Indiana University Indianapolis

NIH Research Projects · FY 2026 · 2024-12

ABSTRACT Hematopoietic stem cells (HSCs) generate all mature blood cells while reproducing themselves in a process, namely self-renewal. Their presence in the bone marrow (BM), mobilized peripheral blood, and cord blood (CB) has allowed their utility in the treatment of both malignant and non-malignant hematopoietic diseases via transplantation. However, the rarity of HSCs and the difficulty of expanding them can be a limitation for their applications. Thus, a better mechanistic understanding of HSC regeneration that might contribute to their homing, engraftment, and self-renewal is vital to improving HSC-based therapies. In the bone marrow, HSCs reside in a microenvironment where the oxygen tension is low (1~5%). Previous work showed that the low oxygen tension (hypoxia) provides a metabolic niche to facilitate HSCs’ functionality in vivo or enhances HSC expansion in vitro. Our recent work from mouse BM cells revealed a significant loss of phenotypical and functional HSCs if harvested in ambient air (normoxia). In contrast, HSCs collected and processed under hypoxia demonstrated higher engraftment and self-repopulating capacity. While these findings are interesting, mechanisms behind this distinction are still poorly understood but have the potential to improve hematopoietic cell transplantation (HCT) and gene therapy. The central goal of this proposal is to understand the signaling and metabolic alterations in HSCs under hypoxic conditions and to explore the potential to manipulate these processes in normoxic conditions to improve HSC functionality. We hypothesize that hypoxia alters the expression of genes involved in regulating lysosomal function, mitochondrial integrity, and iron metabolism to support self-renewal and prevent oxidative damage or differentiation. We have three major aims. In Aim 1, we will determine how hypoxia activates TFE3, a master transcriptional factor for lysosomal biogenesis, to facilitate self-renewal. We will specifically focus on the upstream regulatory elements, such as amino acid availability, and the function of a few TFE3 targeted genes in this process. In Aim 2, we will determine the role of Rho-associated kinase 1 (ROCK1) in regulating mitochondrial fission, a process that is suppressed by hypoxia to prevent oxidative damage in HSCs. In Aim 3, we will explore the role of iron metabolism in driving oxidative damage and differentiation of HSCs, and how a hypoxic environment prevents these processes. In this aim, we are particularly interested in exploring iron chelator as a potential treatment to improve HCT.

NIH Research Projects · FY 2026 · 2024-12

Project Summary/ Abstract Hematopoietic cell transplantation (HCT) is a life-saving treatment for disordered hematopoiesis, including ma- lignant and non-malignant diseases. Umbilical cord blood (CB) is a promising source of hematopoietic stem cells (HSCs) for HCT; however, CB HCT is limited by the low HSC numbers present in single CB units. Thus, there is a critical need to improve the functional competency and/or number of functional HSCs in each CB unit, and treatments to enhance HCT must be improved or supplemented with new treatments. The applicant's long-term goal is to establish himself as an independent investigator at an outstanding academic research in- stitution where his studies will focus on identifying ways to improve HSC function for enhanced HCT, with the goal of discovering novel therapeutic modalities to improve treatment for disordered hematopoiesis for im- proved patient outcomes. The investigator’s immediate career objectives are to successfully complete his post- doctoral training and transition to an independent investigator position. He will accomplish this by seeking guid- ance from expert mentors, improving his technical scientific skillset and developing career skills important to an independent investigator position. As an independent investigator, he will seek to provide insight into manners to improve HSC function using omics approaches supported with cell biology, biochemistry and mouse model- ling of HCT approaches. The goal of the associated research plan is to elucidate molecular programs that can be targeted to enhance CB HSC function and to generate sufficient preliminary data to submit a competitive R01 application in the final year of the award. Specifically, the research plan proposes examining combinations of treatments known to enhance HSC function for HCT, such as Dipeptidyl peptidase 4 (DPP4) inhibition and physioxic isolation of HSCs, to determine if these combinations are additive, synergistic, or have no further ef- fect. The proposed study will examine transcriptomic and epigenomic changes induced by treatments known to improve HSC function and the transcriptome of recently homed and early engrafted HSCs in a mouse model of HCT will be analyzed compared with the pool of cells transplanted to recipient mice. This data will yield insights into common and unique gene programs that are important for HSC function and engraftment. This study will provide candidate genes that can be targeted by inhibition or stabilization to improve HSC function and engraft- ment and will examine their importance using mouse models for HCT. Taken together, this approach will iden- tify new potential treatment modalities to enhance HCT. The applicant’s strong career development plan, guid- ance from his outstanding mentoring team, the environment at the internationally recognized top tier research institution of Indiana University School of Medicine, and completion of the proposed aims in the research ap- proach will prepare him for a productive and highly impactful career managing an academic lab focused on molecular mechanisms that can be exploited to improve patient outcomes for disordered hematopoiesis.

NIH Research Projects · FY 2026 · 2024-12

PROJECT SUMMARY CHARGE syndrome is a congenital disorder characterized by dysmorphic features of inner ear structures, and caused primary by sporadic mutations in CHD7, a gene encoding an ATP-dependent chromatin remodeling enzyme. Results from our recent study revealed distinctive phenotypic differences between CHD7 KO and S834F mutant inner ear organoids, suggesting that CHD7 regulates inner ear development through multifaceted and context-dependent mechanisms. To investigate the chromatin remodeling-independent function of CHD7, we will establish human embryonic stem cell lines harboring the D1812G mutation in a putative WDR5-binding domain or the G1982W mutation in the SANT domain. Gene and protein expression profiles of D1812G and G1982W cochlear organoids will be compared with those of S834F organoids with abolished remodeling activity. Our previous results also revealed down-regulation of multiple deafness genes in CHD7-null otic progenitors. To address if down-regulation of known deafness genes could account for sensorineural hearing loss in individuals with CHARGE syndrome, we will test if these deafness genes are downregulated in hair cells of cochlear organoids carrying defective CHD7. Ultrastructural and functional properties of mutant hair cells will be compared to those of wild-type hair cells. Additionally, cell lineage tracing will be performed to characterize drifted otic lineage specification in mutant organoids. To elucidate the mechanisms underlying CHD7-dependent transcriptional regulation during inner ear development, we will determine CHD7-binding loci, their transcriptional activity states and chromatin accessibility in human otic progenitors through CUT&RUN, scATAC-seq and scRNA-seq. A potential role for CHD7 in posttranslational histone modifications will be assessed by immunoprecipitation and ChIP-PCR. To assess possible contributions of CHD7-dependent cell non-autonomous factors to otic progenitor differentiation, we will employ a proteomic analysis of organoid conditioned media and test if some of the identified proteins can rescue defective otic differentiation in CHD7 KO organoids. The results obtained from the proposed experiments are expected to increase our understanding of the biological functions of CHD7 and will shed valuable insights into the etiology of inner ear pathology in individuals with CHARGE syndrome.

NIH Research Projects · FY 2026 · 2024-12

PROJECT SUMMARY This application is in response to the NIDCD Program Announcement PAS-24-058: Advancing Hearing and Balance Research Using Auditory and Vestibular Organoids. Mechanosensitive hair cells in the cochlea are essential for hearing, but are vulnerable to damage by genetic mutations and environmental insults, resulting in irreversible sensorineural hearing loss. The paucity of human cochlear tissues makes it difficult to study cochlear hair cells and innervating sensory neurons. We recently established a new organoid system that recapitulates cochlear differentiation. These so called human cochlear organoids contain functional hair cells, supporting cells and sensory-like neurons. However, the low efficiency of sensory cell derivation and the lack of mature hair cells and sensory neurons are major limitations for broad pre-clinical applications. We propose a series of experiments to better understand the genetic regulatory network underlying sensory differentiation in human cochlear organoids and improve the efficiency of hair cell differentiation and maturation. In Aim 1, to define the developmental trajectories of key sensory cell types, we will perform a longitudinal scATAC-seq & scRNA-seq multi-omics analysis of otic lineage cells isolated from human cochlear organoids at various time points. Additionally, we will develop a new bioinformatics analytical tool tailored for better integration of temporal multi-modal data by adopting the graph-based neural networks. In Aim 2, we will test if expanding intermediate otic cell populations by genetic programming can increase the number of hair cells arising in human cochlear organoids. Our analysis of longitudinal scRNA-seq data with RNA velocity has identified OTX2 and KLF5 as top candidate driver genes for intermediate otic cells and nonsensory cells, respectively. We will perform CUT&RUN to identify direct target genes for OTX2 in otic progenitors. Additionally, we will test if inducible expression of OTX2 or rapid degradation of KLF5 increases the number of otic intermediate cell populations, leading to a larger number of hair cells and supporting cells in human cochlear organoids. In Aim 3, we will test if augmentation of thyroid hormone signaling accelerates maturation of hair cells and sensory neurons as well as pre-/post-synaptic specification, using a combination of transcriptomic profiling and morphometric analyses. The results obtained from the proposed experiments will advance our holistic understanding of cell fate specification in the human cochlea, offer an improved computational tool for analyzing multi-modal biological data and establish a valuable human in vitro model system for preclinical applications on sensorineural hearing loss.

NIH Research Projects · FY 2026 · 2024-09

PROJECT SUMMARY / ABSTRACT There is no effective clinical treatment to prevent relapse to psychostimulants, although extensive studies have been done focusing on the medial prefrontal cortex and nucleus accumbens. Breakthroughs in the clinical settings rely heavily on a better understanding of brain adaptations, which may explain both chronic drug seeking and taking behaviors. Persistent drug seeking behaviors after a prolonged withdrawal period, as the most accepted laboratory animal model in predicting the risk of relapse, is not exclusively motivation-driven. Here we propose that by the passage of the withdrawal period, the motor/habit components of drug seeking behaviors increase and gradually take over the initial motivational components. The cortical pyramidal neurons (CPNs) in the Supplementary Motor Cortex (M2) will be the central focus of this proposal. Our hypothesis is that high excitability in M2 CPNs and enhanced glutamate release at the M2 CPN terminals in the striatum lead to high drug seeking behaviors at later withdrawal stages after cocaine exposure. Three specific Aims are proposed to, first, examine the excitability of CPNs in the M2 region (Aim 1) and the excitatory transmission in the M2-DLS projection (Aim 2), and then to prevent cocaine seeking (Aim 3) in Sprague Dawley (SD) rats with a history of intravenous self-administration of cocaine. Mechanisms of relapse will be investigated in the motor CPNs in the M2 area and their target dorsolateral striatal synapses. Our hope is to pave the way not only to provide an original perspective in our understanding of addiction-like behaviors, but also to propose novel targets for clinical management of drug relapse.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT Genetic studies have identified novel Alzheimer’s Disease (AD) protective and risk associated genes and genetic variants. PLCG2 is one such gene with protective (P522R) and risk (M28L) variants making it a useful protein to target for the treatment of AD. However, the particular phenotype to be targeted through small molecule modulation of PLCG2 is unknown. Without this knowledge, the validation of PLCG2 as a target for the treatment of AD will be greatly impeded. Therefore, we hypothesize that activation of PLCG2 will mimic the protective effect of PLCG2P522R by inducing molecular signaling changes to PLCG2WT in a manner similar to the protective variant. We will address this hypothesis through the following Specific Aims: Aim 1: Determine how PLCG2 variants associated with AD risk and protection affect PLCG2 related signaling, in vitro. We hypothesize that these variants cause their respective effects (protection and risk) by modulating the preference of PLCG2 to be recruited to the cell membrane, its phosphorylation state, its preferred interaction complexes, and its turnover rate. We will test this hypothesis by stimulating human induced pluripotent stem cell (iPSC) derived microglia like cells (MGLs) expressing either wildtype (WT), protective, or risk variants of PLCG2, and then use both mass spectrometry and traditional biochemical methods to measure these molecular effects. Aim 2: Determine how modulation of PLCG2 activity affects PLCG2WT microglia, in vitro, compared to protective and risk PLCG2 variants. We hypothesize that activation of PLCG2 will produce an activation profile that overlaps with PLCG2P522R microglia. We will test this hypothesis by comparing pAKT levels and Aβ uptake between WT MGLs treated with already developed, novel, small molecule modulators of PLCG2, and vehicle treated MGLs bearing the protective and risk variants. With respect to outcomes as a consequence of the work proposed, we expect to identify changes in the phosphorylation state, subcellular localization, interactome, and turnover rate of PLCG2 caused by the protective and risk variants. We also expect to identify the effect of small molecule modulation of PLCG2 activity on pAKT levels and Aβ uptake with reference to the protective and risk variants. These results will advance our understanding of how PLCG2P522R protects from AD and provide insight into the pharmacological requirements of a commercial PLCG2 modulator as a potential AD therapeutic. Additionally, we expect that this study will guide the design of therapeutic interventions that mimic the protective functions of the PLCG2P522R variant. Finally, this study will contribute to the continued understanding of the role of microglia in response to AD pathology. Overall, this study will have a positive impact because it will provide a biological basis for understanding the pharmacology of novel therapeutics targeting PLCG2 for the treatment of AD.

NIH Research Projects · FY 2024 · 2024-09

TITLE: Targeting P2RX7 Signaling as a Treatment for ADRD ABSTRACT Recent work indicates neuroinflammation is a key driver of cellular dysfunction in Alzheimer’s disease and related dementias (ADRD), where purinergic receptors play a key role. These receptors are cell surface plasma membrane proteins that communicate between neuronal and glia cells, and are sub-divided into G-coupled protein and ligand-gated ion channel receptors, and are evoked by extracellular adenosine 5′-triphosphate (eATP). Subtype-7 P2X receptors (P2X7R), which are expressed on microglia, astrocytes, oligodendrocytes, and hippocampal neurons, are considered “silent” due to inactivity; however, release of eATP, or downregulation of ectonucleosidases CD39 and CD73, results in beneficial channel opening to ions, mobilizing microglia to the site of injury for phagocytosis. Sustained activation of P2X7R via eATP results in Ca2+ overload, NLRP3 activation, superoxide formation, conversion and release of IL1β and IL18 cytokines, recruitment of pannexin-1 pores, auto- induction, NFkB transcription, caspases activation, and cell death of surrounding microglia and neurons. Provided this, P2X7R has been implicated in ADRD and neurodegenerative diseases. The majority of this work have been conducted with mice which retain the P451L loss-of-function mutation, which significantly reduces the affinity for eATP. Moreover, models retaining this SNP show limited human relevant neurodegeneration with age, and work has solely focused on amyloid clearance. Moving beyond amyloid, we will test the hypothesis that P2X7 receptors play a central role in neuroinflammation, and that inhibiting this pathway will prevent neurodegeneration through the use of genetically diverse mouse strains which retain functional P2X7R. In Aim 1, we will characterize WSB.APP/PS1 and CAST.APP/PS1 mice (Aim 1.1), and then determine whether temporally controlled genetic global ablation of P2X7R leads to prevention of neurodegeneration in our mouse models (Aim 1.2). Cohorts will be assessed at 4, 8, and 14 mos, where neuroinflammation (microglial and astroglial), synaptic density, and neuronal health will be assessed via translationally relevant PET/CT imaging. In addition, short term memory will be measured via the spatial novel recognition task (Aim 1.3). Blood and tissue will be collected for biochemical, immunopathology, and molecular analyses (Aim 1.4). To understand the spatial and temporal role of P2X7R signaling, select brain regions determined by PET/CT will be subjected to MALDI- IMS for differential protein expression (Aim 1.5). In Aim 2, we will determine a safe and effective dosing strategy for GSK1482160, a selective and potent P2X7R allosteric antagonist, in WSB.APP/PS1 and CAST.APP/PS1 at 14 mos via PK/PD modeling (Aim 2.1). We will then titrate GSK1482160 (0, 0.4, 4, 40 mg/kg) as a tool compound to specifically inhibit P2X7R activity, and assess its impact on neurodegeneration via PET/CT and behavior (Aims 2.2-2.3), biochemical, immunopathology, and molecular analyses (Aim 2.4), and MALDI-IMS methodologies (Aim 2.5) described in Aim 1. Importantly, all readouts will be conducted in the same cohorts of mice and derived tissues, thus elucidating the role of P2X7R signaling in neurodegeneration across spatial and temporal scales.

NIH Research Projects · FY 2025 · 2024-09

Alzheimer’s Disease (AD) is an irreversible neurodegenerative disorder characterized by progressive impairment in brain structures and functions. Early biomarkers are believed crucial to AD, making it possible to identify and treat AD patients before evident symptoms. Established AD biomarkers are grouped into β amyloid deposition (A), pathologic tau (T), and neurodegeneration (N), including neuroimaging and cerebrospinal fluid (CSF). However, they fall short in explaining the heterogeneity of individual clinical trajectories. In this project, we aim to develop novel computational approaches to identify genetic biomarkers related to AD progression. Leveraging the multi-omic genetic data and multi-modal brain imaging data in AD (e.g., AMP-AD, ADNI), We will 1) identify stage-specific genetic biomarkers of AD with known downstream effect on transcriptome and proteome layers, and 2) identity genetic biomarkers that can help differentiate distinct phenotype trajectories. These results can help with candidate screening in clinical trials and provide stratified risk groups to facilitate the development of therapeutic intervention. Alzheimer’s Disease (AD) is an irreversible neurodegenerative disorder with a long prodromal phase and no clinically validated cure. Detecting when and how molecular and phenotype marker develop along AD progression will provide a template for understanding the underlying etiology of clinical syndromes and for improving early diagnosis, clinical trial recruitment and treatment assessment. Established AD biomarkers can be grouped into β amyloid deposition (A), pathologic tau (T), and neurodegeneration (N), captured from neuroimaging and cerebrospinal fluid (CSF). Despite some applications in early detection, the ATN framework relies on the dichotomous classification of individuals and cannot capture the full spectrum of AD-related pathologies. It could be supplemented with the addition of stage-specific markers or a severity staging scheme. In this project, we will develop novel computational approaches for subject-level stage-specific markers and severity staging scores. We will leverage major multi-omic genetic data and multi-modal brain imaging data in AD, and propose the following two aims, 1) Identify subject-level stage-specific disease modules using multi- omic data, and 2) subject-specific severity staging with longitudinal imaging data based pseudotime. These methods and tools will have considerable potential for improved understanding of disease progression and discovery of associated neuroimaging and genetic markers. These results can help with candidate screening in clinical trials and provide stratified risk groups to facilitate the development of therapeutic intervention.

NIH Research Projects · FY 2025 · 2024-09

There is currently a dearth of FDA-approved medications developed specifically for women with alcohol use disorder (AUD). This represents a major public health concern for several reasons. First, robust sex-related differences set women at a marked disadvantage for progressing to severe AUD more quickly than men (telescoping). Second, alcohol consumption levels in women have been steadily moving towards parity with men. Third, women with AUD remain challenging to recruit into research trials, as many have caregiving commitments and/or feel stigmatized. Together these factors compound the need to develop new relapse prevention medications tailored to women, using methods that encourage women into clinical trials. Hence, a 12-week multicenter clinical trial is proposed, using a validated remote platform, which has been successful at increasing both recruitment and compliance in substance using women. Overarching aims are to examine whether 3mg/d of guanfacine extended release (GXR) Vs placebo (PBO) can reduce drinking severity in AUD women by targeting alcohol craving and emotion regulation during stress. Prior research suggests that these relapse-related processes may represent risk factors particularly pertinent to women and unique to the pharmacodynamics of guanfacine. N=70 women with AUD will be recruited at the primary site (Stony Brook) and N=60 at the secondary site (Rutgers) into a 12-week, randomized, double-blind, placebo-controlled clinical trial. Momentary reports of drinking severity, stress, craving, mood, arousal, anxiety, and emotion regulation will also be collected throughout the day and evening via three, 2-week bursts of Ecological Momentary Assessment (EMA), across the 12-week trial (weeks 1 and 2; weeks 5 and 6; and weeks 9 and 10). All data will be collected using smartphone technology. Participants will also take part in twice weekly remote visits for the entire 12 weeks to assess safety, vitals, collect urines, monitor alcohol use, and receive weekly Medical Management. It is anticipated that GXR will decrease the number of positive urines across the 12 weeks and the % number of days spent drinking (H1a, primary outcomes), as well as the mean number of drinks consumed per occasion and mean number of binge drinking sessions (H1b, secondary outcomes) compared with PBO. It is also expected that GXR Vs PBO will attenuate momentary ratings of stress-induced drinking and alcohol craving (H2a), as well as momentary ratings of stress-induced emotion dysregulation, negative mood, arousal, and anxiety (H2b) such that women taking GXR will have greater decreases in craving and use trajectories over time (H2c). The efficacy of using a remote platform will additionally be explored by ascertaining attendance, compliance and medication adherence rates. Findings will help determine the efficacy and underlying mechanisms of a potentially well-tolerated medication developed specifically for AUD women. The multi-site design will better assess feasibility of telehealth platforms while increasing the generalizability of patient samples.

NIH Research Projects · FY 2026 · 2024-09

The goals of this project are to identify strategies to improve the integration of genomic testing, reporting, and use into the clinical workflow of patient care. Over the last decade, we have acquired the infrastructure and expertise to support genetic testing, reporting, alerting, educating, and returning results. We will now leverage our past accomplishments and our genomics-enabled learning health system (gLHS) to share implementation strategies, conduct two selected intervention projects network-wide and evaluate their impact, and share gLHS tools and resources that can be broadly adopted. To facilitate this work, we are proposing two pragmatic randomized stepped wedge clinical trials: one focused on implementing DPYD testing and one on APOL1 testing. In patients receiving fluoropyrimidine chemotherapies, DPYD testing and genetic-guided dose adjustments reduce the severe toxicities caused by these drugs. In many other countries, DPYD testing is standard of care; in the US, it is beginning to be used in some clinics, but needs additional leadership and effective strategies to overcome the barriers and facilitate broad adoption. APOL1 variants contribute to the development and progression of chronic kidney disease and disparate outcomes in patients with African ancestry. APOL1 testing, together with joint decision making with the patient and provider, helps to manage risk factors that contribute to poor chronic kidney disease outcomes. These trials represent two distinct genetic tests and disciplines of medicine, are tests that are ready for broad implementation, can be implemented in most health care systems, and address disparate healthcare outcomes. The two proposed clinical trials will serve as a platform to implement innovative solutions to drive genomic medicine uptake. We propose that an initial set of solutions would reasonably include EHR-based clinical decision support, educating providers, simplifying genetic test ordering, and streamlining processes to improve insurance reimbursement rates. Following the initial implementation, we will evaluate the success of the strategies and refine and integrate additional improvements in two subsequent rounds of iterative implementation. In order to fully accommodate the selected projects, our implementation team includes experts in genetics, counseling, informatics, pharmacology, bioethics, learning health systems, economics, implementation science and many disciplines of medicine that have a productive track record of working together. Our previous implementation work has led to the identification and resolution of many barriers to genomic medicine. The insights gained from our learning health system, provider and patient feedback, informatics analyses, workflow logistics, and laboratory challenges provide initial basis for our initial interventions; they will also provide immediate strategies for other health care systems to learn from our experience. Upon completion of these trials, we expect to have developed more effective strategies that will expand genomic medicine adoption from the current boutique clinics out to mainstream and diverse clinics and will minimize disparate outcomes across many life-threatening diseases.

NIH Research Projects · FY 2025 · 2024-09

Abstract Cirrhosis is the end-result of chronic liver disease mainly viral hepatitis, metabolic dysfunction-associated steatohepatitis, and alcohol-related liver disease. The only life-saving treatment for decompensated cirrhosis is liver transplantation (LT). Despite the critical need for this lifesaving therapy, there remain significant barriers to accessing LT. While a myriad of disparities exist in the LT care pathway, the largest number of patients in the pathway are in the community awaiting the first step-referral. Previous studies have revealed disparities in LT referrals, with lower odds for Black individuals (OR, 0.19), the uninsured (OR, 0.40), and specific hospital sites (OR, 0.40). Nationally, the waitlist capture rate for decompensated cirrhosis was 0.4 for White individuals and 0.3 for Black individuals; in Indiana, the ratio was only 0.2, ranking in the bottom 5% of all states. At Indiana University Health (IUH), the sole LT center in Indiana, only 5.8% of LT referrals were Black individuals, and less than 1% were Hispanic ethnicity. Many patients experiencing health disparities seek medical care at community gastroenterology (GI) practices where clinicians may not have access to transplant hepatologists and may lack knowledge of specialized evidence-based protocols and policies. This is exacerbated by factors like limited provider time, complexities in navigating the healthcare system, and differing levels of patient literacy and unmet social needs, all of which impede referral. Furthermore, there is variability among community GI practices, including size (i.e. small vs. large) and location (i.e. rural vs. urban), directly impacting the resources available and the capacity to support both providers and patients through the referral process. There is an urgent and unmet need to develop an intervention that addresses this variability but also acknowledges the structural and social barriers faced by patients and caregivers, ultimately improving access to referral. There is currently no multilevel intervention addressing disparities in access to LT. The study team proposes to adapt LT-CARE, a multi-component referral toolkit to address barriers at multiple socioecological levels to improve referral. The study team will use community-based research methods to adapt a usable, acceptable, and feasible toolkit for community GI practices, their patients, and caregivers.

NIH Research Projects · FY 2024 · 2024-09

PROJECT SUMMARY The goal of this work is to identify pain-induced adaptations in the prefrontal cortical dynorphin / kappa-opioid receptor system and the role this system plays in pain-induced mal-adaptive behavior and nociception. Chronic pain drives adaptations in prefrontal cortical circuits and is hypothesized to increase bias towards aversive experiences. Our preliminary data demonstrates that inhibitory and excitatory populations of prefrontal cortical dynorphin neurons are activated and release dynorphin neuropeptide in response to aversive stimuli. However, it is unclear whether prefrontal cortical dynorphin neuron activity and ensuing dynorphin release is impacted by pain. To address this knowledge gap, we will perform ex-vivo electrophysiological procedures and in-vivo single cell calcium imaging in freely moving mice to determine how pain influences activity of excitatory and inhibitory prefrontal cortical dynorphin neurons and their responsivity to noxious stimuli and aversive stimuli. Moreover, we will manipulate the activity of prefrontal cortical dynorphin neurons and release of dynorphin neuropeptides or fast transmitters to determine whether enhanced prefrontal cortical dynorphin neuron activity and transmitter release controls nociception and mal-adaptive affective behavior induced by pain. Taken together, this work will identify how genetically-defined sub-populations of prefrontal cortical neurons and the opioid peptides or fast excitatory and inhibitory transmitters they release influence negative affect induced by pain from the cellular to in-vivo level of analysis. This work is of broad relevance as it elucidates basic principles by which neuropeptide- expressing neurons in prefrontal cortical circuits control behavioral plasticity and delineate potentially new therapeutic targets for the treatment of mal-adaptive affect and increased responsivity to noxious and aversive stimuli associated with pain states.

NIH Research Projects · FY 2025 · 2024-09

ABSTRACT An individual’s phenotypes related to their health conditions are associated with the complex interplay between the individual’s biological, behavioral, social, and environmental processes, including phenomena that occur both within (e.g., genetics, emotion, cognition) and external (e.g., social, built, and natural environments) to the organism. Behavioral and social sciences research (BSSR) at the National Institutes of Health (NIH)—the systematic study of behavioral and social phenomena relevant to health—is key to understanding how these internal and external processes interact to alter health, and for developing efficacious interventions. Nevertheless, BSSR of health face substantial challenges in part due to its broad and complex research landscape, but also because of the “inconsistent use of terms and classification systems making it challenging to integrate findings from individual studies and in turn to cumulatively build bodies of knowledge even in domains that are consistently studied.” Ontologies provide a way to address these challenges in BSSR. Recognized and sponsored by the NIH and various other agencies and professional societies, the National Academies of Sciences, Engineering, and Medicine (NASEM) formed a multidisciplinary committee to study ways to improve the development and use of ontologies in the BSSR domains and produced a comprehensive consensus report identified barriers, opportunities, and recommended approaches to advancing BSSR ontologies important to health. Motivated by the Report, the NIH aims to create a BSSR ontology development network with research projects (PAR-23-182) covering a wide range of disciplines related to BSSR across multiple NIH Institutes/Centers, with a Dissemination and Coordination Center (DCC; PAR-23-181) to (1) facilitate collaboration and cross-project learning; (2) provide ontology-related technical, computational, and informatics expertise and support; (3) facilitate dissemination of resources and training to support ontology expansion, development, and use; and (4) provide active outreach and coordination with relevant stakeholders to increase understanding of and demand for BSSR ontology-related tools and resources. In response to PAR-23-181, we established a multidisciplinary team with the necessary subject matter expertise to carry out the functionalities of the DCC, leveraging existing ontology resources and tools (e.g., Protégé) that we have developed, and the long-standing ontology community (e.g., BiPortal) we have built and sustained. Our Specific Aims are to (1) provide administrative and logistical support for the U01 Research Network; (2) develop a common technical framework with standard operating procedures (SOPs) to guide the development of BSSR ontologies in the U01 Research Network; and (3) develop a “toolbox” of resources through which the products of the Network can be shared with, and adopted for use, by all the relevant communities.

NIH Research Projects · FY 2026 · 2024-09

PROJECT SUMMARY Millions of children globally, including thousands of US children and nearly 15 million African children, are HIV-exposed and uninfected (CHEU). Despite advances in understanding health outcomes of CHEU, neurodevelopmental impacts have not been well characterized. Several small studies conducted in high income countries (HIC) including the United States suggest a higher prevalence of autism in CHEU. CHEU may be at elevated risk for autism due to HIV exposures, which are associated with adverse birth outcomes, iron-deficiency anemia, and maternal and child infectious morbidity. These risks have been shown to be associated with autism outcome in the United States and other HIC. However, large, well-characterized, longitudinal cohorts of CHEU are needed to identify mediating biological and environmental risks on the causal pathway between HIV exposure and autism outcome and inform the selection of modifiable targets for precision detection and intervention strategies. This research has the strong potential to inform the development and validation of neurobehavioral biomarkers and scalable diagnostic tools, including those leveraging eye tracking (ET) technology, for early detection of autism in resource-constrained settings of the United States and across the globe. The objective of this proposal is to examine autism diagnostic outcome and ET biomarkers in CHEU. We will accomplish this by leveraging a longitudinally assessed cohort of CHEU and children who are HIV unexposed and uninfected (CHUU) with well characterized HIV-related and contextual exposures (R01HD104552) and synergistically expanding our team’s ongoing neuro-health capacity building efforts (R21MH127570). Consistent with NIH priorities to leverage real-world, integrated data resources, this study combines longitudinal clinical, behavioral, and neurobiological measures to enable rigorous, reproducible investigation of autism risk and outcomes. In Aim 1, we will compare autism diagnostic outcomes between young CHEU and children who are unexposed and uninfected (CHUU) across a large cohort (N=850) of Kenyan children that has been studied since the prenatal period. In Aim 2, we will determine whether neurobehavioral ET markers predict autism outcome in CHEU and CHUU. Through a series of tiered training and mentorship activities integrated with the proposed research activities, in Aim 3 we will build global scientific partnerships and upskill a Neuro-Health Collaborative, which will generate scalable training and care models that will strengthen workforce capacity and improve autism detection and outcomes in the US and across the globe. This study is innovative as it will be the first to apply rigorous diagnostic and ET methodologies to investigate autism outcomes in CHEU. By leveraging a large, exposure-enriched cohort and integrating multimodal longitudinal data, this project aligns with emerging NIH efforts to use data science approaches to identify contributors to autism etiology and improve the scalability and effectiveness of diagnostic strategies.

NIH Research Projects · FY 2025 · 2024-09

ABSTRACT Alcohol use disorder (AUD) is a major U.S. health problem with 12-month and lifetime prevalence being 13.9% and 29.1%, respectively. AUD is partly caused by genetic factors with the estimated heritability being about 50%. Therefore, identification of genes related AUD can help us elucidate the genetic mechanisms of AUD and develop novel prevention and treatment strategies. While recent large-scale genome wide association studies (GWAS) have identified multiple AUD-associated genes, these genes only explain a small portion of heritability, indicating that many additional genes with small effects remain to be discovered. AUD is a heterogeneous disorder with multiple interacting genetic and environmental influences. Therefore, two individuals could receive the same AUD diagnosis, but have multiple different symptoms. One strategy to interrogate heterogeneous disorders is the inclusion of more homogenous endophenotypes, which deconstruct diagnostic categories into component features that may be more amenable to genetic research and provide a cost-effective way to increase statistical power without dramatically increasing the sample size. Alcohol sensitivity and tolerance are two important contributing features to the development and maintenance of AUD in humans, can be effectively measured in mice, and are known to be at least partly genetically mediated in both species. In this study, we propose a systematic and translational strategy to identify and validate variants and genes related to alcohol sensitivity and tolerance using both human and Diversity Outbred (DO) mice. In Aim 1, we will perform GWAS of alcohol sensitivity and tolerance in both European and African ancestry populations after adjusting for the effects of alcohol metabolism genes. Next, we will use high throughput reporter assays in multiple cell lines representing four major brain cells to identify functional variants with sub-threshold P values. We will then use a Mendelian randomization framework to identify target genes of those functional variants. In Aim 2, we will perform QTL, eQTL, and gene expression-trait correlation analyses to identify genetic variants associated with ethanol sensitivity and tolerance in DO mice. DO mice are genetically heterogeneous with large amounts of recombination that provide high mapping resolution approximating that of a human GWAS, but with a much smaller sample size. In Aim 3, we will employ our newly developed VariantGraph database to identify variants and genes with shared regulatory effects across mouse and human. The findings will advance our knowledge of the genetic architecture of alcohol sensitivity and tolerance and will shed light on AUD etiologies.

NIH Research Projects · FY 2025 · 2024-09

Cystic fibrosis (CF) is a genetic disorder that negatively affects young people across the globe. Pulmonary exacerbations (PEx), episodes of decreased lung function accompanied by coughing, increased sputum production, and weight loss, are a hallmark of CF lung disease. Besides negative health impacts, PEx comes with a burden to healthcare costs in the United States and beyond ($10K-$40K USD per episode). Although PEx can be treated with antibiotics, patients still experience decreased quality of life and ultimately reduced survival. A point-of-care, rapid, and accurate test to identify impending PEx that would benefit from treatment could have an impact on families of children with CF and their clinicians through reducing misdiagnosis and overtreatment. Breath testing, through identifying volatile organic compound (VOC) biomarkers, holds great promise for the development of home-based/clinical testing solutions for PEx. The goal of this research is to develop a hand-held smart sensor system that can detect exhaled VOC biomarkers for PEx noninvasively in real-time. To accomplish this, machine learning will be utilized to identify a breath-based biosignature of PEx (Aim 1). In parallel, the team will design/test a nanosensor array to detect the biosignature (Aim 2) and develop a user-friendly smartphone app to be used at-home or in the clinic (Aim 3). Ultimately, this research will further the development of diagnostic solutions for PEx, and advance knowledge in a multi-faceted fashion across disciplines including basic science, chemistry, engineering, medicine, biotechnology, and health informatics. The technological solution is highly disruptive and challenges the current paradigm of how PEx is diagnosed. From an engineering perspective, sensors are at the cusp of being translated into biomedical devices, and this research aims to overcome challenges in selectivity that can also be leveraged for VOC-based diagnosis of other heart and lung diseases beyond CF. The interdisciplinary team has vast experience in their respective fields, and their collaboration ensures successful completion of the research. A diverse set of resources/equipment from the team's laboratories, along with others on campuses, will be leveraged to support research activities. RELEVANCE (See instructions): The research addresses

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT Alcohol use disorder (AUD) is a prevalent global health problem with substantial social, economic, and public health costs. Progression from initial alcohol exposure to AUD often begins in adolescence and emerging adulthood with impulsive personality traits (IPTs) being critical developmental risk factors. However, there is limited understanding of the dynamic longitudinal interplay between IPTs and alcohol involvement from childhood to adulthood and the role neurobiological and genetic vulnerability play in this etiology in part because: 1) longitudinal studies of IPT-AUD associations to date typically characterize specific developmental epochs (e.g., late adolescence), rarely comprehensively examining these associations across the full developmental span; 2) studies of neuroanatomical variation associated with IPT-AUD pathways are primarily based on small, cross- sectional datasets and restricted subsets of brain regions, despite accumulating evidence that large samples sizes are necessary to reliably characterize brain-phenotype associations and the importance of neurodevelopmental trajectories on complex behavior; and 3) large-scale genome-wide association studies of IPTs have been primarily conducted in older adults of European ancestry, despite the clear developmental context underlying IPT-AUD pathways and need for further incorporation of diverse ancestral populations into genomics research. This K01 proposal addresses these gaps by: 1) Aim 1: characterizing developmental associations (childhood to middle adulthood) between distinct IPTs, heavy alcohol use, and AUD progression through data harmonization and longitudinal integrative data analysis of 16 extant primarily longitudinal study samples (N=85,862); 2) Aim 2: examining whether IPTs represent plausible mechanisms through which brain structure is linked to alcohol involvement using a subset of 9 neuroimaging studies (N=17,297) spanning childhood to young adulthood; and 3) Aim 3: identifying developmentally specific genetic influences on IPTs, alcohol involvement, and the links between them to assess genetic stability and/or uniqueness across development using trans-ancestral genetic data. This K01 would provide the applicant with training to facilitate transition to career independence by enabling him to: 1) acquire expertise in longitudinal data harmonization and integrative data analytic approaches; 2) build a foundational neuroimaging data analysis skill set and developmental addiction neuroscience knowledge base; 3) expand genomic analytic expertise to encompass trans-ancestral and longitudinal genome-wide association study approaches; and 4) cultivate professional and career development toward an independent research career. Collectively, this proposal would prepare the applicant to lead an innovative research program into the genetic and biological etiology of AUD development as an independent investigator and would provide novel insights into the developmental etiology and neurogenetic sources of variation underlying IPT-AUD pathways.

NIH Research Projects · FY 2025 · 2024-09

Musculoskeletal disease and disorders affect 1 out of 2 Americans and continue to be responsible for the majority of physician/hospital visits and medical costs. Addressing this serious health issue requires teams of investigators and clinical scientists with training and experience in studying the tissues and organs of the musculoskeletal system as well as their interactions with other body systems. The Indiana Center for Musculoskeletal Health, ICMH, was established in 2017 to address this need. The over 100 ICMH members, from 5 schools and 26 departments, have formed focused research teams to investigate specific musculoskeletal health issues such as post-traumatic osteoarthritis, osteoporosis, sarcopenia, and fracture healing, as well as the musculoskeletal impacts and contribution in systemic disease processes such as aging, diabetes, cancer cachexia, renal disease and others. To identify targetable cellular and molecular mechanisms involved or responsible for these conditions, investigators collaborate in areas as diverse as periodontal disease, muscle/bone crosstalk, osteosarcopenia, and osteocyte mechanobiology and function. The goal of the ICMH Resource Cores, ICMH-RC, is to promote and support advancements in the field of musculoskeletal research through understanding muscle/bone interactions, mechanobiology, and the roles of the musculoskeletal system beyond movement. To accomplish this goal, it is imperative individual investigators and research teams have access to core facilities that provide services and cutting-edge technology specific to bone/cartilage/connective tissue and muscle. The ICMH-RC will be composed of the Admin/Enhancement Core, Core A and three resource cores: Core B: Histology and Histomorphometry core, Core C: Systems Biology and Multi-Omics core, and Core D: Mechanobiology/3D Bioprinting Core. The Admin Core will not only be responsible for efficient and productive operations of these cores but also for the selection of promising pilot applications proposing novel yet feasible concepts in the study and treatment of musculoskeletal disease. Core A will also place high priority on the training not only of faculty, postdoctoral fellows, and graduate students, but also will introduce high school students and faculty to the field of musculoskeletal research to ensure that the next generation of critical musculoskeletal scientists, engineers, and physicians will comprise an eager, well-trained workforce. New cutting-edge technologies such as 3D tissue and organ bioprinting will be further developed. The ICMH-RC will interface with the clinical cores of the ICMH Clinical Research Center, ICMH CRC, to promote transfer of discovery to clinical application, using the “bench-to-bedside” translational approach to achieve the ultimate goals of musculoskeletal research.*

NIH Research Projects · FY 2025 · 2024-09

Project Summary/Abstract Poor oral health is a national symbol of social inequity, disproportionately affecting vulnerable populations. Despite its profound impact on overall well-being, oral health remains disjointed from medical care delivery. As a result, vulnerable adults often resort to emergency departments (ED) for treatment of non-traumatic dental conditions (NTDCs), leading to suboptimal care, clinician frustration, increased ED crowding, and high health costs. The purpose of this proposal is to use electronic health record (EHR) data to better identify adults likely to seek treatment in EDs for NTDCs. Aim 1, Leverage multi-health systems’ EHR data to predict ED use for NTDCs, will use EHR data from multiple hospital systems and novel social determinants of health data (measured at the individual- and community-level) in a series of advanced machine learning (ML) prediction models. This aim will identify the value of medical and social determinants data in predicting oral health needs. Aim 2, Extract oral health predictors from the clinical notes of medical encounters that occur prior to ED use, will apply natural language processing techniques to create novel data reflective of medical care providers’ documentation of oral health. This aim will create new data for use in prediction modeling. Aim 3, Assess the feature importance within, and the predictive performance of, ML prediction models built on structured and unstructured EHR data, will test the contributions of unstructured and structured data source in prediction models designed to identify adults at risk of using the ED for NTDCs. Furthermore, the mentorship and rigorous training plan proposed in this career development award application will enable Dr. Heather Taylor to become an independent investigator capable of using advanced health informatics approaches to enhance the delivery of oral health services, particularly for underserved populations. To achieve her career goals, Dr. Taylor seeks 1) technical training in the application of machine-learning, including natural language processing, 2) knowledge development in health disparities research, and 3) enhanced professional development and grant writing skills. Dr. Taylor has assembled a diverse, interdisciplinary panel of mentors to foster her proposed career development and research plan. She is supported by a resource-rich institutional environment with access to one of the nation’s oldest and largest clinical EHR data repositories. This study is innovative because it utilizes data collected outside of traditional dental care settings to provide health systems with actionable information about adults in their community who need support navigating the oral-medical divide. The anticipated products of this research include the development and validation of risk prediction models that identify patients who experience poor oral health and require targeted support. After completion, Dr. Taylor plans to transition to research independence and submit an R01 focusing on optimizing risk predictions for ED use related to NTDCs within health systems and connecting identified adults to definitive dental care.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT Firearm injury and death is a public health epidemic that spreads through communities and social networks in the United States much like an infectious disease. Individuals who survive a firearm injury (i.e., nonfatal survivors), experience increases in the risk of adverse health outcomes, such as posttraumatic stress disorder (PTSD), anxiety, and depression. Family members of homicide victims, nonfatal firearm injury survivors (i.e., secondary survivors), and communities more broadly are also affected. Due to methodological limitations, we have very little data on what trauma networks (defined as individual(s) present at a shooting event and/or family member(s) of a fatal victim/nonfatal survivor) look like and much less on the long-term health effects of being in the network of a firearm injury. This is a critical question because these trauma networks – people exposed but not injured – are likely at high risk of adverse health outcomes, particularly children, adolescents, and young adults (CAYA) and are currently being missed in firearm injury focused interventions. The proposed study will use an existing data repository (used in prior work) of individually linked records from healthcare and police data between 2016 - 2022. We specifically aim to 1) develop and describe trauma networks of all individuals (i.e., victims, witnesses per police data) present at a shooting event and/or all family members of firearm injury victims/survivors per Medicaid data and 2) examine clinical utilization, firearm injury, and mortality outcomes among CAYA network members, due to the high risk of adverse health outcomes in the 5-years following firearm injury event. This study will demonstrate defining trauma networks to identify CAYA exposed to firearm injury is feasible (Aim 1) and estimate prevalence rates of mental health outcomes, firearm injury, and mortality among network members (Aim 2) in order to identify future opportunities for interventions. SIGNIFICANCE: Consistent with the NICHD strategic plan, our methodology, data infrastructure, and composition of our research team ensures that our research will significantly extend our understanding of the mechanisms and impacts of firearm injury events beyond the individual directly involved to improve health across the lifespan. Regardless of the results, this will lead to a R01 application addressing the health needs among CAYA within trauma networks and inform interventions that may mitigate adverse and likely debilitating effects among CAYA and will have implications for broader public health significance.

NIH Research Projects · FY 2025 · 2024-09

SUMMARY Surgeon-scientists are an underrepresented population in academic medicine, and federally funded physician- scientist-led research has been in decline over the past decade. Substantial challenges are faced by surgical trainees seeking research training. Importantly, surgical resident training programs with ample protected time for dedicated experiential research training have effectively contributed to the “pipeline” of successful, independently funded surgeon-scientists. These researchers have meaningfully impacted surgical oncology patient care and scientific knowledge. The proposed Surgical Oncology Research Training at Indiana (SORTI) program brings together the extensive multidisciplinary oncologic expertise at the Indiana University School of Medicine (IUSM) and the Simon Comprehensive Cancer Center to create a multidisciplinary, two-year research training program consisting of (1) a Health Services and Outcomes Research (HSOR) Track and (2) a Basic Science and Translational (BTS) Research Track. The SORTI program will draw upon the Department of Surgery’s 30-year experience of training in BTS oncology research and the 15-year surgical oncology HSOR resident training collaboration between the Surgical Outcomes and Quality Improvement Center (SOQIC), the Regenstrief Institute, and the American College of Surgeons (ACS) Cancer Programs. Since 2011, SOQIC has trained 44 surgical research trainees who have gone on to oncology-focused academic careers. During a two-year, intensive training period (100% protected time), trainees in the SORTI program will be immersed in multidisciplinary, experiential research training, using a team science approach, with tailored mentorship and focused didactic teaching (Master’s degree for those without prior advanced degrees). Each trainee will have an experienced and diverse Mentor Team (content and methodological experts), tailored to their training needs. The SORTI program core curriculum will include courses in study design and analytic approaches, best practices and ethics in research, communicating research, and career development. Trainees will participate in the workshops, seminars, and research training opportunities at Indiana University and at relevant surgical and oncologic professional societies. Trainees will design and execute research projects that culminate in presentations at major national conferences and publications in high-impact journals. Continuous monitoring and iterative program improvement will be achieved by engagement of a highly accomplished External Advisory Committee and robust evaluation by Indiana University’s Center for Evaluation, Policy, and Research. The ultimate goal of the SORTI program is to increase the number of investigators who successfully achieve academic careers as independently funded surgical oncologic scientists. Indiana University School of Medicine is uniquely positioned to assure the success of the SORTI program and continuing to develop future surgical oncology scientist leaders and mentors.

NIH Research Projects · FY 2025 · 2024-09

The proposed project, NEXT STEPs (Nursing Home EXplanatory Clinical Trials: Supporting Transformation by Enhancing Partnerships) Network, creates a national infrastructure that will address systemic barriers to conducting research in nursing homes (NHs). NHs care for people with serious, complex medical illnesses and are a critical site of care for people with Alzheimer’s Disease and Related Dementias (ADRD). The proposed infrastructure is needed to identify and overcome the multiple challenges to conducting research in this setting of care, which include navigating regulatory and recruitment challenges for people with ADRD. The NEXT STEPs Network aims to provide integrated support for investigators to conduct equitable, explanatory trials in nursing homes. NEXT STEPs will create a collaborative community of NH leaders, staff, residents and care partners, clinicians, and researchers to identify research priorities for the field and develop action plans to address them. This will be accomplished via project Cores including Central Coordination; Recruitment and Retention; Methods, Measures, and Data; and Projects and Training. Consensus-based best practices for NH clinical trials will be used to create toolkits and research guidance and foster collaborations. A pilot projects program and research mentorship will create increased national capacity to conduct NH clinical trials. A key component of this project is the bi-directional collaboration with the Long-Term Care Data Cooperative (LTCDC), which has created integrated datasets for hundreds of thousands of nursing home residents. The NEXT STEPs Network Real World Data Scholars and pilot grant awardees will utilize the LTCDC for proposed projects. The development of experiential learning models will provide embedded opportunities for collaboration and learning for researchers in the nursing home setting. Research and Industry/Advocacy Advisory Panels and Partner Workgroups will represent key viewpoints in nursing home care and inform all aspects of the project. The project team will focus on disseminating best practices, pilot project findings and hosting annual meetings, presenting at national conferences and convening regional partner meetings to ensure effective dissemination of key findings. The NEXT STEPs Network will enable researchers from different disciplines to collaborate, with a goal of increasing the number and quality of clinical trials conducted in NHs, including those that focus on prevention, medical therapeutics, behavioral, health services interventions, and the care of special populations including people with ADRD. NEXT STEPs will create strong linkages between researchers, industry, and other crucial parties, fostering evidence-based innovations on key priorities to optimize resident care and quality of life for those living, supporting loved ones, and working in NHs.

NIH Research Projects · FY 2025 · 2024-09

ABSTRACT The overarching goal of this project is to identify tissue-specific genetic variants and genes that are associated with cellular and tissue morphology in normal tissues. We plan to apply advanced computational pathology methods based on machine learning and computer vision to analyze normal tissue histology H&E images from Genotype-Tissue Expression Project (GTEx), extracting interpretable image features as quantitative traits. Next, we will apply bioinformatic and statistical genetic methods to identify the morphological traits that are correlated with eQTLs in donor population, thus generate image QTLs (a.k.a. imQTLs) for all normal tissue types in GTEx with over 100 samples. In addition, we will apply advanced functionally informed GWAS (FiGWAS), which has been successfully applied to eQTL research and significantly boosted the detection of rare variants in genomic association study, to further investigate the association of non-eQTL genetic variants with the interpretable quantitative morphology features described above and to generate supplementary imQTLs. Neither of these two approaches has previously been applied to identify imQTLs. The workflow will initially focus on cell type morphological features, then expand to features related to tissue development and cell-cell interactions. The identified imQTLs (or the genes/image traits associated with them) will be further tested in histopathological images in corresponding tissues from The Cancer Genome Atlas (TCGA) for any difference in terms of imQTL presence, abnormality in the associated image traits, or expression in the associated genes. The identified imQTLs will not only generate new insights about the tissue differentiation, development, and morphological variations in the normal population, but also will provide a solid basis for comparing pathological changes in many types of diseases and help quantify the level of the corresponding histopathological changes. The resulted image features and imQTLs will be made available through a web portal called PathoGenome Viewer for general public query and use.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY An ideal "universal flu vaccine" should have the ability to trigger broadly protective antibody and humoral memory responses against both existing and future influenza viral strains. However, a major obstacle in achieving this goal is immunodominance in B cell responses, which tend to prioritize dominant epitopes at the expense of conserved yet subdominant epitopes. In the case of the influenza hemagglutinin (HA), the variable globular head domain is highly immunodominant over the stem epitope that is much more conserved. To address this challenge, a Toll-like receptor 7 (TLR7) agonist-based nanoparticle (NP) adjuvant (TLR7-NP) has been developed to induce prolonged and localized TLR7 signaling in the draining lymph nodes. In contrast to TLR7 agonist formulated with alum (TLR7-alum), the TLR7-NP adjuvant has demonstrated a remarkable ability to overcome immunodominance and stimulate a broad antibody response targeting the subdominant HA stem epitope. There is a critical need to understand the precise mechanisms controlling B cell immunodominance and the breadth of humoral responses. The goal of this research is to gain mechanistic insights into how adjuvant formulation influences B cell intrinsic and extrinsic factors that shape immunodominance and the breadth of humoral responses. The overarching hypothesis is that the interplay between B cell intrinsic and extrinsic factors, prompted by prolonged TLR signaling through nanoparticle formulated adjuvant, facilitates the recruitment, selection and effector differentiation of germinal center (GC) B cells specific for the subdominant epitopes. This hypothesis will be tested through two specific aims. Aim 1 will determine how TLR7-NP impacts immunodominance, breadth and tolerance across different stages of B cell memory development. The goal of this aim is to identify the “bottleneck” that has the greatest influence on the immunodominance and breadth of the humoral response. Aim 2 will elucidate the interplay between extrinsic factors and B cells underlying the enhancement of antibody breadth by TLR7-NP. In this aim, we will use genetic models to manipulate the surrounding cell types as well as the microenvironment to investigate how these specific pathways contribute to the altered B cell immunodominance, a crucial mechanism underlying the breadth induced by TLR7-NP. This research proposal is innovative because it makes a conceptual advance in understanding the mechanisms governing the breadth of humoral responses, linking it to the spatiotemporal regulation of signaling through adjuvant formulation. This research proposal is significant because these insights will not only advance our understanding of how vaccine adjuvants work but also will empower us to refine vaccine strategies to precisely manipulate the breadth of B cell response. Ultimately, this will unlock new opportunities for the rational design of universal influenza vaccines. 1

NIH Research Projects · FY 2024 · 2024-08

Recent policy shifts in the Midwest of the United States have abruptly restricted access to pregnancy-related care for millions of women. Since June 2022, Midwestern states reinstated 150+ year old laws banning types of care, legislated the first new ban in the country, and saw widespread clinic closures. To understand the barriers faced by pregnant women seeking pregnancy-related care in the Midwest amidst these rapid policy changes and to quantify the impact of these changes on women’s health and lives, unbiased, locally relevant data are urgently needed. Yet, most reproductive healthcare access research has recruited participants from clinics—participants who, by virtue of having gotten to a clinic, represent a population that was able to overcome many barriers to care, and further, excludes women who use telemedicine and those who continue their pregnancies. A clinic-based sampling mechanism thus induces selection bias, resulting in research that misses or underestimates barriers to care. To address these methodological limitations, prospective research is needed that (1) expands recruitment to the full population of women seeking pregnancy-related healthcare, not just those who make it to a clinic, (2) sensitively measures barriers and facilitators to care, (3) captures the full set of possible pregnancy pathways (including miscarriage; birth; and clinical, telemedicine, and self-managed pregnancy care) and their sequelae over time, and (4) is sufficiently powered to answer nuanced questions at the regional level. To address this time-sensitive need in an underserved region, we propose four linked aims in a two-phase study. In the first year of the R61, we will recruit 500 pregnant women searching for information online and receiving support from community-based organizations to identify (via surveys; Aim 1) and characterize (via 30 in-depth interviews; Aim 2) the barriers women face while seeking pregnancy-related care in all 12 Midwest states. In the second (R33) phase of the study, we will follow these participants forward in time with additional surveys at 4, 10, and 18 months to measure pregnancy outcomes and the implications of their pregnancy outcomes on broader physical, mental, and socioeconomic health across states with different policy environments (Aim 3). Finally, we will implement a human-centered design approach to engage community-based organization leaders and Midwesterners who have sought pregnancy-related care to develop and test messages to address knowledge and information barriers (Aim 4). Findings from this study will drive more informed, geography-specific interventions to address urgent access challenges in the Midwest.