a significant portion of patients develop castrate-resistant prostate cancer (CRPC). Clinical studies showed that CRPC is often enriched with AR splice variants lacking the ligand binding domain (LBD), thereby being constitutively active in an androgen-independent manner and escaping from the antiandrogen enzalutamide-mediated AR inhibition. The goal of this RO1 application is to develop and characterize novel therapeutics that effectively degrade (remove) AR proteins in PCa. Technical innovations include the development of a novel therapeutic platform for AR protein degradation using autophagy-targeting chimera (AUTOTAC). We successfully generated AR- targeted AUTOTACs, which are bifunctional molecules, composed of enzalutamide (an AR ligand binding domain inhibitor) or VPC-14449 (an AR DNA binding domain inhibitor) as a target binding ligand, linked to YTK-6-2 as a ligand of the autophagic cargo receptor p62/Sequestosome-1/SQSTM1. AR-targeted AUTOTAC brings the target protein AR to p62, forming a ternary complex. YTK-6-2 induces p62 self-oligomerization, resulting in AR-p62 oligomeric complexes. YTK-6-2-activated p62 facilitates AR-p62-LC3 interaction on autophagic membranes and promotes autophagosome biogenesis and degradation of AR proteins. Using clinically relevant animal models that reflect the relevant tumor microenvironment and tumor heterogeneity, we will test our novel hypothesis that AUTOTAC-mediated degradation (removal) of AR and its variants is a more effective approach than the currently available therapeutics including anti-androgen therapy. We also hypothesize that unlike anti-androgens which result in therapy-resistant PCa cells involving activation of noncanonical AR signaling programs, the AR-targeted AUTOTAC platform is unlikely to result in noncanonical AR signaling-associated malignant phenotypes. Aim 1 will characterize the molecular actions of AR LBD- targeted AUTOTAC (Enz-AUTOTAC) and AR DBD-targeted AUTOTAC (VPC-AUTOTAC), and evaluate their therapeutic efficacies in prostate cancer cell models. Aim 2 will evaluate the therapeutic efficacies of Enz- AUTOTAC and VPC-AUTOTAC in vivo using PTEN knockout and patient-derived xenograft (PDX) mouse models of prostate cancer. Activation of potential oncogenic pathways in therapy-resistant tumors will be examined in relation to the expression levels of AR and the status of AR variants. In addition to the transcriptome analysis of tumor cells, tumor heterogeneity, cancer cell plasticity and tumor microenvironments will also be assessed by single cell RNA-seq analysis before and after drug treatments. These data may lead to the identification of additional therapeutic target(s) in castrate-resistant PCa. Successful completion of the proposed study will develop a revolutionary drug platform of AR degraders for patients with CRPC.

NIH Research Projects · FY 2026 · 2023-08

Abstract Sepsis is a common condition with significant morbidity, mortality, and annual costs-of-care in the billions of dollars. Despite innumerable studies on the causes of, and therapies for, sepsis, the mortality rate has not changed substantially in the last 20 years. Treatments remain generic, with current guidelines recommending the same approach for all patients, regardless of the obvious litany of differences that exist at baseline. Moreover, the blanket administration of 30cc/kg of intravenous fluid (IVF) to all patients is recognized as being directly harmful to some. Patient-level heterogeneity in prior sepsis trials is recognized as a substantial contributor to all of these problems, yet no prior investigation has attempted to identify septic phenotypes, a necessary first step towards precision care. This project will address these critical knowledge gaps by systematically characterizing profiles of patients with sepsis using a combination of sonographic and biomarker variables. This will begin at the time of presentation to the Emergency Department and continue at specified time points during hospitalization, thereby creating a rich longitudinal database from which identification of unique phenotypes will be possible. Our central hypothesis is that central venous hypertension: (i) is deleterious to the function of the lungs, liver kidneys, and vascular endothelium; (ii) is worsened by cardiac dysfunction and IVF administration; and (iii), contributes to adverse organ-specific and overall outcomes. Cardiac function will be assessed with echocardiography while congestion in the lungs, and kidneys will be assessed using previously validated sonographic markers of congestion. Biomarkers for each organ will be collected concurrently thereby increasing the fidelity of our phenotypic profiles by pairing indicators of macro- and microscopic stress and dysfunction. Classical and machine learning approaches will be used to analyze our large data-stream and develop a rule-based system to identify distinct sub-populations of sepsis patients who have greater risk/likelihood of both organ-specific and overall adverse outcomes. Completion of the project will result in generation of the first comprehensive description of septic phenotypes at the level of individual organs and the organismal level, including the effects of resuscitative interventions. The combination of longitudinal imaging and laboratory data is novel and offers potential short-term (outcome-based guidance of IVF administration and prognosis via sonographic data) and long-term (development of biomarker panels and targeted therapeutics) patient-oriented benefits.

NIH Research Projects · FY 2024 · 2023-08

Abstract Immunotherapy has made an enormous impact in the treatment of multiple types of cancer, however most patients fail to benefit and methods to monitor response are lacking. To bridge this gap, we have developed non-invasive positron emission tomography (PET) tracers to interrogate immune activity in the tumor microenvironment. Immune cell release of interferon-γ (IFN-γ) is a hallmark of CD8+ cytotoxic T cell (CTL) and Th1-mediated immune activity, both of which contribute to anti-tumor immunity. Our results thus far show that antibody-based immunoPET tracers targeting IFN-γ can detect anti-tumor immunity after administration of immunotherapy, which correlates to treatment outcomes in preclinical tumor models. The current proposal will advance these studies by examining the potential of IFN-γ PET as a pre-treatment predictor of immunotherapy response. We will utilize novel Collaborative Cross recombinant inbred mouse models developed during the course of this project, which exhibit a wide range of response rates to immune checkpoint inhibitors (ICI) due to select variants in their genetic background. These models mimic the diversity of the human population, yet limit tumor heterogeneity as a variable by utilizing genetically identical tumor lines, allowing us to focus on the role of host immunity. We find pre-existing intratumoral IFN-γ expression correlates to ICI outcomes in these models, and we will now test whether IFN-γ PET imaging prior to therapy is predictive of ICI response. Positive results will support the use of IFN-γ PET as part of the formulation of patient treatment strategy. In a necessary step for clinical translation, we will also prepare to submit an investigational new drug (IND) application. Dosimetry, stability studies, and toxicology will be performed. Collectively, the proposed studies will support the clinical use of IFN-γ PET to address critical unmet needs, including predictive biomarker discovery and treatment monitoring technology for cancer immunotherapy. While this proposal focuses on oncology, immune monitoring technologies may also have additional application in multiple inflammatory and autoimmune conditions.

NIH Research Projects · FY 2025 · 2023-08

Project Summary Epithelial cells carry out secretory and absorptive functions that require the polarized distribution of transporters, receptors and adhesion molecules at the apical and basolateral plasma membranes. Acquisition and maintenance of apical-basolateral polarity involves sorting of plasma membrane proteins along the biosynthetic and recycling pathways. The question of how apical and basolateral proteins in transit to the plasma membrane are sorted in endosomal compartments has been at the center of research effort for decades. An important mechanism of protein sorting is mediated by the clathrin adaptor protein AP-1B, which is a feature of epithelial cells. AP-1B recognizes cargo at discrete sorting motifs and it was initially identified as mediator of basolateral sorting. However, recent research by the applicant and others has determined that AP-1B also mediates apical sorting. Moreover, some epithelia like the kidney proximal tubule and the retinal pigment epithelium constitutively lack AP-1B. Therefore, additional mediators of polarized sorting must exist in these epithelia. This fact, and our fluid understanding of AP-1B-mediated trafficking warrants revisions of the traditional models of polarized protein sorting in epithelial cells. Filling these gaps in knowledge will reveal mechanisms that can be exploited to understand and treat pathologies where epithelial physiology is compromised. The applicant’s research program aims to characterize mechanisms of polarity in AP-1B-deficient epithelia and their role in tissue organization, homeostasis and disease. The focus of the current R35 application will be on epithelial cells of the kidney proximal tubule and retinal pigment epithelium, which lack AP-1B. We will study the roles of an endosomal sorting protein previously characterized in non-polarized cells, sorting nexin-27 (SNX27) and vesicle-associated membrane proteins (VAMPs) in polarized trafficking. We will also employ proteomics and high throughput approaches to identify additional molecular mediators of polarized trafficking. We will test whether the identified mechanisms mediate apical delivery of Megalin, a physiologically relevant surface receptor expressed in kidney proximal tubule and retinal pigment epithelium. The R35 funding mechanism is an excellent fit for achieving these goals, since it is intended to support the pursue of ambitious and flexible ideas with the intent of developing the applicant’s research and training program, rather than an individual project. At the same time, completing this proposal will lay out foundations for future advances in diagnosis, treatment, and prevention of diseases, therefore directly contributing to the mission of the National Institute of General Medical Sciences.

NIH Research Projects · FY 2025 · 2023-07

Anxiety affects nearly one in three adolescents and contributes to substantial burden on both individuals and society. Although evidence-based interventions for adolescent anxiety exist, treatment response is modest and relapse rates are unacceptably high. Outcomes are even worse among low resource and racial/ethnic minority populations. Recent studies have pinpointed neurodevelopmentally-informed targets that are relevant to current evidence-based treatments for adolescent anxiety; namely, exposure-based cognitive behavioral therapy (CBT), which relies on principles of fear extinction. Our group and others have shown that fear extinction and frontolimbic circuitry change dynamically across the first two decades of life, and is modulated by the endocannabinoid (eCB) system. Further, our exciting preliminary data show that acute exercise is associated with lower anxiety and elevated eCB signaling in youth, and is therefore a promising approach for optimizing efficacious treatments for adolescent anxiety. However, these advances have not yet translated to improved therapeutic outcomes for youth. The proposed project will leverage a multi-modal experimental therapeutics approach to test whether acute exercise modifies hypothesized targets that are relevant for the pathophysiology and treatment of anxiety in youth. One hundred and twenty adolescents will be recruited from a diverse population at elevated risk of anxiety and randomized into either an acute moderate-intensity aerobic exercise or sedentary control condition, performed immediately after a fear extinction paradigm (i.e., during the memory consolidation phase). Our hypothesis is that acute exercise will boost eCB signaling, which will result in increased fear extinction recall and enhanced frontolimbic activation and coupling. Concurrent neuroimaging, psychophysiological recordings, self-reported fear and anxiety, and circulating biomarkers will allow us to evaluate target engagement at several levels; specifically, we will test fear extinction, frontolimbic circuitry, and eCB signaling as targets for exercise’s effects on fear extinction and anxiety risk. This project is ideally suited for the NIMH BRAINS award because it will support the development of a productive early-stage investigator in innovative, high-impact research. Results of the proposed project will demonstrate that a relatively low cost and low risk (compared to pharmacotherapy, for example) behavioral intervention may be used alone or in conjunction with current treatments to improve outcomes for youth. This significant and timely study is an essential first step in a continuum of research that will ultimately lead to efficacious treatments for adolescent anxiety, and novel preventive interventions for at-risk youth. This work will also further our understanding of how fear is acquired and regulated in the adolescent brain. These outcomes are highly aligned with the NIMH goals of delineating brain mechanisms (Goal 1), understanding risk factors and biomarkers of illness and treatment response (Goal 2), improving prevention (Goal 3), and strengthening the public health impact (Goal 4), given our group’s ties to mental health providers and school programs.

NIH Research Projects · FY 2024 · 2023-07

Per- and polyfluoroalkyl substances (PFAS) are a class of ubiquitous man-made chemicals utilized for their surfactant properties in industrial and consumer products (cookware, clothing, carpets) as well as in firefighting foams. PFAS are highly resistant to degradation, leading to their bioaccumulation in the environment and in humans. Because of this, PFAS are currently circulating in humans at blood concentrations that are magnitudes higher than other legacy contaminants. Exposure to PFAS have been associated with increased risk factors for cardiometabolic disease (i.e., increased circulating cholesterol), or major clinical outcomes related to atherosclerosis that include stroke and heart attack. However, no studies have yet reported on whether exposure to PFAS can induce the development of atherosclerosis. The overarching objective of this proposal is to determine if PFAS exposure induces atherosclerosis and to characterize the underlying mechanisms leading to PFAS-induced development of atherosclerosis risk factors. We have shown in our preliminary studies that Low Density Lipoprotein Receptor deficient mice (Ldlr -/-) fed an atherogenic diet and exposed to a simple mixture of 5 environmentally relevant PFAS (PFOS, PFOA, PFNA, PFHxS, and GenX) for 7 weeks resulted in increased circulating cholesterol and bile acids as well as decreased bile acid excretion. We also observed that PFAS exposure results in induction of ileal bile acid transporters, especially the ileal reuptake apical sodium dependent bile acid transporter (ASBT). Therefore, the central hypothesis of this proposal is that PFAS exposure induces atherosclerosis through mechanisms related to ASBT-mediated bile acid transport and excretion. To test this hypothesis, I will utilize Ldlr -/- mice, which is the gold standard for mechanistically investigating atherosclerosis because this genetically modified model has cholesterol profiles that closely mirror those seen in humans and is a model sensitive to dietary manipulation. Experiments proposed in Aim 1 will investigate the development of atherosclerosis in hyperlipidemic mice by monitoring lesion development in the aorta and aortic roots, quantifying lipid, sterol, and bile acid levels, and transcriptomic profiling. Aim 2 will investigate mechanisms underlying the PFAS-mediated increases in circulating cholesterol and atherosclerosis. Experiments proposed in Aim 2 will explore how ASBT inhibition modulates circulating cholesterol and bile acids, as well as the development of atherosclerosis. Collectively, these data will describe new mechanisms linking exposure to PFAS and increased risk of atherosclerosis. This proposed work will have broad implications for the use of well-tolerated pharmaceuticals as effective interventions against PFAS-mediated toxicity.

NIH Research Projects · FY 2024 · 2023-07

Drug-resistant, focal epilepsy impacts millions of children, and treatment often requires invasive evaluation. This process entails identification of intracranial electroencephalography (iEEG) biomarkers such as spike-and-wave discharges (SWDs), to guide surgical removal of the presumed epileptogenic zone responsible for generating habitual seizures. In pediatric patients, extra-operative iEEG evidence indicated that a SWD proxy measuring coupling between delta wave phase and high-frequency oscillation (HFO) amplitude – Modulation Index (MI) – accurately tracks epileptogenicity. Since MI does not detail the causality of neural propagations, it’s powerful to also consider iEEG Transfer Entropy (TE), which measures effective connectivity, and dynamic tractography to provide plausible propagation pathways. Granted, capturing adequate interictal and ictal epileptic events often requires days of extra-operative iEEG recording, and this procedure is grueling, expensive, and replete with major risk factors. Thus, development of intra-operative techniques for induction and reliable measurement of epileptic iEEG biomarkers is imperative to avoid the above pitfalls and help expand utility of one-stage procedures. Pertinently, sevoflurane anesthesia reversibly activates spike activity, but there is much debate over its specificity. Preliminary iEEG results in children suggest that sevoflurane may intra-operatively augment both MI and HFO effective connectivity (TE) in seizure foci. However, large cohort studies are needed to validate this finding, as it is unknown how sevoflurane impacts intra-operative MI and HFO-TE in healthy versus epileptogenic brain areas and if these signals spread via major white matter tracts. Thus, the main aims of the current proposal are to: 1) build normative atlases of intra-operative MI and HFO-TE, at varying concentrations of sevoflurane, and 2) determine if sevoflurane-induced modulation of these features can localize seizure foci and predict seizure outcomes. To accomplish these aims, the trainee will map patient electrodes to 3-dimensional magnetic resonance brain images, quantify the intra-operative iEEG metrics at stepwise increases of sevoflurane, and combine iEEG effective connectivity with white matter tractography (i.e. dynamic tractography). Characterizing the endogenous distribution of MI and HFO effective connectivity, with and without sevoflurane, will provide critical baseline reference for iEEG interpretation. In addition, understanding how sevoflurane impacts these metrics in epileptic networks is expected to improve interictal localization efforts during surgery, reduce invasive diagnostic burden by mitigating the need for extra-operative recording, optimize treatment cost effectiveness, and ultimately improve seizure outcomes. Through this project, the trainee will: (1) gain insight into and help refine clinical epilepsy treatment, (2) enhance scientific understanding of how neural oscillatory coupling and effective connectivity respond under sevoflurane, and (3) will undergo a training regimen to foster development of critical thinking and hypothesis-driven, ethical research design. The cumulative effect of the research and didactics will provide a critical foundation for the trainee’s career as a physician-scientist.

NIH Research Projects · FY 2026 · 2023-07

Black adolescents with type 1 diabetes (T1D) face disparities in diabetes-related health outcomes such as higher risk for suboptimal glycemic control, which can lead to diabetes complications. Given the critical protective role played by families in the health of adolescents with T1D, family-based interventions have commonly been used as a strategy to promote optimal adolescent health. However, despite the extensive literature documenting health disparities, few randomized clinical trials, including those testing family-based-interventions, have included adequate samples of Black adolescents with T1D. Moreover, rigorous, adequate powered clinical trials testing behavioral interventions specifically designed and tailored for Black adolescents with T1D and their families are lacking. Primary caregivers of youth with T1D experience elevated levels of stress, depression and anxiety, and caregivers of Black youth are at higher risk for such distress. Family-based interventions for Black adolescents with T1D hold the potential to improve the health not only of the individual youth but of the caregiver as well, as improved family interactions can have spillover effects for caregiver mental health. eHealth interventions have shown promising effects in a number of areas of behavioral health care and may also increase the accessibility of behavioral health interventions to Black families. Our group has developed and tested a culturally tailored, brief eHealth intervention design to promote optimal parenting practices for primary caregivers of Black adolescents with T1D. Results of our recent clinical trial where the intervention was delivered during diabetes clinic visits showed promising results, as adolescents whose caregivers received the intervention had improved glycemic control and families reported reduced diabetes-related family conflict. However, findings also suggested the need for further refinement of the intervention, including the development of additional content to help parents support their adolescent with diabetes management. The proposed study will include an intervention adaptation phase where we will convene community advisory boards to develop new material designed to optimize diabetes-related family interactions that can be integrated into the existing intervention. Subsequently, we will test the efficacy of the new intervention to improve youth glycemic control, improve diabetes-related family relationships and reduce caregiver diabetes-related diabetes distress in a multi-center, randomized controlled trial. The new intervention will be delivered via a mobile health approach to optimize its accessibility. 216 Black adolescents with T1D and their primary caregiver will be recruited from two clinical sites and receive the intervention or an attention control intervention during a six-month window. Dose effects and intervention cost-effectiveness will also be evaluated. If successful, the intervention has the potential to improve health outcomes in a vulnerable population of youth and their family members.

NIH Research Projects · FY 2026 · 2023-07

PROJECT SUMMARY Inter-organelle membrane contact sites (MCSs) are emerging as critical hubs that regulate metabolic pathways which enables cellular adaptation to external perturbations and environmental stress. Because of their roles in cellular metabolism, dysregulation of MCSs has been linked to numerous metabolic and neurodegenerative disorders as well as aging. Numerous studies to date have linked MCSs to non-vesicular lipid transfer and spatial organization of lipid metabolism. Using yeast as a model system, we found that contact sites between the Endoplasmic reticulum (ER) and the lysosome recruit lipid enzymes to spatially organize lipid synthesis and storage as a response to lipotoxic stress. Lipid biosynthesis is intimately linked to methionine metabolism. Specifically, S-adenosyl methionine (SAM) which is a direct product of methionine, is an essential methyl donor required for membrane lipid biosynthesis. However, how MCSs regulate the availability and utilization of non- lipidic metabolites such as SAM is currently unknown. Surprisingly, we found that deletion of ER-lysosome tethers in yeast significantly alters SAM levels and sensitizes yeast to methionine-deprivation. In addition to providing methyl groups for phospholipid methylation, SAM is involved in other cellular functions such as nutrient signaling, epigenetic regulation, and maintaining redox balance. Therefore, lowering SAM levels by perturbing ER-lysosome contact sites will have direct consequences on these essential cellular pathways. Based on these results, we propose that lipid synthesis at ER-lysosome contact sites regulates SAM availability, which serves as an important driver for regulating downstream cellular pathways. The central goal of this proposal is to test this hypothesis by 1) elucidating the mechanisms by which ER-lysosome tethers regulate phospholipid synthesis; 2) determining the functional consequences for altered compartmentalization of phospholipid methylation; 3) defining molecular and structural basis for lipid binding and transport by ER- lysosome tethers; and 4) identifying conserved mechanisms for SAM regulation by ER-lysosome contact sites. Collectively, the results of our studies will provide insight into the organizational aspects of SAM regulation, how this alters the lipidome, and its relationship to aging and metabolic diseases.

NIH Research Projects · FY 2026 · 2023-06

ABSTRACT The ultimate goal of this project is to discover and validate the gene Х heavy metal (GXM) interactions in human livers and to understand their role in nonalcoholic fatty liver disease (NAFLD). NAFLD is the most common chronic liver disease affecting over 30% of the U.S population, resulting in a heavy social burden. NAFLD is characterized by a spectrum of histological changes with multiple cells involved. Currently, no approved drug treatment is available for NAFLD. Therefore, it is an urgent need to identify both genetic and environmental risk factors to facilitate the development of new diagnostic, preventive, and therapeutic strategies. NAFLD is a typical complex disease involving gene-environment (GXE) interactions. Over the past decade, while GWAS for NAFLD have identified numerous genetic risk alleles, a growing body of research has demonstrated that exposure to heavy metals (Pb, Cd, Hg, As, etc.) are associated with increased NAFLD risk. However, there is no compelling study thus far to assess the correlation between various naturally accumulated metals in human livers and the NAFLD histology, especially in clinically defined patient cohorts. More importantly, there lacks critical knowledge about how naturally and chronically accumulated metals interact with the liver genome and together confer risks for NAFLD. Our preliminary studies in human liver tissues have successfully demonstrated that multiple metals are indeed correlated with NAFLD. By leveraging our previously collected multi-omics data, we have preliminarily identified numerous metal-response genes (MR-genes), expression quantitative traits loci (eQTLs), and allele-specific expression loci (ASEs), which are further enriched to NAFLD and its related pathways. We aim in this study to further expand our study to a large-scale, highly detailed, and integrated analysis to thoroughly understand the role of GXM interactions in NAFLD in humans. To this end, our team has been collaborating to establish collections for human liver tissue, cells and NAFLD patient cohorts. We have also developed various technical platforms e.g. bulk/single cell (Sc) RNAseq and ATAC-seq, as well as multiple bioinformatics and statistical tools for GXE data analyses. With these preparations, we specifically aim: 1) To profile heavy metals in frozen human liver tissues (n=560), identify MR-genes, eQTLs/ASEs, and test their associations with NAFLD; 2) To treat the primary liver cell populations with various metals and to elucidate how the genome of different liver cells respond to metals with Sc-RNAseq and ATAC-seq, and 3) To validate the association between GXM interactions and NAFLD histology severity in a large clinically defined NAFLD patient cohort (n=1313). Our study will for the first time evaluate the role and mechanism of GXM interactions in NAFLD and will provide the scientific community important data to open new avenues to NAFLD research, drug discovery, and beyond.

NIH Research Projects · FY 2026 · 2023-06

Project Summary/Abstract Retinal neovascularization (RNV) is a debilitating complication of advanced diabetic retinopathy, which despite the use of anti-VEGF and laser treatments continues to cause blindness. Less is known as to why RNV develops only after patients have had diabetes for decades. Although endothelial cell (EC) angiogenic activation is a hallmark of this transition, how ECs adapt their metabolism to sustain such activation remains a significant gap in our knowledge. Our long-term goal is to determine the bioenergetic mechanisms of RNV. The Warburg effect is a metabolic shift from mitochondrial oxidative phosphorylation (OxPhos) to hyperglycolysis that was first found in cancer cells. This metabolic shift not only produces ATP faster than OxPhos, albeit less efficiently, but also provides precursors required for lipid, protein, and nucleotide synthesis during cell proliferation. Recently the Warburg effect was rediscovered as a key contributor in various endothelial-related diseases; however, its role in diabetic retinopathy is not well-defined. In this application, the overall objective(s) are to define the role of the Warburg effect in diabetic retinopathy and to identify its underlying mechanisms. Here, we propose that multiple hits are needed to cooperatively alter EC metabolism to fulfill biosynthetic demands of transforming a quiescent EC into an angiogenic cell. Tissue hypoxia is the most common risk factor associated with advanced diabetic retinopathy. Thus, our central hypothesis is that diabetes primes quiescent ECs to be angiogenic (first hit) and that hypoxia (second hit) is necessary for angiogenic switch via the metabolic adaptation of the Warburg effect. Aim1 will test the hypothesis that persistent activation of the energy sensor, AMP-activated protein kinase (AMPK) sustains the Warburg effect to mediate EC angiogenic activation. Our approach is to use a two-hit model of diabetes and hypoxia in AMPKα1 endothelial-specific conditional knockout (AMPKα1End-/-) mice and in AMPKα1 silenced human retinal ECs (HRECs) to achieve this aim. We will also use vitreous samples from patients with proliferative diabetic retinopathy to test the correlation between the development of RNV and the Warburg effect- associated metabolites. Aim2 will investigate the role of endoplasmic reticulum (ER) stress in mediating the Warburg effect-induced EC angiogenic activation. We hypothesize that activation of Inositol-requiring enzyme (IRE)1, a unique ER-stress sensor protein with kinase and RNase activities, is a key mediator for the Warburg effect-induced EC angiogenic activation. We will use CRISPR/Cas9 to test the effect of inhibiting downstream signaling of IRE1 kinase and RNase activities on the Warburg effect-induced EC angiogenic activation. We will also test our hypothesis in vivo using a two-hit model of hypoxia and diabetes and selective pharmacological inhibitors. Overall, this new-investigator initiated R01 capitalizes on the interdisciplinary expertise of a biochemist, a mitochondrial biologist, an ER biologist, and a clinician to use a novel two-hit model of advanced diabetic retinopathy to gain mechanistic insights into the bioenergetic basis of RNV. Understanding the role of the Warburg effect will reveal novel targets in the treatment of diabetic retinopathy.

NIH Research Projects · FY 2026 · 2023-04

Incarcerated individuals have higher rates of chronic disease than the general population, and disease burden will likely increase as the prison population continues to age. Despite this, health care services in prison primarily focus on infectious diseases, mental illness, and substance abuse, largely neglecting chronic physical health conditions. The Chronic Disease Self-Management Program (CDSMP) is an evidence-based program developed to support self-management for people with chronic illnesses. At least 10 randomized trials conducted in non-correctional settings have linked CDSMP to disease-related improvements and reduced healthcare expenditures. CDSMP has been used in state corrections systems via 3 approaches: 1) bringing community agency CDSMP leaders into the correctional setting to lead the program, 2) training correctional staff as program leaders, and 3) training incarcerated individuals to serve as peer leaders. To date, however, there has been little research into efficient and effective strategies for scaling up the intervention within state corrections systems using any of these approaches. Scale-up (i.e. deliberate efforts to increase the impact of successfully tested health innovations to benefit more people and promote sustainability) is an understudied concept in implementation science, with few existing empirical studies that explicitly focus on the process of scale-up. Assessing scalability, however, is crucial for ensuring sustainability of complex interventions within resource-poor settings. The research objective of this K01 is to evaluate and maximize the scalability of CDSMP among older adults in state correctional systems. Guided by the Scaling up Management Framework, we will use a mixed methods research approach to query community agency leaders, staff, and incarcerated individuals about ways to maximize the scalability of CDSMP within state correctional settings and develop and refine CDSMP scale-up strategies for these settings. Our goal is to develop scale-up strategies to be evaluated in a subsequent randomized implementation trial. The training objectives of this K01 will add expertise in implementation science and justice-involved research to my existing knowledge base in biopsychosocial models of disease, social gerontology, quantitative analysis, and chronic disease management, to reach my goal of becoming an independent researcher who uses implementation science approaches to improve health outcomes among justice-involved populations. As the prison population continues to age, the burden of chronic disease within correctional systems will continue to increase, which contributes to skyrocketing correctional costs. Understanding how to expand evidence-based chronic disease programs within correctional systems is crucial for reducing disease-related morbidity and mortality among incarcerated individuals and for reducing costs. This line of research will identify and test scale-up strategies for chronic disease management in prisons.

NIH Research Projects · FY 2026 · 2023-04

Project Summary/Abstract: The uniform lethality of glioblastoma (GBM) with a survival of less than 2 years despite best available therapy is attributed to treatment resistance due to DNA repair mechanisms that drive disease relapse and tumor heterogeneity. One prognostic factor identified as a reliable biomarker for GBM sensitivity to temozolomide (TMZ) and radiotherapy (RT) is the overexpression of O6-methylguanine-methyl- transferase (MGMT) enzyme. Patients with active MGMT were found to receive little benefit from TMZ and RT and represent a group of great unmet need with no treatment options that significantly improve survival. Recently, several preclinical and clinical studies suggest that alcohol aversion drug, disulfiram (DSF), inhibited MGMT and improved the efficacy of TMZ in GBM when combined with copper (Cu). However, phase II trial showed that there was no survival benefit from oral Cu/DSF. Nevertheless, the major limitation of oral Cu/DSF has been delivery of fragile DSF within the in vivo system. We have developed 2-hydroxypropyl beta cyclodextrin (HPßCD) encapsulating Cu complex of DSF metabolite, diethyldithiocarbamic acid (DDC), Cu(DDC)2 delivery system that addresses major drawbacks of the Cu(DDC)2: easy degradation in the blood and non-specific interactions with cells and serum proteins and lack of tissue specific delivery. HPßCD providing stability of Cu(DDC)2 is identified. In vitro cell culture study revealed that HPßCD-Cu(DDC)2 inhibited MGMT through the ubiquitin-proteasome pathway. Inhibition of MGMT activity in cell cultures vastly increased the alkylation-induced DNA double-strand breaks, cytotoxicity, and the levels of apoptotic markers like -H2AX, JNK-P and cleavage of PARP-1. Preliminary intravenous delivery of HPßCD- Cu(DDC)2 in combination with TMZ in an MGMT-positive patient derived orthotopic xenograft (PDOX) model demonstrated tumor size regression with prolonged survival. HPßCD-Cu(DDC)2 targets MGMT-145-cysteine and its unique cytotoxic mechanism circumvents MGMT-mediated chemo- and radiation resistance. The present work aimed at the development and application of HPßCD-Cu(DDC)2 for targeted delivery of drug to GBM. Chlorotoxin (CTX), a peptide reported to bind selectively to glioma cells while showing no affinity for non- neoplastic cells, will covalently be coupled to functionalized HPßCD encapsulating Cu(DDC)2. We have shown that PDOXs without MGMT expression are sensitive to RT. Therefore, we hypothesize that the combination of GBM targeting CTX-HPßCD-Cu(DDC)2 with TMZ and RT will overcome TMZ+RT resistance and show synergistic cytotoxic effect in PDOXs which will be monitored by MRI studies. Our objectives of the proposed research are A) To incorporate targeting ligand CTX with HPßCD-Cu(DDC)2 to obtain an active brain drug delivery system, B) To determine the efficacy and safety of oral DSF/Cu versus intravenous CTX-HPßCD- Cu(DDC)2 alone or in combination with TMZ in PDOX, and C) To determine the efficacy and safety of CTX- HPßCD-Cu(DDC)2 in combination with TMZ and fractionated radiation in MGMT upregulated PDOX models.

NIH Research Projects · FY 2026 · 2023-03

Inositol is essential for the viability of eukaryotic cells. Myo-inositol is the precursor of all inositol compounds, which play pivotal roles in cell signaling and metabolism. Consistent with its importance, perturbation of inositol homeostasis is associated with pathologies as diverse as neurological and psychiatric illnesses, myopathies, cancer, and diabetes. Furthermore, inositol depletion is a hypothesized therapeutic mechanism of action of drugs used to treat bipolar disorder, a devastating psychiatric illness that affects ~2% of the population. In this light, it is striking that very little is known about the regulation of inositol homeostasis in human cells. Our research seeks to determine how inositol synthesis is regulated and how inositol deprivation affects essential cellular functions in human cells. Toward this end, we identified the first negative transcriptional regulator of inositol synthesis in mammalian cells – inositol hexakisphosphate kinase 1 (IP6K1). IP6K1 represses expression of ISYNA1, the gene coding for the rate-limiting enzyme of inositol synthesis, myo-inositol-3-P synthase (MIPS). Further, we demonstrated that binding of IP6K1 to phosphatidic acid (PA) is required for nuclear localization of IP6K1 and repression of MIPS expression. Our first project will rigorously dissect the mechanism of regulation of ISYNA1 by IP6K1. To elucidate the consequences of inositol deprivation, we constructed an ISYNA1 knock-out (ISYNA1-KO) human cell line, which cannot synthesize inositol. Inositol-deprived ISYNA1- KO cells exhibit profound alteration of lipid homeostasis and expression of genes involved in stress signaling. Specifically, we observed an increase in ceramides and increased expression of genes that mediate the unfolded protein response (UPR), a stress response that is activated by ceramides. Our second project will test the hypothesis that inositol deprivation results in activation of the UPR pathway by upregulating ceramides. Intriguingly, inositol-deprived cells exhibited increased levels of monolysocardiolipin (MLCL), which is the biochemical hallmark of the mitochondrial disorder Barth syndrome. Consistent with mitochondrial dysfunction, we observed increased activation of ERK, a promoter of mitochondrial fission, in inositol-starved cells. The third focus of our studies will test the working hypothesis that inositol deprivation perturbs mitochondrial function. Successful completion of the proposed studies will lead to the first molecular model of regulation of inositol synthesis in mammalian cells and demonstrate how inositol deprivation affects the cellular stress response and mitochondrial function. This knowledge will have important implications for understanding inositol homeostasis as well as the therapeutic mechanisms of inositol-depleting mood stabilizing drugs.

NIH Research Projects · FY 2025 · 2023-03

Growing evidences suggests that exposure to volatile, very volatile, and semi-volatile organic compounds (collectively abbreviated VOCs) during vulnerable life windows of susceptibility is an important determinant of maternal-fetal health, with implications for preterm birth, children sensitivity to infections, asthma and other adverse health outcomes. The central theme of this application focuses on deciphering the signaling pathways by which exposure to VOCs during pregnancy have an impact on early life growth and development with a focus on the fetal immune system. Our central hypothesis is that inflammation in the placenta and decidua due to maternal exposure to VOCs, alters the programming of the fetal immune system, which results in an aberrant post-natal immune response to respiratory viral infections. Our preliminary studies suggest that although the fetus may be protected against microbial infection, the outcome of maternal exposure, protective or deleterious, depends on the nature of the immune response and the severity of the inflammatory process at the implantation site (placenta-decidua interface). The mechanisms underlying the response of the fetal immune system and how indirect training by the maternal inflammation takes place is unclear and understudied. Our specific aims are: Aim 1. To determine the effect of VOCs maternal exposure on placental and fetal inflammation. Aim 2. To determine the impact of VOCs exposure on TLR signaling responsible for the homing and differentiation of T and B cells. Aim 3. Characterize the signals from the placenta responsible for the susceptibility to respiratory viral infections of the offspring. Upon completion of these aims we will have a better understanding of the outcomes associated with the impact of VOCs exposure on the placental/decidua unit and its consequent influence on fetal programing and its potential effects on the development of an appropriate neonatal immune responses. Adequate response to infection is the result of a delicate balance between an efficient immune response against pathogens and its quick resolution preventing widespread over-activation. The cellular and molecular components of this regulatory balance are determined during fetal development.

NIH Research Projects · FY 2026 · 2023-01

Abstract In the kidney, the thick ascending limb (TAL) of the loop of Henle is critical for NaCl homeostasis and blood pressure regulation. In humans and animal models of salt-sensitive hypertension, NaCl absorption is abnormally increased in the TAL, where NaCl absorption depends on the renal transporter NKCC2, an apical Na+/K+/2Cl- co-transporter. We showed that the presence of NKCC2 at the TAL apical membrane controls NaCl absorption in this epithelium. The molecular mechanisms that control apical membrane NKCC2 levels involve endocytosis, recycling and exocytic insertion. Inhibition of endocytic retrieval causes NKCC2 accumulation at the membrane and increased NaCl absorption. Any gene or protein affecting NKCC2 endocytosis could potentially influence NKCC2 activity and renal salt transport but only few proteins are known to bind NKCC2. Using a targeted proteomics screen, we identified ALMS1 (Alström Syndrome 1) and ACTN4 (alpha-actinin 4) as interacting partners of NKCC2. We also found that ALMS1 and ACTN4 interact with each other, raising the possibility that these proteins form a complex. Single nucleotide polymorphisms in ALMS1 and ACTN4 are associated with hypertension and decreased kidney function. We found that ALMS1 knockout rats have higher surface NKCC2 and high blood pressure. We found that ACTN4, a protein involved in podocyte biology, is also expressed throughout the nephron, including the TAL. The roles of ALMS1 and ACTN4 in renal NaCl handling by the TAL and their role in blood pressure regulation are unknown. We hypothesize that ALMS1 controls surface NKCC2 levels and NKCC2-mediated NaCl absorption by binding the carboxyl-terminus of NKCC2 and ACTN4 to mediate NKCC2 endocytosis from the apical membrane. A decrease in ALMS1 or ACTN4 expression in the TAL increases surface NKCC2, NKCC2-mediated NaCl reabsorption, tubulo-glomerular feedback (TGF) sensitivity and leads to salt-sensitive hypertension. Our long-term goal is to increase our understanding of the role of ALMS1 and ACTN4 in kidney NaCl transport.

NIH Research Projects · FY 2026 · 2022-12

Abstract A critical gap exists in understanding how nitrative stress, which has been effectively targeted to inhibit cell death in other models, alters cochlear protein signaling to induce apoptosis in cisplatin-induced ototoxicity. Continued existence of this gap represents an important problem for the 40%-80% of cisplatin-treated cancer patients who suffer with significant and in some cases permanent hearing loss as a result of cisplatin use. Until the underlying nitrative stress mechanism is delineated the promise of this new interventional target for mitigating a dose-limiting side-effect of cisplatin likely will remain unrealized. The long-term goal is to better understand the functional as well as mechanistic role of cochlear nitrative stress in acquired hearing loss. The objective is to delineate signaling pathways by which cisplatin-induced nitrative stress, particularly nitration of cochlear LMO4, facilitates ototoxicity, because cisplatin treatment nitrates and downregulates LMO4 protein. LMO4 is a transcriptional regulator that controls pathways regulating cell survival and cell death. The central hypothesis is that cisplatin-induced nitrative stress downregulates cochlear LMO4 and compromises STAT3- mediated anti-apoptotic signaling to facilitate ototoxicity. Understanding the mechanisms whereby nitrated cochlear LMO4 promotes cisplatin-induced ototoxicity is likely to contribute to the development of strategies to prevent this debilitating adverse effect. Guided by strong preliminary data this study will pursue three specific aims: (1) establish the causal link between cisplatin-induced LMO4 nitration and ototoxicity; (2) determine the effects of cisplatin-induced LMO4 nitration on JAK/STAT signaling; and (3) determine the otoprotective efficacy of pharmacological inhibition of nitration. In Aim 1, cisplatin-induced apoptosis will be analyzed after blocking nitration of LMO4 by site-directed mutagenesis and inhibiting proteasomal degradation of nitrated-LMO4 by lactacystin. The link between LMO4 protein levels and cisplatin-induced ototoxicity will be ascertained by testing cochlear apoptosis/hearing loss in LMO4 knockout and overexpressing mice. In Aim 2, cisplatin- induced changes in protein-protein interactions of cochlear LMO4 will be analyzed using a mass spectrometry- based proteomics approach while JAK/STAT related apoptotic and inflammatory signaling will be analyzed using targeted gene arrays. In Aim 3, the otoprotective efficacy of SRI110, a peroxynitrite decomposition catalyst, will be assessed using CBA/J mice; potential interference of SRI110 with anti-cancer activity of cisplatin will be analyzed using SCID mice. This innovative research departs from the status quo by shifting the focus from oxidative stress to the pivotal role of nitrative stress in cisplatin ototoxicity. Significantly, outcomes are expected to vertically advance understanding of how nitrative stress regulates cochlear apoptosis in cisplatin-induced ototoxicity. Findings will have important translational applications in mitigating cisplatin- induced hearing loss and preventing other otopathologies where nitrative stress plays a crucial role.

NIH Research Projects · FY 2025 · 2022-09

Evidence suggests that exposure to volatile organic compounds (VOCs), such as benzene, toluene, ethylbenzene, and xylene (BTEX), trichloroethylene (TCE), and tetrachloroethylene (PCE), is an important determinant of maternal-offspring health, with implications for preterm birth (PTB) and associated adverse health outcomes. VOCs emanate from landfills, brownfields, and Superfund sites, contaminating shallow soils and groundwater below residential, commercial, and industrial properties, leading to exposures via vapor intrusion. The Center for Leadership in Environmental Awareness and Research (CLEAR) is dedicated to understanding and mitigating this serious environmental health problem with a focus on post-industrial urban centers. Headquartered in Detroit, Michigan, CLEAR will use Detroit as a study site, which has the highest PTB rate in the country (15.2%) and is located in a state where 37 of the 67 Superfund sites must manage VOC contamination. CLEAR hypothesizes that VOC exposure through vapor intrusion during early life incites inflammatory responses in maternal tissues and/or the developing offspring that re-program the developing immune and other critical systems, setting the stage for PTB and/or associated adverse health outcomes. Five integrative environmental science and engineering and biomedical research projects (E1, E2, B1, B2, B3), supported by five cores, will investigate toxic mechanisms, exposure pathways, biomarkers, and strategies to prevent exposures and improve public health outcomes by: (1) developing and testing new detection methodologies, including phytoscreening (E1); sensor technology that integrates Internet of Things and edge computing for real-time contaminant detection and rapid-response, mitigation, and remediation (E2); and a controlled toxicity bioassay using WSU-designed sealed chambers for evaluating reproductive, neurological, behavioral, immunological and multigenerational responses in zebrafish (B1); (2) studying mechanistic effects of VOC exposure in a pregnant mouse model (B2); (3) applying epidemiologic methods for estimating exposure effects via human biological specimen analysis (B3); applying advanced chemical analysis, statistical approaches and visualization tools to obtain and integrate project data (Chemical Analysis, and Data Management and Analysis Cores), thereby establishing the impact of VOC exposures on PTB and associated adverse health outcomes; (4) creating an innovative model for transdisciplinary education by engaging new trainees to solve complex environmental health problems (Research Experience and Training Coordination Core); (5) engaging stakeholders and the community to inform our inquiry/analysis, participate in sampling, and employ health interventions (Community Engagement Core); and (6) uniting around an Administrative Core that conducts targeted research translation to ensure a legacy of scientific awareness and supports the Superfund Research Program to improve public health in urban centers impacted by environmental contamination to protect affected communities.

NIH Research Projects · FY 2025 · 2022-09

C ABSTRACT Myc Transcription Factor Inhibitor Design: Integrating Atomic and Mesoscale with Semi-Supervised Gen- erative Deep Learning Models Inhibition of master regulators such as Myc have considerable interest due to the reversal of the oncogenic state evoked by their removal. Adding to the mystique is the technical challenge in targeting a protein which possesses large regions of disorder. Though widely considered “undruggable”, the library of hits that disrupt Myc function continuously grows. The chemical features of a hit are difficult to deduce besides high molecular weight, aro- maticity, rigidity, and hydrophobicity. Understanding the more specific features of a protein-protein interaction (PPI) inhibitor is considerably difficult. In order to circumvent answering this question, machine learning methods have been applied to expand the library of experimentally determined hits in hopes of finding an improved inhibitor nearby in chemical space. Recently, the natural application of generative deep learning techniques to this prob- lem have been reported. This proposal explains a protocol for a semi-supervised expansion of small molecules which inhibit various reactions in the Myc transactivation pathway. The PPI inhibitors from three publicly available databases make up the training set (n=9516) while the known Myc inhibitors are the test set (n=100). In order to surpass the effectiveness of the test set, all known Myc inhibitors are removed from the training set. A number of latent variables which suffice to recreate the training set are solved. These variables represent the general struc- tural properties of PPI inhibitors, which may be associated with activities at various binding sites. The efficient calculation of activities is crucial to obtaining good performance. Therefore, a well-tempered ensemble of target configurations is pre-calculated at the all-atom resolution. Additionally, in order to incorporate the population level behavior of multiple Myc molecules into inhibitor design, mesoscale coarse-grain simulations in various sol- vents which drive liquid-liquid phase separation are performed. To identify interactions which correlate with phase response, various points in coarse-grain phase space are converted to all-atom resolution, further refined, and converted into contact maps. When evaluating a new lead, ensemble-based docking calculations are used, which calculate an average of averages of a ligand in different poses binding to different conformations randomly drawn from the ensembles. Reinforcement learning is applied to significantly reduce the time spent docking batches of leads while maintaining confidence in the result. Once new molecules are generated, these new leads are also optimized using absolute and relative free energy of binding methods. Ultimately, this study will test the limits of generative models to integrate data across multiple scales and develop inhibitors which evoke potent inhibition of intrinsically disordered proteins.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT Caregivers of persons living with dementia (PLwD) do not act in isolation and are typically involved in broader networks of family and friends who also assist with care, and yet this expanded caregiver network is largely neglected in research. Theories of mutual influence and empirical work suggest that caregivers’ stress and mental health have a negative impact on both caregivers and PLwD, yet recent reports and meta-analyses suggest disappointingly small effects of care-related behavioral interventions. Given that PLwD typically have broad networks involved in care, an exclusive focus on a primary caregiver may be missing the mark. This study aims to fill this immense gap by 1) Classifying dementia caregiver networks into profiles based on compositional and psychosocial facets of the care network, 2) Examining the association between dementia caregiver network profiles on PLwD’s well-being and important healthcare utilization, and 3) Identify the association between changing care needs of PLwD and how dementia caregiver networks change over time. We will first identify a sample of PLwD in the nationally representative National Health and Aging Trends Study (NHATS), which conducts annual interviews with age-eligible Medicare beneficiaries by pooling unique data from the 2011, 2015, and 2017 waves with additional waves forthcoming (2021 and moving forward annually). To characterize care network profiles, we will use information about caregivers from NHATS and additional rich detail from the associated National Study of Caregiving (NSOC), which interviews up to 5 caregivers per eligible NHATS participant. Latent class analysis models will be used to characterize and cluster networks based on similar patterns of compositional and psychosocial characteristics. Measures of between and within- network variability (e.g., average care stress in network, variability of stress within network) will be included. We will then explore the association of care network profiles, and individual components of network profiles, with key health outcomes for PLwD including sleep quality, depression and anxiety symptoms, and important health care utilization (potentially preventable hospitalization, Emergency Department visits, and long-term care stay) through linked NHATS respondent-level Medicare claims. Finally, taking advantage of the longitudinal nature of NHATS and NSOC, we will follow care networks over time to examine the association of network changes with change in PLwD’s self-care needs. This work is significant as results will substantially improve understanding of these caregiver networks and how they may increase risks for negative outcomes for PLwD such that future interventions may target those networks with potential for adverse influence. In line with NIA priorities to improve assessment of caregiving processes and understand connections between family dynamics and patient health and well-being, this understanding of care networks can fundamentally shift the paradigm of caregiving research by establishing the importance of considering family and network systems for research and intervention development, moving beyond the standard focus on a sole, primary caregiver.

NIH Research Projects · FY 2026 · 2022-09

Project Summary Regulated transcription of genes is essential for cell proliferation, differentiation and viability. Any misregulation may lead to a disease or a developmental abnormality. The transcription cycle of eukaryotic genes consists of multiple steps. The accomplishment of each of these steps requires a number of accessory factors. The initiation of transcription by RNAPII requires gene-specific factors and the general transcription factors (GTFs). The conventional view is that the general transcription factors (GTFs) operate exclusively at the initiation step of transcription. Many recent studies have challenged this dogma. The successful execution of the transcription cycle requires some GTFs to function at the termination step as well. Evidence from my laboratory and others has demonstrated that the general transcription factor TFIIB has an evolutionarily conserved role in termination of transcription. My laboratory has further implicated TFIIB in promoter directionality. Our preliminary results strongly suggest that the mechanistic basis of TFIIB action is, in both cases, dependent on its ability to interact with the termination factors and facilitate their recruitment on the gene. An investigation into the role of TFIIB in termination and directionality is critical to understanding of transcriptional regulation in eukaryotes, and may provide clues into the role TFIIB plays in several diseases. The overall objective of this application is to determine the prevalence and the molecular basis underlying termination of transcription and promoter directionality by TFIIB. The central hypothesis of this application is that TFIIB is involved in termination and promoter directionality of at least a subset of genes. We propose that these apparently unrelated processes rely on the ability of TFIIB to recruit termination factors. To accomplish the objectives of the proposal, we propose the following Specific Aims: (1) Genomewide analysis of TFIIB functions in termination and promoter directionality; (2) Elucidate the molecular basis underlying role of TFIIB in termination and promoter directionality; (3) Purification and characterization of the holo-TFIIB complex. The successful completion of this proposal will enable us to comprehend the role of transcription factors in the broader context, and will serve as a paradigm for understanding the coordination among steps of the transcription cycle in yeast and mammalian systems.

NIH Research Projects · FY 2026 · 2022-08

Opioid use disorder (OUD) is a global epidemic affecting a high proportion of child-bearing women. The drastic 4-fold increase in OUD among pregnant women from 1999-2014 has promoted opioid maintenance therapies (OMT) such as buprenorphine (BUP) to mitigate effects associated with opioid abuse. BUP is a mixed (partial mu-receptor agonist/kappa-receptor antagonist) semi-synthetic OMT that produces better infant outcomes after gestational treatment as compared to other OMTs (i.e. methadone). However, effects of BUP on pregnant women transitioning to motherhood is not well understood. Endogenous opioids play a significant role in orchestrating neuronal adaptations within the maternal brain network (MBN) for the successful ‘switch’ from a nulliparous brain to a maternal brain to incentivize nurturing maternal behaviors from the dam. It is well known that exogenous opioid exposure to morphine (full mu-agonist) during gestation can alter the endogenous opioid system and disrupt maternal care. However, there is a knowledge gap about the effects of BUP on the endogenous opioid system during the transition to motherhood and thus on maternal care behavior in the context of OUD during pregnancy. The proposed studies aim to address this knowledge gap by implementing a translational rodent model to evaluate the effects of BUP compared to morphine on the maternal brain, behavior and the microbiome well as on the long-term outcome of the offspring. Female rats will be exposed to BUP and/or morphine in clinically relevant paradigms starting before conception and continued throughout early postpartum. Effects of gestational BUP and morphine exposure on neurochemical and activation pattern changes in the maternal brain will be evaluated using state of the art imaging techniques. We will also evaluate the effectiveness of oxytocin as a potential intervention to increase maternal-offspring interaction in opioid- exposed dams and investigate the role of the microbiome in the long-term health outcome of exposed offspring. Our proposal will be able to add to the preliminary human findings on altered functional connectivity in buprenorphine-exposed mothers by using ‘whole (half) brain activity mapping’ which will allow us to simultaneously illuminate activity at cellular resolution in several brain areas of the MBN and compare patterns between opioid-exposed and control dams and investigate associations with changes in maternal behaviors. We expect that perinatal exposure to BUP inhibits the neuronal ‘switch’ from aversive to rewarding perception of pups that is necessary to initiate appropriate maternal behavior thus subsequently influencing offspring survival. The scale of the opioid epidemic requires an urgent response in order to promote treatment of OUD in pregnancy within an already marginalized population. We hope to advance science on the consequences of opioid drug use/therapy during gestation and to apply these outcomes toward clinical knowledge to improve public health via effective translation, implementation, and dissemination of our scientific research findings.

NIH Research Projects · FY 2024 · 2022-08

Drug use trends change rapidly among youth, leaving intervention experts struggling to respond to emerging drugs promptly. We have a critical need to advance implementation strategies to optimize system responsiveness to these emerging issues. COVID-19 has increased the urgency for implementation science to facilitate rapid, equitable responses using existing treatment and prevention efforts. Tier 1 evidence-based interventions (EBIs), such as the Michigan Model for HealthTM (MMH) lend themselves to addressing emerging trends. Our overall objectives are to 1) Improve the responsiveness of school-based EBIs in addressing urgent issues and 2) Find ways to support teachers in implementing updated EBIs, attending to unique considerations of low-resource schools. We will use After Action Review (AAR), to guide the systematic design of RAPD, Rapid Adaptation to Prevent Drug use, a novel bundle of implementation strategies. AAR is a reflective process focused on improving public health systems’ rapid response capacity. Key phases of AAR include: 1) Objective observation; 2) Analyze gaps/best practices, 3) Identify and test suitable implementation strategies to improve responsiveness to the next event (e.g., COVID-related drug use escalation). Sustainable strategies are central to achieving our objectives. Thus, we will also conduct a preliminary stakeholder-focused cost analysis. The rationale for this research is that designing and testing RAPD will enhance schools’ capacity to respond to urgent drug issues sustainably, and provide up-to-date, relevant resources for effective and equitable prevention. The proposed research will address the following aims: Aim 1: Identify implementation gaps and best practices in responding to urgent drug use events. We will use AAR to review the statewide response to the vaping crisis using the MMH using rapid qualitative analysis and identify gaps to be addressed with RAPD. Aim 2: Design and pilot test RAPD implementation and effectiveness. We will design a RAPD to optimize responses to urgent drug use events and pilot test in schools serving low-income students to assess equity. We will assess implementation outcomes using a convergent mixed methods design, and effectiveness via a group RCT comparing RAPD and standard MMH implementation. Aim 3: Assess costs and benefits of RAPD from multiple stakeholder perspectives. We will use an exploratory sequential mixed methods design to understand and identify key costs and outcomes from key stakeholder perspectives. We will develop a costing guide and conduct a preliminary cost analysis to inform an economic evaluation for a larger trial. The proposed research is innovative because to date implementation strategies have not been designed to support systems in responding to changing public health trends with attention to equity. In addition, this study focuses on implementation strategies to reduce the health impact of emerging drugs and provide an infrastructure to make future adaptations that can be applied in other contexts. The proposed research is significant because it will advance applying implementation science to achieve access, cost and equity for drug use prevention.