University Of California At Davis

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT Inflammation is a biological response in many diseases and organ systems, including Graft-versus-Host Disease (GvHD) and inflammatory bowel diseases (IBD). Toward alleviating the detrimental impact of acute or chronic inflammation, there has been rapid development of cell therapies such as mesenchymal stem cells (MSCs). While these have shown significant success in animal murine models for IBD and GvHD, dose efficacy and mechanism of action remain open questions for research. Further, these therapies have seen mixed success when evaluated in human patients, indicating a disconnect between preclinical and clinical evaluation of therapeutic efficacy. Experimental therapeutics in animal models are traditionally assessed on histopathology via pathologist scoring, but this has been shown to suffer from subjective interpretation and variability. Correspondingly, the field of digital pathology has had a transformative impact in biomedical research including significant advances in automated and computational evaluation of digitized slides from human specimens. However, preclinical research has not yet seen the benefits of rigorously developed digital pathology tools that are specifically optimized for slides and data from animal models. More accurate quantification of the underlying aspects of inflammation in preclinical data could help establish a more rigorous understanding of the therapeutic efficacy of cell therapies for inflammatory conditions occurring across multiple organs and diseases. Toward addressing these issues, this project will develop and validate a specialized suite of inflammation Digital Pathology Tools (iDPT) which will comprise new pathomics and machine learning models tailored for use in preclinical models of IBD and GvHD. iDPT modules will quantify domain- and data-specific characteristics of disease and treatment effects, thus facilitating deeper mechanistic insights, accelerating the identification of novel therapeutic targets, and ultimately contributing to the development of more effective treatments. This will be accomplished via three Aims. Aim 1 will construct iDPT modules for pre-processing, annotation, and quantitative assessment of inflammatory components on a pre-existing cohort of N=700 digitized hematoxylin and eosin (H&E) slides from preclinical models of IBD and GvHD. iDPT modules will then be evaluated through two distinct use-cases: (i) Aim 2 will evaluate the therapeutic efficacy of MSCs in a preclinical IBD model to confirm dosing and cell viability for suppressing inflammation, and (ii) Aim 3 will evaluate the use of MSCs in preventing the development of xenogeneic GvHD and mediating inflammatory response as a result of CAR-T therapy. Our project is built on the principles of open access and multidisciplinary collaboration to ensure broad impact for its outcomes, toward enabling innovation and discovery across the scientific community. Our long- term objective is to establish a new paradigm for preclinical inflammatory disease studies, offering standardized, scalable, and reproducible methods with broad impact while enabling discovery across the scientific community.

NIH Research Projects · FY 2025 · 2025-08

This application is in response to the Notice of Special Interest (NOT-DE-25-038): Basic and Translational Oral Health Research Related with HIV/AIDS. As stated in the NOSI, oral malignancies in people living with HIV (PLWH) are associated with enhanced local and systemic inflammatory states and closely linked to other viruses, such as Kaposi’s sarcoma-associated herpesvirus (KSHV). It has been known that a significant amount of KSHV virion is identified in saliva and KSHV-linked KS tumors in the oral cavity. Our application aims to understand the association between inflammatory tissue environment and KSHV replication. Our hypothesis is that cellular inflammatory signaling support the activation of a KSHV gene enhancer, which helps to maintain reactivatable latent chromatin. We will focus on viral IL-6 (vIL-6) functions to dissect the contribution of inflammatory signaling in KSHV replication and reprogramming of infected cells. Because the oral cavity is constantly stimulated with inflammatory signaling by oral bacteria, we hypothesize that such tissue environment allows KSHV to maintain epigenetically active latent chromatin. Trained immunity is a recently recognized feature of immune regulation, in which immune cells respond more quickly and robustly to subsequent exposure to similar triggers via transcription memories. Accumulating evidence suggested the broad benefit of trained immunity for normal host defense when cytokine expression is tightly controlled. However, studies also linked the transcriptional memory with chronic inflammatory disease when cellular responses continue to be overly reactive. In cells infected with the KSHV, an inflammatory viral cytokine is strongly expressed from viral gene independent of the tightly controlled host immune signaling networks. A critical question also pertains to why KSHV evolutionarily maintains multiple cytokine homologues that activate host inflammatory responses. KSHV infection is indeed associated with inflammatory diseases, including Kaposi's sarcoma and KSHV inflammatory cytokine syndrome (KICS). KSHV genome is also frequently maintained (detected) in inflammatory tissues such as the oral cavity and KS tumor. Here, we propose to study a hypothesis that KSHV utilizes a host cell trained immunity function by inducing cellular inflammatory signaling for its replication with vIL-6. We will study how KSHV vIL-6 re-programs viral and host gene expression by activating respective genomic enhancer domains to form transcription memories. We will also reveal whether the inflammatory tissue microenvironment is associated with maintaining active (re-activatable) latent chromatin. Completing this study should increase our understanding of the vIL-6 function in viral replication and association with KSHV pathogenesis.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Bioactive compounds produced by native or transient members of the gut microbiota are finding increasing use as therapeutics for a variety of complex human diseases, including obesity, diabetes, cardiovascular disease, and neurological disorders. Once these compounds cross the blood-brain-barrier, some can result in beneficial (e.g., anti-inflammatory, neuroprotective) or detrimental (e.g., pro-inflammatory, neurodegenerative) effects on the central nervous system (CNS). Specific cell types in the CNS are involved in this process, yet how the bioactive compounds interact with them remains poorly understood. Cell culture models offer better experimental control and scalability than animal models and avoid the limits of working with different species. However, most in vitro models suffer from low biological relevance in part due to not being able to simultaneously contain the critical cell types (i.e., neurons, astrocytes, microglia). There is a need for scalable in vitro models that can capture phenomenological outcomes of bioactive compound-CNS interactions and alterations to neural function observed in vivo and thus allow further mechanistic studies, bioactive compound discovery, and translation to human use. To address this critical need, we will employ a novel primary rat cortical cell tri-culture (primary neuron, astrocyte, microglia) model of neuroinflammation and integrated extracellular recording electrodes. In contrast to the co- culture of just neurons and astrocytes, the tri-culture model more faithfully captures neurotoxic and neuroprotective features observed in vivo. Since the tri-culture model is maintained simply by including IL-34, TGF-β, and cholesterol supplements in the conventional co-culture media, it is amenable to scale-up and screening studies in multi-well formats. In the proposed project, we will use the tri-culture model to simulate neuroinflammation that is present in many disorders that range from cancer to neurodegeneration. Microglia plays a particularly important role in neuroinflammation, where various phenotypic changes are observed, including impaired phagocytic capacity. Here, we will use a lipopolysaccharide (LPS)-induced neuroinflammation model. By introducing the bioactive molecules, before, during, or after LPS treatment, we will further mimic scenarios such as the normal presence of gut microbiota bioactive molecules (e.g., prevention) vs. their post- symptom introduction (e.g., therapeutic). Across two aims, we will reveal the effects of bioactive compounds on neuroinflammatory responses via morphological, and proteomic read-outs, as well as on cellular function by evaluating microglial phagocytic capacity and neuronal electrophysiological activity. We expect that this project will create a scalable in vitro tool to study the influence of gut microbiome-derived bioactive compounds on neuroinflammation. This tool can then be used broadly for mechanistic studies and for discovering new bioactive molecules that inform regular diet or prescribed as therapeutics for improving and maintaining neurological health.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Perinatal asphyxia is the leading cause of mortality in term newborns globally. Persistent pulmonary hypertension of the newborn (PPHN) is reported in about a quarter of the neonates with perinatal asphyxia is often secondary to meconium aspiration syndrome (MAS) and is a significant contributor to mortality. Majority of critically ill newborn infants with asphyxia and PPHN undergo therapeutic hypothermia and have systemic hypotension requiring vasopressors. However, commonly used vasopressors in newborns have variable effects on systemic and pulmonary vascular beds. Non-selective increase vascular tone in both systemic and pulmonary circulations in response to vasopressor agents may exacerbate PPHN. However, persistently low systemic blood pressure can lead to prolongation of a right-to-left shunt and worsen hypoxemia. The ideal vasopressor that is selective to systemic circulation and increases the ratio of systemic to pulmonary vascular resistance (SVR/PVR ratio) and enhances vital organ perfusion is not known. Additionally, the vascular and cellular mechanisms of vasopressor-induced changes in systemic and pulmonary circulations, in the setting of increased pulmonary vasoconstriction from PPHN and exposure to supplemental oxygen therapy remain unknown. In this K08, I will perform a randomized trial comparing the effect of use of dopamine, norepinephrine, epinephrine and vasopressin on SVR/PVR ratio, ventricular function, and cardiac output in a perinatal term ovine model of meconium aspiration, PPHN, therapeutic hypothermia and systemic hypotension. I hypothesize that use of norepinephrine and vasopressin will selectively increase SVR resulting in higher SVR/PVR ratio compared to dopamine and epinephrine that will non-selectively increase SVR and PVR without affecting the SVR/PVR ratio. Furthermore, I will perform in-vitro vascular reactivity testing with the vasopressor agents on systemic and pulmonary arteries from control and PPHN lambs ventilated with 30% and 100% O2 respectively and investigate vasopressor receptor expression. My training goals include acquiring hands-on experience in performing bedside targeted neonatal echocardiography, attaining practical experience in testing in-vitro vascular reactivity in response to vasopressors, interpreting vasopressor receptor expression in PPHN and hyperoxia, and enhancing my knowledge of biostatistics while developing professional and leadership skills necessary for executing development, that are in line with my research aims in this K08. These four key training goals along with preliminary data generated from this K08 will prepare me in applying for an R01 to investigate optimal blood pressure management in PPHN including dose-escalation of vasopressors.

NIH Research Projects · FY 2025 · 2025-08

Project Summary Spermatogenesis produces fertilization-competent sperm that ensure the continuity of life. The 3D chromatin structure in male germ cells changes dynamically during spermatogenesis. However, the relationship between the 3D genome and transcription in the male germline remains largely unknown. To elucidate how the 3D genome regulates the transcriptional program during mouse spermatogenesis, I performed high-resolution Hi-C in the male germline. I found that CTCF-mediated 3D genome formation in undifferentiated spermatogonia serves as an epigenetic memory, initially establishing a gene expression program that is subsequently used to direct gene expression during late spermatogenesis. My central hypothesis is that the 3D genome predetermines a transcriptional program that directs subsequent spermatogenic differentiation. Elucidating the molecular mechanism by which this 3D genome is formed will provide insight into how the transcriptional program is established to ensure unidirectional spermatogenic differentiation. Aim 1 will determine whether the 3D genome is established in prospermatogonia by capturing Hi-C and CTCF profiles from PGCs to undifferentiated spermatogonia. Aim 2 will elucidate molecular mechanisms underlying formation of the 3D genome by determining how specific CTCF-binding sites are selected from PGCs to undifferentiated spermatogonia. Completion of these aims will elucidate how the transcriptional program for male germline differentiation is established from the perspective of the 3D genome during the period from PGCs after sex determination to conversion to spermatogonia, thereby providing mechanistic insight into how male germ cell identity is established. This research will contribute to the treatment of male infertility and the improvement of human reproductive success in the future. So far in my postdoctoral career in the Namekawa lab at UC Davis I have received training in experimental and analytical skills for comprehensive epigenomic studies, which is the basis for this research proposal. In the K99 phase, I will receive training in analysis techniques specific to 3D genomes and develop and refine my future directions. With the training plan and support of the K99/R00 award, this project will provide me with a strong foundation for success as an independent investigator pursuing important biological questions in germ cells.

NIH Research Projects · FY 2025 · 2025-08

Project Summary/Abstract Post-acute sequelae of SARS-CoV-2 infection (PASC) is a persisting health challenge characterized by a range of symptoms affecting multiple organ systems, which continues to impact approximately 10% of COVID-19 survivors. Multiple, potentially overlapping, mechanisms have been identified that may play a role in PASC. However, with no effective preventative measures or treatments, there is a critical unmet need for understanding the pathophysiology of PASC; as previous studies, often limited by focus on peripheral blood biomarkers only or confined to single organ systems, have not sufficiently and quantitatively investigated the multisystemic and immune-related complexities of this condition in non-blood tissue. The long-term objective of this project is to bridge this knowledge gap by providing insight into the immune and systemic manifestations of PASC, through the innovative use of total-body dynamic positron emission tomography (PET) with the 18F-AraG radiotracer, which particularly offers selectivity towards activated T cells. To achieve this, we will use the dynamic PET images obtained from a high-sensitivity total-body PET scanner to develop, optimize, and validate a kinetic model for 18F-AraG in different anatomical sites and tissue types for multi parametric quantification of uptake. We expect that this will not only improve the quantification accuracy compared to standard static imaging, but also can shed light on the underlying mechanisms of uptake. The multi parametric imaging will be firstly used to identify sites of immunological perturbation in PASC patients, offering a total-body view of tissue-level manifestations of PASC. For this, we will compare the kinetic parameters of different tissues between symptomatic PASC participants and a control group consisting of individuals with a complete COVID-19 recovery. Second, we will integrate the multiparametric imaging data with peripheral blood assays, aiming to assess the correlations between certain 18F-AraG kinetic parameters and biomarkers of inflammation, immune dysregulation, and endothelial dysfunction in peripheral blood. Particularly, to identify vascular alterations in tissue and their association with endothelial markers in blood, we will use vascular permeability modeling to estimate the blood flow in different tissues from the early frames of the kinetic data. Third, we will employ a longitudinal design to quantify changes in 18F-AraG kinetic parameters and correlate them with evolving PASC symptom profiles over time. We will include two follow-up scans of the PASC participants at 4 months and 8 months after the baseline scans with systematic symptom assessments, focusing on individual patient trajectories. Through this, we expect to establish a direct and meaningful connection between molecular imaging data and clinical manifestations. In summary, the incorporation of cutting-edge imaging technology with quantitative modeling techniques for non- invasive evaluation of total-body immune response, combined with the longitudinal design of the study promises to provide unprecedented insights into this complex condition and would extend well beyond the confines of the PASC condition, offering frameworks and tools that could as well be used for other post-viral conditions.

NIH Research Projects · FY 2025 · 2025-07

Project Summary/Abstract Chikungunya virus (CHIKV) is an emerging mosquito-borne alphavirus endemic in 110 countries that causes febrile illness and arthritic disease. In November 2023, FDA approved the first CHIKV vaccine, a live attenuated vaccine (LAV) marketed as IXCHIQ. Although there are many important human pathogenic alphaviruses with a near global distribution, IXCHIQ is the first alphavirus vaccine licensed for humans. No studies have evaluated the ability of IXCHIQ to protect against infection, viremia, and disease after exposure to other alphaviruses that also occur in Latin America in areas with CHIKV or determined how prior heterologous alphavirus infection impacts IXCHIQ efficacy. This information is important to know as it will identify impacts of IXCHIQ campaigns in contexts where the vaccine is administered to populations with or without prior alphavirus exposure. As a LAV, IXCHIQ produces human viremias up to 5 log10 genomes/ml that peak at 3 days and last 1 week. These viremias exceed infection and transmission thresholds for CHIKV vector mosquitoes Aedes aegypti and Aedes albopictus in studies we and others performed with the same or nearly identical CHIKV strains as the backbone used for IXCHIQ. However, no studies have examined whether IXCHIQ is capable of transmission by mosquito vectors. If IXCHIQ is transmitted by Aedes, it could be spread by mosquitoes infected from viremic vaccinees, leading to infection of children, immunosuppressed, and pregnant people for which the vaccine is not approved. The goals of this project are to understand alphavirus circulation dynamics in the novel landscape of IXCHIQ rollout. These goals will be accomplished via the following 3 project Aims: 1) Determine cross-protective efficacy of IXCHIQ- induced immunity against heterologous alphavirus species and define the role of cross-reactive antibody in protection, 2) Define infectivity and transmissibility of IXCHIQ in Ae. aegypti and Ae. albopictus mosquitoes and determine protective efficacy of mosquito-delivered IXCHIQ, and 3) Determine the impact of prior infection with a heterologous alphavirus on IXCHIQ-induced immune responses and efficacy against CHIKV and define the role of cross-reactive heterologous antibody in protection or disease enhancement. We will use 4 heterologous alphavirus species that occur in Latin America where IXCHIQ rollout is expected as well as CHIKV in established mouse and non-human primate models, including a new Mayaro virus rhesus macaque model we developed. This project will identify consequences of IXCHIQ rollout in contexts where the vaccine is administered to people with or without prior alphavirus exposure and it will define the potential for mosquito-borne IXCHIQ spread in areas with Aedes, which can inform recommendations for recent vaccinees to prevent mosquito exposure. If IXCHIQ protects against disease caused by other alphaviruses, vaccine rollout could reduce incidence and burden of other alphaviruses in addition to CHIKV.

NIH Research Projects · FY 2025 · 2025-07

Project Summary/Abstract Exposures to aeroallergens and air pollution remain prevalent in young children, whose lungs are rapidly devel- oping and growing. Such exposures may increase the susceptibility to developing asthma and other lung dis- eases. Previous studies in non-human primate models of early life exposure, asthma and recovery showed that co-exposure to ozone and house dust mite (HDM) between 1 month to 6 months of age leads to hallmarks of asthma phenotypes in infant rhesus macaques (6-month cohort). Interestingly, following removal of exposures, exposed animals continued to have abnormal lung growth and airway hyperresponsiveness when they reached adolescence (36-month cohort). These long-lasting effects of early life exposure could be due to the epigenetic imprints of exposures on gene expression. Although relationships between early-life exposure, epigenetic mech- anisms, and persistent phenotypic changes in the lung have been supported by studies in humans and animal models, direct evidence supporting causal relationships from a model resembling human early life development is lacking and remains as a significant research gap. Our long-term goal is to understand the respiratory health effects of air pollution and aeroallergen and design mechanistically-driven novel therapeutics for lung diseases. The objective of this proposal is to characterize the impact of postnatal early-life exposure to HDM and O3 on the lung epigenome and explore a mechanistic link between these changes and alveolar development and air- way remodeling. Leveraging collected samples from established models and cutting-edge comprehensive epige- nomic analyses, we will test the hypothesis that postnatal early-life exposure to ozone and HDM alters the pulmonary epigenome and leads to changes contributing to persistent airway remodeling and abnormal lung development. Aim 1 will determine the epigenetic mechanisms responsive to early-life exposure to ozone and aeroallergen in the 6-month cohort of rhesus monkeys. Epigenetic changes induced by exposures and associ- ated with phenotypic changes will be identified from lung tissues collected following the last exposure in rhesus monkeys aged 6 months, and gene regulatory mechanisms underlying these changes will be inferred and pre- dicted. Aim 2 will determine the epigenetic mechanisms underlying continued abnormal lung function and de- velopment following early life exposures in the 36-month cohort. Lung tissues will undergo the same analyses as in Aim 1, and differences between the exposed and control groups will be determined. Data from Aims 1 and 2 will also be integrated to provide a better understanding of the longitudinal impact of early exposures. Comparative analyses to human asthma and mouse models of asthma may futher support the utility of non- human primate models of asthma. Leveraging existing tissues from established models, this exploratory and developmental R21 will provide new insights into the mechanisms of postnatal early-life lung development and the impact of early-life exposure on airway and lung remodeling and may provide new preventative and thera- peutic targets for lung diseases with an early life origin.

NIH Research Projects · FY 2025 · 2025-07

Project Summary/Abstract The flavivirus St. Louis encephalitis virus (SLEV) infects humans to cause febrile illness and rare fatal encephalitis. In California (CA), although human SLEV cases were detected from the 1940s-2003, none were reported from 2004-2015. Since 2015, SLEV reemerged and spread to cause more cases in CA and the Western US, including in areas with concurrent West Nile virus (WNV), a related flavivirus that invaded CA in 2003 that uses the same Culex vectors and avian hosts as SLEV. The reasons for the 11-year disappearance of SLEV from CA and its expansion since 2015 are unclear. Spread of WNV across the US was facilitated by augmented avian infection and enhanced vector competence but no studies have assessed host or vector phenotype for reemerging SLEV. Our genetic tracing studies show that post-2015 (contemporary) SLEV in the Western US likely originated in South America and is genetically distinct from pre-2003 (historic) SLEV. Our experimental studies show that birds inoculated with historic SLEV one month after WNV inoculation, a time when WNV neutralizing antibody titers are high, do not mount SLEV viremias, suggesting that SLEV after 2003 was displaced by extensive WNV avian herd immunity. Antibody-mediated neutralization is a hallmark of protection from flavivirus viremia and disease and antibody therapies are used in severe human cases of WNV and SLEV. Antibody is assumed to protect WNV immune birds from SLEV viremia and to prevent reinfection from sequential SLEV>SLEV infection but has never been experimentally confirmed. Our preliminary data show that serum from birds inoculated with historic WNV poorly neutralize some contemporary SLEV strains in vitro. Reduced antibody mediated cross-neutralization of contemporary SLEV by contemporary WNV could explain reemergence and persistence of SLEV in avian hosts. Augmented avian or mosquito infection may also contribute to SLEV spread. We observe increased infectivity of reemerging SLEV isolated in successive years since 2015 in duck embryonic fibroblast cells, but whether this pattern also manifests in avian reservoirs and in mosquito vectors is not known. To understand drivers of reemergence, the goal of this project is to identify how changing SLEV-vector-avian host interactions and cross-protection by WNV promote SLEV reemergence. This will be accomplished via 3 project Aims: 1) Determine transmission competence and fitness of contemporary versus historic SLEV in Culex, 2) Define avian fitness, antibody kinetics, and antibody-mediated protection for contemporary versus historic SLEV, and 3) Evaluate avian cross-protection conferred by prior WNV for contemporary versus historic SLEV. This project is significant in that it will define SLEV transmission dynamics in the context of sequential invasion and concurrent spread of 2 Culex-borne flaviviruses endemic to the US.

NIH Research Projects · FY 2025 · 2025-07

Abstract Entamoeba histolytica is a pathogenic amoeba and the causative agent of amoebiasis in humans. Despite its impact on human health, E. histolytica is dramatically understudied. The species name (histo-: tissue; lytic-: dissolving) derives from the ability to destroy host tissues. E. histolytica trophozoites (“amoebae”) invade the large intestine, causing ulceration and can enter the bloodstream, and they can disseminate to disseminate to other tissues, causing fatal abscesses. Little is known of the mechanisms that allow E. histolytica to damage the intestine. We established a new paradigm by discovering that amoebae attack and kill human cells by biting off and ingesting human cell fragments, which we named “amoebic trogocytosis” (trogo-: nibble) (Ralston, et al., Nature, 2014). Since trogocytosis occurs in many other eukaryotes, this process may be fundamental to eukaryotic cell biology. However, the mechanistic differences between eukaryotic trogocytosis and phagocytosis (ingestion of entire cells) are unclear, as are the signals that initiate each process. When amoebae are incubated with live human cells, they take bites, but when amoebae are incubated with dead human cells, they eat the entire cells. This suggests that either live human cells play an active role in trogocytosis, or that physical properties of live cells, such as deformability, are necessary. Building on the discovery of amoebic trogocytosis, we propose to further delineate the interplay between the amoeba and human cell, and what about this interplay leads the amoeba to perform trogocytosis vs. phagocytosis. Beyond E. histolytica, trogocytosis is a burgeoning theme with has far-reaching applications to eukaryotic biology. Several microbes use trogocytosis to kill other cells, including the “brain-eating” amoeba Naegleria fowleri. In multicellular eukaryotes, trogocytosis is used for cell-killing, cell-cell communication and cell-cell remodeling, and it plays roles in the immune system, the central nervous system, and during development. An improved understanding of the mechanism and biology of E. histolytica trogocytosis will apply both directly to the pathogenesis of amoebiasis and broadly to eukaryotic trogocytosis in general. This work is significant and high-impact as it will improve understanding of trogocytosis, a process that is changing the paradigm for amoebiasis pathogenesis. New understanding of the interplay between cells during trogocytosis will also apply broadly to eukaryotic trogocytosis in general.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY The long-term goal of this study is to increase our understanding of the immune mechanisms involved in the pathogenesis of allergic disease and asthma. Despite the identification of elevated Macrophage migration inhibitory factor (MIF) levels in asthmatic patients, the contribution of MIF to the pathogenesis of asthma remains elusive due in parts to the considerable heterogeneity of asthma endotypes and lack of mechanistic insights on the signaling of MIF in immune cells. Group 2 innate lymphoid cells (ILC2) are innate lymphocytes crucial in asthma, with their activation influenced by various external factors, including cytokines. Our pilot studies show that MIF is induced in the lungs after acute IL-33 and Alternaria alternata-driven lung inflammation, with activated lung ILC2s rapidly upregulating MIF receptor CD74. Our protein and transcriptomic analysis further suggest that the MIF/CD74 axis is associated with higher ILC2 activation, proliferation and metabolic activity in the lungs. Understanding how MIF controls ILC2 function can therefore provide a molecular and cellular framework for appropriate therapeutic approaches to target the pathophysiological root of asthma rather than symptom relief, meeting the pressing demand for innovative, mechanism-based therapies. Based on existing evidence and preliminary data, we hypothesize that the modulation of the MIF/CD74 axis in ILC2s has the potential to reprogram the intrinsic metabolic activities of ILC2s. This, in turn, could directly influence cellular function and contribute to the subsequent development of airway hyperreactivity (AHR). In the first part, we will identify the role of MIF on ILC2 function. Aim 1 will assess the expression patterns of CD74 in the lungs and measure the effects of MIF or CD74 inhibition on ILC2 activation and development of AHR in different in vivo murine models of ILC2-dependent asthma. The role of MIF on ILC2 metabolism will be characterized in the second part. Aim 2 will analyze how MIF affects specific ILC2 metabolic pathways fueling the activity of the mitochondria and provide potential metabolic approaches for the modulation of ILC2 activity. The proposed studies are conceptually and technically innovative because we will be the first to test the effects of the MIF/CD74 pathway in ILC2-driven asthma. In particular, our comprehensive combined cellular, molecular and metabolism approach utilizing 3 different MIF/CD74 inhibitors and genetically modified mice was not addressed in any other immune cell population. Our studies will therefore provide pre-clinical novel data on a previously unrecognized role of this pathway in ILC2 regulation. To conduct these studies, we have assembled an outstanding team including experts in the fields of asthma and immunometabolism. The convergence of the two aims in this project will address the mechanism through which MIF regulates ILC2-driven immune responses. Overall, the proposed project therefore holds the promise of uncovering novel mechanism-based targets that could significantly advance the treatment of allergic diseases and asthma.

NIH Research Projects · FY 2025 · 2025-07

Fragile X-associated Tremor/Ataxia Syndrome (FXTAS) is a rare, inherited, X-linked, late-onset, progressive neurodegenerative disease characterized by intention tremor, cerebellar gait ataxia, parkinsonism, and cognitive decline. Pathologically, it features brain white matter degradation, ubiquitin-positive intranuclear inclusions, widespread astrogliosis, and general atrophy. FXTAS is caused by a trinucleotide (cytosine- guanine-guanine or CGG) expansion between 55 and 200 repetitions within the fragile X messenger ribonucleoprotein 1 (FMR1) gene. Although there are no proven treatments for FXTAS, ongoing studies demonstrate that therapeutic strategies will be available soon in the form of antisense oligonucleotides (ASOs) that target the repeat-containing mRNA, agents that suppress cellular stress cascades, and molecules that lower neuroinflammation and reduce repeat associated non-AUG (RAN) translation. However, without fully validated fit-for-purpose clinical outcome assessments (COAs) and sensitive markers of treatment response, these future pivotal trials may not come to fruition and fail to provide conclusive, clinically meaningful, and impactful results for this devastating and rare disorder. In response to PAR-22-184, “Clinical Trial Readiness for Rare Neurological and Neuromuscular Diseases,” the overall purpose of this Research Cooperative Agreement (U01) application, “Clinical Outcome Assessment and Treatment Monitoring Markers for Clinical Trial Readiness in FXTAS” is to finalize refinement and validation of our primary COA – The FXTAS Rating Scale (FXTAS-RS) and a discrete set of neurocognitive, motor and neuroradiological outcome measures suitable for detection of treatment response. This will be accomplished by the study of 100 men and women with FXTAS who will be assessed at multiple time points over one-year, mimicking procedures that will be used in future clinical trials, including assessments of motor function, cognition, brain structural changes using MRI, and patient reported outcome measures (PROMs). We will document critical properties of the measures such as reliability, validity, sensitivity to change and clinical meaningfulness, and an Advisory Committee including patient self-advocates, clinicians, researchers and drug industry representatives will guide the work. We will develop a training program for clinical trial site teams to use the FXTAS-RS and harmonization of other measures will occur across the sites. By the end of this 5-year project, we will have 1) a validated fit-for- purpose primary COA and a discrete set of key cognitive and motor secondary endpoints for FXTAS clinical trials; 2) neuroradiological markers that are sensitive to FXTAS disease state and progression; and 3) a rigorous multi-center harmonized protocol and infrastructure that is ready to conduct Phase 2/3 trials for FXTAS.

NIH Research Projects · FY 2025 · 2025-07

Project Summary: The central nervous system contains a myriad of different cell types that, in the right numbers and at the right positions, receive and integrate information from the environment to generate the appropriate biological responses. Failure to produce the right composition of cells can result in several mental and physical diseases that range from cognitive disorders to severe brain malformations. In the neocortex, a specialized pool of neural progenitors -called radial glial cells- gives rise to all types of projection neurons in a conserved temporal sequence. The textbook’s view has been that the ability of the neural progenitors to produce different subtypes of neurons is controlled by an intrinsic cascade of transcription factors that restrict progenitor competence over time. However, there are reasons to question this model and in fact, the molecules responsible for the synchronization of fate acquisition with developmental time remain elusive. Importantly, published and preliminary data indicate that microRNA (miRNA) levels also affect neuron fate determination in a cell- autonomous manner, highlighting the previously overlooked importance of posttranscriptional mechanisms in cortical development. Here, we propose to unbiasedly uncover the dynamics of miRNA expression and the mRNAs targeted by miRNAs pairs in neural progenitors over time using miR-eCLIP. We will further unveil relevant molecular mechanisms regulated by miRNA-targeted mRNAs, and, in particular, their effects on RNA methylation and how such RNA modification affects PN fate specification. Overall, the proposed research will define the miRNAs and their target mRNAs involved in cortical cell fate acquisition and shed light on the molecular mechanisms regulated by this important post-transcriptional mechanism.

NIH Research Projects · FY 2025 · 2025-07

Epilepsy is a common neurologic disorder that cannot be controlled with medication in a third of patients. The most common form of epilepsy, and the most difficult to control, is medial temporal lobe epilepsy. Patients with epilepsy and their caregivers note substantial cognitive difficulties that are not well correlated with neuropsychological testing. Emerging evidence suggests this discrepancy is due to the lack of neuropsychological tests for core deficits in epilepsy. These deficits are primarily related to episodic memory. Emerging psychological theory places 'scenes' - integrated spatial and object representations - at the core of episodic memory, recollective experience, and imagining future scenarios. These memory processes appear to be at the heart of cognitive difficulties in epilepsy and overlap with disturbances in recognition memory that encompasses both familiarity judgments, including hyperfamiliarity and recollection. Our long-term goal is to delineate core memory circuits, particularly in relation to the deficits in epilepsy and after epilepsy surgery, and then develop better means to test these processes and avoid morbidity. To this end, we have updated a unique paradigm that now includes photorealistic dynamic scenes that can independently assay familiarity and distinguish this from validated recall. This paradigm addresses core features of memory and memory disturbances by including objects, spatial features, and temporal dynamics; it can be used to assay familiarity, recollection, and potentially memory generalization. We hypothesize that coordination between two distinct memory networks for encoding is required to perform this task during study scenes. During test scenes, recollective experience is necessary for the task and can overlap with encoding. This coincidence of recollective experience and encoding is a possible feature of deja vu, which, interestingly, can be elicited by this paradigm. We will use this approach and other behavioral measures to study a unique and large cohort of patients with epilepsy surgery-related focal lesions of pertinent brain regions. We will also prospectively test patients before and after surgery. This is followed by electrophysiological studies with paradigm modifications, including event- related potentials (in control subjects) and human intracranial electrophysiology (in pre-surgical epilepsy patients). These approaches will enable us to delineate the precise timing and neural circuits underlying these phenomena. Our rationale is that through these studies, we can design better tests for memory problems, avoid surgical injury to memory, and ultimately contribute to developing new memory treatments. We anticipate that our proposed studies will have impacts that extend beyond epilepsy in the clinical sphere and be highly informative for psychological theory and computational memory models. These studies will, therefore, significantly impact our understanding, classification, scoring, and treatment of memory disorders. Innovative approaches are employed in these studies, and we expect impacts on theory and memory models.

NIH Research Projects · FY 2025 · 2025-07

Project Summary The economic cost of traumatic injuries accounted for approximately 20% of the US economy in 2019, and were a top ten cause of death in the United States.1 More than one half of this cost ($2.4 trillion) was among working- aged adults (aged 25–64 years).1 Musculoskeletal injuries, including fractures, are by far the most commonly involved body system injured in multisystem trauma.2 This interaction of systemic trauma and fracture healing has profound effects on patient recovery, quality of life, and societal resource utilization. The field of bone regeneration has been primarily focused on healing fractures and treating bone loss in the context of isolated musculoskeletal injuries. Most closed, isolated fractures heal well in contrast to fractures in polytrauma patients which exhibit impaired healing in greater than 30% of cases.3,4 Clinical evidence suggests there may be distinct physiologic, cellular, and molecular mechanisms contributing to fracture repair in polytrauma. The underlying mechanisms contributing to fracture repair in polytrauma environments are poorly studied. This proposal is structured to interrogate the role of polytrauma on fracture repair. Leveraging my combined orthopaedic surgeon clinical experience and training in mechanistic research, I will investigate the role of inflammation and immunomodulation in a polytrauma model of fracture healing, focusing specifically on the response of mesenchymal stromal cells (MSCs). (1) I will first characterize the upregulation of inflammatory cytokines and innate immune cells both systemically and locally at the fracture site in the presence and absence of polytrauma. (2) I will then characterize the anti-inflammatory and therapeutic effects of MSCs on fracture healing when delivered systemically versus locally to the fracture site in a murine polytrauma model. My career goal is to conduct clinically relevant mechanistic research as an orthopaedic traumatologist. This K08 proposal will allow me to focus my efforts on acquiring the skills necessary to become a successful, independent clinician-scientist. My prior experience in the lab has afforded me the deep appreciation for the rigors of scientific inquiry, experimental design, and data analytics. I am already quite familiar with many techniques and through this career development grant, I will expand my mechanistic skillset. This award will provide a solid foundation for ongoing rigorous study design, execution, troubleshooting, and data management that will launch an independently funded, clinically relevant, research career. My advisory committee is comprised of highly accomplished and diverse group of mentors who will help catalyze my continued growth into a competitive researcher in the field of bone injury and healing. I am fortunate to be part of a world-class trauma unit with clinical partners who will support my clinical load and protect my time for the intensive demands of executing rigorous science. By harnessing my unique position, I will focus my research endeavors to combat the ubiquitous societal scourge of musculoskeletal trauma and advance fracture care with a goal of enabling patients to recover more effectively.

NIH Research Projects · FY 2025 · 2025-07

This R13 application requests funds to help support the 27th International Workshop on Kaposi’s Sarcoma Herpesvirus and Related Agents in 2025. Our annual conference will bring together researchers and clinicians working on the oncogenic human herpesvirus, Kaposi’s sarcoma herpesvirus (KSHV), and other closely related pathogens. KSHV is the etiology agent of Kaposi’s sarcoma (KS), which is still one of the most prevalent cancers in Africa, especially in HIV-1 infected individuals and other forms of immunosuppression, including treatments for organ transplant, malaria, and autoimmune disease. Many of these virus-associated malignancies are endemic to specific geographical regions and likely involve complex genetic predispositions and/or high-risk environmental co-factors. New and better therapies are needed for treating and preventing these virus-associated malignancies. The funds will be used to waive the conference registration costs for outstanding trainees (pre-/postdoctoral and early career researchers), who study oncogenic DNA viruses and the cancers associating with these viruses. The goals of these meetings are consistent with the mission statements of the NIH, NCI, NIAID, and NIDCR, namely, to advance and promote the pace of research on infections associated with human cancer and other diseases, including in the setting of HIV-AIDS. The main KSHV conference will be held in Redondo Beach, California, from the afternoon of June 29 through July 2, 2025. There will be a training workshop to engage both established researchers and trainees, clinicians, and basic researchers, to interact and develop collaborations on translational research projects. All remaining costs for the conference will be raised from registration fees paid by the conferees and contributions from host institutions, foundations, and pharmaceutical and biotechnology companies. The major focus of the KSHV meeting is the biology of oncogenic herpesviruses and associated human diseases, with specific emphasis on viral pathogenesis, viral latency and reactivation, viral gene expression and replication, host responses to infection, epidemiology, vaccine development, therapeutic intervention, clinical and translational research. In addition to KSHV, studies related to herpesvirus saimiri (HVS), murine herpesvirus-68 (MHV-68), and rhesus rhadinovirus (RRV) will be presented. The 27th International Workshop on Kaposi’s Sarcoma-Associated Herpesvirus and Related Agents will mark a new quarter century of research on KSHV since the discovery of this virus in 1994, a new century to engage clinicians and researchers.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY Salmonella enterica serovar Typhi (S. Typhi) is the causative agent of the systemic illness known as typhoid fever that affects 21 million people each year resulting in an estimated 200,000 deaths. Unlike other Salmonella serovars, S. Typhi is restricted to the human host where it causes a chronic systemic infection characterized by fever and flu-like symptoms. Due to the strict host-specificity of the bacteria, the mouse models available to study typhoid fever are limited to costly humanized mice or the use of the related bacterium Salmonella enterica serovar Typhimurium (S. Typhimurium). While S. Typhimurium and S. Typhi share 89% of their genomes, both serovars contain unique genes with over 600 S. Typhi specific genes, highlighting one of the limitations of using S. Typhimurium to model typhoid fever. Furthermore, in the humanized mouse model, mice are only susceptible to S. Typhi infection through parenteral routes (intraperitoneal or intravenous). For these reasons there is limited knowledge on how S. Typhi uses its virulence factors to invade the gastrointestinal tract and disseminate to systemic sites. Our preliminary results characterize a novel mouse model of typhoid fever in which mice are permissive to S. Typhi infection through a more natural, oral route. This important advancement allows research into the mechanisms of S. Typhi-specific invasion and dissemination from the gastrointestinal tract to be performed. We propose that S. Typhi uses its invasion associated type III secretion system-1 (T3SS-1) and/or flagella to invade the gastrointestinal epithelium and then evades host immune detection via the virulence polysaccharide capsule, Vi. To test this hypothesis, we will combine techniques such as bacterial genetics, antibody mediated neutralization of select cell types, and flow cytometry to assess the host immune response. The proposed research will generate robust and informative information on the mechanism of S. Typhi invasion that can help direct future research on typhoid fever treatment and vaccine development.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY/ABSTRACT This NIH Mentored Clinician Scientist Development (K08) proposal describes a comprehensive 5- year training plan designed to establish Geoanna Bautista, M.D. as an independent physician- scientist focused on mechanosensitive signaling pathways that contribute to neonatal intestinal dysmotility. Prematurity is the leading cause of death and disability in infants in the United States, and intestinal dysmotility resulting from an immature GI system is a significant yet understudied complication. Yet, no effective therapies address prematurity-related intestinal dysmotility due to critical knowledge gaps in understanding the development of GI motility. To address this foundational gap, Dr. Bautista will investigate mechanosensitive regulation driving intestinal maturation, which is important for establishing normal GI motility. Based on strong preliminary data supporting a critical role of the mechanosensitive ion channel, Piezo1, in normal intestinal motility and development, Dr. Bautista will test the hypothesis that Piezo1 is developmentally regulated and has a distinct role in the modulation of SMC Ca2+ activity during intestinal maturation. She will employ her previously established conditional knockout murine model to selectively deplete Piezo1 in SMCs during the neonatal period, combined with cutting-edge techniques such as super-resolution microscopy, live single cell and in situ Ca2+ imaging with and without stretch, to rigorously test this hypothesis. Specifically, in Aim 1, she will define the precise distribution and role of Piezo1 activation in SMCs of the developing intestine, and in Aim 2, she will determine the developmental consequences of Piezo1-deficient SMCs for intestinal contractility. The data to be obtained will provide new knowledge and mechanistic insights into the mechanical regulation of SMC [Ca2+]i signaling during intestinal development. More importantly, Dr. Bautista will gain training from the pioneering expertise of her diverse mentorship team in an excellent scientific environment. Dr. Bautista’s long term goals are to: 1) lead and perform rigorous mechanistic research to test clinically relevant models of intestinal mechanobiology during gut development, and 2) utilize these findings to explore innovative therapeutic strategies to address the burden of intestinal dysmotility in neonates. These goals will form the foundation of her independent research program, aligning closely with the NIDDK’s strategic plan and overall mission.

NIH Research Projects · FY 2026 · 2025-07

PROJECT SUMMARY Molecular imaging with hybrid positron emission tomography (PET) and x-ray computed tomography (CT) has been widely used in clinical applications. CT in PET/CT is typically operated with a single x-ray energy to provide anatomic localization and attenuation correction for PET. A major limitation of single-energy CT is its limited ability to characterize tissue compositions quantitatively. Dual-energy CT (DECT) fills this gap by employing two different x-ray energies to allow quantitative material decomposition. Combining DECT with PET may enable PET/CT for several applications that have been hitherto impossible or less feasible due to the lack of tissue composition information, for example, (i) for simultaneous PET and contrast-enhanced CT imaging where the presence of contrast media may compromise attenuation correction, and (ii) for PET imaging of bone marrow where neglecting the trabecular bone volume may underestimate tracer uptake in true bone marrow. Despite all the clinical potential, integration of DECT with PET/CT would require either a costly scanner hardware upgrade or a significant increase in radiation dose and scan cost. As a result, application of combined PET and DECT imaging has been largely hampered in molecular imaging with PET/CT. The PI and team have proposed a PET- enabled DECT imaging method. This method does not require a change of existing PET/CT scanner hardware or increase the radiation dose but exploits the inherent annihilation-photon attenuation property of a radiotracer to derive a 511 keV high-energy gamma-ray CT (gCT) image from a standard time-of-flight PET/CT emission scan. The enabling technique is the simultaneous reconstruction of the gCT image from the existing PET emission data, and the combination with low-energy x-ray CT (usually ≤140 keV) to form dual-energy imaging. Our preliminary work has demonstrated the feasibility of this PET-enabled DECT method for quantitative material decomposition using computer simulation and physical phantom studies. The goal of this R01 grant proposal is to further develop, validate, and translate this method in the context of PET/CT molecular imaging. We propose three specific aims to fulfill this goal: (1) Develop the technical approaches for gCT imaging with PET; (2) Evaluate the PET-enabled DECT method for material decomposition in human subjects; (3) Apply the method for metabolic PET quantification of bone marrow. Successful completion of these specific aims will establish an innovative PET-enabled DECT imaging method with broad applicability. The enabled or enhanced ability for quantitative material decomposition on PET/CT will open new avenues to improve molecular imaging. This project would make a significant impact in numerous clinical applications where the characterization of tissue composition matters, such as for bone-marrow PET/CT imaging or contrast-enhanced PET/CT in cancer, heart disease, and aging.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY Roughly 15% and 8% of adults in the United States experience a substance use disorder or major depressive disorder, respectively. The psychedelic class of medicines with agonism at the serotonin 2A receptor has shown promise for treatment of each of these; however, the often concomitant hallucinatory effects diminish their clinical utility, regulatory approval, and widespread adoption. A major question in the psychedelic field is whether the hallucinations can be separated from the antidepressant effects, or if they themselves confer therapeutic effects. The lab of Dr. Olson, a co-sponsor of this proposal, has developed a non-hallucinogenic serotonin receptor agonist, tabernanthalog (TBG), that shows preclinical antidepressant effects. The mechanisms of action of this small molecule are not completely understood. TBG has affinity for multiple receptors and pleiotropic actions within the brain. The availability of hallucinogenic and non-hallucinogenic compounds with shared therapeutic effects enables studies that to address a major question in the field: what are the commonalities and differences in drug-induced cellular and molecular responses between these classes of drugs. I will use paired snRNAseq and snATACseq as well as complementary histology and epignenomics assays to characterize the transcriptomic and epigenomic changes mediated by TBG and a hallucinogenic psychedelic, 5-Methoxy-Dimethyltryptamine (5-MeO-DMT) 24 hours after administration. I will test for cell type specific effects that are shared or differ between drugs and test if cellular responses are correlated with expression of the presumed relevant receptor protein, the serotonin 2a receptor. Specifically, I am focused on the synaptic and plasticity related genes that may drive the persistent therapeutic effects. There have been no reported studies applying these methods to non-hallucinogenic psychedelics, and most previous genetic studies focus on one or a limited set of genes. Finally, using paired snRNA and snATACseq will enable both cell-type specific resolution and linking transcriptomic and epigenetic changes. I will focus on synaptic/plasticity gene loci and attempt to distinguish signaling aspects that are common to both drugs (and thus potentially driving therapeutic effects). Additionally, I will test spatial organization of the responsive cells using RNA-FISH and perform histone CUT&RUN to test for chromatin state changes associated with accessibility and transcriptional regulation. This work will expand understanding of the mechanisms of action and the target cell types of these pleiotropic small molecule psychedelic drugs, and do so with multiomic modalities and systems-biology level of comprehensive analysis not previously attained. This proposal represents a cross-disciplinary effort linking pharmacology and genomics that brings powerful approaches to understanding basic cellular and molecular activities underlying promising psychedelic compounds for the treatment of MDD and other disorders, and these results will have further value towards development of next generation therapeutics.

NIH Research Projects · FY 2025 · 2025-07

Tuberculosis (TB) remains the leading infectious cause of death worldwide. A quarter of the human population is exposed to TB of which 5-15% will progress to active disease. Despite its extreme prevalence, prediction of disease progression is poor. To address this, our proposal integrates whole genome sequencing and single-cell RNA sequencing (scRNA-Seq) on the entire repertoire peripheral blood mononuclear cells (PBMCs) at three crucial TB disease states: latent TB infection, recent Mtb infection, and post-TB treatment completion. We will be the first to leverage this unique study design across TB states, for expression quantitative trait loci (eQTL) mapping. Our study population in South Africa resides in a TB-endemic area where we have over a decade of established research infrastructure, enabling us to efficiently capture these critical TB phenotypes at a relatively low cost. Previous TB eQTL mapping studies have been limited by inadequate phenotyping (e.g., samples from TB cases months- years after clearing infection), bulk RNA-seq (aggregating cell-type specific effects), or scRNAseq on one cell type. We are generating CITE-seq profiles from PBMCs, a cutting-edge technology that enables simultaneous profiling of gene expression and cell surface protein composition at the single-cell level [funded by CZI, co-PI Suliman]. This approach allows us to capture the fine-scale heterogeneity of immune cell states. To identify the genetic variants that regulate these identified cellular and transcriptomic changes, we propose to generate whole genome sequencing data paired with the transcriptomic data for eQTL fine-mapping. South African populations exhibits high levels of genetic heterozygosity and are multiway admixed, amplifying statistical power for discovering eQTL variants. To characterize the unique genetic diversity of our population we have optimized state-of-the-art ancestry estimation methods. Outcomes of this grant include multiomic data from 225 individuals across three TB states and eQTL identification of ancestry- and cell-specific variants which affect gene expression in early TB infection.

NIH Research Projects · FY 2025 · 2025-06

PROJECT SUMMARY / ABSTRACT Mycobacterium tuberculosis (Mtb) remains a threat to human health, and treatment of tuberculosis (TB) is arduous, requiring multiple, potentially toxic antibiotics administered for 4 months or more. Thus, there remains a critical need to understand the mechanisms enabling Mtb to survive prolonged exposure to lethal antibiotics. Mtb, like many bacteria, has the ability to form antibiotic persister cells ,which can survive for prolonged when exposed to bactericidal antibiotics. The goal of this proposal is to apply a combination of genetic and proteomic approaches to identify the key pathways enabling the survival of antibiotic persister cells in Mtb. In Aim 1 we propose to rigorously identify the genetic determinants of persister cells to identify genes needed for persister cell formation in multiple contexts, enabling an identification of core pathways contributing to persistence across multiple contexts. In Aim 2 we will use proteomics to define the signaling events associated with persister formation which will help both to prioritize factors studied in vivo, and to form clear biochemical hypotheses. If successful, these studies would provide insights that could be leveraged therapeutically to develop “persistence inhibitors” that act cooperatively with traditional antibiotics to hasten Mtb eradication in patients.

NIH Research Projects · FY 2026 · 2025-06

More frequent and intense wildfires in recent years have increased concerns about potential health impacts of wildfire exposures and the need to understand how to mitigate their effects, especially in vulnerable populations like pregnant people and their children. Prenatal ambient air pollution and particulate matter (PM) exposures are linked to adverse pregnancy and child neurodevelopmental outcomes. Epidemiologic evidence supports similar impacts of wildfire smoke events on pregnancy outcomes, but little is known about the potential mechanisms or their association with downstream child neurodevelopmental and behavioral outcomes. Pregnancy is a period vulnerable to environmental exposures like air pollution that can disrupt immune regulation and induce inflammatory responses. Further, given strong neuro-immune connections, maternal immune dysregulation and changes in cytokines and chemokines can disrupt fetal brain development. Non-human primate studies linked early gestational wildfire exposure to increased pregnancy loss, and greater inflammation, blunted cortisol, memory impairment and behavioral differences in offspring. Early-life wildfire smoke exposure also led to longterm changes in macaque nasal epithelium methylome over genes impacting the immune and nervous systems, including synaptogenesis, with related expression differences. As the closest animal model of human neurodevelopment and function, these macaque studies suggest developmental impacts of wildfires, with an early critical exposure period, immune and neuro-biologic pathways impacted, and cognitive and behavioral consequences. Preliminary findings from the B-SAFE cohort suggest similar impacts of prenatal wildfire PM2.5 on differentially methylated regions (DMRs) in baby nasal epithelium enriched for neuronal and immune related pathways and associated with bivalent chromatin marks of developmental genes, as well as novel evidence that prenatal wildfire PM2.5 is linked to significantly increased child behavioral problems. We also show associations with maternal inflammation. Wildfires can induce emotional and psychological distress. Combined exposure to contaminants and maternal stress could impact overlapping biologic pathways, including inflammatory responses, and induce serious long-term developmental consequences. We propose to be among the first to assess these links in a prospective cohort of 544 women and their children who were exposed perinatally to some of the highest PM ever recorded in Northern California wildfires from 2017 through 2021. The goals of this R01 are to examine whether wildfire PM2.5 exposure during pregnancy increases maternal immune dysregulation and altered epigenetic programming in children, whether these modifications mediate downstream increases in risk for adverse child neurodevelopmental and behavioral outcomes, and whether maternal stress modulates these responses. Enhanced understanding of immunologic and epigenetic pathways associated with prenatal wildfire exposures and their relevance to child neurodevelopmental outcomes could highlight strategies to minimize the adverse health effects of future wildfires and other environmental exposures.

NIH Research Projects · FY 2025 · 2025-06

Abstract Neurotrauma, such as traumatic brain injury (TBI) and spinal cord injury (SCI), impacts over 60 million people per year globally with an estimated fiscal burden of $400 B/year. The National Neurotrauma Society (NNS) is committed to the promotion of neurotrauma research by enhancing communication, providing a forum for scientific exchange, and increasing national and international support for neurotrauma research and clinical advances. Our Annual Symposium provides a forum for researchers, clinicians, and trainees from around the world to meet and discuss the latest breakthroughs to improve the lives of individuals living with TBI and SCI related disability. The 42nd Annual Neurotrauma Symposium, jointly sponsored by the NNS and the American Association of Neurological Surgeons/Congress of Neurological Surgeons (AANS/CNS) Joint Section on Neurotrauma and Critical Care will be held in Philadelphia, PA, June 15 - June 18, 2025. This meeting will focus on showcasing the latest technological innovations in the neurotrauma field while celebrating the foundational biomedical research that continues to influence emerging knowledge and tools to improve neurotrauma research and clinical care. Broad objectives of the Symposium include: 1) updating attendees on current and timely topics in basic and clinical neurotrauma, 2) facilitating exchange of information among participants, 3) building new scientific collaborations, with a foundation in rigor, reproducibility, transparency, and translation, and 4) fostering active engagement of trainees and young investigators through our travel awards, poster competition, data blitz oral presentations, workshops and activities developed and supported by the Training, Education and Mentoring (TEAM) Committee. Recent NIH R13 funding has enhanced our ability to train, mentor and facilitate attendance for graduate students, postdoctoral researchers, residents, and young investigators. This application seeks funding to support 20 travel awards for pre-and-postdoctoral trainees, and an additional 10 travel awards to pre- and-postdoctoral trainees who meet the NIH definition of historically underrepresented in the biological sciences. These awards will allow trainees to present their ongoing work, attend scientific sessions, and to network and build collaborations with peers and faculty. In addition, we request support for programming for trainee and young investigator development sessions, sponsored by the TEAM Committee. Finally, we request support to continue our unique Local Scholars Program, successfully initiated in 2022, which provides support for undergraduate students who come from under-served backgrounds and who attend local universities to attend the NNS Symposium and receive specific training to promote interest in careers in neurotrauma and neuroscience research. Along with showcasing the best of our research over the last year, our goal is to create an inclusive and equitable environment, where trainees and faculty alike feel like they belong. In this way we hope to be able to better recruit, better train, and better retain the next generation of translational neuroscientists in the NNS.

NIH Research Projects · FY 2025 · 2025-06

There are ongoing clinical trials premised on the idea that systemic treatments that increase Nicotinamide Adenine Dinucleotide (NAD+) will increase baseline metabolism within retinal ganglion cells (RGCs) which will make them more resilient to increases in intraocular pressure or other insults that might result in the characteristic pattern of axon and vision loss that defines glaucoma. However, much of our understanding about how NAD+ mediates neuroprotection derives originally from studies of Wallerian Degeneration Slow (WldS) mice, which after traumatic injuries have not only a ten-fold delay in axonal degeneration but also greatly delayed inflammation. Very recently, using a novel optic nerve crush (ONC) model in young Xenopus laevis (X. laevis) tadpoles, we reproduced the strong axon protective effect of the NAD+ boosting interventions and similarly found a large inhibition of myeloid cell recruitment to both the optic nerve and the main brain innervation target of RGCs. However, we then demonstrated that pharmacogenetic ablation of myeloid cells fully recapitulates the axon preservation afforded by the NAD+ boosting interventions. Our studies thus suggest that at least some of the neuroprotection afforded by NAD+ boosting interventions may be due to their anti-inflammatory activity. Here, we will determine whether NAD+ based interventions also work in a less traumatic, more glaucoma relevant, novel axon injury model that is based on conditional expression in RGCs of a rare glaucoma-associated Optineurin (OPTN) variant that produces extensive axonal loss. We will characterize the immune reaction in this model and determine whether immune cells are similarly consequential to the axonal degeneration. And then, using another novel axon degeneration model based on conditional expression in RGCs of a constitutively active form of the NAD+ catabolic enzyme Sterile Alpha and toll/interleukin-1 resistance (TIR) motif containing 1 (Sarm1), we will directly test the hypothesis that the timing of axon destruction due to NAD+ consumption within RGC axons in vivo is necessarily dependent on the consequent immune invasion. We will also test a corollary hypothesis, namely that injured axons damage healthy axons through the immune cells they recruit. In the process of answering these mechanistic questions about how axons may be damaged in glaucoma, this proposal will develop and characterize two novel axon injury models that we hope to use in the near future for large-scale genetic and pharmacological screens.