Washington University

NIH Research Projects · FY 2025 · 2023-09

Hodgkin lymphoma (HL) accounts for ten percent of all lymphomas in the western world and remains a substantial clinical problem. Relapsed and refractory (R/R) HL presents a particularly critical unmet clinical need because approximately 25% of HL patients will be refractory to the standard of care or will relapse while receiving treatment, and the overall survival of R/R HL patients is ~50%. New targeted treatment options and improved risk assessment are needed to improve the therapeutic options for these patients. HL is defined by a rare malignant B cell (Hodgkin-Reed-Sternberg , HRS cells) that occupies a small fraction of the tumor microenvironment. Despite many advances in next generation sequencing technologies, the identification of somatic variants in cancers characterized by rare cell populations remains technically and analytically challenging. These obstacles have resulted in a limited number of studies that have used high throughput genome-wide technologies to characterize the genomic landscape of newly diagnosed HL, and far fewer have attempted to use these technologies to address the genomic landscape of R/R HL. Here, we address this shortfall by asking three questions: 1) Can ultra-deep genome-wide sequencing, paired with analysis and filtering strategies optimized for low VAF variants identify somatic variants driving treatment resistance in R/R HL? 2) Can we use somatic mutation status in a large cohort of newly diagnosed and R/R HL to identify genes that are predictive of response to therapy? 3) Can somatic mutations identified in genomic and/or cell free DNA describe clonal heterogeneity among HRS cells and can cell free DNA serve as biomarker of HL at diagnosis or relapse? This research will be broadly impactful because: 1) it will describe the genomic landscape of R/R HL and will identify biomarkers that are predictive of poor response to treatment; 2) it will begin to address the relationship between clonal architecture in HL, disease outcome, and prognosis; 3) it will further define cfDNA as a suitable target for HL disease monitoring By completing this proposal Dr. Felicia Gomez will gain advanced training in genomics, bioinformatics, lymphoma biology, and cancer biology in general. This proposal takes advantage of the cutting-edge genomics and cancer research at Washington University School of Medicine (WUSM). WUSM is an ideal location to complete the experiments and training described in this proposal. When Dr. Gomez transitions to independence she will continue to focus on cancer genomics and will broaden her research to address the interactions between somatic and germline variation and how this interaction affects the development and progression of lymphoma and other malignancies.

NIH Research Projects · FY 2026 · 2023-09

Project Summary Amyloid-beta and tau are hallmarks of mild cognitive impairment (MCI)/Alzheimer’s disease (AD). The relationships of in-vivo amyloid-beta, tau, and neurodegeneration with cognitive, clinical, and genetic markers are not well understood. Patients with AD pathology exhibit heterogeneity in their clinical symptoms and illness course. Understanding the underlying neurobiological heterogeneity mechanisms of AD and improving the outcomes have been the central goals. This proposal leverages complementary information of in-vivo amyloid- beta positron emission tomography (amyloid PET), tau PET, structural magnetic resonance imaging (sMRI), cognitive, clinical, and genetic measurements via advanced machine learning methods and investigates the relationships among these measurements in patients with MCI/AD relative to normal controls. The proposal will study the data from the Alzheimer Disease Neuroimaging Initiative (ADNI; N = 898) and the Washington University’s Knight Alzheimer Disease Research Center (Knight ADRC; N = 1,121). This study will be the first to examine regional amyloid PET, tau PET, and sMRI markers and their relationships with cognitive, clinical, and genetic phenotypes using machine learning predictive modeling and heterogeneity analytics in AD research. The proposal will quantify regional PET outcomes as distribution volume ratio (DVR) and sMRI as the volumes and investigate their associations with cognitive [Mini-mental state examination (MMSE)], clinical [clinical dementia rating sum of boxes (CDR-SB) and CDR], and genetic [polygenic risk scores (PRS) and apolipoprotein E (APOE)] measurements. Aim 1 will develop machine learning modeling methods to study the relationships of amyloid PET, tau PET, and sMRI with cognitive and clinical phenotypes and test the hypothesis of whether regional brain-based imaging measurements exhibit multivariate predictive associations with cognitive and clinical phenotypes in MCI/AD patients and controls. Aim 2 will study the regional heterogeneity of amyloid PET, tau PET, and sMRI outcomes via semi-supervised machine learning methods. The study will compare the imaging outcomes between identified subgroups of patients or controls vs. each subgroup of patients to test the hypothesis of whether imaging markers differ between subgroups of patients. Aim 3 will examine the relationships of amyloid PET, tau PET, and sMRI heterogeneity signatures with cognition and genetics to test whether imaging signatures associate differentially with cognition and genetics in the subgroups of MCI/AD relative to controls. Overall, this innovative proposal will yield critical information on AD heterogeneity mechanisms, and contribute to precision medicine of diagnosis and treatment of AD. 1

NIH Research Projects · FY 2025 · 2023-09

Project Summary/Abstract Within the years 2013 to 2016, 6.2 million patients were diagnosed with heart failure in the United States. Heart failure has multiple causes, and many fatal cases include patients being predisposed to arrhythmias. One mechanism for triggering a pro-arrhythmic state are changes cardiac action potential such as the prolongation of the depolarization. The main contributor of this rapid depolarization is the ionic current supplied by the Nav1.5 voltage gated sodium channel within the human heart. In addition, there are multiple subunits that interact with the channel in a physiological setting including the subunits of intracellular fibroblast growth factor (FGF) 12A, FGF12B, and calmodulin (CaM). In preliminary data, it has been shown that FGF12A is upregulated in the left ventricle in failing hearts and that FGF12B is the most prevalent FGF isoform in the human heart. Both FGF12A, FGF12B and CaM have shown to alter the inactivation of the Nav1.5 channel through modulation of the DIII and DIV voltage sensing domains (VSD). However, there is no research as to the combined effects of these subunits and their potential to synergistically modulate the Nav1.5 VSDs. As the DIII and DIV VSDs are modulated by these proposed subunits, it can be hypothesized that the efficacy of class 1b anti-arrhythmic drugs are also affected by the proposed subunits. These preliminary findings confer to the two hypotheses: (1) combinations of modulating subunits bound to Nav1.5 can collectively alter gating to disrupt activation and inactivaiton and (2) the subunits of FGF12A, FGF12B, and CaM will alter the interaction of efficacy of class 1b anti-arrhythmic drugs. To support these hypotheses, three aims were created. Aim 1 will focus on determining the biophysical changes the combined subunits have on the Nav1.5 VSDs. The aim will be accomplished with the use of voltage clamp fluorometry to examine the changes in the activation of the individual VSDs. Aim 2 will develop a machine learning model to decipher how alterations to the VSD activations change the overall ionic current. Aim 3 will conclude the proposal by looking at changes in the effectiveness of the class 1b anti-arrhythmic drugs lidocaine, mexiletine, and ranolazine in response to varying levels of each subunit. This proposal has implications that stretch both at the biophysical understanding of the Nav1.5 channel to the clinical application in the use of specific anti-arrhythmic drugs. The overall application will provide rigorous and exemplary training for the applicant to successfully become a translation research scientist.

NIH Research Projects · FY 2024 · 2023-09

PROJECT SUMMARY/ABSTRACT Alcohol use is a global health problem that often begins in adolescence/young adulthood. Despite extensive research efforts, effective preventative and treatment measures remain elusive. While bi-directional associations between alcohol use and brain function form the cornerstone of prominent neurobiological theories of addiction, the reciprocal relationship between alcohol use and brain function remains underexplored. This 5-year K99/R00 proposal is designed to address this fundamental gap in knowledge, through the novel application of cutting- edge techniques which overcome many of the limitations of between-subject cross-sectional designs, including low reliability and small effect sizes. It is hypothesized that neurocognitive mechanisms of decision making which underly elevated risk taking in adolescence (i.e., elevated reward processing relative to lower cognitive control) contribute to alcohol use initiation in adolescence, as well as escalation and maintenance in young adults, and that the consumption of alcohol in turn influences these same mechanisms, thereby promoting future use. Crucially, this proposal is designed to support the applicant’s transition to independence through: 1) technical training in advanced longitudinal and machine learning analysis, and the collection and analysis of precision functional mapping (PFM) data, 2) training in assessments of alcohol use behavior in young adults, and 3) professional and career development, including leadership and mentorship, grantsmanship, professional networking, publication, and scientific presentations. Aim 1 (K99) will assess prospective long-term associations between reward, cognitive control, and alcohol use in two large extant datasets of adolescents and young adults (IMAGEN: ages 14-22, N=2000; ABCD: ages 9-18, N=11,000). Analyses will use multivariate neural signatures of these processes, which are more reliable and sensitive to individual differences than regional estimates. Aims 2&3 (K99/R00) will collect the novel ALC-21 sample – a sample of college students who engage in heavy episodic drinking (N=24), who will be scanned repeatedly (8-10 scans each) over a semester, immediately following periods of elevated use (e.g., 21st birthday) and reduced use. The use of precision functional mapping (PFM) – studying an effect with large within-subject variation by acquiring large quantities of imaging data per subject – maximizes the neuroimaging signal, spatial resolution, and measurement reliability. PFM results in robust and reliable within-subject brain-behavior associations. A subset of the full sample (N=5) will be collected for Aim 2, and will be used to test the short-term effect of heavy episodic drinking on reward and cognitive control. The remainder of the sample will be collected during the R00 portion for Aim 3, which will assess bi- directional short-term associations between reward, cognitive control, and alcohol use. Together, these studies will provide fundamental insights into the reciprocal relationship between alcohol use and brain function, both over the long-term and in the short-term. Further, completing this training plan will prepare the applicant to lead an innovative research program as an independent investigator.

NIH Research Projects · FY 2025 · 2023-08

The Washington University Specialized Program of Research Excellence (SPORE) in Pancreatic Cancer is a highly translational cancer research program focused entirely on the deadliest form of the disease, pancreatic ductal adenocarcinoma (PDAC). Our outstanding investigators have complementary expertise in basic and clinical studies, and our teams leverage individual expertise in immunology, cancer biology, drug development, genomics, and clinical care to develop novel diagnostic and therapeutic approaches for Pancreatic Ductal Adenocarcinoma (PDAC). This SPORE application includes three research projects, an administrative core (Core A), two shared-resource cores [Biospecimen Core (Core B) and Biostatistics and Bioinformatics Core (Core C)], and developmental research (DRP) and career enhancement (CEP) programs. In collaboration with our External (EAB) and Internal (IAB) Advisory Boards, we intentionally selected projects with the greatest potential to impact the outcome for patients with PDAC. Basic research at WUSM and UPenn has led to the development of the following three research projects, all of which feature innovative investigator-initiated therapeutic trials for PDAC. All projects deemed to have the most potential for translational impact and share an immunologic theme in their approach. This immunologic theme leverages institutional strengths and highlights our team’s expertise and leadership in this field. Thus far, the immunologic and metabolic therapies that have been increasingly successful for controlling other cancer types have not been impactful in PDAC. Our project leaders have made noteworthy contributions to elucidate the mechanisms underlying this immunologic resistance and drug resistance in PDAC and developed several exciting strategies with potential to overcome these obstacles. The three projects in this application are designed to have significant potential to change clinical practice within 5 years. Project 1. Employing CD11b-agonists to render PDAC responsive to immunotherapy Project 2. Mechanisms of Resistance to Neoantigen Vaccines in PDAC Project 3. Targeting Stress-induced MK2 as Novel Strategy in Pancreatic Cancer Our long-term goals are to improve the survival and the quality of life of patients diagnosed with PDAC. To achieve this goal, we will promote institutional and inter-institutional collaborative research, with an emphasis on translation. We anticipate that no singular approach will solve PDAC and fully commit to supporting the development of novel research concepts as well as highly promising young investigators. Our SPORE will provide access to pancreatic cancer–specific resources to facilitate this mission.

NIH Research Projects · FY 2026 · 2023-08

Abstract Losing weight can be life saving for people with obesity. Although many people with obesity lose weight with diet and exercise, the vast majority regain this lost weight over time. Multiple pharmacological treatments have been approved by the US Food and Drug Administration (FDA) to help patients lose weight. However, in each case the effect of these treatments is temporary, and weight is regained when the treatment is stopped. This means that patients must take these medications for the rest of their lives to realize their health benefits. In addition to the financial cost of life-long medication, patients must contend with side effects and medication resistance. As such, there remains an unmet need for new medical interventions that produce weight loss without lifelong treatment. In this proposal we aim to investigate how dopamine function is altered as animals gain weight. Reductions in dopamine function have been linked to obesity, and drugs that decrease dopamine function (such as antipsychotic medications) cause weight gain. This leads to our central hypothesis that obesity reduces dopamine release in the nucleus accumbens (NAc), which causes overeating and weight gain. A corollary of this hypothesis is that boosting dopamine release would lead to weight loss. Indeed, either direct stimulation of dopamine neurons or administration of drugs that increase dopamine release (such as amphetamine) cause weight loss. However, due to side effects and abuse liability, dopaminergic stimulants are not useful therapeutics. For this reason, our primary Aim is to understand the cellular changes in dopamine function that occur as animals become obese, to inform the design of neuromodulation approaches to permanently reverse these changes to drive lasting weight loss without life- long medication. To test this hypothesis, we propose to compare in vivo dopamine release in the nucleus accumbens of control vs. obese mice (Aim 1). We will also employ ex vivo electrophysiological approaches to determine the cellular mechanisms underlying changes in dopamine neuron activity in obese mice (Aim 2). And finally, we will engage neuroplasticity in dopamine neurons to chronically boost dopamine release, to test whether this causes long-lasting reductions in food intake and body weight (Aim 3). This research will provide a critical foundation to advance efforts for modulating food seeking and intake, to inform and improve weight loss outcomes in people with obesity.

NIH Research Projects · FY 2025 · 2023-08

PROJECT ABSTRACT Sub-Saharan Africa (SSA), a region dominated by low-resource communities and relatively poor families, is experiencing rising HIV prevalence among adolescents and youth (AY). Household economic hardships heighten the risk for AY’s engaging in health-compromising behaviors and their poor engagement with care. This increases their risk for contracting and transmitting HIV and non-adherence to ART treatment. Economic empowerment (EE) interventions have demonstrated substantial promise in reducing HIV-related risk-taking behaviors, and improving ART treatment adherence and mental health outcomes. Based on 10+ years of research utilizing savings-led EE interventions focused on HIV prevention, care and support for AY affected by HIV [AYaAIDS] (including AY living with HIV [AYLHIV]; and AY orphaned by AIDS [AYoAIDS] in SSA, our group has demonstrated the effectiveness of a multi-component EE intervention, Bridges, in four NIH-funded randomized control trials (RCT) in Uganda (R01 HD070727, R01HD074949, R34MH081763, R01MH113486), and one foundation-funded study in Kenya. Bridges involves: 1) financial literacy training (FLT) and mentorship; 2) family income-generating activities (IGA); and 3) incentivized savings via a matched Youth Development Account (YDA). Bridges has demonstrated robust effects on HIV-related risk-taking behaviors, ART adherence, mental health, psychosocial outcomes, educational achievement, family economics, and family cohesion. Yet, scaling EE interventions has been a challenge, signaling the need to identify and test scale-up strategies and examine determinants of implementation and sustainment. In Bridges2Scale, we will compare two multifaceted strategies (standard vs enhanced) for scaling Bridges in a two-arm Hybrid III effectiveness-implementation cluster RCT. The standard implementation strategy has been applied in our prior RCTs and involves educational meetings that prepare staff to deliver Bridges with minimal disruption to site workflow. This will be compared to an enhanced strategy that will be developed using Implementation Mapping. We will use the public school system to recruit 1440 AYaAIDS (ages 13-17 years) from 48 schools in the Greater Masaka region of Uganda, a region with 11.7% HIV prevalence. Schools will be the unit of randomization (n=24 schools per arm; n=720 students per arm). Four specific aims guide our study: Aim 1. Compare the implementation effectiveness of the standard implementation strategy vs. an enhanced implementation strategy; Aim 2. Determine the clinical effectiveness of Bridges implemented via a standard vs. enhanced implementation strategy; Aim 3. Explore implementation processes, mechanisms, and determinants; and Aim 4. Compare the cost and cost-effectiveness of the two implementation strategies. The study will address a critical challenge: how to best support the implementation, scale-up, and sustainment of EE interventions, which have been proven to be highly efficacious in improving youth-focused HIV prevention, care, and support outcomes, but are yet to be widely scaled up.

NIH Research Projects · FY 2025 · 2023-08

The Route 66 Endometrial Cancer SPORE brings together interactive research teams from three institutions to create a dynamic translational research program aimed at developing and testing new strategies to prevent and treat endometrial cancer. This SPORE includes three research projects; an administrative core; a biostatistics and bioinformatics core; a biospecimens, metabolomics, and pathology core; and developmental research and career enhancement programs. The three projects, chosen and refined with extensive input from our internal and external advisory boards, are designed to have significant potential to change clinical practice within five years. Project 1: HSPA Proteins in Advanced and Recurrent Endometrial Cancer Therapy. Project 2: Inhibiting AXL to Improve Treatment Response in Endometrial Cancer. Project 3: Improving Primary Prevention and Uterine Preservation in Premenopausal Women with Obesity and Endometrial Hyperplasia. Projects 1 and 2 are clinical/translational projects, whereas Project 3 is an early detection, prevention, or population science project. All three projects include clinical trials and represent carefully chosen marriages between selected endometrial cancer research priorities and the strengths of Washington University in St. Louis and our collaborators at the University of New Mexico and the University of Oklahoma Health Sciences Center. The overall aims of our SPORE are to 1) Test promising new therapies to treat or prevent endometrial cancer; 2) Elucidate the key biologic processes that drive endometrial cancer and develop novel biomarkers to predict development of endometrial cancer and response to therapies; 3) Leverage and enhance capacities of shared research resources; 4) Recruit and mentor new investigators and support innovative ideas in translational endometrial cancer research; 5) Facilitate collaboration of those interested in endometrial cancer research; and 6) Ensure fair enrollment in clinical trials and involvement of many community members and investigators in research. In completing the work proposed here, we will test three new strategies to prevent or treat endometrial cancer. Future work can be directed at moving the most promising treatment approaches into larger trials. Additionally, we will obtain an unprecedented level of molecular, cellular, immunologic, and metabolomic detail regarding endometrial cancer and response to treatment, which will likely lead to development of additional novel clinical trials. Finally, by developing new ideas, investigators, and collaborations, we will expand the breadth and depth of research aimed at treating or preventing endometrial cancer.

NIH Research Projects · FY 2026 · 2023-08

ABSTRACT Autoimmune diseases affect more than 20 million people in the United States, and the worldwide prevalence is rising. Patients usually suffer from a lifetime of deteriorating illness because no cures are available for most autoimmune diseases, including type 1 diabetes. Addressing this formidable challenge requires a better understanding of the pathogenic elements eliciting autoreactive responses. Although extensive studies have examined the role of native antigens, whether neoantigens/neoepitopes function to drive the autoimmune process remains poorly understood. The identity of neoantigens, especially post-translational modifications (PTMs) responsible for generating immunogenic neoepitopes, requires a critical examination. Endocrine tissues are commonly targeted by autoimmunity. Diverse endocrine cells use the crinophagic pathway to dispose of excessive amounts of secretory proteins to maintain cellular homeostasis. In this process, the regular secretory granules are directly fused to lysosomes. The resulting vesicle, crinophagic bodies, or crinosomes, are enriched with catabolized peptide segments. In type 1 diabetes, a deteriorating autoimmune disease targeting the insulin-producing β cells in pancreatic islets, crinosomes function as an antigen source providing native peptides for recognition by pathogenic CD4 T cells. Our recent examination by immunopeptidomics uncovered diverse PTMs in crinosome-derived peptides, indicating that crinosomes are a specialized repository of potential neoantigens and neoepitopes related to type 1 diabetes. A novel neoepitope family, C19S (cysteine-to-serine conversion in the insulin B-chain), became progressively prominent along with the development of the diabetic autoimmune process. Most importantly, we identified highly compatible sequences of C19S in mice and humans with T1D. In mice, C19S can be recognized by previously unidentified CD4 T cells distinct from those reactive to the native epitope. It is therefore necessary to determine the pathogenicity of the autoreactive T cells targeting C19S. Furthermore, as a representative neoepitope family, C19S provides a setpoint for delineating how β-cell crinophagy functions as a novel pathogenic component in T1D. C19S may initiate a "feed-forward" loop in T1D pathogenesis by linking β-cell stress with autoimmunity. We also propose to assess translational applications of C19S in human crinophagy and T cell pathobiology. These analyses may lead to a broad identification of neoepitopes generated by human crinophagy, which may serve as valuable targets for T1D pathogenesis and therapeutics. Analysis of potential C19S-reactive T cells in human PBMCs may extend our knowledge of autoimmune pathogenesis beyond the current paradigm defined by native antigens.

NIH Research Projects · FY 2024 · 2023-08

Project Summary Diarrheal diseases account for 1 in 10 child deaths worldwide. Rotavirus (RV) and enterotoxigenic Escherichia coli (ETEC) are major etiological causes of acute gastroenteritis and severe diarrhea worldwide, together resulting in approximately 300,000 deaths each year, mostly in children under the age of five. Current RV vaccines have limited efficacy in endemic countries and no direct antivirals are available. No vaccine is licensed for ETEC. Our overall objectives are to better understand the biology of RV and ETEC and to use that information to develop therapeutic interventions to alleviate diarrhea and sequelae. In preliminary studies, we utilized an improved RV reverse genetics system developed by our lab and generated a recombinant murine RV that encodes the protruding domain of the human norovirus VP1 protein. We found that this RV-based viral vector induces a robust antigen-specific serum IgG and fecal IgA responses in inoculated mouse pups. Based on these data, we constructed several new recombinant RVs that express ETEC heat-labile and heat-stable toxins. We hypothesize that one or more novel RV-ETEC dual vaccine candidates will induce a protective humoral immune response in mice and reduce pathogen burden and pathogenesis from subsequent RV and ETEC infections. To test this hypothesis, we have developed a highly tractable murine RV reverse genetics method, disease relevant neonatal and adult mouse models, and several innovative primary human small bowel organoid cultures, which will provide an unprecedented resolution of understanding of the immunogenicity and protective efficacy of our vaccine candidates. In Aim 1, we will characterize RV replication and define the immunological responses of our dual vaccines in neonatal mice. In Aim 2, we will examine the efficacy of the recombinant RV- ETEC dual vaccines in protecting immunized mice from RV and ETEC challenges in adult mice. Collectively, we expect our proof-of-principle study to start to establish the utility of RVs as an innovative live-attenuated mucosal vaccine platform to encode foreign antigens and broadly protect against common enteric pathogens.

NIH Research Projects · FY 2024 · 2023-08

Rotavirus (RV) is a major etiological cause of severe gastroenteritis and diarrhea worldwide, resulting in the death of more than 200,000 children each year. Current RV vaccines have limited efficacy in endemic countries and no direct antivirals are available. Our overall objective is to better understand the mechanism by which human RVs enter human small bowel intestinal epithelial cells (IECs). The head region (VP8*) of the RV outer capsid spike protein VP4 is responsible for viral entry. However, the identities of host factors that interact with VP8* and their roles in human RV entry is unclear. In preliminary studies, we conducted a flow cytometry-based genome-scale CRISPR/Cas9 knockout screen and identified several transmembrane host proteins as putative attachment factors. In Aim 1, we will use CRISPR/Cas9 gene editing and classic adsorption assays to functionally examine the role of these candidates in human RV entry into HT-29 cells and primary human IECs. In Aim 2, we will employ chemical labeling and proteomic approaches and identify the VP8*-interacting cellular partners during human RV entry. Using host genetic screens, RV reverse genetics, and human intestinal organoid cultures, we expect that our study will elucidate the molecular mechanisms by which host molecules facilitate the early replication of human RVs. We ultimately aim to harness that knowledge to develop countermeasures to impede infection and protect against diarrheal diseases.

NIH Research Projects · FY 2025 · 2023-08

Project Summary Abstract The economic, physical and emotional harms borne by AUD-affected families are great. 7.5 million U.S. children live with an AUD-affected parent and have increased risk for poverty, abuse and neglect in addition to heightened genetic risk for alcohol problems. Remission from AUDs is common, but this is seldom acknowledged in research on the costs and consequences of AUDs. Up to 50% of individuals with lifetime AUDs experience remission, many within 14 years of AUD onset and many during prime child-bearing and child-rearing years. Our broad goal for this project is to comprehensively probe the remission phenotype and its role in the intergenerational transmission of AUDs. We will use family-based data from the Collaborative Study on the Genetics of Alcoholism, a study ongoing since 1989 that recruits families with heightened risk for AUDs and more than 15,000 ever-drinkers. Because of COGA's high-risk design, there are sufficient numbers of AUD- affected individuals (N=7724, 49%), and therefore available for remission, to permit this examination of remission within families and its effect on offspring outcomes. In Aim 1 we will use survival analysis and person-centered longitudinal methods to characterize the course of AUD and remission (chronic AUD, stable or relapsing remission, movement through different types of remission [abstinent, non-abstinent]) and identify demographic and behavioral antecedents and sequalae of remission and relapse (marital status, children, employment, income, education, co-occurring substance and psychiatric disorders, treatment). In exploratory analysis, we will construct a measure of family density of remission and test its association with AUD and remission. Because the genetic and environmental factors that influence AUD and remission do not entirely overlap, we expect this measure to have a small but significant association with the probability of not developing AUD and with the likelihood of remission in individuals with AUD, independent of polygenic risk (PRS) for AUD. In Aim 2, we use biological parent-offspring pairs to characterize the familial environment of adolescent offspring (household income, parental marital status, childhood trauma) and variation in adolescent and adult offspring alcohol use and AUD/remission as a function of parental AUD/remission. Sibling comparisons will delineate for whom parental remission is likely to have the greatest impact, while providing rigorous control for potential genetic and environmental confounders shared by siblings. The proposal is innovative in its focus on resilience, rather than risk, in individuals and families; in its extension of the influence of parental AUD/remission into young and mid- adulthood; and in its use of a genetically-informed approach to understanding the role of remission in the intergenerational transmission of AUDs. Results can provide leverage for clinicians to encourage recovery in patients who are or plan to become parents and will contribute to improved prevention and treatment efforts to reduce the intergenerational transmission of AUDs and associated problems.

NIH Research Projects · FY 2024 · 2023-08

Project Summary/Abstract (30 lines) We seek to understand what if any relationship exists between initiation of transmucosal buprenorphine and poor dental outcomes in a retrospective study among a cohort of patients followed at the VA with a diagnosis of opioid use disorder. Prior data suggest that dental problems have arisen after initiation of buprenorphine, however a simple pre-post evaluation of buprenorphine initiation would be insufficient to answer the question of an association, due to confounding. Dental disease worsens with age and over time, an example of time- varying confounding, and patients with OUD, once in treatment, may be more likely to seek routine healthcare that was deferred due to active addiction, an example of healthy user or adherer bias. As a result, we seek to use Aim 1 to characterize longitudinal patterns of MOUD prescribing, as well as patterns of preventive and acute dental care services over time. To rigorously understand the association of transmucosal buprenorphine and poor dental outcomes, we will require an adequate control group with minimal confounding. To achieve this, in Aim 2 we will compare patients with OUD on transmucosal buprenorphine (our main exposure group) to patients with OUD receiving injectable or implantable buprenorphine, methadone, and naltrexone using negative control outcomes. Once we have determined appropriate control groups based on our analyses from Aim 2, in Aim 3 we will report the cumulative risk and risk difference of poor dental outcomes among patients on transmucosal buprenorphine as compared to other MOUD. We will use the VA Corporate Data Warehouse to obtain both prescribing and dental data among patients with OUD diagnosis between 2001 and 2021. Our study team is uniquely suited to employ our proposed methodologies. Drs. Durkin and Roy (Co-PIs) have experience using large prescription datasets to understand trends in utilization. Dr. Butler (Co-I) has extensive expertise in the use of negative control outcomes to reduce bias in comparative effectiveness studies. Dr. Suda (Co-I) has expertise in the VA Corporate Data Warehouse and the use of large datasets to understand dental healthcare utilization patterns. Without rigorous study, the FDA warning may have the unintended consequence of deterring patients from initiation of buprenorphine. In light of the opioid overdose epidemic and known benefits of MOUD, every effort must be made to reduce barriers to initiating buprenorphine. This study will be the first step in understanding the association between transmucosal buprenorphine and dental outcomes.

NIH Research Projects · FY 2025 · 2023-08

The overarching goal of this Research Scientist Development Award (K01) is to provide Dr. Hannah Szlyk with the training and research activities needed to become an independent researcher who develops, tests, and implements cutting-edge digital therapeutic interventions that can promote recovery from opioid use disorder (OUD) plus commonly occurring mental disorders (COMD). This application outlines an integrated training and research plan that will provide Dr. Szlyk with the skills needed to achieve her career goal. Under the mentorship of experts in clinical trials, addictions medicine, implementation science, user-centered design, and OUD research, Dr. Szlyk’s training will focus on the following key areas: (1) clinical trial methodology (Dr. Esther Lu; (2) implementation science and user-centered design (Drs. Patricia Cavazos-Rehg and Emily C. Williams); and (3) recovery from addictions and mental health (Dr. Darrell Hudson). Training in these three areas will be achieved through didactic and applied activities, participation in national conferences and institutes, and mentoring meetings. Dr. Szlyk will apply the skills acquired during the training activities to conduct a research project to appropriately adapt and test an mHealth intervention for residents of medium-sized midwestern cities with OUD plus COMD. Rates of opioid-related deaths continue to rise among residents of such communities in the United States, like St. Louis, Missouri, where this research is set. Yet, few interventions are developed to fit the unique needs of these clients; none address their heightened overdose risk often compounded by COMD which decreases likelihood for recovery success. Also, these clients experience significant barriers to life saving OUD treatments, like medication for OUD (MOUD). To address this gap, the K01 research project will integrate public health praxis, implementation science, and user-centered design strategies to appropriately adapt an mHealth intervention for residents of a medium-sized midwestern city with OUD plus COMD. The central hypothesis is that adaptations to the mHealth intervention will improve engagement and usability among participants, improve adherence to MOUD, and impact decreases in COMD. This research will address key gaps in treatment by 1) adapting an mHealth intervention for Individuals with OUD plus COMD; 2) identifying barriers and facilitators to implementing an mHealth intervention among stakeholders who serve clients with OUD plus COMD; 3) examining the preliminary efficacy of an adapted mHealth intervention. These data will inform the design of a larger RCT for a future R01 application. Dr. Szlyk’s training and proposed research will be carried out in the outstanding intellectual environment of Washington University School of Medicine, which will provide her with a multitude of resources to meet her goals.

NIH Research Projects · FY 2026 · 2023-08

Project Summary/Abstract The goal of this mentored career development award is to provide a robust course of training in the biology of neurodegeneration and aging for Dr. Eteleeb, Ph.D., a candidate with extensive experience in genomics, omics, and machine learning, to enable his transition to research independence. Washington University School of Medicine is a nationally recognized leader in medical research and provides an outstanding environment for the candidate’s training with world-renowned figures in the fields of Alzheimer’s disease (AD) and aging. This proposal will be conducted under the mentorship of an excellent interdisciplinary team of leaders with extensive and complementary sets of expertise in AD, aging, neuroscience, genetics, and cross-omics who are dedicated to support Dr. Eteleeb in completing his research and training goals proposed in this award. With the guidance of this team, Dr. Eteleeb will pursue a rigorous training program to address gaps in his knowledge and allow him to accomplish the aims of this K25 award. The training objectives will focus on Dr. Eteleeb’s transition into the field of AD and aging and include 1) acquire a strong foundation in neurology, clinical, and neuropathology aspects of AD and related dementias, 2) learn and employ novel ways to identify molecular subtypes of AD and specific biomarkers associated with cognitive function, followed by validation techniques in humans and model organisms, 3) gain in-depth understanding of the genetic and epigenetic factors affecting AD molecular subtypes, and 4) develop leadership and professional skills for leading an independent lab focused on transitional research in AD and aging. These objectives will be accomplished through courses, workshops, seminars, journal clubs, conferences, and feedback from the advisory committee. The primary objective of the proposed research is to identify and characterize molecular subtypes of AD associated with cognitive function by employing an innovative approach that combines cross-omics and machine learning. AD is a heterogeneous neurodegenerative disorder affecting over 50 million individuals worldwide. One critical and often overlooked factor impeding development of effective treatment for AD is the clinical and molecular heterogeneity among AD patients. Cross-omics approaches integrate heterogeneous molecular profiles to study not only how these profiles change in AD, but also uncover relationships and correlations between biological molecules. The specific proposed research aims are 1) identify and characterize cross-omics AD molecular subtypes associated with cognitive function conserved across cohorts and brain regions, 2) Determine whether molecular subtypes are specific to AD or present in other neurodegenerative disorders, and 3) identify CSF/blood-based biomarkers from AD molecular subtypes. The results will reveal insights into AD subpopulations and identify AD molecular subtypes, pathways, and biomarkers, which could lead to new pathways for implementing precision medicine and developing novel therapeutics for AD. Successful completion of this award will facilitate future independent funding to leverage cross-omics to deepen our understanding of the molecular mechanisms of AD progression.

NIH Research Projects · FY 2025 · 2023-08

Imbalance of myocardial oxygen supply and consumption precipitates a cascade of physiological changes resulting in ischemic pathology. While assessment of myocardial perfusion alone may allow accurate assessment of myocardial oxygen consumption in some pathophysiological conditions, the perfusion-oxygen consumption relationship is derailed in several conditions, including: myocardial ischemia and infarction, hypertrophic and dilated cardiomyopathies, heart failure, valvular heart disease, and septic cardiomyopathy. Importantly, this oxygen supply/perfusion mismatch occurs early – before mechanical dysfunction. Therefore, evaluation of myocardial consumption independent of perfusion is of importance for early diagnosis and monitoring of these pathophysiological conditions. Myocardial oxygen extraction fraction (mOEF), which relates the biologic coupling of myocardial blood flow (oxygen supply) to oxygen consumption, may provide a more accurate assessment of this balance. For example, in ischemic cardiomyopathy, adequate myocardial perfusion is commonly reduced by high grade epicardial coronary artery stenoses. To avoid ischemia-caused injury, mOEF is likely to be increased to compensate for decrease in myocardial perfusion and oxygen delivery to myocytes. Consequently, the affected myocardial regions drop into a so called “hibernating” state that is effective in the short term. In this respect, an accurate mOEF assessment is a unique tool, with the potential to determine the likelihood of cardiac functional recovery after reperfusion. To date, the reference method for non-invasive quantification of mOEF in vivo is Positron Emission Tomography (PET). We recently developed a novel contrast- free cardiovascular magnetic resonance (CMR) acquisition method to quantify mOEF in vivo that has several advantages over PET: our CMR method has better spatial resolution, shorter acquisition time, does not expose the patient to ionizing radiation, and could be more widely available than PET. The overall objective of this study is to leverage our expertise in CMR imaging to refine and rigorously validate this new mOEF method. In Aim 1. This technique will be developed with assistance of a novel deep learning approach for artifact-free images and then validated using large animal models with and without induced coronary artery disease. Invasive catheter-measured and non-invasive PET-MRI-measured mOEF will be used as reference. In Aim 2, the CMR mOEF method will be validated in a small cohort of patients with hibernating myocardium in vivo, with the availability of 18F-FDG-PET as reference. Although we will study this mOEF technique in hibernating myocardium, this imaging method can be applied to the diagnosis and evaluation of treatments in a wide range of cardiomyopathies. Given the capability of CMR for the comprehensive assessment of myocardial function, tissue characterization, and viability, successful completion of CMR mOEF validation will provide a ‘one-stop shop’ evaluation of metabolic, functional, and structural abnormalities in patients with cardiomyopathy. Furthermore, the advantage of fast imaging without using an intravenous contrast agent is well-suited for repeat quantitative mOEF measurements needed to guide the effects of novel therapeutic interventional strategies.

NIH Research Projects · FY 2025 · 2023-08

PROJECT SUMMARY/ABSTRACT Youth from resettled refugee families are at elevated risk of developing serious mental health challenges compared to their U.S. native peers. Studies have found that refugee youth living in high income countries have high prevalence of trauma-associated mental disorders like PTSD, (19% to 54%) and depression (3% to 30%). Additionally, youth belonging to these communities have high rates of substance use, school dropout, incarcerations, violence including gang-related homicides, with suicides emerging as a serious public health threats. The purpose of this K01 is to provide the candidate with formal training and mentoring in: (1) systematically adapting and implementing community-based interventions; (2) collection, management, and analysis of qualitative and mixed methods data; and (3) advanced implementation science research, including the application of theories and frameworks, methods for identifying and prioritizing implementation determinants, and systematic approaches to developing and testing implementation strategies. This training will prepare the candidate for independent research conducting multidisciplinary research that develops and tests community-based interventions aimed at addressing the myriad of mental health challenges among youth in resettled refugee communities. Accordingly, the candidate will conduct a type 1 hybrid effectiveness-implementation trial to test a combination intervention designed to improve intergenerational trauma-related mental health symptoms among second generation refugee children (SGRC). Guided by Social Action and Family Systems theories, and applying them to the ITT framework, the proposed combination intervention consists of: family strengthening (FS) model delivered through multiple family groups (MFG) + peer mentoring program called TeenAge Health Consultants (TAHC) adapted for delivery in virtual environment (Virtual TAHC). The intervention will have the following specific aims: (1) systematically adapt an evidence-based family strengthening (FS) model delivered through multiple family groups (MFG) combined with a peer mentoring program (Virtual TAHC) (Goal 1); (2) assess preliminary short- and long-term impact of the combination intervention (MFG + Virtual TAHC) on behavioral emotional disorders (aggressive behavior, antisocial behaviors, anxiety, depression, and PTSD) related to intergenerational trauma among SGRC in the trial (Goal 2); and (3) utilizing mixed methods and applying the Exploration, Preparation, Implementation, Sustainment (EPIS) framework, examine implementation strategies, facilitators, and barriers of the RRF4H intervention (Goal 3). Given the transdiagnostic application of the intervention, this study is significant because it has potential to advance our understanding on how to mitigate the intergenerational impact of war trauma among children who have not directly experienced war trauma.

NIH Research Projects · FY 2024 · 2023-08

Abstract Inflammasomes are multi-protein signaling scaffolds that form in the cytosol upon stimulation by pathogen and damage signals to activate caspase-1. Canonical inflammasome sensors are mainly nucleotide-binding domain (NBD) and leucine-rich repeat containing (NLR) proteins, such as NLRP1, NLRP3 and NLRP6. Once inflammasome is activated, caspase-1 is recruited to the platform and activated through proximity-induced autoproteolysis. Activated caspase-1 processes pro-interleukin (IL)-1b, pro-IL-18, and the pore-forming protein gasdermin D (GSDMD), resulting in the maturation and release of these cytokines, as well as pyroptotic cell death. NLRP6 plays versatile roles in host defense. It is highly expressed in the intestine and the liver. The inflammasome function of NLRP6 has been reported to protect the host from pathogen evasion and injury-induced tissue damage. On the other hand, the excessive NLRP6 inflammasome activation and the subsequent hyperactive IL-18 signaling may exacerbate the tissue damage and cause chronic inflammatory diseases such as inflammatory bowel disease (IBD). Whether there is an intrinsic cellular signal that tunes the activity of NLRP6 inflammasome becomes an intriguing question. The question is critical to understanding how our body maintains tissue homeostasis by harnessing this important inflammasome and to provide targeted therapy for certain diseases. Nuclear import is an essential cellular process in innate immune defense by translocating activated transcription factors into the nucleus for interferon production. Plant NLRs themselves have been reported to enable host defense through pathogen effector-mediated nuclear translocation. In our preliminary study, we found that importin-b1, a common nuclear import receptor, inhibits dsRNA-induced liquid-liquid phase separation of NLRP6 in-vitro, which implicates the potential link between nuclear import and NLRP6 inflammasome signaling. In this application, we will investigate the potential role of importins in regulating the NLRP6 inflammasome by tackling two questions: 1) molecular basis of the importin-NLRP6 interaction (cryo-electron microscopy and biochemistry), and 2) the roles of importin-b1 in negatively regulating NLRP6 inflammasome upon viral infection (inflammasome cell biology).

NIH Research Projects · FY 2026 · 2023-08

Hypertrophic cardiomyopathy (HCM) is the most common heritable cardiac disease. Individuals with HCM have adverse clinical outcomes, including heart failure, arrhythmias, and sudden cardiac death (SCD) It is estimated . that, in the U.S., 6 out of 7 individuals with HCM are unaware of their diagnosis. There are recognized racial disparities in the diagnosis of HCM. While Black individuals have been observed to have more clinical and ECG signs that should promote consideration of the diagnosis of HCM, HCM is underdiagnosed in Black patients. Furthermore, even once diagnosed, there are well-recognized disparities between Black and White patients with HCM; Black patients have lower rates of referral to HCM Centers, referral for genetic testing, referral for SCD risk stratification, and referral for interventions such as septal reduction therapy and implantable cardioverter- defibrillator (ICD) placement. The lower rates of referral are in direct opposition to the data that shows that, compared to Whites, Black individuals with HCM have approximately 2-fold the risk of SCD and development of class III or IV (moderate or severe) heart failure. There are several potential contributors to these racial disparities. HCM is typically diagnosed using routine echocardiography and clinical genetic testing is used to help diagnose ambiguous cases. Black individuals with HCM are more likely to have ambiguous presentations and clinical testing is less useful as, compared to Whites, Black patients with HCM are ~40% less likely to have a pathogenic or likely pathogenic (P/LP) sarcomeric variant identified and more than 40% more likely to have variants of uncertain significance (VUS). Furthermore, hypertension (HTN) is more prevalent in Black individuals and physicians may attribute the patient’s left ventricular hypertrophy to HTN and not consider HCM. In addition, recent evidence suggests that HTN may be an environmental modifier /”trigger” of worse disease in patients with HCM. Dr. Huebsch, Co-investigator on this proposal, created an in vitro micro-heart (μHM) model system from cardiomyocytes derived from human induced pluripotent stem cells (iPSC) harboring single sarcomere mutations and has used this model system to help tease apart the contributions of afterload and genetics in HCM. We hypothesize that the presence of (1) multiple sarcomeric variants and (2) HTN contribute to the worse outcomes observed in Black HCM patients. AIM 1. To assess adverse outcomes in Black HCM patients with (vs without) (A) more than one variant (i.e. 2 P/LP variants or 1 P/LP variant + 1 or more VUS) and (B) HTN, in 2 real-world U.S. cohorts of HCM patients. An exploratory analysis using Somascan assays to identify novel plasma proteomic biomarkers of adverse outcomes will also be performed. AIM 2. To characterize the effects of multiple variants on cardiac physiology and fibrosis in our novel in vitro μHM model subjected to increased afterload. Combining patient clinical data with powerful in vitro tools of iPSC technology, genome editing and tissue engineering, will allow us to gain important insights and understanding of potential triggers and mechanisms contributing to worse outcomes in Black HCM patients.

NIH Research Projects · FY 2024 · 2023-08

COVID-19 pandemic highlights the impact of emerging infectious diseases and the global socioeconomic impact. Alphaviruses such as the chikungunya virus (CHIKV) has re-emerged along with many arthropod-borne viruses (arboviruses) and these viruses continue to pose a significant threat to global human health especially with recent vector expansion into non-endemic regions. CHIKV belongs to the Togaviridae family and is transmitted mainly by Aedes albopictus and Aedes aegypti mosquitose. The bent-up posture caused by CHIKV fever disease and the virus can be classified into three different lineages with distinct genotypes corresponding to their respective geographical origins. CHIKV infections cause high serum viral loads, and therefore, high viremia also contributes to rapid spread. Like other alphaviruses, there are no treatments for CHIKV and related viral infections. In recent exploratory studies, we have identified two inhibitors that inhibit CHIKV and related alphavirus Mayaro virus (MAYV). While prior studies have shown a potential role for HSP90 inhibitors, a major limitation associated with further development of HSP90 inhibitors is that pan-HSP90 activity results in toxicity. HSP90 family consists of 4 different paralogs, two cytosolic forms HSP90α (encoded by HSP90AA1), HSP90β (encoded by HSP90AB1 gene), Grp94, an endoplasmic reticulum resident HSP90 (encoded by HSP90B1) and mitochondrial Trap1. While pan-HSP90 activity provides the initial insights, they have many clinical liabilities. In our proposed studies, we will develop HSP90β specific inhibitors with reduced toxicity and enhanced efficacy as a host-directed anti- alphavirus therapeutic lead. We expect to move one or more scaffolds from hit-to-lead with selectivity index >200 and determine the mechanism of action (MOA). Collectively, we will fill a critical gap in the field.

NIH Research Projects · FY 2025 · 2023-08

PROJECT SUMMARY Innate immune antiviral pathways are upregulated in Alzheimer’s disease (AD) and are thought to drive AD pathogenesis. The specific mediators of maladaptive antiviral responses in AD are still not understood. Innate immune system utilizes a series of nucleic acid sensors to detect viral genetic material to activate interferon expression. The central DNA sensor in the cell is cyclic GMP-AMP synthase (cGAS). cGAS signaling has emerged as an important player in many neurodegenerative diseases like Parkinson’s disease and amyotrophic lateral sclerosis. The role of cGAS signaling in tau-mediated neurodegeneration remains unstudied. We discover that cGAS signaling is hyperactivated in the brains of mice expressing mutant tau (P301S mice) and human AD patients. Using behavioral assays, electrophysiological readings and single nuclei RNA sequencing, our preliminary results indicate that cGAS activity modifies both the cellular responses to and the functional deficits caused by tauopathy. We observed that partial or complete loss of Cgas rescued tauopathy-associated spatial learning and memory deficits. cGAS is highly enriched in immune cells including microglia. We find that tau induces a cGAS-dependent interferon signature by triggering mitochondrial DNA stress in microglia and show that Cgas deletion alters microglial disease transformation, characterized by reduced expression of interferon genes in disease microglia. It is not known how tau can alter mitochondrial and nuclear dynamics to activate cGAS and how cGAS activation enhances neurotoxic effects of tau. This proposal will investigate the mechanisms and consequences of cGAS and nucleic acid sensing pathways in tauopathy. I aim to dissect the molecular mechanism of cGAS activation in response to tau by focusing on tau dependent cellular processes that induce mitochondrial stress and microglial senescence (Aim 1) and investigate how cGAS mediates maladaptive microglial responses and neuronal damage in tauopathy (Aim 2). Finally, since antiviral responses could be species-specific, there is strong rationale to extend mouse studies of innate nucleic acid sensing to human models. Human and mouse cGAS share only 60% amino acid similarity and have different activation requirements. I will establish human pluripotent stem cell (hPSC)-based platforms to study broader nucleic acid sensing pathways and perform unbiased CRISPR-screens to identify novel regulators of human nucleic acid sensing in tau toxicity (Aim 3). The experiments outlined in this proposal will lead to a better understanding of the role of nucleic acid sensors in tauopathy and AD which might prove translatable to humans.

NIH Research Projects · FY 2025 · 2023-08

PROJECT SUMMARY/ABSTRACT Current immunotherapies, including immune checkpoint blockage therapy and adoptive T cell therapy, have resulted in promising results in certain types of cancer patients. However, these immunotherapies have so far been insufficient to reproducibly eliminate tumors. It is clear that tumor-reactive T cells are suppressed and dysfunctional in the suppressive tumor microenvironment that is a major obstacle for successful tumor immunotherapy. Thus, dissecting the distinct mechanisms responsible for T cell dysfunctional states within the suppressive tumor microenvironment should provide novel avenues for tumor immunotherapy. We discovered that induction of T cell senescence is an important T cell dysfunctional state and a novel suppressive mechanism utilized by both human naturally occurring and tumor-derived regulatory T (Treg) cells in the tumor microenvironment. In fact, significant accumulation of senescent CD8+ T cells has also been found in the tumor-infiltrating T cells (TILs) from various types of cancer patients. Importantly, we found that these senescent T cells are functionally suppressive and molecularly distinct from anergic and exhausted T cellsand that they are a critical mediator and amplifier for immune suppression within the tumor microenvironments. Therefore, a better understanding of this novel suppressive mechanism and the molecular processes in responder T cells suppressed by Treg cells is essential for the development of effective strategies to treat human cancers. Cellular energy metabolism directs T cell survival, proliferation and their specific functions. Different T cell subsets have different metabolic profiles. We have more recently identified that Treg-induced senescent T cells exhibit active lipid metabolism, resulting in upregulation of lipid metabolic enzymes and secretory lipid species, and accumulation of lipid droplets (LDs). The central hypotheses of this proposal are that: 1) Excessive lipid metabolism is critical for senescence development and immunosuppression of effector T cells mediated by Treg cells; 2) Senescent and dysfunctional tumor-specific T cells can be rejuvenated via lipid reprogramming for enhanced anti-tumor immunity. Specific Aim 1 seeks to identify whether the excessive lipid metabolism is involved in senescence development and immunosuppression of T cells induced by Treg cells. Specific Aim 2 will explore the novel concept and develop effective strategies to overcome senescent and exhausted tumor-specific T cells via lipid metabolism reprogramming combined with selective checkpoint blockage therapy of anti-PDL1 for enhanced anti-tumor efficiency in the adoptive T cell transfer therapy tumor models. The positive outcome of these studies should lead to novel strategies to reprogram lipid metabolism and effector functions of tumor-specific T cells for cancer treatments.

NIH Research Projects · FY 2025 · 2023-08

ABSTRACT High-throughput functional genomics of variants in genes linked to substance use disorders Understanding of the genetic basis of substance use disorders has advanced significantly in the recent past; the generation of a reference human sequence in the early 2000’s enabled Genome Wide Association Studies (GWAS), which in turn led to the identification of thousands of genetic variants that are statistically significantly associated with substance use and substance use disorders. The number of these clinically relevant variants continues to grow proportionally with the increase in sample sizes for GWAS and genome sequencing studies of substance use disorders. Model organism studies have also led to the identification of genes and pathways that are linked to these phenotypes. However, one persistent bottleneck in this field has been the lack of experimental testing of nearly all (>99%) of these variants in genes and regulatory regions to determine which are functionally significant. Determining the functional effects of genetic variants is crucial to acquiring a molecular understanding of the basis of substance use disorders, and this deeper understanding will enable future development and testing of targeted and rationally designed therapeutic interventions. This forward- thinking application proposes to address this gap by applying highly innovative high-throughput functional genomics approaches, including Massively Parallel Reporter Assays (MPRAs) and Deep Mutational Scan (DMS) techniques, that were recently deployed for the study of rare Mendelian disorders, to the study thousands of both noncoding and coding variants associated with substance use disorders. These approaches have not previously been applied to the genetics of substance use disorders, although they have enabled significant advancements in other fields, such as the genetics of neurodevelopmental and neuropsychiatric disorders. Furthermore, this early-stage investigator’s multidisciplinary training, including MD/PhD degrees, Pediatrics residency, a postdoctoral fellowship, and four years of dedicated K08 Early Career Development work that was focused on acquiring expertise in the use of functional genomics techniques, has established him as a rising leader in the field of human genetics. His prior paradigm-altering and team-based accomplishments include establishment of an exome sequencing testing pipeline for children with autism in 2015, ongoing coordination of the Model Organism Screening Center for the NIH-funded Undiagnosed Diseases Network, and leadership of a $1.5 million institutionally-funded, multi- investigator functional genomics project that resulted in deployment of critical infrastructure and delivered a 10:1 return on investment in subsequent grant funding. Although this talented investigator has not previously focused on substance use disorders, he has established a collaboration with a world expert in this field, Dr. Laura Bierut, who has agreed to advise him in this research program and connect him with other relevant investigators in the field. In sum, this very high impact and extremely innovative proposal from a particularly well qualified early- stage investigator will significantly advance the understanding of the genetics of substance use disorders.

NIH Research Projects · FY 2024 · 2023-08

Project Summary/Abstract: An antigen test for Neisseria gonorrhoeae (Ng) that is accurate, simple, scalable, non-invasive, rapid, and effective at the point-of-care (POC), would transform public health decisions. Although antigen detection rapid diagnostic tests (Ag-RDT's) for Ng are available, they have poor sensitivity (50% or less) compared to gold standard nucleic acid amplification tests (NAAT). A NAAT based POC test was recently cleared by the US FDA, however its use in resource-poor settings is limited by its high cost. To overcome these barriers for the first time, we will harness ultrabright fluorescent nanoconstructs, called plasmonic-fluors, a recent breakthrough from our labs that enables such antigen testing with sensitivity that matches NAATs. The objective of this proposal is to develop an ultrasensitive plasmon-enhanced lateral flow assay (p-LFA) that detects the lipoprotein H.8 antigen present on Ng surface. The rationale underlying this proposal is that a highly sensitive Ag-RDT that can match the accuracy of NAAT could meet the critical need for diagnosing Ng infection at the POC and in resource-limited settings. We will achieve this by pursuing two specific aims: 1) Develop and optimize the p-LFA for detecting Ng lipoprotein H.8 antigen in urine samples obtained from healthy people. 2) Determine the diagnostic accuracy of the Ng antigen test compared to gold standard NAAT in urine specimens from patients with suspected Ng infection. The proposed approach is innovative in that it harnesses plasmonic-fluor as a fluorescent nanolabel in a LFA as opposed to the conventional gold nanoparticles, which provide only a weak colorimetric signal. With over 1000-fold improvement in sensitivity, this approach has the potential to transform antigen testing. The proposed research is significant as the novel p-LFA will meet the critical needs for widespread Ng testing scalability, speed, and low cost. The expected outcome of this work is a simple, non-invasive, Ag- RDT for Ng infection, implemented as a LFA that can replace NAAT by virtue of its accuracy and low cost in resource-limited settings. This test will have a tremendous positive impact immediately as it will be applicable to large scale Ng testing with an inexpensive test strip and minimal battery-powered portable equipment, without relying on skilled personnel. Furthermore, this test will serve as a prototype for ultrasensitive antigen tests applicable to many other infectious diseases in addition to the possibility of incorporating multiple antigens on a LFA for diagnosing other sexually transmitted infections such as chlamydia and trichomoniasis in addition to Ng.

NIH Research Projects · FY 2025 · 2023-08

Project Summary Children facing adversity are at high risk for poor socio-emotional and cognitive outcomes and later psychopathology. However, there is increasing evidence that these negative developmental trajectories can be significantly improved by enhancing nurturing caregiving early in development. Despite numerous empirically supported early interventions, these programs are not readily accessible to the majority of US children most in need. Based on this, there is an urgent need to make these interventions feasible and readily available to these communities. To do this we propose to test the effectiveness of a brief 6 session parent-child preventive intervention conducted by video conference in the family's home and study two implementation methods within 3 high-risk school districts. The intervention, entitled “THRIVE,” is a previously tested early parent child intervention piloted in a St. Louis county district which proved feasible, acceptable and appeared effective. Caregiver-child dyads aged 4.0-6.11 meeting inclusion criteria will be randomized to a THRIVE condition or an established online parenting education of comparable length. Although the use of schools as a service delivery platform will increase accessibility, this system presents unique and complex challenges for implementation of a mental health prevention. Therefore, we will use a hybrid effectiveness-implementation design to test the effectiveness of THRIVE compared to Parenting Wisely, while also assessing the impact of two forms of implementation (coaching vs. no coaching) on study outcomes. Assessments of key outcome measures, including child behavior, social, and emotional functioning, child psychopathology, parenting stress, optimism, and depressive symptoms, and changes in parenting and the parent-child dyadic relationship (observational and neural using functional near infrared spectroscopy' fNIRS) will be measured at baseline, post-treatment (P1), and 12-weeks post-treatment (P2). We will also test how the preventive intervention mechanistically targets the quality of the caregiver-child relationship by enhancing caregiver responsiveness and sensitivity via baseline, mid-treatment and post-treatment (P1) assessments. This project provides the first test of a brief parent-child early prevention accessed through schools and delivered in home by video conference with minimal therapist training to enhance access to care. It further examines the cost-effectiveness of therapist coaching as a means to improve implementation and clinical outcomes.