Washington University

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Post-traumatic joint contracture (PTJC) causes debilitating loss of motion following joint injury and is particularly impactful in the elbow. Clinical treatment is limited due to a poor understanding of key mechanisms leading to motion loss, making treatment targets elusive. This study will use a validated preclinical animal model of PTJC to identify the key aspects of physical- and biological-based interventional strategies that best limit PTJC following injury. Early joint remobilization improves range-of-motion (ROM); however, clinical practice requires a period of joint immobilization following injury to reduce instability and prevent joint overloading. The parameters of active therapy (i.e., initiation, duration, intensity) that best limit PTJC after an initial immobilization period without destabilizing or overloading the healing joint remain unknown. In addition, while studies have shown that modulation of the inflammatory response can improve healing after joint injury, and that T-cell-mediated signaling might represent a particularly effective target, protocols guiding inflammation-based therapeutic approaches for PTJC remain poorly defined. Overall objective: identify fundamental aspects of physical and biological treatment strategies (i.e., initiation, duration, intensity, synergy) that prevent the development of PTJC using a preclinical animal model and multi-modal, machine learning (ML)-based analyses. Aim 1: Identify parameters of voluntary active physical therapy that are most critical to minimizing PTJC while promoting healing after joint injury. This study will determine the optimal implementation of active physical therapy protocols to best preserve ROM yet limit load-induced damage. Image-based ML algorithms will be used to automate/accelerate spatial analysis of joint tissues and advance clustering analyses to elucidate cell- and tissue-level responses to physical treatments. Hypothesis: moderate intensity/duration physical therapy will maximize motion and limit joint damage, with additional benefit achieved by implementing a slightly staged increase in intensity after joint remobilization. Aim 2: Develop biological strategies to reduce PTJC using anti-inflammatory intervention and targeted modulation of the T cell mediated immune response following joint injury. Anti-inflammatory prevention strategies will be developed and strategically combined with physical therapy to target multiple phases of immune-mediated biological activity. ML algorithms will combine multi-modal experimental data to explore spatial relationships in PTJC pathophysiology. Hypotheses: (i) reducing inflammation in the post-injury and post-remobilization periods will help preserve ROM; (ii) improved outcomes from blocking T cell activity will demonstrate a key mechanism of PTJC etiology; (iii) ML-driven data analysis will determine that abrogation of capsule fibrosis, reduced remobilization-induced ligament hypertrophy, and limited T cell activity will be most predictive of preserved joint function. While results obtained using an animal model aren’t directly translatable to human care, this study will greatly advance understanding of PTJC pathophysiology and elucidate key principles of physical and biological interventional strategies that can be leveraged to inform future treatment of PTJC.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT Endometriosis affects 10-15% of women, causing musculoskeletal pain, bowel and bladder problems, mood disorders, and infertility. Chronic pelvic pain is the most common symptom affecting 57.2% of patients. Endometriosis is responsible for an estimated $69.4 billion per year in excess health expenditures, and significant loss of workplace and household productivity. Current strategies to treat endometriosis, such as hormonal therapy or surgery, often fail to prevent recurrence and the associated pain. Therefore, we must develop, test, and implement new strategies to treat this condition. Given the broad impact of endometriosis- associated pelvic pain on multiple domains of life, we developed the Peer Empowered Endometriosis Pain Support (PEEPS) intervention that is comprised of three evidence-based approaches: interdisciplinary and integrated care delivered in a group care model. The PEEPS intervention was adapted from Chao et al.’s Centering Chronic Pelvic Pain group care program by our team of a physical therapist, yoga instructor, clinical psychologist, and endometriosis specialist. Adaptation was informed by implementation science and community engaged research methods. Through this process, we optimized PEEPS to an 8-session group care program that incorporates education on endometriosis, physical therapy, mindfulness, yoga, nutrition, and strategies to cope with chronic pain. We hypothesize that PEEPS will lead to decreased endometriosis-associated pain interference, improved quality of life, and decreased pain catastrophizing. To determine PEEPS preliminary effectiveness, I propose to perform a pilot single-site randomized controlled trial of PEEPS to education. I hypothesize that patients in PEEPS will show greater decrease in pain interference in daily activities than those in the education arm. With 30 patients in the PEEPS arm and 30 in the education arm, this pilot trial may be powered to detect an effect size of 0.8 based on the effectiveness of the intervention components. In parallel to the randomized controlled trial, we will evaluate barriers, facilitators, and implementability of PEEPS. Most research and evidence-based interventions never reach patients and clinical practice. As PEEPS is comprised of several effective interventions that have a high probability of synergistically improving participant symptoms, we will use an implementation science approach in order to understand how to most effectively translate PEEPS to real-world practice. I will use the data and experiences I gain in conducting this single-site randomized controlled trial to optimize PEEPS and apply for R01-level funding to scale up the intervention and perform a multi-site randomized controlled trial.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY The proper folding of proteins is essential for all life, making aberrant protein folding severely problematic. The protein homeostasis (proteostasis) network is a tightly regulated system that ensures that proteins are folded and degraded as necessary. However, misfolded proteins can overwhelm proteostasis. Misfolding is associated with numerous devastating neurodegenerative, heart, and kidney disorders, as well as forms of cancer. Protein disaggregases can engage misfolded substrates and dissolve them, promoting their return to proper fold and function. Disaggregases may serve as the final defense against collapse of proteostasis. Our central hypothesis is that disaggregases play key roles in maintaining cellular health, but are vulnerable when the proteostasis network becomes overwhelmed in disease. Therefore, technologies that enhance disaggregase activity may be therapeutically useful. However, disaggregases are the least characterized branch of the proteostasis network. My research program focuses on advancing our understanding of how disaggregases counter misfolding, both in health and under stress. We seek to elucidate how cells maintain proteostasis, how proteostasis fails, and how protein disaggregases might ultimately be applied to prevent or reverse collapse of proteostasis. During the first period of this award, we focused on engineering tailored Hsp104 variants, better understanding mechanisms of misfolding, and have begun characterizing human disaggregases. During the next phase, we will continue and expand our efforts to better understand the roles disaggregases play in safeguarding against misfolding. We will focus on two primary themes: (1) we will develop finely-tuned Hsp104 variants and use these variants as probes to test the effects of disaggregation in different systems. These studies will also reveal insights into the mechanism of Hsp104, and we aim to better understand how Hsp104 recognizes and distinguishes among substrates. Further, because Hsp104 is a member of the AAA+ family of proteins, which serve crucial roles across biology, our findings can be broadly applied. (2) We will characterize and develop approaches to modulate human disaggregases. Here, we will focus on understanding the structure, mechanism, and substrate repertoire of human disaggregases. We also aim to develop approaches to modulate the activity of human disaggregases. Beyond their possible therapeutic utility, finely-tuned disaggregases can be used as probes to test the hypothesis that misfolded species are toxic, and that restoration of proteins to their native folds and functions can reverse disease phenotypes. Ultimately we aim to apply the findings from these studies to develop new strategies to treat protein-misfolding diseases. This is especially important because, despite intense efforts, there are no effective therapeutics available to treat numerous protein-misfolding disorders.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Therapeutic restoration of protein function is a key goal for many neurodevelopmental disorders (NDDs) caused by genetic haploinsufficiency. MYT1L syndrome is a recently identified, understudied NDD caused by heterozygous loss of function mutations in the MYT1L gene, characterized by global developmental delay, particularly in motor and language development, intellectual disability, highly penetrant obesity and hypotonia, and a significant comorbidity of autism spectrum disorder (ASD) and/or attention-deficit/hyperactivity disorder (ADHD). Mice and human neurons with a MYT1L stop-gain mutation only show ~30% decreased transcript and protein levels, yet display profound cellular, molecular, and behavioral anomalies, indicating that MYT1L levels need to be tightly controlled for normal function. However, it is unknown how MYT1L levels are regulated. Understanding this regulation is a key step to identifying clinically relevant strategies for upregulating MYT1L as a therapy for MYT1L syndrome. An emerging protein upregulation strategy is to use antisense oligonucleotides (ASOs) to block elements that normally destabilize mRNA. Thus, ASOs could be used to increase protein expression from the mRNA of the remaining healthy allele. MYT1L has a conserved and longer than average 3’ untranslated region (UTR), which contains many regulatory elements responsible for transcript stability and translation efficiency, properties critical for protein synthesis. Using bioinformatic analyses, existing microRNA (miRNA) binding data, and a preliminary screen for active regulatory elements using a massively parallel reporter assay (MPRA), we have identified regions that may reduce stability of MYT1L transcripts, which I have therefore termed MYT1L Negative Regulatory Elements (MNREs), and discovered several candidate ASOs that increase expression. MNREs contain several predicted miRNA response elements (MRE) and Pumilio (PUM) response elements (PRE), which both induce translational repression or transcript degradation. This project seeks to study MYT1L post- transcriptional regulation (PTR) and its potential to be harnessed in translational therapies like ASOs for MYT1L syndrome. Aim 1 will test the importance of specific MRE and PRE sequences in MYT1L mRNA stability and screen for additional MNREs using an MPRA in iPSC-derived neurons to capture PTR in an appropriate cellular context. ASOs will both be used as tools and potentially be deliverables. Aim 2 will test the efficacy of a candidate ASO targeting a conserved MRE in restoring MYT1L protein levels in MYT1L haploinsufficient mice and rescuing associated transcriptomic and behavioral phenotypes. These findings will begin to explore MYT1L PTR and provide clinically relevant insights for rescue of MYT1L haploinsufficiency by direct targeting of MYT1L. This project will also provide the applicant the opportunity to train in RNA therapeutics, computational genomics, animal behavior to prepare for a future career as a physician-scientist studying neurogenetic disorders.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY / ABSTRACT: Cognitive deficits are, arguably, the most prevalent, the most disabling, and the most economically costly symptom of schizophrenia (SZ). There are, however, no FDA-approved medications to improve cognitive function in SZ. Computational models have proposed that cognitive deficits in SZ arise from disruptions to cortical excitation:inhibition (E:I) balance, and these models have inspired new outcome measures for randomized control trials (RCTs), as well as new model paradigms for SZ. Nevertheless, it is unclear whether computational models of E:I balance in SZ will spur innovations in cognitive-enhancing therapeutics. Conventional approaches to modeling of E:I balance in SZ are limited in several ways, which may negatively impact their ability to improve the assessment and treatment of cognitive deficits among those with SZ. The following F31 NRSA proposal seeks to translate a recently-validated modeling approach, which does not exhibit these limitations, to the study of E:I balance in SZ. Mesoscale Individualized Neurodynamic (MINDy) modeling, which was created by members of this NRSA proposal team, is an approach for inferring the biophysical parameters of an individual’s brain using functional magnetic resonance imaging (fMRI) data. MINDy models provide a proxy-measure for E:I balance that is, unlike conventional computational models of E:I balance in SZ, both statistically reliable (Test-Retest R > 0.7) and quick to compute for every region of interest in the brain (~1min compute time). The following NRSA proposal leverages these advantages to investigate disruptions in brain-wide E:I associated dynamics among those with SZ using a wide range of clinically-relevant study designs. For aim one of this NRSA, we will use MINDy to investigate brain wide disruptions to E:I-associated dynamics among individuals with SZ by using resting-state fMRI data from the Human Connectome Project – Early Psychosis (HCP-EP) study. While conventional approaches have demonstrated that SZ is primarily associated with disruptions to E:I balance in association cortical areas such as the prefrontal cortex, there is some evidence that E:I balance may also be disrupted in sensory cortical areas. This has not been explored with computational models of E:I balance. We hypothesize that MINDy will uncover disruptions to E:I-associated dynamics in both association and sensory cortical areas among those with SZ. For aim two of this NRSA, we will use MINDy to investigate genetic contributions to individual differences in brain-wide E:I balance among a sample of approximately 900 monozygotic and dizygotic twin pairs from the Adolescent Brain and Cognitive Development Twin (ABCD-Twin) study. While existing work suggests that general changes in gene expression across the neocortex are related to differences in E:I balance between cortical areas, aim two of this NRSA proposal will be the first study to investigate whether the distribution of E:I balance across the brain is related to individual differences in heritable genetic factors. Lastly, aim 3 of this NRSA proposal will combine MINDy with a computational model of decision-making, called the drift-diffusion model, in order to investigate links between brain-wide E:I balance and the efficiency of evidence accumulation during perceptual decision making. The contribution of E:I disruptions to cognitive deficits have primarily only focused on the association cortex and also have only focused on working memory paradigms. In order to address both of these gaps, this F31 NRSA proposal will use MINDy modeling to investigate relationships between brain-wide E:I disruptions in SZ and perceptual decision making.

NSF Awards · FY 2024 · 2024-09

This project will develop theory and methods to address network-level uncertainty in the control of large-scale networked systems. In such systems, the communication topology is described by a directed graph, whose nodes represent the agents in the system and the edges represent the communication links between them. To meet the modelling demands of realistic multi-agent settings, it is known that one needs to account for process and observation noise. The goal of this project is to explore what lies beyond these requirements and account for uncertainty at the level of the communication topology. To this end, the project will integrate the fields of random graph theory, and particularly graphon theory, with structural system theory to develop the needed theoretical tools to model and understand network-level uncertainty in control systems. Graphons are relatively new models in the landscape of random graph theory. They generalize many existing random graph models, such as the Erdös-Rényi model, by allowing for heterogeneous edge densities. A major research goal of this project is to characterize completely how a given structural system property behaves under network uncertainty. The main research problem of the project is the following: Given a desired system property (e.g., controllability and stability), what is the probability that a graph sampled from a graphon can sustain the property? The problem is by nature combinatorial and probabilistic. To tackle these challenges, this project will rely on tools from analysis and geometry to develop a new set of ideas geared toward the computation the aforementioned probabilities in the asymptotic regime, where the size of the random graph goes to infinity. The intellectual merits of this project lie in the use of methods from graphon theory, probability, and combinatorics to understand control system properties. More specifically, the project will (1) formulate new problems at the intersection of structural system theory and graphon theory, (2) develop a new toolbox for analyzing structural properties for network systems drawn from graphons, and (3) establish new theoretical results and algorithms that may have impacts on both areas and beyond. A major novelty of the proposed approach is that the project leverages tenets from graphon theory to circumvent the complexity of combinatorial problems, leading to the use of analytical tools and geometric approaches to provide complete solutions. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-09

Program Summary Amyotrophic lateral sclerosis (ALS) is a devastating neurodegenerative disorder characterized by loss of motor neurons that leads to weakness, respiratory failure, and death within 3-5 years of symptom onset. The importance of prognostic and pharmacodynamic biomarkers in therapeutic development is highlighted by the emergence of neurofilament light (NfL) and phosphorylated neurofilament heavy (pNfH) as potential neurodegenerative biomarkers for ALS. Neurofilaments (NFs) are represented by three subunits: NfL, neurofilament medium (NfM), and NfH that complex with -internexin in the central nervous system (CNS) or peripherin in the peripheral nervous system (PNS). NFs undergo extensive post-translational modifications (PTMs) (i.e. phosphorylation, O-glycosylation) that regulate neurofilament assembly, transport, and function and are known to form pathologic aggregates in ALS. An antisense oligonucleotide to SOD1, tofersen, was recently granted accelerated approval for hereditary SOD1-ALS based on its ability to lower NfL and pNfH by immunoassay in serum and CSF by ~60% at 12 weeks, long before clinical improvement was observed at one year. However, immunoassay methods are vulnerable to non-specific signals and are unable to discriminate between alternative isoforms or PTMs that may occur with disease. We have developed a proteomic assay for NfL that has indicated NfL exists only as truncated fragments in ALS CSF and have found that Coil 1 domain peptide species correlate best with ALS disease progression. We have also developed reagents and methods to extend analysis to NfM and NfH. By comparing neurofilament (NF) species in ALS and control biofluids, we anticipate that we will identify NF isoforms and PTMs unique to ALS. We will also measure NF isoforms pre- and post- tofersen treatment in blood and CSF from SOD1-ALS participants and compare their performance to existing NfL and pNfH immunoassays. We recently demonstrated that stable isotope labelling kinetics (SILK) can be safely employed in ALS participants and showed that mutant SOD1A5V protein turnover is faster than its wild-type counterpart. In this study, we will examine the effect of SOD1 lowering therapy on neuronal proteins, tau and NfL, and perform proteomic analysis to assess changes in protein expression pre- and post-treatment. We propose that in-depth proteomic and protein kinetic analysis of biofluids from the tofersen treated SOD1-ALS population provides an unparalleled opportunity to uncover biomarkers related to clinical improvement in ALS.

NIH Research Projects · FY 2025 · 2024-09

ABSTRACT The long-term goal is to understand how basal forebrain (BF) circuits underlie cognitive functions like sustained attention. Traditionally, BF was thought to mediate attention through its cholinergic neurons and cortical acetylcholine release. However, recent recordings suggest that cholinergic signaling reflects arousal and reinforcement learning instead. If not cholinergic neurons, what alternative BF circuits support attention? The objective of this proposal is to determine the circuit and behavioral roles of an understudied BF population: parvalbumin-expressing GABAergic neurons (BF-PV) that project to the cortex. Our previous work using a sustained attention-demanding auditory detection task revealed that BF cholinergic neurons respond to reinforcement surprises, independent of temporal fluctuations of attention. This discovery prompted an investigation into alternative BF neuron types. Our preliminary studies suggest that BF-PV neurons have key attributes relevant for sustained attention: their activity predicts detection accuracy and reaction time on a trial-by-trial basis, and their optogenetic stimulation enhances performance. Using quantitative behavior, cell-type-specific recordings and manipulations, viral tracing, and computational modeling, we aim to explore the link between BF-PV neurons and sustained attention. Aim 1 will map BF-PV cortical projections and investigate whether they produce cortical gain control through disinhibition. Aim 2 will record BF-PV neuron activity to assess how its dynamics predict momentary attention levels during sustained attention tasks and use optogenetics to evaluate their necessity and sufficiency. Aim 3 seeks to determine if BF-PV neurons convey motivational salience signals that guide attention allocation. This work will elucidate how long-range, cortex-projecting BF-PV neurons support sustained attention, distinguishing their contributions from known cholinergic effects. Defining the computations and connectivity underlying sustained attention will provide fundamental insights into basal forebrain circuits for normal cognition. Illuminating this poorly understood pathway may also reveal new therapeutic targets for attention disorders and BF-related dementias like Alzheimer's and Parkinson's disease.

NIH Research Projects · FY 2026 · 2024-09

PROJECT ABSTRACT Sensory over-responsivity (SOR), or strong negative reactions to and avoidance of innocuous sensory stimuli, affects about one in five school-age children and about two-thirds of children with autism spectrum disorder (ASD) and several other common neurodevelopmental disorders. Children with SOR experience considerable short- and long-term distress and impairment including fear and anxiety, poor sleep and nutrition, isolating social difficulties, and increased risk of mental illness. The cost of SOR in childhood is compounded by its disruption of developmentally appropriate social and situational experiences and its deleterious effects on family functioning. Despite its prevalence and impact on health and wellbeing, the causes of SOR are poorly understood and existing treatment approaches have met with limited success. Identifying the specific neural mechanisms that are disrupted in SOR could provide insights into its etiology and suggest promising approaches for developing effective interventions. Studies of typical sensory processing have revealed basic neural mechanisms that promote adaptive sensory responses, highlighting a powerful new translational approach to investigating the neural bases of SOR. The goal of this K99/R00 Pathway to Independence Award is to provide the applicant with the training needed to test if these neural mechanisms are disrupted in children with SOR and to support her continued success as an independent investigator. To achieve this goal, the applicant has assembled a committee of exceptional mentors and experts who will provide her with training in clinical presentations and assessments of ASD and SOR (Drs. Constantino, Sylvester, Green, and Pruett), administering functional MRI scans to children with and without ASD and SOR (Drs. Green, Sylvester, Dapretto, Pruett, and Dosenbach), applying multilevel models to complex datasets (Dr. Jackson), and developing skills for success as an independent investigator at a major research institution (all committee members). The proposed training will allow the applicant to test predictions about the relationship between one neural mechanism (suppression) and SOR in children using existing data and to pilot a functional MRI task to assess a second neural mechanism (surprise) in children during the K99 phase. Results from this work will inform the R00 phase, which will entail testing whether three neural mechanisms (adaptation, suppression, and surprise) are attenuated in sensory and fronto-limbic brain areas of children with SOR, both with and without ASD. This innovative research approach will clarify whether predictive mechanisms are disrupted in children with SOR and localize disruptions to specific brain areas, advancing scientific understanding of SOR and highlighting promising targets for interventions to be tested in a R01. Collectively, the proposed training and research will provide the applicant with the data, tools, and skills needed to launch a successful career at a top-tier research institution.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT The dynamics of typical visual input challenge the brain: movement of objects or caused by navigation shifts visual information over the retina, and complicates the brain’s goal of understanding visual input as an organized collection of objects. Visual scene segmentation occurs in the ventral - ‘what’ - stream in the primate brain, hence prior studies have studied these circuits mostly under static conditions, since motion is thought to be mainly processed elsewhere, in the dorsal – ‘where’ – stream. These prior studies found that the earliest surface segmentation signal in the primate brain is selectivity for border ownership. Border ownership neurons respond differently to an identical border in their receptive field depending on which side of the border is owned by foreground. It is poorly understood how these circuits contribute to the processing of the dynamic, complex visual scenes in everyday environments. Recently the applicants found that border ownership units with similar properties as those in the brain emerge in an artificial neural network trained to predict the next frame in natural videos, even though the network was not trained to segment objects. This indicates that these units aid in predicting future input. The overarching goal of this proposal is to understand how border ownership units contribute to this objective. Specific aims are to understand 1) the stimulus diet that drives the emergence of border ownership units in artificial neural networks (Aim 1); and 2) how these units benefit the prediction of future input in natural videos (Aim 2). To this end the applicants will perform experiments in artificial neural networks (in silico) as well as in the non-human primate brain. This exploratory research may thus lead to a paradigm shift in the understanding of scene segmentation circuits, which traditionally have been assumed to perform a function typically considered under static conditions: segmenting objects from background. This research addresses several research needs recognized by the National Eye Institute, including exploring the connections between biological measurements and theoretical models of vision processes, and understanding processing in higher brain areas to inform the design of next-generation cortex prostheses. Moreover, it may lead to better diagnostic and therapeutic tools for disorders characterized by disrupted perception of complex visual input, such as visual agnosias and schizophrenia. This project is a collaboration between Dr. Franken (PI), an electrophysiologist with expertise in border ownership circuits in the primate brain, and Dr. Wessel (co- I), a neurophysicist with expertise in studying visual processing (including natural videos) in a variety of systems, including in artificial neural networks. The experiments will break new ground and extend previous work in a new and promising direction. Our ultimate goal is to understand why neural signals that always had been assumed to segment objects emerge in networks that have a fundamentally different objective – predicting future input. Thus this research is high-risk high-reward. For these reasons, the proposal is closely aligned with the purposes of the R21 funding mechanism.

NIH Research Projects · FY 2025 · 2024-09

Project Summary Human genetic studies have linked autism and related neurodevelopmental disorders (NDDs) to disruption of genes encoding epigenetic factors, but a significant number remain poorly understood. Defining the functions of these genes and their associated epigenetic pathways in the brain will provide insights into the molecular pathology underlying brain dysfunction. An epigenetic modification with emerging connections to NDD mutation is histone H2B ubiquitination (H2Bub), a chromatin mark involved in gene regulation. Mutation of the adapter protein necessary for recruitment of H2Bub deposition factors to chromatin, WW domain containing adapter with coiled coil (WAC), is causative to the recently described NDD known as DeSanto-Shinawi Syndrome (DESSH). Although the pathology of DESSH is primarily neurological, the role of WAC and H2Bub in the brain remains entirely unstudied. Recent evidence outside the nervous system suggests a role for H2Bub in regulating inducible transcription, specifically the shut-off following induction. This may be particularly relevant to the brain, as tuning of neural connectivity during development and plasticity in neurons requires precise control of the activity- induced gene expression program. Upon depolarization, this suite of genes is rapidly transcribed then silenced to provide a burst of synaptic regulatory proteins that facilitate circuit development and plasticity. Our preliminary studies have revealed dynamic changes in H2Bub at these response genes and a requirement for WAC expression in their proper shut-off, the exact mechanism of this regulation remains unclear. This proposal will determine the extent and molecular mechanism by which WAC and H2Bub regulate neuronal activity dependent transcription and explore the functional consequences of their disruption in a DESSH model. In Aim 1, an orthogonal depletion of H2Bub along with structural manipulation to WAC will be used to parse the key function of WAC in H2Bub deposition and activity-dependent transcription in neurons. The molecular mechanism will then be dissected through integration of transcriptional and epigenetic disruption upon a loss of WAC in neurons. This will define the mechanism by which WAC functions in activity-dependent transcription and establish H2Bub as a novel regulator of this pathway. Aim 2 will investigate the molecular and functional consequences of a DESSH-relevant heterozygous loss of WAC in murine learning and memory, which is well established to depend on activity-dependent programs. This analysis will begin to uncover molecular drivers of neurocognitive phenotypes such as intellectual disability observed in DESSH. Overall, these studies will define a novel function of WAC and H2Bub in neurons and provide insight into how their disruption can drive neurodevelopmental disease.

NIH Research Projects · FY 2025 · 2024-09

Program Objectives and Goals: This proposal aims to provide recent college graduates with an intensive two-year mentored research experience in diabetes, endocrinology, and metabolism, and tailored professional development coursework to successfully guide scholars through the graduate school application process and ensure their success in completing a research-focused biomedical degree program (Ph.D. or MD/PhD). Through our proposed recruitment efforts and multi-layered mentoring approach, this proposal will build and enhance the biomedical research workforce in NIDDK mission areas by focusing on strengthening the research excellence of program scholars. Program Design: This program is designed for college graduates who have graduated within 36 months prior to the start of the program and are not currently enrolled in a degree program. Eight scholars in total will participate in the program. In year 1, we will recruit four scholars for two years of intensive training in biomedical diabetes, endocrinology, and metabolism research and professional development by leveraging the robust physical, administrative, and educational infrastructure and resources available at WashU. In year 3, an additional four scholars will be recruited to participate in the program. Our Program Directors (PDs) have an extensive collaboration record in designing, organizing, and recruiting for diabetes, endocrinology, and metabolism-related trainee programs. They lead highly successful summer and year-round programs for undergraduate and graduate scholars at WashU to support long-term research pathways. We will use data-driven recruitment strategies and will leverage our strong relationships in the field of endocrinology and metabolic disorders to recruit scholars into the program. The Research Training Plan will include: 1) State-of the-art, structured, yet individualized research training by performing diabetes, endocrinology, and metabolism-related scientific projects; 2) Personalized selection of seminars, workshops, and coursework for scholars to develop skills necessary for acceptance and completion of top-tier doctoral degree programs and expertise in diabetes, endocrinology, and metabolism research; and 3) Engagement with other WashU post-bac and graduate-level scholars.

NIH Research Projects · FY 2025 · 2024-09

Project Summary/Abstract A core function of the human brain is object recognition, which underlies many higher-order functions like memory and emotion. Deficits in recognizing objects are associated with various neurological disorders, such as visual agnosia, schizophrenia, and Alzheimer’s disease. However, the neural mechanisms for object recognition remain elusive. Specifically, little is known about how the brain translates visual inputs of objects into meaningful semantics. Previous studies have proposed a hierarchical neural network for object processing, critically including the visual temporal cortex (VTC) and the downstream medial temporal lobe (MTL). While recent single- neuron studies in non-human primates evidenced an axis-based visual feature encoding in the VTC, human MTL neurons have long been characterized to carry a sparse and selective code for individual exemplars (exemplar- based coding). Yet the process by which visual feature representations in the VTC are transformed into semantic representations of abstract labels in the MTL remains unknown. This project aims to address this question by examining the neural computations and dynamics within the VTC- MTL neural network during object recognition. We will utilize intracranial recordings at both individual neuron and neural-circuit levels across different brain areas, coupled with sophisticated computational algorithms. Three distinct neural coding models will be surveyed across the VTC and MTL, including the axis-based feature model, the exemplar-based model, and the region-based feature model (a novel model proposed in our recent studies; K99 AIM 1). VTC-MTL interactions and dynamics that are critical to object recognition will be identified (K99 and R00 AIM 2). The derived results and analysis pipeline from the K99 phase will then be utilized to investigate how different neural models transition from one to another along the VTC-MTL pathway to achieve the representation transformation (R00 phase). The central hypothesis is that different coding models are employed at different stages of object processing, and the novel region-based feature coding serves as an intermediate step that bridges the axis-based coding in the VTC and the exemplar-based coding in the MTL. This proposal will be conducted at Washington University in St. Louis (WUSTL), a top-rank research university that offers excellent scientific support and career training. A team of established mentors will provide necessary training in: iEEG data recording and analysis (Drs. Brunner and Willie), inter-areal interaction analyses (Dr. Rutishauser), neural networks and cognitive neuroscience more broadly (Dr. Hershey), computational modeling and statistics (Dr. Wang), and career development (all mentors). While the primary aims of this proposal is to provide new insights into object recognition in humans across multiple scales, the anticipated outcomes have the potential to inspire new therapeutic interventions for disorders involving impaired object recognition. This K99 award will critically facilitate the success of the proposed research and the applicant’s transition to independence. 1

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY Type 1 diabetes (T1D) is the second most common disease of childhood which results in substantial morbidity and mortality. This autoimmune disease is characterized by the infiltration of CD4 and CD8 T cells into the islets of Langerhans in the pancreas, where they ultimately deplete insulin-secreting  cells. Recent studies have shown that conventional type 1 dendritic cells (cDC1) are required for the development of Type 1 diabetes in the murine NOD model, which has many parallels to human disease. cDC1 are unique in their ability to efficiently cross-present  cell antigens to and prime autoreactive CD8 T cells. Furthermore, cDC1 are potent producers of IL-12 and may facilitate Th1 differentiation of CD4 T cells. A therapy that specifically eliminates cDC1 may therefore be expected to prevent the development of T1D by blocking autoreactive CD4 Th1 development as well as the presentation of self-antigens to autoreactive CD8 T cells. Our preliminary data show that a chimeric antigen receptor (CAR) T cell targeting XCR1, a chemokine receptor expressed by cDC1, is successful in depleting cDC1 in the spleen and pancreatic lymph node of NOD mice. Furthermore, cDC1 depletion by this CAR T cell also successfully inhibited the proliferation of a cDC1 dependent, self-reactive CD4 T cell in vivo. Thus, the central premise of this proposal is that XCL1 CAR T cells may be useful for the prevention of T1D. To address this, we will assess the ability of this CAR to prevent diabetes in NOD mice. Experiments proposed in Aim 1 will validate the specificity of the CAR for cDC1s and address the durability of cDC1 depletion in vivo. Aim 2 will assess the functional effects of cDC1 depletion by CAR T cells on autoimmune diabetes by assessing CD4 and CD8 T cell numbers and phenotypes within the islets as well as the rate of spontaneous diabetes incidence in NOD mice. Collectively, these studies may lead to the development of a novel preventative treatment for human Type 1 diabetes and establish a paradigm for CAR T cell mediated immunomodulation via selective targeting of DC subsets.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY. Candidate: Dr. Zellers is a physical therapist (Doctor of Physical Therapy, Columbia University) with a PhD in Biomechanics and Movement Science (University of Delaware). She is Assistant Professor in Physical Therapy and Orthopaedic Surgery at Washington University School of Medicine in St. Louis, having completed her postdoctoral fellowship (Washington University School of Medicine). Dr. Zellers is a translational, tendon researcher with expertise in in vivo and ex vivo assessment of human subjects and tissues to elucidate person- and tendon-specific characteristics influencing patient treatment and outcomes. Mentor and Advisory Committee: Dr. Farshid Guilak will be primary mentor, bringing extensive experience as a principal investigator in orthopaedics and specific expertise in leveraging the mouse models included in this study to answer similar questions regarding the effect of obesity in the context of osteoarthritis. Drs. Alayna Loiselle and Spencer Lake are co-mentors imparting tendon-specific expertise including mechanical, histological, and molecular biological outcomes assessment. Drs. Gretchen Meyer and Simon Tang are co- mentors with experience leveraging similar animal models to investigate questions relating to fat-muscle crosstalk and collagenous soft tissue healing, respectively. Training Plan: In this Career Development Award, Dr. Zellers will gain training and experience in leveraging animal models (murine models of obesity, lipodystrophy, and tendon injury) to investigate mechanisms underlying observations gleaned from her clinical, human studies research line. The proposed training plan emphasizes building skills in incorporating animal models into her translational research program. Training goals also incorporate biological approaches to tendon assessment, specifically histological and molecular biological techniques, which will carry over to Dr. Zellers’s study of human tendon tissue. This award would provide the training and experience for Dr. Zellers to grow as a translational researcher with a comprehensive toolkit of in vivo and ex vivo techniques spanning preclinical models to human tissues to human subjects. Research: Obesity presents both mechanical and biochemical effects on tendon tissue homeostasis and healing that have not been well elucidated. The proposed study leverages murine models of obesity and lipodystrophy to determine the effects of high bodyweight and metabolic dysfunction on tendon tissue. We hypothesize that metabolic dysfunction, more so than mechanical load, promotes tendon degeneration (Aim 1) and impairs healing (Aim 2), evidenced by abnormal histological appearance, altered gene transcription, disrupted collagen organization, and impaired tendon function at the tissue and functional performance levels. Institutional Commitment to the Candidate: Dr. Zellers holds a tenure-track faculty position and has been provided bench and clinical laboratory space, start-up package, and trainee support.

NIH Research Projects · FY 2025 · 2024-08

1 PROJECT SUMMARY 2 Sepsis caused by Streptococcus pneumoniae (Sp; the pneumococcus) causes significant morbidity and 3 mortality, despite vaccines, antimicrobials, and supportive care. Sepsis is defined as “life-threatening organ 4 damage due to a pathogen-driven, dysregulated immune response.” The immune-centric model posits that 5 imbalances between pro- and anti-inflammatory immune mediators create a perfect septic storm of hyper- 6 inflammation, tissue damage, and immuno-suppression. However, no immunomodulatory therapies have yet 7 shown efficacy in improving mortality of sepsis. Our novel model of Sp sepsis prompts us to reconsider this 8 immunocentric model. We found a novel single nucleotide variant (SNV) in the gene encoding co-enzyme Q6 9 (COQ6) to be associated with susceptibility to Sp disease in an at-risk human population. The variant converts 10 an aspartate (D) to a tyrosine (Y) residue, and so we call the variant “DY” (COQ6-DY). We created the “DY 11 mouse” line, homozygous for the homologous Coq6-DY variant, which suffer increased mortality after Sp lung 12 infection. We then made chimeric mice to separate immune from non-immune cell dysfunction. Surprisingly, 13 recipient DY mice were more susceptible to Sp sepsis, despite reconstitution with wild-type (WT) immune cells. 14 Reconstitution of WT mice with DY immune cells improved survival after Sp infection. Our data reveal that non- 15 immune factors drive mortality during Sp sepsis, prompting re-evaluation of the immunocentric model of sepsis. 16 To explore why Coq6-DY increases Sp susceptibility, we tested how Coq6-DY disrupts COQ6 function. 17 Located in the inner mitochondrial membrane, COQ6 biosynthesizes the lipid ubiquinone (Q). Q serves as an 18 electron acceptor in the electron transport chain (ETC) during oxidative phosphorylation (OXPHOS). In health, 19 most tissues rely on OXPHOS for ATP generation. Infection induces metabolic remodeling and OXPHOS 20 downregulation, necessary to generate and sustain an anti-pathogen, pro-inflammatory response. Abnormal 21 metabolic remodeling may drive sepsis. In DY tissues, Q biosynthesis was intact, but OXPHOS downregulation 22 was accelerated in DY heart muscle after Sp challenge. Our proposal tests the hypothesis that aberrant 23 metabolic remodeling in DY mice causes hyperinflammation and increases organ damage in Sp sepsis through 24 three aims. First, we define the inflammatory and physiological derangements in Sp-infected DY recipients. 25 Second, we analyze metabolomic and single cell RNASeq profiles of infected DY recipients for predicted 26 correlations between metabolic abnormalities and pro-inflammatory transcriptional profiles. Finally, we test if 27 therapies chosen to counteract metabolic abnormalities in DY recipients improve survival in Sp sepsis. As an 28 Infectious Disease physician-scientist with expertise in cellular immunology, I am uniquely qualified to lead this 29 research. Our validated mouse model and compelling preliminary data assure that proposal completion will fill 30 key knowledge gaps in current sepsis models, and will accelerate discovery of therapies for unmet clinical needs.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY/ABSTRACT We propose to broadly disseminate and extend intuitive, powerful cloud-based resources for optical brain mapping that facilitate efficient, accurate, and standardized processing that will harmonize the emerging set of optical measurement strategies within the growing ecosystem of network level analyses used throughout the greater brain mapping community. The neuroimaging community faces numerous challenges in data collection, preprocessing, estimation of brain connectivity, and analyses of relationships between brain connectivity and behavior. An ever-expanding community of researchers are employing optical methods based on functional near infrared spectroscopy (fNIRS) in order to infer pathophysiological state of tissue, such as inflammation and metabolism for detection/characterization of disease or cerebral hemodynamics for understanding human brain health, development, and aging. Recent developments of high-density diffuse optical tomography (HD-DOT), a silent, flexible, and scalable technology have demonstrated dramatically improved anatomical specificity and image quality over traditional fNIRS. Further, recent developments in wearable HD-DOT, even using frequency domain and time resolved strategies, open the door to unconstrained mapping of naturalistic human brain function with superior image quality than previously possible. Given the growing worldwide adoption of fNIRS and HD-DOT methods and further developments of next-generation optical brain mapping methods via the BRAIN Initiative, there is an urgent and present need for standardized, accessible and flexible tools that directly support workflows from optical tissue parameter recovery to functional brain mapping to relating variance in brain function to behavior and outcome. To address these needs, our teams have developed and validated computational tools including NIRFAST, NeuroDOT, and Network Level Analyses (NLA), for tissue parameter recovery, optical brain mapping, and model-based connectome-wide association studies of brain function and behavior, respectively. While these tools each support growing user communities, the tools are based in Matlab, which significantly limits accessibility and adoption. Additionally, much of these analyses are computationally intensive and expensive, limiting full use to institution-based, server-level resources. Further, extant widely available software packages for fNIRS are limited in scope and do not support the full set of pipelines for end- to-end analyses that together NIRFAST, NeuroDOT, and NLA provide. We therefore propose herein to utilize funding from RFA-NS-23-026 to address this unmet need for data resources with (1) greater dissemination and training for our tools, (2) cloud deployment of our software to increase scale and accessibility, while easing the computational burden for the user, and (3) expanded utility of these powerful, flexible tools to meet the evolving needs of users at the forefront of optical imaging technology development. This proposal executes BRAIN Initiative goal 7 that seeks to integrate new technological and conceptual approaches to discover how dynamic patterns of neural activity are transformed into cognition, emotion, perception, and action in health and disease.

NIH Research Projects · FY 2025 · 2024-08

Project Summary/ Abstract Exogenous oxytocin (OT) is widely used for the induction of labor in pregnant women. Though induction of labor with OT confers many maternal and neonatal benefits, its use is not without adverse effects. Epidemiological studies suggest an association between OT use and neurodevelopmental disorders, but they are conflicting and do not provide mechanistic information. This knowledge gap is concerning because signaling through the OT receptor (OTR) is critical for the establishment of social behavior in mammalian species. With the rising popularity of elective induction of labor, where labor is induced for convenience and not for maternal or fetal indications, it is important to comprehensively understand the impact of maternally administered OT on the developing brain. Though OTR dysregulation in the developing brain has been suggested as a possible mechanism, mechanistic studies are lacking because of the lack of a high-fidelity animal model that mimics labor management in pregnant women. To address this, we successfully created and validated a pregnant rat model for labor induction by continuously delivering an escalating dose of intravenous OT through a microprocessor-controlled precision pump. In new preliminary studies, we adapted this high-fidelity model to mice with minor technical modifications and confirmed that induced birth with OT was associated with a substantial decrease in OTR gene expression in the neonatal mouse cortex. The fundamental objective of this proposal, therefore, is to chart the developmental trajectory of OTR and OT expression in the postnatal brain after induced birth with OT. To accomplish this, we will induce labor with OT in transgenic reporter mice (OTRvenus/+) and leverage the use of high-resolution brain- wide imaging techniques (serial two-photon tomography and light sheet fluorescent microscopy imaging) to comprehensively visualize the developing OTR-OT system in the postnatal brain (Aim 1). Motivated by our observations of early communicative delay and impaired empathy-like behaviors in male rat offspring after induced birth with OT, we will extend these behavioral studies to mice to enable mapping of dysregulated behavior to brain-wide changes in the OTR-OT system (Aim 2). Successful completion of these experiments will (i) provide a comprehensive perturbation map of how perinatal OT exposure alters the developmental trajectory of the OTR-OT system, and (ii) delineate the neurobehavioral impact of such perturbation. Because these studies are ethically inconceivable in human subjects, combining our mouse model for labor induction with innovative whole brain imaging techniques provides unparalleled opportunities to comprehensively answer this important question. Regardless of the outcome of our research, our findings will have a significant impact — be it a paradigm shift in current dogma or providing scientific reassurance to potential mothers.

NIH Research Projects · FY 2025 · 2024-08

Abstract Interstitial cystitis/bladder pain syndromes (IC/BPS) are a debilitating painful condition with unknown etiology. The cells and the circuits in the spinal cord that process non-noxious (bladder function) and noxious (intense bladder pressure or discomfort) sensory information from the bladder is not well established. What specific roles different spinal anatomical substrates play in processing pathological bladder sensations and voiding dysfunction is unclear. In this proposal, we will test how distinct spinal cord cells that are activated by cystitis are necessary for maladaptive micturition and bladder nociception. We will use chemogenetics and optogenetics approaches to determine the precise roles the cystitis activated spinal cord cells play micturition and bladder nociception. Furthermore, we will determine how this information is streamed to the distinct brain regions. Finally, we propose to perform molecular description of genetically classified spinal cell populations using transcriptomics and fluorescent in situ hybridization. These studies will lead to a cellular and functional characterization of cystitis activated spinal cord cells and to a better understanding of how different symptoms of cystitis are regulated in the spinal cord. These efforts will advance our understanding of spinal circuits in cystitis and may enable development of new therapeutic strategies for the treatment of IC/BPS.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY Approximately 70% of preterm births are preceded by preterm labor, which is primarily associated with non- infectious etiologies. However, little is known about the non-acute-inflammatory causes of preterm birth. This project builds on extensive published and preliminary data indicating that T cells and the cytokine IL-9 at the maternal-fetal interface play an important role in preterm labor and birth. The overall goals of this project are to define the mechanisms by which the cytokine IL-9 at the maternal-fetal interface promotes preterm birth and negative consequences for offspring, to determine the mechanisms by which vitamin D can prevent these outcomes, and to identify new T cell subtypes that contribute to preterm labor in humans. Aim 1 is to elucidate the mechanisms by which IL-9 promotes preterm labor and birth. This aim tests the hypothesis that IL-9 targets both hematopoietic and non-hematopoietic cells at the maternal-fetal interface in mice. Aim 1 will also explore whether IL-9 signaling is required within the maternal or fetal tissues to promote preterm birth. Additionally, the mechanisms whereby vitamin D prevents IL-9-induced preterm birth in mice will be determined. Aim 2 is to define the impact on offspring of IL-9 at the maternal-fetal interface. This aim tests the hypothesis that maternal IL-9 that enters the amniotic cavity causes adverse fetal and neonatal outcomes. Specifically, this aim will examine the fetal lung, intestine, and placenta and neonatal airway and gut immunity. Additionally, the extent to which prenatal vitamin D supplementation can mitigate the impact of in utero IL-9 exposure will be investigated. A hypothesis-generating Aim 3 will identify T-cell subsets at the maternal-fetal interface in women with preterm labor and birth. This project will make use of an advanced mouse model of IL-9-induced preterm birth, imaging mass cytometry, spatial transcriptomics, single-cell RNA sequencing of human placenta samples from term and preterm pregnancies, and functional testing on T cells isolated from patients with term or preterm birth. The innovative fusion of immunology and omics approaches used here will unveil non-infectious causes of preterm labor and birth and may lead to development of a treatment – such as vitamin D, which is FDA-approved during pregnancy – to reduce the risk of preterm birth. Additionally, the new immune targets identified could lead to development of novel strategies to reduce preterm birth and its sequelae. This project is directly relevant to the call for research “exploring the contribution of immune dysregulation to gestational disorders and adverse pregnancy outcomes” to improve reproductive health.

NIH Research Projects · FY 2025 · 2024-08

Project Summary Neuromodulators including dopamine and serotonin have profound effects on spinal circuits for locomotion. A wealth of pharmacological manipulations has shown that drugs mimicking or blocking these neuromodulators can change the properties of rhythmic motor output in the isolated spinal cord. However, these studies often conflict and cannot capture the normal range of behaviors expressed in vivo. Furthermore, it is entirely unknown whether neuromodulators are released onto different spinal targets across different behaviors. Finally, neuromodulatory neurons are highly branched, making it difficult to disambiguate the spinal vs supraspinal consequences of their action. We will leverage new tools for imaging and manipulating neuromodulator signaling, combined with the transparency and accessibility of the young zebrafish, and a quantitative modeling approach, to understand the effects of dopamine and serotonin on genetically defined components of the spinal locomotor circuit in vivo. First, we will measure the activity of neuromodulatory axons during three distinct behaviors, testing whether dopamine and serotonin axons differentially participate in these behaviors. Next, we will quantify neuromodulator release during these behaviors directly, both in the whole spinal cord and in genetically defined populations of neurons with distinct contributions to locomotion. We will then test the significance of descending neuromodulatory influence on spinal circuits by targeted axotomy that will allow disambiguation of the spinal and supraspinal consequences of neuromodulator release. Finally, using newly developed chemogenetic approaches, we will selectively block neuromodulatory receptors in motor neurons and measure the consequences on the three distinct behaviors in freely moving animals. Throughout the project, we will use experimental data to develop both single-segment and multi-segment computational models of neuromodulatory action, and in turn use these models to make testable predictions about circuits and behavior. Together, these experiments will reveal for the first time when and where dopamine and serotonin are acting in the spinal locomotor circuit, and how their actions influence behavior in vivo.

NIH Research Projects · FY 2024 · 2024-08

The goal of the Society for Heart and Vascular Metabolism (SHVM) conference is to bring together scientists and trainees from around the globe to present and discuss cutting-edge research in the field of cardiac and vascular metabolism. This year’s meeting in St. Louis, Missouri, September 2024, will focus on the latest research regarding the role of interorgan crosstalk in the pathogenesis of cardiovascular disease (CVD). Most prevalent cardiovascular pathologies, such as heart failure and ischemia, have underlying metabolic derangements directly involved in disease progression, but recent studies have highlighted the relevance of interorgan crosstalk as a new frontier for cardiovascular research with the ultimate goal of ameliorating CVD. To that end, SHVM’s 2024 meeting is designed to foster ‘crosstalk’ between the investigative fields of heart and vascular metabolism and other key organ systems. Despite new investigations of interorgan communications, such as exosome trafficking as well as novel hormonal axes that affect the heart and vasculature, cross-disciplinary research has unique funding and operational barriers. This is not surprising given the significant challenges of creating multidisciplinary infrastructure and developing novel techniques evaluating heart and vascular metabolism in the context of manifold interorgan signals. Some scientists/trainees (PhD students, MD and postdoctoral fellows) may not feel equipped to tackle the unique barriers to performing cross-disciplinary research. To address these issues, this conference aims to provide a comprehensive and wide-ranging forum in which investigators from multiple disciplines will interact and discuss recent research studies that have resulted in meaningful outcomes relevant to cardiovascular metabolism. SHVM meetings have always featured strong participation of diverse trainees from many different regions in the United States and from around the globe. Moreover, SHVM has an established track record for fostering productive collaborations and career advancement. This year’s meeting will offer trainee workshops directed by top experts in metabolic imaging. Titled, “Cutting edge tools for the evaluation of myocardial metabolic metabolism and function,” the trainee workshop will facilitate interaction between trainees with junior and established investigators. The 2024 meeting will feature 2 keynote speakers, 19 plenary speakers, 4 speakers for Continuing Medical Education talks, 4 Early Investigator winner talks, and 9 other short talks selected from submitted abstracts, in addition to 2 workshops, and 2 poster sessions. The sessions will have strong representation from early-stage investigators, women, and underrepresented minorities. We are requesting funds to offset costs associated with trainee, speaker, and keynote speaker participation to continue the tradition of SHVM meetings, which are considered the premiere conferences in cardiac metabolism.

NIH Research Projects · FY 2025 · 2024-08

SUMMARY Emerging and re-emerging neurotropic viruses, including flaviviruses and alphaviruses, generally cause fever, malaise, and related symptoms. However, in some patients, these viruses enter the central nervous system (CNS) and cause severe illness and disease resulting in meningitis, encephalitis, and death. Several cellular mechanisms have been described that allow viruses to bypass the blood-brain barrier. However, to date, there have been no anatomical routes identified that enable direct passage of virions in the periphery into the CNS. This highly collaborative and interactive proposal between the Diamond and Kipnis laboratories will advance our mechanistic understanding of how meningeal structures function in the regulation of and response to neurotropic virus infections. Our overarching hypothesis is that neurotropic viruses can directly enter the CNS through arachnoid cuff exit (ACE) points, a newly described meningeal structure linking the CNS and periphery. We hypothesize that neurotropic virus infections early-in-life alter the development of meningeal immunity, and this increases the severity of and vulnerability to heterologous neurotropic virus infections later in life. We will address our hypotheses in four specific aims: (1) We will define the role of arachnoid cuff exit (ACE) points in neurotropic flavivirus entry into the CNS and provide a detailed transcriptional profile of meningeal and brain responses to neurotropic flavivirus infection at different ages (e.g., in utero, neonatal and adult); (2) We will determine how early-life neurotropic virus infection and IFN-g signaling modulates the development of meningeal lymphatic vessels; (3) We will test the contribution of IFN-g signaling in meningeal stromal cells to ACE point development in the context of early-life neurotropic virus infection; and (4) We will define the effects of early-in-life neurotropic virus infection (with Zika virus [ZIKV], a TORCH pathogen) on the neuropathogenesis of a second, heterologous neurotropic virus in adulthood. Conceptually, we will define the role of newly described meningeal structures in viral entry into the CNS. Mechanistically, we will explore how virus-induced IFN-g signaling modulates the development of meningeal immunity and its subsequent impact on neurotropic virus infection. Our suite of relevant neurotropic flavivirus and alphavirus mouse models (e.g., ZIKV, WNV, VEEV, WEEV), novel inducible Cre-driver mouse lines enabling targeting of specific cell types in the meninges (e.g., arachnoid barrier cells, Dpp4-CreERT2; dural border cells, Slc47a1-CreERT2) and state-of-the- art immunological assays of meningeal structure and function will enable us to define how meningeal immunity is affected by and modulates the neuroinvasion and neuropathogenesis of virus infection. Together, our team will elucidate the mechanisms and reciprocal relationships between CNS invasion by neurotropic viruses and meningeal immune responses, which could provide new targets for therapeutic intervention against neurotropic viruses.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY Survival from cervical cancer is stagnating in the United States. Despite many intervening clinical trials, the standard of care (SOC) definitive chemoradiation therapy (CRT) has remained unchanged since the addition of concurrent platinum-based chemotherapy to radiation in the 1990’s. Reasons for this are several-fold, and include a patient population that is disproportionately affected by barriers to care, inherent radioresistance of gross disease as evidenced by local recurrence after a mean primary tumor dose of >200Gy delivered with brachytherapy plus EBRT, and insufficient systemic control, with distant failure contributing to two thirds of cervical cancer deaths. A suboptimal immune response at the time of definitive CRT is associated with local, regional, and distant recurrence, as well as death from cervical cancer. We have previously shown that enrichment of immunosuppressive cells and high expression of squamous cell carcinoma antigen (SCCA), known as SERPINB3, are associated with higher risk of recurrence after SOC CRT in cervical cancer, and that CRT induces further infiltration of tumor permissive myeloid derived cells. Preliminary data suggest that tumor associated macrophages, myeloid-derived suppressor cells, and regulatory T cells are increased in mid- treatment tumor specimens from patients undergoing standard CRT. Using preclinical models, we find that brachytherapy stimulated expression of immune-stimulatory signals to a greater degree than an equivalent dose of EBRT. Finally, SERPINB3 directly promotes expression of chemokines that recruit immune-suppressive cells, particularly myeloid-derived sub-populations, blunting the T-cell anti-tumor response in cervical cancer. We hypothesize that brachytherapy alone delivered to the primary tumor prior to regional lymph node EBRT will safely minimize patient trips, further stimulate the immune system, and potentiate the efficacy of immunotherapy. Two aims are proposed to directly test this hypothesis: Aim 1 will determine if accelerated brachytherapy-forward chemoradiation therapy (ABC-RT) is a safe and effective approach to shorten overall treatment time, and maximize anti-tumor immune response through a phase I/II clinical trial for patients with locally advanced cervical cancer. Aim 2 will determine if ABC-RT potentiates the anti-tumor activity of immune checkpoint therapies and/or the myeloid-cell inhibitor CCR2i using a preclinical murine tumor model with a novel intracavitary brachytherapy system developed for this proposal. Secondary endpoints to validate candidate biomarkers SERPINB3, and post therapy FDG-PET as predictors of recurrence after this experimental approach are proposed to precisely stratify patients for subsequent trials incorporating drug-ABC-RT combinations. Success of these aims will provide the preliminary data to support randomized trials of ABC-RT based regimens compared to the SOC and ultimately a paradigm shift in the definitive treatment of cervical cancer. The proposed translational studies will provide a template for integration of therapies that synergize with CRT and the immune response. The overall goal of this proposal is to improve survival for patients with locally advanced cervical cancer.

NIH Research Projects · FY 2025 · 2024-08

As the world ages, Alzheimer’s Disease (AD) and related dementias are quickly becoming some of the most pernicious and burdensome diseases around the globe. This burden is particularly high for Latino older adults, who have a higher risk of developing AD, and exhibit age-related cognitive decline earlier and with more severe dementia symptoms than non-Latino Whites. Despite these known disparities, the Latino community continues to be significantly under-represented in AD research. Toward this end, preliminary evidence suggests that mindfulness-based programs (MBPs) – which refer to training of mindfulness meditation skills in group-based settings – may help prevent AD in at-risk Latino individuals. Indeed, MBPs have been found to improve cognitive health and psychological well-being by addressing major AD risk factors (e.g., chronic stress, depression), and as such, have significant potential as a low-cost and scalable lifestyle intervention to reduce AD vulnerability. However, a significant challenge to further research in this area is that the traditional curricular format of MBPs may not be applicable, relevant, or accessible to Latino older adults, due to language and cultural barriers, as well as disparities in access to health care. The goal of this R61 proposal is to address this challenge by evaluating the feasibility and effectiveness of a culturally-adapted MBP that is sensitive to the specific needs of the Latino older adult community. To achieve this goal, we fully leverage the complementary expertise of our investigative team, drawing on the extensive experience and strong community partnerships forged by co-PI Parra Perez in adapting mindfulness-based stress reduction (the most widely utilized MBP) for use with Spanish-speaking immigrant communities, by further refining the curriculum to be targeted towards Latino older adults. We rigorously examine the MBP intervention, in terms of effectiveness and key mechanisms of action, drawing upon cutting-edge advances in the cognitive neuroscience of aging and mindfulness science, by utilizing a powerful new experimental design strategy developed by co-PI Braver with post-doctoral fellow Lin. Specifically, we utilize a longitudinal EEG protocol to test whether enhanced engagement of a focused attention mindfulness state is selectively related to neurocognitive benefits, while monitoring the translation of mindfulness into daily-life contexts through an Ecological Momentary Assessment (EMA) protocol delivered via a customized mobile application designed specifically for use with Spanish-speaking Latino older adults. Finally, we test for the role of preclinical AD and related neurodegeneration in moderating MBP-related benefits, using state-of-the-art blood plasma-based biomarkers to assess AD risk with low cost and participant burden. This proposed project is a direct response to recurrent calls for prioritizing the creation and dissemination of culturally informed research approaches and programs for Latino populations. The study will provide a rich set of preliminary pilot data that can be used to inform a subsequent early-stage clinical trial at reducing AD prevalence/burden within the Latino community.