University Of Southern California

NIH Research Projects · FY 2026 · 2024-06

Project Summary Reliable, reproducible estimates of the costs of Alzheimer’s disease (AD) and AD related dementias (ADRD) are critical for stakeholders and decision makers to fully understand the size of the economic and financial disease burden and to inform priority setting. Dementia cost estimates typically include medical costs and the cost of long term services and support and some indirect costs such as the value of the time of unpaid caregivers for persons living with dementia. Yet, dementia cost magnitude, when based on a limited set of costs, is likely a significant underestimate of costs, particularly as it relates to costs associated with unpaid care provided by family and friends. Time spent caregiving may result in productivity losses and future income and wealth losses. Caregivers may experience health and quality of life impacts and medical costs associated with the challenges of caregiving. Additionally, participation in interventions or clinical trials may impose costs to both persons living with dementia (PLWD) and their care partners and caregivers. Vulnerability to financial and physical abuse is more common among persons living with dementia than for persons without dementia, with important financial and non-financial costs associated with both. We propose to provide national, annual, comprehensive cost estimates of dementia from the societal perspective. Concurrently, we will build the infrastructure to greatly increase national research capacity for generating reliable, reproducible cost estimates and for utilizing innovative methods for analyzing what impacts these costs, how and for whom. We have brought together dementia experts from across the nation and the experience in convening lived experience panels through partnership with the Alzheimer’s Association. We will draw on the significant expertise of our multidisciplinary team in dementia cost estimation using nationally representative survey and administrative data sources. We will build upon decades of investment in validated and sophisticated dynamic microsimulations models for estimates and projections of population health and spending, the Future Elderly Model (FEM) for middle age and older US adults and the ADRD FEM for dementia estimates, and Future Adult Model (FAM) for young to middle- aged adults. We will innovate on these models to build the U.S. Costs of Dementia Model (USCDM) that will fill gaps in cost estimates and facilitate reproducibility and use by the research community. Utilizing accessible platforms for data and code sharing, and a comprehensive pilot program with innovative mentoring, we will expand the capacity of the research community to produce comprehensive, robust and replicable estimates of the costs of dementia in the US over time, and to use dynamic microsimulation and modeling tools to quantify how changes in population health, demographics, care models and systems, public and workplace policies, and new treatments impact costs of dementia today and in the future.

NIH Research Projects · FY 2025 · 2024-05

Abstract Alcoholic liver disease (ALD) is one of the most prevalent chronic liver diseases, accounting for more than half-million deaths worldwide each year. To date, there exist no effective medical interventions; thereby, posing a serious threat to public health. ALD represents a broad-spectrum hepatic disorder from alcoholic fatty liver (AFL), alcoholic steatohepatitis (ASH) to cirrhosis, the development of which is largely determined by the duration, quantity, and pattern of substance abuse. AFL, the 1st stage of ALD, is characterized by the massive accumulation of macrovesicular lipid droplets in hepatocytes without histological features of inflammation. AFL develops as soon as 2 weeks of alcohol abuse and can also undergo a rapid and complete resolution upon abstinence. However, if abstinence cannot be achieved, AFL, in turn, serves as the foundation for the development of advanced stages of ALD such as ASH and cirrhosis, which are associated with extremely high mortality and morbidity rate as well as poor reversibility even with long-term abstinence. With the opportunity to efficiently mitigate the disease burden of ALD, AFL represents an attractive therapeutic target. To this end, furthering our understanding of AFL pathophysiology serves as a critical milestone. In this study, we established a novel in vitro study model of AFL with a highly physiological culture system of terminally differentiated human hepatocyte (HH), then applied for the elucidation of hepatocyte-intrinsic response to the metabolic stress inflicted by AFL. Cellular response to metabolic stress is a coordinated adaptation process obligatory for homeostatic maintenance; thus, the failure leads to cell injury. While the induction of transcriptome changes is an undoubtedly important process in stress response, the synthesis of respective proteins is ultimately required to execute the designated gene functions. Consequently, we conducted an integrative analysis of polysome profiling and mRNA-sequencing to elucidate the concordance between transcriptome change and mRNAs translated by ribosomes. Our analysis revealed that AFL induces a significant dysregulation of translatome, suggesting that the transcriptome change is discordantly processed at the protein translation machinery. This observation led to our hypothesis that “AFL dysregulation of translatome results in a global alteration of the proteome, which acts in concert to impair the stress response and leads to the cytotoxicity.” Accordingly, this exploratory proposal is designed to elucidate the mechanism and significance of translatome alteration as the critical pathophysiology of AFL through the following specific aims: Aim 1: Determine the molecular mechanism of dysregulated translatome in HH of AFL, and Aim 2: Define the correlation between transcriptome, translatome and quantitative proteomics of AFL HH. The successful completion of the proposed studies will provide unique and novel insights on AFL pathophysiology and a framework for further investigation with in vivo models as well as translational studies involving clinical specimens.

NIH Research Projects · FY 2026 · 2024-05

The relationship between education and Alzheimer’s Disease and related dementias (ADRD) is well-established; however, inequalities in access to education are a fundamental social disparity, resulting in differential cognitive outcomes in diverse contexts. Country, birth cohort, and male/female differences in access to education and occupation can be a source of natural experiments to reveal trends and mechanisms for the relationship between education and cognitive outcomes such as ADRD, including how genetic propensities both manifest and change across diverse historical and social settings. We propose to leverage the Interplay of Genes and Environment across Multiple Studies (IGEMS) consortium of longitudinal twin studies of development and aging to clarify the role of educational inequalities for ADRD. The IGEMS consortium is a collaboration involving 18 longitudinal twin studies of adult development and aging. Established in 2010, the consortium has developed standard processes for collaboration, data sharing, data harmonization, and data analyses. The current application proposes a new direction for the IGEMS consortium to support novel investigations of the education-ADRD association in the context of country, birth cohort, and male/female differences in access to educational opportunities, integrating multiple indicators of education. First, in addition to harmonized International Standard Classification of Education (ISCED) scores across countries and birth cohorts, IGEMS includes polygenic scores for education (PGSED). Second, parental education measures allow for a study of intergenerational mobility and clarifies potentially spurious relationships between education and ADRD that result from family background effects. Third, we will examine the impact of birth cohort and country, comparing access to educational and occupational opportunities using indexes of educational inequality (GINIED) as well as measures of social differences. Dynamic male/female differences that vary over context and time will also help us to explain established male/female differences in the relative protective effects of education and occupation for cognition including ADRD. The twin structure of our data permits the use of established twin methods to test hypotheses on the nature of gene-environment (GE) interplay, while genotyping allows us to confirm and extend these twin analyses through analyses of PGSED and polygenic scores for dementia. Importantly, IGEMS is a multidimensionally diverse sample that spans multiple countries and historical periods, allowing us to determine whether models of GE interplay established at the micro-level (i.e., individual) might vary at the macro-level (i.e., country/historical period). We propose the following research aims. AIM 1: Investigate mechanisms of educational influences on cognitive functioning and ADRD risk at multiple levels: genetic (PGSED), individual (ISCED), inter-generational (parent ISCED), and environmental (GINIED). AIM 2: investigate the impact of women’s differential educational and occupational opportunities across cohorts and countries on education-cognition and ADRD relationships and male/female differences in ADRD risk.

NIH Research Projects · FY 2025 · 2024-05

ABSTRACT Herpesviruses are ubiquitous human pathogens worldwide. Though herpesvirus infections are often asymptomatic in normal individuals, they cause significant morbidity and mortality in immune-compromised individuals. Antivirals have been successfully developed to target the viral thymidine kinase for the past nearly half-century. In the presence of such antivirals, drug- resistance of herpes simplex viruses (HSV), including HSV-1 and HSV-2, is rapidly emerging, particularly in those immune-compromised individuals such as AIDS patients. Thus, new antivirals are desperately needed to cope with the dire situation. We have previously identified the HSV-1 UL37 deamidase that deamidates and inactivates cytosolic RIG-I and cGAS, thereby muting host innate immune defense. The enzymatic activity of UL37 represents an “Achilles heel” that can be targeted to restore host immune defense. Continuing our original discoveries on protein deamidation, we synthesized series of glutamine analogues to identify inhibitors of selected protein deamidases. In this study, we have identified three lead compounds that selectively inhibit UL37-mediated protein deamidation, but not those mediated by cellular deamidases. Ground on these preliminary results, we will characterize the mode of action of these UL37 inhibitors in protein deamidation and innate immune response during HSV-1 infection. We will further employ a combination of mass spectrometry, structural and pharmacological analysis to improve these lead compounds via iterative synthesis and functional assays. The resulted best inhibitors will be assessed by pharmacokinetics and pharmacodynamic studies as well as antiviral efficacy using mouse model. This pilot study will not only provide proof-of-concept to counteract HSV-1 infection via inhibiting a viral deamidase and restoring host immune defense, but also develop important tools that enable further investigation into protein deamidation in fundamental biological processes.

NIH Research Projects · FY 2025 · 2024-05

Project Summary/Abstract Neurodevelopmental disorders (NDDs) comprise of a group of disorders associated with abnormal brain development, leading to abnormal brain function of language, motor, behavior, memory and learning. NDDs, e.g., attention-deficit/hyperactivity disorder (ADHD), autism spectrum disorder (ASD) and intellectual disability, affect approximately 15% of children in the United States and pose a significant public health burden on society. The causes of NDDs are still largely unknown. A growing body of evidence support that most NDDs are result from a combination of genetic, biological, psychosocial and environmental risk factors rather than any single origin. Furthermore, there are high rates of co-occurrence between different NDDs, as well as similarities in functional impairment and clinical features, which make the diagnosis and treatment of NDDs difficult. Hemispheric asymmetry is a core element of the brain’s usual organization required for optimal functioning and a key landmark of brain development. We have recently revealed that brain asymmetry is closely associated with NDDs and their genetic/environmental risk factors and clinical features, though the causal interacting relationships are unclear. Here we hypothesize that the biological process of brain asymmetry development is influenced by complex genetic and environmental contributors for NDDs, and different dysregulations of this multifactorial dynamic system may mediate the emergence of different clinical symptoms/behaviors in NDDs. Previous studies of brain asymmetry in NDDs are limited by the case-control and cross-sectional study designs, and the factors contributing to NDDs have yet to be studied using an integrative method. This proposal seeks to overcome the limitations of previous studies by applying our novel integrative analytic algorithms and a transdiagnostic approach to several existing public and internal longitudinal (and cross-sectional) datasets (total N~16,000), which are designed for studying brain development and related health issues and comprise extensive data of demographics, medical records, behavioral/clinical assessments, cognitive tests, genetics, and multimodal brain imaging. The proposed analyses seek to address the following aims: 1) Model the developmental mechanisms of brain asymmetry involving spatiotemporal interactions between multimodal brain asymmetry components, genetic and environmental influences and this system’s impact on children behavior/cognition in typically and atypically developing children and adolescents over time, and 2) characterize within-diagnoses trajectories and cluster biologically similar participants cutting across conventional diagnostic boundaries to understand both shared and distinct developmental trajectories of brain asymmetry and behavior/cognition in NDDs. Our study has the potential to provide definitive neurobiological models of NDDs, with practical implications for helping in reformulating current diagnostic categories of NDDs with novel disorder definitions rooted in the biology of neurodevelopment processes, and in identifying predictive biomarkers for diagnosis and the right time window for proper intervention/treatment.

NIH Research Projects · FY 2025 · 2024-05

PROJECT SUMMARY There is a growing sector of modern tobacco-free oral nicotine pouches (NPs) that are federally regulated as non-medicinal nicotine/tobacco products. While NPs employ marketing approaches that may attract current tobacco users, such as marketing themes connoting minimal harm, information on the long-term health effects of NPs is lacking. NPs may appeal to young adults because they are available in similar product characteristics (e.g., nicotine concentration, protonated nicotine) that many young people prefer to use in e-cigarettes. In addition, NPs may be of particular interest to young adult e-cigarette users because these products can be used discreetly where vaping is not allowed, which may translate into an increased likelihood of becoming dual users of e-cigarettes and NPs. Indeed, approximately 15% young adults who used EC in the past 30 days were past 30-day NP users. Manufacturers of modern NPs use acid additives to lower pH, which changes nicotine from a free-base to a protonated nicotine, resulting in improved appeal and sensory experience and higher abuse liability. Thus, nicotine concentration and pH in modern NPs should be focal targets for regulatory policies. Evidence is also lacking on mechanisms mediating differences in product appeal and abuse liability of NPs across products varying in nicotine concentration and pH level. The scientific objective of this research plan is to assess the effect of variation in nicotine concentration in NPs and its interaction effect with pH level on three proximal outcomes of relevance to FDA regulation: (1) sensory attributes, (2) product appeal, and (3) abuse liability among young adult dual users of NPs and e-cigarettes. This innovative project proposes to conduct double-blind within-subject randomized clinical trials in which participants (N≈156 [Aim 1 N = 72; Aim 2 N = 84]) will administer NPs varied by nicotine concentration (e.g., 4 vs 2 mg) and pH (e.g., ≥9 vs ≤8, ≥1 point difference) across two flavors (e.g., mint, fruit) to achieve the project aims: to evaluate the effects of nicotine concentration, pH, and flavor on subjective product appeal and sensory attributes of NPs (Aim 1); to assess the effects of nicotine concentration and its interplay with pH on abuse liability of NPs (Aim 2); to estimate the extent to which sensory attribute mediates the pH-moderated effect of nicotine concentration on product appeal and abuse liability (Aim 3). The findings of this proposed research will provide the FDA with new evidence necessary to inform regulatory restrictions on product characteristics and constituents of NPs, which may put young adults at risk of using a novel class of oral nicotine products. A career development plan that complements the research plan builds on the applicant’s background in behavioral epidemiology and quantitative analytic skills, which outlines new training in three areas: (1) clinical trial experiments testing the effects of exposure to tobacco products on behavioral responses (e.g., abuse liability), (2) product characteristics and constituents of emerging oral nicotine products, and (3) within-subject data and causal mediation analyses. The combined research and training plan will prepare the candidate as an independent investigator with expertise in tobacco regulatory science.

NIH Research Projects · FY 2025 · 2024-05

ABSTRACT Hydrogels play a key role in tissue engineering, providing a supporting structure to mimic a native extracellular matrix (ECM) micro-environment for cell adhesion, proliferation and migration. As polymer networks infiltrated with water, hydrogels are similar with human body, constituting most of their cells, extracellular matrices, tissues and organs due to hydrophilic polymer networks infiltrated with water so that they have been widely applied for diabetic wound healing, bioadhesive sealant and wearable device interface in biomedical applications. An ideal engineered biomedical hydrogel's elastic properties should match well with the native tissue that it is being integrated. In biomedical applications, mechanical interactions with dissimilar mechanical stiffness between target tissues/organs and hydrogels can cause impaired functional efficacy such as foreign- body response, tissue damage or scar formation. In tissue engineering, the mechanical stiffness of hydrogels is dynamic because numerous features can influence their elastic properties such as cell adhesion, proliferation, migration, levels of polymer molecular weights, levels of cross-linked collagens and cell density. Therefore, characterizing hydrogel mechanical stiffness over time is critical. Mechanical tests such as dynamic mechanical analysis (DMA) are the standard method to characterize hydrogel mechanical stiffness. However, the major limitations of mechanical tests are that they are destructive measurements, multiple replicate samples are required for studies with multiple time points and only global elasticity measurements are provided. These limitations lead to difficulties for exploring the optimization of mechanical stiffness of hydrogels to host tissues in biomedical and tissue engineering applications. In this proposal, 1) we will develop an ultrasound-based elasticity measurement technique named two-dimensional acoustic force elastography microscope (2D-AFEM), which can address current difficulties of mechanical tests and can repeatedly measure 2D (x, y) Young's modulus of thin-layer cell encapsulated hydrogels. 2) To study 2D-AFEM performance, we will test the 2D-AFEM elastography resolution, 2D Young's modulus of homogeneous and heterogeneous hydrogel scaffolds, and validate experimental results by numerical simulation and DMA. 3) To study how living cells affect stiffness changes of hydrogel scaffolds to obtain a desired functionality, we will use 2D-AFEM to explore elastic changes of cell encapsulated scaffolds and explore the relationship between elastic changes and biochemical markers over time for longitudinal studies. The proposed 2D-AFEM will be a promising modality to repeatedly and quickly evaluate 2D Young's modulus of living hydrogels over time to explore the best match of the elasticity to host tissues for various applications in biomedical, tissue engineering and biomaterial fields.

NIH Research Projects · FY 2025 · 2024-05

Project Summary/Abstract Development of a functional jaw requires precise shaping of the underlying skeleton and proper integration with the musculature through tendons and ligaments. In the jaw, both the skeleton and connective tissues such as tendons and ligaments derive from multipotent cranial neural crest-derived cells (CNCCs). My project focuses on two novel mechanisms by which skeletal and connective tissue fates are balanced in the vertebrate jaw. In a previous study, we revealed that the nuclear hormone receptor Nr5a2 functions to promote connective tissue at the expense of skeletal fates in both the mouse and zebrafish jaw, yet the mechanisms by which Nr5a2 represses jaw skeleton formation were unknown. By analyzing single-cell sequencing data of nr5a2 mutants, I have identified the bile acid synthesis enzyme Hsd3b7 and the transcription factor Foxp2 as two potential targets of Nr5a2 that repress Hedgehog-dependent induction of lower jaw cartilage. In Aim 1, I test that Hsd3b7 functions as a jaw-specific repressor of Hedgehog signaling by depleting the Hedgehog co- factor cholesterol from the cell membrane of tendon-forming CNCCs. In Aim 2, I test that Foxp2 competes with the Hedgehog target Foxc1, with Foxp2 closing cartilage enhancers in the tendon-forming region of the jaw that would normally be opened by Foxc1. My research will therefore show novel and jaw-specific mechanisms by which two Nr5a2 target genes function upstream and downstream of Hedgehog signaling to pattern the region of the developing jaw generating tendons and other connective tissues. These findings will shed light on how alterations to Hedgehog regulation and chromatin accessibility may underlie human birth defects affecting jaw development. The research project and training plan for the fellowship period have been designed to lay the groundwork for my long-term goal of obtaining a position as a tenure-track professor at a top-tier academic research institution. I will receive mentorship from Dr. Gage Crump, a leading scientist in zebrafish craniofacial development. The experiments in this proposal will take place on the Health Sciences Campus of the University of Southern California, which hosts vibrant communities of craniofacial and stem cell scientists.

NIH Research Projects · FY 2025 · 2024-04

Neurodegenerative dementias, such as Alzheimer's disease (AD), frontotemporal lobar degeneration, and Lewy body dementia, are caused by abnormalities in proteostasis and accumulation of misfolded prion-like proteins in the brain. Mutations in the Leucine-rich repeat kinase 2 (LRRK2) gene result in diverse neuropathology, including tauopathy, synucleinopathy, TDP-43 proteinopathy, and AD pathology. Patients with LRRK2 mutations frequently develop clinical dementia, and nearly half of patients with LRRK2-driven neurodegeneration develop tau or other pathology instead of synucleinopathy. LRRK2 is therefore a key regulator controlling protein aggregation and neurodegeneration, and defining the mechanisms by which LRRK2 mutations drive such varied prion-like pathology may lead to novel targeted therapeutics. The proposed study aims to characterize the proteostatic mechanisms upstream and downstream of LRRK2 using patient-derived neurons and rare postmortem LRRK2 patient brain tissue. The study has three goals. First, to define the role of LRRK2’s subcellular localization in its cellular functions and degradation, with a particular focus on LRRK2 microtubule association and endolysosomal activation. Second, to test the extent to which newly identified LRRK2 protein degradation pathways rescue neurotoxicity in patient-derived aged neurons and other disease relevant systems. Third, to define the mechanisms by which LRRK2 mutations can drive both synucleinopathy and tauopathy using real-time quaking-induced conversion (RT-QuIC), detailed neuropathology, and cell-type specific transcriptomics on a rare cohort of LRRK2 patient brain tissues. Overall, these studies should begin to define the mechanisms by which LRRK2 drives myriad neuropathology and provide insight into the relevance of LRRK2 as a therapeutic target for tauopathies.

NIH Research Projects · FY 2026 · 2024-04

PROJECT SUMMARY / ABSTRACT Diabetic kidney disease (DKD) develops in more than 50% of youth with type 2 diabetes (T2D) as they transition to young adulthood. Early identification of youth at risk of DKD, either due to environmental or biological factors, has the potential to inform clinical care and alter the course of disease. Despite the substantial burden of DKD in youth with T2D, however, modifiable risk factors and effective therapies remain limited. Emerging evidence links per- and polyfluoroalkyl substances (PFAS), a group of ubiquitous artificial chemicals used for more than 60 years in consumer and industrial products, with impaired kidney function, worse glucose regulation, and higher levels of uremic toxins, key risk factors for DKD in youth with T2D. This study proposes to perform the first prospective study to examine the association of PFAS exposure with DKD risk in youth with T2D and to investigate potential biological mechanisms by integrating key information on metabolites and protein levels. The project will build upon existing health and omics data (metabolomics and proteomics) in the Treatment Options for Type 2 Diabetes in Adolescents and Youth (TODAY) study, a longitudinal study of youth with T2D who underwent annual measures of kidney function for an average of 10.2 years. Using archived plasma samples, this project will measure levels of 10 common PFAS and up to 250 novel and emerging PFAS at baseline. Due to the long biological half-lives of most of these chemicals, single PFAS measures are a good proxy for medium to long-term PFAS exposures. Analyses will examine associations between individual PFAS and PFAS mixtures with risk and progression of DKD using survival analysis and mixture methods. To examine the potential biological pathways that link PFAS exposure with DKD, analyses will then examine associations of PFAS (individual and mixtures) with metabolites and proteins potentially associated with DKD risk. Further, a novel latent unknown clustering approach will be applied to comprehensively analyze PFAS exposure, multiomics, and clinical data to identify subgroups of youth with T2D at high risk of DKD. These research findings will contribute to the body of evidence needed to inform PFAS regulation and develop innovative environmental health interventions to detect and prevent DKD in youth with T2D. The culmination of my scientific training has prepared me to successfully carry out the proposed K01 research. Through a multifaceted training plan supported by a strong transdisciplinary mentoring and advisory team, this K01 will provide the necessary support for me to further develop as an independent environmental health researcher. In addition to the proposed research project, a robust yet attainable training plan incorporating didactic instruction, seminars, conferences, and mentorship will prepare me to successfully complete a future NIEHS R01 to examine personalized DKD prevention strategies in young individuals, when potential lesions may still be reversible, to promote healthier lives in this at-risk population.

NIH Research Projects · FY 2026 · 2024-04

Abstract – Molecular basis of complement anaphylatoxin receptor activation, regulation, selectivity and signaling bias The complement system is a vital part of the immune system, involved in promoting inflammation and clearance of pathogens. Dysregulation of this system has been linked to various diseases, including autoimmune disorders, neurodegenerative conditions, and cancer. Understanding the molecular mechanisms underlying complement activation and signaling is crucial for therapeutic development. This study aims to investigate the molecular basis of complement anaphylatoxin receptor activation, regulation, selectivity, and signaling bias. As part of our studies, we will obtain valuable insights through structural determination and characterization of these receptors, revealing unique binding pocket topologies and details of receptor activation, and signaling. The specific aims of the study are: 1) elucidating the mechanisms of receptor activation and beta-arrestin recruitment, 2) identifying molecular determinants of signaling bias and receptor selectivity, and 3) investigating immune response regulation and antagonism of complement receptors. The research strategy involves cryo-electron microscopy, signaling assays, and structural and biochemical techniques. The significance of this work lies in its potential to advance our understanding of complement receptor biology, provide frameworks for drug design, and offer new insights into immunomodulatory roles of these receptors. The findings could lead to the development of novel compounds and contribute to the understanding of complement receptor functions in physiology and disease contexts. This study represents one of the most comprehensive investigations of any GPCR subfamily to date and aims to address long-standing questions regarding ligand selectivity, signaling bias, regulation, and antagonism. The results will have implications for the development of tool compounds and therapeutic interventions targeting complement receptors and have the potential to impact a wide range of diseases associated with complement system dysfunction.

NIH Research Projects · FY 2026 · 2024-04

ABSTRACT Anorexia nervosa (AN) has one of the highest mortality rates of all psychiatric conditions due to long-term physical complications and a suicide rate over 50 times higher than the general population. The mortality rate encompasses a combination of the medical sequelae of underweight and death by suicide – and increases by around 5% with each decade of illness. Around one-third of people with AN develop a chronic disorder. If AN is not treated, serious complications such as heart conditions and kidney failure can arise and eventually lead to death. Despite the severity of AN, the neurobiological mechanisms that influence risk and treatment outcomes in AN are poorly understood; AN is severely understudied compared to other psychiatric conditions. Little is known about the alterations in brain circuitry and function in AN, and which brain changes predict future remission or relapse. Findings to date often raise questions related to the acute state: What are the acute and lasting effects of starvation? Which brain differences are related to the illness rather than undernutrition, and which are reversible with treatment? Which clinical measures, risk factors, and brain abnormalities are most important in understanding risk for AN and longer-term illness courses? To address these knowledge gaps, we launch the ENIGMA Eating Disorders Initiative – a global alliance to bring together clinical and neuroimaging experts in eating disorders worldwide to address pressing questions in AN research. This new international initiative is a coordinated data analysis platform to determine multimodal neuroimaging signatures of AN from a broad range of 31 clinical datasets worldwide. We build on our promising pilot data that shows 1) one of the largest effects on brain structure ever observed among psychiatric conditions, and 2) some of the brain anomalies in AN are reversible after weight restoration treatment in a subset of patients. Identifying circuits and functions that can be restored, and in which patient subgroups, is critical. In the largest worldwide dataset ever analyzed (n~1,600 cases and ~1,700 HCs), we will identify multimodal brain biomarkers of AN, its driving mechanisms, which brain abnormalities can be reversed, and in which patients. To boost statistical rigor, reproducibility and power, our coordinated international alliance will pool a broad range of brain and clinical data internationally using harmonized analytic workflows. We build on the successful launch of our ENIGMA Eating Disorders working group, which published the most highly powered neuroimaging study of AN. Our Specific Aims are to: (1) identify multimodal brain signatures of AN, with the power and range of data to understand which brain signatures are reproducible in patients across the world; (2) disentangle undernutrition effects on the brain from the aberrant brain circuitry that drives the condition and increases risk of relapse; and (3) longitudinally assess relapse and recovery, to better predict remission and risk for AN. Our Precision Medicine approach responds to NIMH’s mission to seek more reproducible, objective biomarkers of disease for individual monitoring and management, to assist prognosis, and discover factors that influence relapse, suicidality, or recovery.

NIH Research Projects · FY 2025 · 2024-04

PROJECT SUMMARY Misfolding and aggregation of mutant huntingtin exon1 (mHTTex1) promotes neurodegeneration in Huntington’s disease (HD). The intrinsically disordered mHTTex1 misfolds into a heterogeneous mixture of assemblies, however, how they form, and which conformers mediate neurotoxicity in vivo remain to be identified. This gap in knowledge is due to a lack of molecular diagnostic tools to study the oligomerization pathway of native mHTTex1 in live neurons and to determine how it may induce neurodegeneration. Conformer-specific nanobodies will enable us to perform these studies. Nanobodies are genetically encoded variable heavy domains (VHH), which are naturally produced by camelids and sharks and display potent specificity towards target antigens. This project will combine the experience of Dr. Langen’s lab in generating defined and stable helical oligomers and protofibrils of recombinant mHTTex1, with the that of Dr. Khoshnan’s lab in nanobody technology and its application to studies on the neurobiology of mHTTex1. We recently reported that among the various assemblies of recombinant mHTTex1, protofibrils and their precursors helical oligomers are neuroinvasive since they can penetrate human neurons, propagate, and induce nuclear damage. Towards identifying similar conformers in vivo, we now have isolated llama nanobodies, which bind to helical oligomers, and protofibrils of recombinant mHTTex1. We plan to investigate the binding properties of these nanobodies and develop diagnostic tools like chromobodies by fusing them to GFP or RFP. These biosensors will be used to study the early steps of native mHTTex1 oligomerization in live human neuronal stem-cell derived neurons, and examine whether helical oligomers and/or protofibrils are neurotoxic. Nanobodies, which may inhibit the oligomerization of mHTTex1, will be tested for ability to prevent aggregation and toxicity in the same model. This project has the potential to make a significant contribution to our understanding of mHTTex1 oligomerization and neurotoxicity in a physiologically relevant model, identify therapeutic modalities, and provide a set of novel molecular diagnostic tools for the broader HD research community.

NIH Research Projects · FY 2026 · 2024-04

Pressing Issues Page 5 of 49 Abstract The use of repeated, momentary, real-world assessment methods known as the Experience Sampling Method and Ecological Momentary Assessment (EMA) have been broadly embraced over the last few decades. These methods have extended our assessment reach beyond lengthy retrospective self-reports as they can capture everyday experiences in their immediate context, including affect, behavior, symptoms, and cognitions. In this paper we evaluate nine conceptual, methodological, and psychometric issues about EMA with the goal of stimulating conversation and guiding future research on these matters: the extent to which participants are actually reporting momentary experiences; respondents’ interpretation of momentary questions; the use of comparison standards in responding; efforts to increase the EMA reporting period beyond the moment to longer periods within a day; training of EMA study participants; concerns about selection bias of respondents and selection bias of moments; the reliability of momentary data; and, for which purposes EMA might be considered a “gold standard” for assessment. Resolution of these issues should have far-reaching implications for advancing the field. 119 Pressing Issues Page 6 of 49 Introduction Behavioral science research has resulted in scores of psychometrically sound self-report instruments about thoughts, opinions, feelings, events, and behaviors that are intended to be summaries of significant time periods (retrospective reports) or to characterize a person’s usual levels and dispositions across situations or over time (trait or global reports). In light of some limitations of these traditional assessment options, researchers have long voiced desire for methods to collect higher resolution data with greater ecological validity (Brunswik, 1941). Higher resolution is desired so that associations between immediate contexts and experiences can be examined and so that dynamic processes occurring over relatively short time periods (minutes, hours, days) can be explored; greater ecological validity is desired so that observed associations and processes are representative of respondents’ everyday lives. These aspirations have yielded a collection of within-day momentary data capture methodologies. Selected examples of, and reviews about, momentary research are presented in the following papers: (Bolger, Davis, & Rafaeli, 2003; Conner & Barrett, 2012; Csikszentmihalyi & Hunter, 2003; Degroote, DeSmet, De Bourdeaudhuij, Van Dyck, & Crombez, 2020; DeVries, 1987; Ebner-Priemer & Trull, 2009; C. D. Fisher & To, 2012; Gorin & Stone, 2001; Hamaker & Wichers, 2017; Heron, Everhart, McHale, & Smyth, 2017; Kirtley, Lafit, Achterhof, Hiekkaranta, & Myin-Germeys, in press; Myin-Germeys et al., 2009; Reis & Gable, 2000; Scollon, Kim-Prieto, & Diener, 2009; S. Shiffman, A. A. Stone, & M. R. Hufford, 2008b; J. Smyth, Juth, Ma, & Sliwinski, 2017; J.M. Smyth & Stone, 2003; Stone & Broderick, 2007; Stone, Obbarius, Junghaenel, Wen, & Schneider, 2021; Stone & Shiffman, 1994; Stone, Shiffman, Atienza, & Nebling, 2007; Trull & Ebner-Priemer, 2020). Several sources provide summaries and discussion of the analytic techniques used for these complex data (Bolger & Laurenceau, 2013; Mehl & Conner, 2011; Schwartz & Stone, 2007; Shiffman, 2014). A core feature of Ecological Momentary Assessment (EMA) methods is the brief periods about which respondents report; this is meant to reduce bias and error attributable to inherent memory limitations and to limit the use of cognitive heuristics in self-reports. Relatively unobtrusive in-the-field data collection as respondents go through their everyday lives is intended to ensure ecological validity. Following a first generation of paper-and-pencil methodologies (DeVries, 1987), advances in smart phone, Internet, and computer-assisted applications have further increased the appeal of 120

NIH Research Projects · FY 2026 · 2024-04

Many high-resolution structures of the cross-β core of amyloid fibrils have been solved using solid-state NMR and cryo-electron microscopy in recent years. However, most fibrils important in neurodegenerative diseases, as for example α-synuclein (aSyn) fibrils found in in Lewy body dementia (LBD) and other synucleinopathies, have sizable intrinsically disordered regions (IDRs) surrounding their ordered cross-β fibril core. These IDRs are part of the fibril surface, where they can interact with fibril-specific binders and be important for fibril toxicity. Therefore, determining the residual structure and dynamics of these IDRs, how they interact with other cellular components, and how they relate to fibril toxicity is the next logical step in the LBD and Alzheimer’s disease related dementia (ADRD) field. The goal of this proposal is to determine the conformational ensemble and dynamics of the IDRs from aSyn fibrils important in LBD, ADRD and other synucleinopathies. The N and especially the C-terminus of aSyn are intrinsically disordered in the fibril. These IDRs are binding sites for fibril-specific interactors such as the co-chaperone DNAJB1. Our central hypothesis is that a specific fibril core structure (polymorph) will determine the residual structure and dynamics of these IDRs and consequently the interaction with fibrils-specific binders. The rationale of this research is that only complete molecular models of amyloid fibrils that include their IDRs will allow us to determine polymorph-specific binding partners, which can explain the difference between LBD and other synucleinopathies. These complete molecular models will not only point to natural interactors, but also to disease-specific biomarkers and therapeutics for LBD as well as ADRD and other synucleinopathies. We will use a combination of solid-state NMR, EPR, and molecular dynamics simulations to test our hypothesis using three specific aims. Aim 1 is to determine the change in residual structure and dynamics of IDRs upon fibril formation. Based on the known core structure and our conformational ensemble of the IDRs, we will create a model of the entire fibril. Aim 2 is to determine the effect of fibril polymorphs on residual structure and dynamics of IDRs. We will show to what degree a specific cross-β core determines the conformational ensemble of its adjacent IDRs and determine the cytotoxicity of different aSyn polymorphs and chimeras. Aim 3 is to determine the effect of the fibril core on aSyn-DNAJB1 interaction. Here, our hypothesis is that fibril formation increases the accessibility of the DNAJB1 binding site and that some fibril cores found in LBD and other synucleinopathies do this better than others. These aims will (i) determine the changes in the IDRs outside the fibril core upon fibril formation and result in a whole fibril model. We will (ii) learn how specific cross-β core structures found in LBD and other synucleinopathies change the conformational ensemble and dynamics of the IDRs, and (iii) we will understand how these changes influence the interaction of specific binders, in our case DNAJB1. Together these advances will facilitate the development of new approaches to diagnose and treat LBD and ADRD.

NIH Research Projects · FY 2026 · 2024-04

Project Summary The functional significance of actin in cellular health and fitness is unquestionable, as the actin cytoskeleton is required for a diverse array of cellular processes as simple as movement and motility. The loss of function of actin is seen in clinical manifestations in many diseases, and specifically during the aging process. However, these studies are limited to the cytoplasm, despite actin being identified in the nucleus over 50 years ago. The lack of studies of nuclear actin during aging is likely owning to the poorly understood mechanistic function and the available tools to study nuclear actin. In fact, even studies of cytoplasmic actin during aging is still a burgeoning field, making the studies of nuclear actin even more elusive. This highly innovative proposal aims to tackle this major black hole in the field, leveraging our expertise in cytoplasmic actin to shift our research direction into studying nuclear actin in DNA repair and transcriptional regulation. Our first goal is to synthesize a robust method to visualize the quality, function, and dynamics of nuclear actin during stress and aging in C. elegans. As a new field, there are still no existing tools to reliably study nuclear actin in perhaps the most powerful genetic model organism for aging research. Therefore, we propose to synthesize multiple methods to reliably stain the numerous variants of nuclear actin. Furthermore, we will leverage diverse genetic methods to directly study how changes to nuclear actin stability, function, and dynamics can impact the aging process. We hypothesize that nuclear actin dysfunction during aging contributes to physiological consequences and perturbing nuclear actin can result in premature aging. Emerging studies have implicated nuclear actin in DNA and RNA regulation, quality control, and function, which is unsurprising considering the contents of the nucleus. Here, we aim to study the hypothesis that nuclear actin can influence DNA repair. Specifically, we hypothesize that nuclear actin filaments form under stress to recruit DNA repair machinery, and breakdown of this process is what leads to genomic instability and decline of organismal health during aging. As an alternative hypothesis, we also propose that nuclear actin may play an important role in transcriptional fidelity. Like DNA integrity, transcriptional fidelity is important to prevent the synthesis of mutated proteins that can aggregate and cause physiological consequences during aging. Thus, we argue that nuclear actin can drive protein homeostasis by preventing transcriptional errors, and the breakdown of this functional process during aging can result in age-related disease. Ultimately, we propose to venture into two new research directions for our lab: nuclear actin and DNA repair. Moreover, we propose to merge these two novel fields into a highly innovative study understanding how nuclear actin impacts transcriptional fidelity and vice versa, creating a unique research proposal perfectly in line with the vision of the Stephen I. Katz Early Stage Investigator Research Project Grant.

NIH Research Projects · FY 2026 · 2024-04

PROJECT SUMMARY Despite the widespread use and success of [18F]FDG-PET imaging in oncology, there are many situations where [18F]FDG cannot be used for diagnosis or treatment monitoring, especially for cancers where there is high background uptake, low tissue density, or slow-growing tumors. Infection or inflammation frequently leads to false positives. Many tracers are being developed that provide improved contrast, sensitivity, and accuracy compared to [18F]FDG. Of particular interest is the nucleoside analog [18F]FMAU, which is incorporated into DNA when tumor cells divide, thus directly measuring increased cell proliferation, a universal hallmark of cancer. Promising clinical data of [11C]FMAU and preclinical data of [18F]FMAU have led to initial clinical studies of [18F]FMAU in cancer patients, and there is high interest in clinical studies of [18F]FMAU PET as a means to assess treatment response in diverse cancer types. However, the radiosynthesis of [18F]FMAU is very challenging, hindering translational efforts such as multicenter clinical trials that are needed to ensure sufficient recruitment of patient populations. Over many years, Dr. Chen’s lab at USC has improved the [18F]FMAU synthesis using a 1-pot process, but it still involves lengthy reaction steps (~3 h synthesis), uses corrosive and unstable reagents that are not compatible with automated synthesis modules, and has a relatively low activity yield (~5%), making adoption of this approach extremely difficult. To widely enable clinical studies, this proposal leverages an innovative droplet-radiochemistry approach developed in Dr. van Dam’s lab at UCLA, and now being commercialized by DropletPharm, in which reactions are performed in 10 µL volumes instead of 1 mL volumes, enabling higher isotope concentration, higher reaction yields and much shorter synthesis time. A preliminary study showed [18F]FMAU could be produced in <1 h. Our central hypotheses in this project are that cell proliferation imaging can provide a superior assessment of treatment response (e.g. earlier response detection and higher specificity to tumor growth) than other tracers, and that droplet technology can facilitate widespread access to [18F]FMAU for clinical translation. Leveraging synergistic academic and industry expertise and capabilities, our multi-PI team will advance the clinical translation of [18F]FMAU via four complementary aims: (1) Develop an optimized droplet radiosynthesis of [18F]FMAU at multi-dose scale; (2) Customize a droplet radiosynthesizer for [18F]FMAU production, then automate and validate the synthesis; (3) Pursue preclinical studies to evaluate [18F]FMAU for assessing treatment response (compared to [18F]FDG and [18F]FLT) to support future clinical trials; (4) Enable wide availability of [18F]FMAU by providing a low-cost self-shielded synthesizer, developing reagent/consumable kits, establishing a user training program, and compiling an IND amendment that can be referenced in future trials. We believe that quantitative [18F]FMAU-PET imaging will provide early and accurate assessment of cancer treatment leading to individually tailored therapeutic plans with improved patient outcomes.

NIH Research Projects · FY 2025 · 2024-04

This is a project to study to a single nucleotide polymorphism (SNP) in the aryl hydrocarbon receptor repressor (AHRR) gene that may influence the lung cancer risk of Black/African American (B/AA) subjects. AHRR is a negative regulator of detoxification responses involved in the metabolism and elimination of tobacco smoke carcinogens. B/AA men have the highest rate of lung cancer death compared to all other groups; they show a 12% higher lung cancer incidence rate and a 15% higher lung cancer death rate than White/European (W/E) men. Elevated AHRR expression has been implicated in lung cancer risk; thus, a SNP that improves AHRR function would likely increase lung cancer risk. We have identified a missense SNP, rs35008248, in AHRR exon 4 that, while rare in W/E subjects, is present one third of B/AA subjects. The SNP was not included in most previously used arrays and has only recently begun to be studied. Preliminary data from GWAS explorer shows no significant associations in W/E subjects but suggests an association with smoking in B/AA males (p=3.87E- 3) and with smoking-related cancers in B/AA subjects (p=0.04927). As B/AA individuals comprise only a small fraction of the subjects (<4%), power to determine the role of SNPs in this group has been limited. AHRR is a negative regulator of the aryl hydrocarbon receptor (AHR). Upon tobacco smoke exposure, AHR binds to polycyclic aromatic hydrocarbon, enters the nucleus, and dimerizes with ARNT. The AHR/ARNT dimer binds to xenobiotic response elements (XREs), switching on genes involved in the inactivation/secretion of xenobiotic compounds. In a negative regulatory loop, AHR/ARNT turns on AHRR, which also binds to ARNT. The transcriptionally inactive AHRR/ARNT complex competes with AHR/ARNT for XRE sites, dampening the detoxification response. The reference allele of rs35008248 (T) encodes a leucine at position 114, while the alternate allele (C) encodes a turn-inducing proline. Combined annotation-dependent depletion (CADD) suggests the SNP might affect AHRR function. In the co-crystal structure of the AHRR/ARNT heterodimer, leucine 114 borders a loop entwining ARNT. We hypothesize that a proline at position 114 constrains AHRR loop conformation, thereby stabilizing AHRR/ARNT interaction, increasing the repressive effect of AHRR/ARNT heterodimers, interfering with the detoxification response, and increasing lung cancer risk of B/AA smokers, 34% of whom carry the minor allele. Here we propose to test for associations between genetic variation at AHRR with lung cancer outcomes using existing large-scale genetic data in B/AA individuals (Aim 1), to use CRISPR-based genome editing to engineer the minor allele into the genome of our unique immortalized human alveolar epithelial cell lines and assess its effects on the response to tobacco smoke in vitro (Aim 2), and to test whether the amino acid at position 114 affects the kinetics of the interaction between AHRR and ARNT (Aim 3).Together, the specific aims of this pilot project will provide insight into the role of a potential functional SNP in AHRR that is common in B/AA subjects and that could help explain their increased lung cancer risk.

NIH Research Projects · FY 2026 · 2024-04

SUMMARY Brain imaging approaches are a promising method for unveiling network abnormalities in living brains of patients with psychiatric disorders; however, understanding these pathologies will only be possible when human specific abnormalities in cell connectivity, network circuit activity and transcriptomes can be directly interrogated and correlated in an experimental system amenable to high-throughput manipulation and phenotyping. Despite the excitement surrounding human brain organoids derived from pluripotent stem cells, scientists are frustrated with the limited physiologically relevant anatomy and maturation of brain organoids. We propose to pioneer a multi-organoid-on-chip (MoC) platform, combining tissue engineering technologies and newer organoid analysis readouts, to address this challenge. As a first proof of concept study, we propose to recapitulate the human visual system, a commonly affected network in multiple neuropsychiatric disorders, by co-developing three (retinal, thalamic, cortical) organoids on a microfluidic device designed to foster functional connectivity, thereby allowing: i) co-development of distinct central nervous system (CNS) structures with distinct anatomical organizations; ii) establishment and mapping of reproducible and functional long-range connectivity between defined CNS structures; iii) activity-dependent maturation of synaptic connections by spontaneous, optogenetically-regulated, and sensory-evoked stimulation of neuronal activity. The synthetic platform will also facilitate high-throughput multimodal analysis with direct correlation between specific circuit connectivity and molecular dynamics at single cell resolution, in healthy and diseased organoids. In addition to improving the understanding and screening of drugs for complex brain disorders, our platform provides a valuable resource for the broader research community interested in modeling circuit-level dysfunctions in other types of neurological disorders, including retinal diseases and optic neuropathy. Finally, we expect that the method and modular technologies developed within this proposal will be foundational and easily adapted to recapitulate connectivity between other brain regions, thereby broadening its future impact and applications.

NIH Research Projects · FY 2026 · 2024-04

Project Summary The complexity and dynamics of protein kinases and epigenetic modifications in cellular processes necessitate the development of precise biosensors for live imaging. This proposal aims to develop single-FP-based, high- performance, ultrasensitive biosensors through directed evolution in mammalian cells. These biosensors will be used in multiplexed imaging and dynamic visualization of signaling activities in situ, with a specific focus on improving chimeric antigen receptor T cells (CAR-T) for cancer immunotherapy. Addressing the limitations of CAR-T therapy, particularly T cell exhaustion in solid tumors, requires a better understanding of the molecular mechanisms involved. Although kinases and epigenetic markers, particularly H3K27me3, play key roles in T cell regulation, our understanding of their spatiotemporal dynamics during cancer-immune interactions and through the course of CAR-T cell rejuvenation remains limited due to the absence of appropriate investigative tools. Thus, parallel examination of these key regulators in cancer-immune interacting environments should reveal new insights into the systematic behaviors and identify essential links for therapeutic manipulation. My hypothesis is that 1) the reversal of CAR-T cell exhaustion involves a rejuvenation of ZAP70 and Lck kinase’s function and a reprogramming of the H3K27me3, transitioning from an exhausted state to a naïve T cell-like state; and 2) transient knockdown of exhaustion-related genes could prevent and reverse exhaustion of CAR-T cells, which can be reflected by the kinase and epigenetics coordinated response patterns. Leveraging the combined expertise of my mentors, I have demonstrated the feasibility of engineering single-FP biosensors for tyrosine kinases, directed evolution of single-FP biosensors in mammalian cells and established a transient gene knockdown system that can be remotely controlled by focused ultrasound (FUS). Building on these achievements, three distinct aims have been further proposed: Aim 1 focuses on engineering novel single-FP prototype biosensors for monitoring tyrosine kinases or epigenetics in CAR-T cells of different phenotypes during cancer cell engagement and establishing a multiplexed imaging platform. Aim 2 involves developing ultrasensitive single-FP biosensors through directed evolution, high-throughput screening, and next- generation sequencing. Aim 3 focuses on the application of these biosensors for multiplexed imaging and manipulation of kinase-epigenome signaling in CAR-T cells using a FUS controllable gene knockdown system. This biosensor engineering platform can potentially be extended to develop any other kinase and epigenetic biosensors for live cell imaging. Similarly, the novel FUS-controllable gene knockdown system could be generalized for broader manipulations of various cellular processes. Successful execution of this project could revolutionize biosensor engineering and kinase imaging, profoundly impacting our understanding and treatment of cancer and other diseases.

NIH Research Projects · FY 2025 · 2024-04

SUMMARY Upper Tract Urothelial Carcinoma (UTUC) is a rare disease that accounts for 5-10% of all urothelial tumors with an estimated annual incidence in Western countries of almost two cases per 100,000 inhabitants. The 5- year specific survival is < 50% for stage 2-3 patients but < 10% for those with stage 4 disease. This poor survival rate demands improvements in preoperative detection. Diagnosis and preoperative risk-stratification of UTUC patients present distinct challenges given the limitations of current available tools. Although urine cytology has an acceptable specificity to detect UTUC, the sensitivity is relatively low, particularly in low-grade carcinomas and pelvicalyceal tumors. The invasive ureteroscopy (URS) biopsy can enhance the diagnostic accuracy, yet under-grading and under-staging rates remain a concern at 32% and 46%, respectively. Nowadays, clinical decision-making and the prognosis of UTUC relies heavily on TNM stage and pathological grade that is not accurately available until radical nephroureterectomy (RNU) is performed. Major improvements in predictive tools have occurred in recent years and unique parameters have been proposed to distinguish between low- and high- risk tumors. DNA methylation alterations have emerged as potential diagnostic and prognostic factors for urothelial carcinoma at the molecular level. For example, bladder cancer-specific DNA methylation changes can be detected in urine sediments and can be used as non-invasive diagnostic or surveillance markers. Thus, a personalized medicine approach for the diagnosis and prognosis of UTUC is urgently needed. The goal of this study is to develop DNA methylation biomarkers to identify aggressiveness of UTUC. In a preliminary study, we identified differential DNA methylation patterns in a small group of patients who had exhibited DNA markers for UTUC aggressiveness. We hypothesize that bladder cancer DNA methylation marker panels can be used to: 1) characterize UTUC-specific and/or aggressive UTUC specific DNA methylation markers in primary UTUC specimens (Aim 1); 2) validate these markers in urine sediments of UTUC patients for monitoring disease diagnosis, prognosis and recurrence (Aim 2). Results from this proposal will support further independent investigations into the role of DNA methylation markers in UTUC diagnosis and prognosis, improve the standard of care in predictive tools and different parameters have been proposed to distinguish between low- and high- risk tumors and provide UTUC-specific DNA methylation profiles that aid physicians’ decisions.

NIH Research Projects · FY 2026 · 2024-03

Project Summary One in every five youth are living with obesity, placing them at increased risk for developing diabetes and cardiovascular disease. Identifying risk factors contributing to obesity is extremely critical so that prevention strategies can be taken early to mitigate the obesity risk. Maternal diabetes in pregnancy is a strong risk factor for offspring obesity. Multimodal neuroimaging has the potential to reveal important mechanistic insights into the link between prenatal exposure to maternal diabetes and offspring obesity, but most existing studies possess major flaws including: 1) cross-sectional design evaluating later developmental periods, introducing uncertainty around the influence of other perinatal exposures; 2) poor accounting for the effects of maternal obesity; 3) low statistical power and overly homogeneous populations; and 4) use of a single imaging modality, limiting our ability to understand the complexity and full scale of brain abnormality associated with exposure to maternal diabetes. By integrating noninvasive structural and functional magnetic resonance imaging (MRI) with developmental neuroscience techniques and targeting the critical period of development, the first 1000 days of life, we aim to test the hypothesis that prenatal exposure to maternal diabetes will be independently associated with altered brain development during very early childhood, and that maternal obesity will further exacerbate these effects. We further hypothesize that these brain alterations will contribute to a higher risk of obesity early in life. To explore this hypothesis, the applicant, and her team plan to leverage longitudinal brain and body weight and length data during the first ~1000 days of life from 8 existing cohorts that participated in NIH-funded The Organization for Imaging Genomics in Infancy (ORIGINs) consortium, part of the Enhancing Neuroimaging Genetics through Meta-analysis (ENIGMA). We aim to examine effects of maternal diabetes exposure either separately or together with maternal obesity on brain metrics at birth (Aim 1) and brain developmental trajectories from birth to 2-3 years of age (Aim 2). Furthermore, we will discriminate exposed vs. un-exposed offspring (Aim 3) with replication by using innovative machine learning algorithms and identify multi-modal imaging markers that predict obesity by age 2-3. This will be the largest and most highly powered neuroimaging study to identify robust multi-modal brain signatures of prenatal exposure to maternal diabetes, thereby enhancing our understanding of etiologic/causal pathways of obesity development.

NIH Research Projects · FY 2026 · 2024-03

PROJECT SUMMARY/ABSTRACT We have recently identified that rifabutin (RBT) is hyper potent against A. baumannii and we have described this unique mechanism of action. We have characterized the mechanism for the hyperactivity and found that RBT hijacks the A. baumannii iron transport protein FhuE, resulting in a Trojan horse-like, active accumulation in the bacterial cell. However, FhuE expression is suppressed in the presence of rich media, containing a high amount of free iron, and is expressed only in nutrient depleted conditions (e.g., RPMI-1640 media and in vivo), explaining why RBT has been previously overlooked. There is clinical interest in the translation of RBT for A. baumannii therapy and BV100 (RBT IV formulation) is currently being studied in a phase I clinical trial (NCT04636983) and phase II clinical trial (NCT05685615). However, significant barriers remain for the adoption of RBT as a treatment for A. baumanniii infections. The goal of this proposal is to further support clinical translation through the rigorous testing of a broad panel of clinical isolates, to develop a preclinical mouse model to model key RBT humanized pharmacokinetics (PK) parameters to support defining MIC breakpoints, and to establish a high-throughput RBT susceptibility testing protocol with low iron media that can be adopted by clinical labs according to CLSI standards. Specific Aim 1: Develop and validate a RBT susceptibility testing method according to CLSI standards. A) Head-to-head comparison of disc diffusion, or iron-depleted MHII and RPMI-1640 broth microdilution for RBT susceptibility testing across independent testing sites. B) Validate quality control (QC) strains for RBT susceptibility testing across independent testing sites. C) Using our optimized testing method, determine MICs for 250 international A. baumannii clinical isolates. Specific Aim 2: Characterize RBT-antibiotic drug combinations in vitro to identify synergy and antagonism. A) Determine RBT-antibiotic interactions for antibiotics using a checkerboard assay. B) Determine bacterial killing and frequency of resistance emergence using a hollow fiber infection model. Specific Aim 3: Characterize RBT-containing therapeutic combinations in murine blood and lung infection models. A) Determine if RBT-containing antibiotic combinations show in vivo synergy when measuring CFUs as an endpoint in murine blood and lung infection models. B) Determine if RBT-containing antibiotic combinations show in vivo synergy when measuring time to moribund condition as an endpoint in murine blood and lung infection models.

NIH Research Projects · FY 2026 · 2024-03

PROJECT SUMMARY Head and Neck Squamous Cell Carcinoma (HNSCC) is an aggressive malignancy derived from stratified squamous epithelium of the oral cavity. Leukoplakia is a premalignant condition and is recognized as the precursor lesion of HNSCC. During leukoplakia-associated neoplastic evolution, benign squamous epithelium first becomes dysplasia, then carcinoma in situ, and finally progresses to invasive SCC. Therefore, leukoplakia serves as an ideal premalignant model for the investigation of the step-wise neoplastic evolution of oral squamous epithelial cells. However, our understanding of the molecular mechanisms promoting leukoplakia- associated neoplasia remains quite limited. This shortcoming is largely due to viable and valid human models representing this unique pathological transition have been lacking. To address this key challenge, we have developed two independent, cross-species organoid systems modeling the step-wise neoplastic evolution of HNSCC. Both metabolic reprogramming and epigenomic deregulation are cancer hallmarks. Here we have mapped a key mechanistic crosstalk between the methionine cycle and epigenomic reprogramming, through a novel cascade involving LAT1-methionine-EZH2. Specifically, we find that HNSCC exhibits the highest activity of this cascade among all human cancers. High LAT1 expression correlates with poor survival of HNSCC patients. Importantly, this cascade is indispensable for the survival and proliferation of HNSCC cells, representing a tumor-specific vulnerability. Notably, this novel LAT1-methionine-EZH2 cascade can be targeted by either pharmacological approaches or dietary intervention. These interesting and significant findings prompt us to hypothesize that the novel LAT1-methionine-EZH2 cascade functionally promotes both early precursor transformation and cancer development, representing a prominent actionable target for cancer prevention and treatment. To test this hypothesis, we will leverage our versatile premalignant and tumor organoid systems as well as animal models, to investigate the biological significance of the LAT1-methionine-EZH2 cascade during both the initial steps of oral neoplastic transformation and fully-established HNSCC tumors. We will establish the molecular basis of this pathway using advanced metabolomic profiling and epigenomic approaches such as Cut&Tag. We will perform preclinical evaluation of pharmacological and dietary blockade of this cascade, to rigorously assess both tumor-prevention and tumor-killing effect of candidate approaches. We will further develop LAT1-high vs LAT1-low organoid models to investigate whether LAT1 serves as a biomarker to predict cellular dependence on the LAT1-methionine-EZH2 axis. This study not only investigates fundamental cancer biology, but also has significant translational implications for both early intervention and treatment of HNSCC.

NIH Research Projects · FY 2026 · 2024-03

PROJECT ABSTRACT Sensorineural hearing loss is a major public health issue; by 2050, over 900 million people will have some form of auditory impairment. The spiral ganglion nerve (SGN) plays a crucial role in hearing by transmitting acoustic signals from the inner ear to the brain. Because the auditory nerve lacks intrinsic regenerative capacity, damage to the nerve leads to permanent deafness. Cochlear implants are ineffective when the SGN is damaged or lost, as observed in auditory neuropathy (AN) cases. Therefore, there is a critical need to develop novel treatments for AN, and regenerative medicine holds enormous potential to restore SGN and treat this condition. The spiral ganglion glial cells support, nourish, and protect the SGN and are critical for auditory nerve function, development, and homeostasis. Direct neuronal reprogramming converts somatic cells to induced neurons by overexpression of neuronal transcription factors (NTFs), bypassing the pluripotent state. The spiral ganglion glial cells are considered a promising source for cellular reprogramming in the inner ear because of their plasticity, proliferative capacity, survival post-nerve damage, and proximity to the nerve. Recent work has shown that neonatal spiral ganglion glial cells can be converted into SGNs by overexpression of NTFs in vitro and in vivo. However, despite administering multiple NTFs, reprogramming spiral ganglion glial cells remains inefficient. Recent cellular reprogramming strategies incorporate neuroregulatory microRNAs (miRNAs) to enhance the conversion of fibroblasts into induced neurons in combination with NTFs. The role of these miRNAs in regulating SGN fate is unknown. Our primary goal in this grant proposal is to convert spiral ganglion glial cells into SGNs via gene therapy and to advance cellular reprogramming in the inner ear. We identify three knowledge gaps in advancing cellular reprogramming in the inner ear: 1) We know very little of the glial cell response after SGN damage, 2) driving exogenous transgenes specifically into glial cells is challenging, and 3) reprogramming of spiral ganglion glial cells into neurons is very inefficient, and it is not clear if this generates the correct neuronal subtypes. We hypothesize that spiral ganglion glial cells can be converted to SGNs in normal and deafened mice with a combination of NTFs and miRNA delivered to the glial cells via an AAV vector. The aims of this project are (1) the characterization of age-related glial cell response to acute SGN apoptosis by creating a neonatal mouse model of AN, (2) to optimize exogenous transgene delivery into cochlear glial cells in neonatal mice, and (3) explore direct reprogramming of normal or damaged glial cells in vitro. The expected results of this study will be (1) an improved understanding of age-related changes in reactive gliosis in mice in response to SGN death, (2) improved targeting of cochlear glial cells with a combination of glial cell-specific AAV serotypes and promoters, and (3) providing experimental evidence of the role of NTFs and miRNA in inducing SGNs from normal and post-damaged glial cells.