Ohio State University

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY Spinal cord injury (SCI) has a devastating outcome in humans, particularly cervical spinal cord injuries, as they result in the patient being severely impaired with significantly diminished (or no) use of their upper limbs. The repair of corticospinal tract (CST) axons is a major obstacle to the recovery of the adult spinal cord after an injury. Historically, CST axons following injury fail to regrow and retract rostrally from the original lesion site; therefore, any therapeutic intervention needs to offer a positive growth milieu and provide meaningful connections to the limbs once innervated. Developing a regenerative therapy for SCI would be transformational. Even small improvements in upper limb function will provide cervical injured patients a chance for increased independence. Establishing an effective cellular treatment for SCI would greatly reduce the significant and ongoing care costs to the US healthcare system. We have developed a reproducible in vitro differentiation protocol to produce deep cortical neurons, as a viable and promising cellular transplantation strategy for cervical SCI. We will employ a rat cervical contusion (C5 level) model of spinal cord injury to measure integration, survival, synaptic circuitry and functional outcomes following deep cortical neuron transplants at a single defined age point (Day 35). Our animals will also undergo additional behavioral/functional testing following deep cortical neurons transplantation. We hypothesize that our deep cortical neurons transplants successfully survive, integrate, and form appropriate functional and anatomical outcomes after a cervical spinal injury. We propose that any observed functional improvement will arise from the transplanted deep cortical neurons forming new connections to host spinal cord circuitry, and in addition, host CST axons will form synaptic connections to those neurons from the deep cortical neuron transplants. Functional tests encouraging the use of the injured forelimb and assessment will also be undertaken in groups to ascertain improved growth and target specificity. We believe our strategies will help patients with cervical spinal cord damage by using techniques that can be matched to the individual patient, reduce dependence on anti-rejection drugs and provide significant recovery capacity.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY In humans, microRNAs (miRNAs) of approximately 22 nucleotides (nt) are key components of mature RNA- induced silencing complexes (RISCs) and play essential roles in silencing specific genes. Our structural studies of human Argonaute proteins (AGOs) have revealed important differences among them, leading to several discoveries, including the identification of tiny guide RNAs (tyRNAs). TyRNAs are defined as guide RNAs of 17 nt or shorter that are associated with AGO proteins. We also uncovered a tyRNA biogenesis pathway driven by specific exonucleases, which generate 14-nt tyRNAs from AGO-associated miRNAs that are 21–23 nt in length. Among these tyRNAs, some, referred to as "cleavage-inducing tyRNAs (cityRNAs)," can catalytically activate AGO3. Despite these breakthroughs, the study of tyRNAs is still in its infancy, with significantly fewer publications compared to research on miRNAs. Meanwhile, more evidence has emerged that the four human AGOs play unique roles beyond conventional gene silencing. This project aims to address these gaps by focusing on five key topics: 1. Both AGO2 and AGO3, when loaded with cityRNA, appear to directly recognize the sequence upstream of the tyRNA target site (UTy), thereby influencing their target mRNAs differently from miRNAs. Our previous study demonstrated that cityRNA-mediated gene silencing relies heavily on target cleavage rather than translational repression or mRNA destabilization. However, the molecular mechanism of target recognition by cityRISCs remains unknown. We aim to address this knowledge gap through structural and functional studies. 2. Poly(A)-specific ribonuclease (PARN) processes miRNAs to approximately 22 nt in length and selectively degrades specific mRNAs, but the molecular basis of these activities is poorly understood. We aim to elucidate these mechanisms by determining the structure of the PARN-AGO complex bound to a miniature mRNA. 3. During duplex loading, AGOs recognize the less thermodynamically stable end of the duplex to capture the 5′ end of one strand as the guide, forming the mature RISC. The molecular mechanism for this asymmetric guide strand selection remains unclear. We will determine the cryo-EM structures of each step in the RISC assembly process. 4. Single-point mutations in AGO1 and AGO2 have been linked to neurodevelopmental disorders (NDDs). Our data show that specific 3′→5′ exonucleases trim miRNAs associated with these NDD-related AGOs into unusually short tyRNAs. To understand the impact of these mutations, we will determine the cryo-EM or crystal structures of NDD-relevant AGO mutants loaded with miRNAs and atypical tyRNAs. 5. Our structural studies have revealed distinct differences among the four human AGOs. To further characterize these differences, we will determine the structures of the four AGOs using the same set of guide and target RNAs. These proposed studies will explore a broad range of cellular events regulated by conventional and novel small noncoding RNAs. The outcomes will provide valuable insights and establish a solid foundation for the development of RNAi-based therapeutics.

NIH Research Projects · FY 2026 · 2026-05

Project Summary/Abstract The overarching goal of this proposal is to explore novel aspects of the actin cytoskeleton organization and function in health and disease. The remarkable functional versatility of the actin cytoskeleton stems from its ability to assemble into a variety of diverse structures – branched networks/meshes and aligned bundles. This architectural complexity is orchestrated by actin-binding proteins, whose activity is delicately regulated in response to internal and external signals. Human plastins are versatile actin-binding proteins that organize actin filaments into higher-order assemblies: bundles and networks. Plastins are involved in cytokinesis, migration, and stabilization of membrane protrusions, but also in the pathogenesis of the following diseases: carcinogenesis, neurodegeneration, and hereditary and infectious diseases. Despite the importance and long- lasting interest of the research community in these proteins, understanding of their interaction with actin and their regulation is superficial. Our first research direction is to contribute to human health and well-being by advancing the understanding of the actin cytoskeleton organization by actin-bundling proteins plastins and their contribution to pathologies (e.g., congenital diseases and metastatic cancers) at the molecular and cellular levels. Playing numerous vital roles in human defense mechanisms, the actin cytoskeleton is a common target for numerous bacterial pathogens, which developed various elegant and sophisticated ways to disrupt and usurp it by producing actin-targeting effectors. By hijacking the actin cytoskeleton, pathogenic toxins disturb cell morphology, cell motility, phagocytosis, epithelial permeability, and antigen presentation. Being constantly adjusted to the host cytoskeleton by co-evolution, they recognize weaknesses in the host defense and represent powerful tools that foster the understanding of the cytoskeleton on molecular and cellular levels. The urge for a thorough understanding of bacterial pathogenicity is further necessitated by the dissemination of multidrug- resistant pathogens that undermine the efficiency of antibiotics. The second research direction of this proposal is to decipher the in-depth molecular and cellular mechanisms of bacterial effectors targeting the actin cytoskeleton to i) enable alternative ways of targeting pathogens and ii) get a deeper understanding of the actin cytoskeleton per se. The current proposal is directly relevant to the NIH mission as it focuses on three families of bacterial effectors, all produced by human pathogens: Vibrio cholerae and Vibrio parahaemolyticus VopF/VopL (1) and VopM/VopV (2), and Legionella pneumophila MavH, RavH, and VipA (3). Furthermore, the proposal is of interest for a general understanding of human physiology as each of the above toxins reveals novel properties of the actin cytoskeleton. Finally, the understanding of the molecular and cellular mechanisms governing the function of actin-bundling proteins plastins contributes to explaining the pathology of plastin-related human diseases.

NIH Research Projects · FY 2026 · 2026-05

Project Summary / Abstract The circadian (24hr) timing system influences a vast array of behavioral and neurophysiological processes. Central to this time-keeping process is the circadian pacemaker located within the suprachiasmatic nucleus (SCN). Through both neuronal and hormonal pathways, the SCN sets the phasing of ancillary clock oscillators in a variety of brain regions, including cortico-limbic circuits, and it is the coordinated effects of the SCN and these ancillary forebrain clocks that impart rhythm modulation on complex behavioral states, including mood, memory and executive function. Further, the disruption of clock physiology (at both a cellular and systems level) is a contributing factor in a number of acquired and congenital disorders of the nervous system. Although recent work has provided important clues regarding the profound and far-reaching functional effects of the circadian timing system in the CNS, we still know little about clocks outside of the SCN. Along these lines, within these cortico-limbic circuits, is the circadian timing system cell autonomous (as it is in the SCN); Is it built around the same canonical core clock feedback circuits that form the basis of the core-clock transcriptional feedback loop (TTFL) in the SCN; Is the relationship between the TTFL and ancillary transcriptional circuits similar to, or different from, the relationship in the SCN? Here, we propose to determine the functional significance and mechanistic underpinnings of clock physiology in cortico-limbic circuits, and to assess how these cortico-limbic clocks gate complex behavioral states over the circadian cycle. For these studies, we will utilize a highly innovative combination of approaches, including novel transgenic mouse models, AAV-based genetic interrogation methods, in vitro and in vivo cellular-level longitudinal profiling and novel cellular imaging methods to characterize the relationship between clocks and cognition. In Aim 1, we will test the fidelity and functional properties of the core clock TTFL within cortico-limbic cells and circuits. Mechanisms of TTFL entrainment and rhythm regulation via the CREB/CRE pathway will also be examined. In Aim 2, we propose to use 2-photon in vivo profiling to examine clock timing and entrainment mechanisms within cortico-limbic circuits. In Aim 3 we propose to test the functional relationship between the circadian clock and plasticity- associated signaling pathways and how clock timing could shape the underlying cell biology that gives rise to memory formation. Given the critical role that clock timing plays in human health, new insights into the molecular-, cellular- and systems-level design principles that underlie clock timing through the CNS is absolutely critical for the development of new, efficiently targeted, chronotherapeutic interventions.

NIH Research Projects · FY 2026 · 2026-05

PROJECT SUMMARY Neuronal communication depends on the precise coordination of synaptic vesicle exocytosis and endocytosis at presynaptic terminals. After neurotransmitter release, efficient retrieval of synaptic vesicle membranes through endocytosis is essential for sustaining neurotransmission. Despite its fundamental importance, the mechanisms linking synaptic vesicle exocytosis and endocytosis remain poorly understood, largely due to the limitations of current imaging technologies. Existing methods lack the spatial and temporal resolution needed to directly visualize these nanoscale processes in living cells. To address this critical gap, this Joint NINDS/NIMH Exploratory Neuroscience Research Grant (PA-25-150) proposal aims to develop a three-dimensional (3D) total internal reflection fluorescence structured illumination microscope (TIRF-SIM). By integrating super-resolution imaging with evanescent field nanometry, this technology will enable unprecedented visualization of exo-endocytic coupling at the nanoscale in living neurons. Our research plan consists of two key objectives: (i) We will iteratively optimize the experimental parameters of 3D TIRF-SIM using well-characterized in vitro and in vivo imaging conditions to maximize spatial and temporal resolution. (ii) Using these optimized parameters, we will evaluate the performance of 3D TIRF-SIM for high-resolution reconstruction of clathrin-coated structures both live cultured cells and tissues of Drosophila melanogaster embryos addressing long-standing questions in endocytosis research. Beyond its technological advancements, this proposal directly addresses a critical need in neuroscience research. By enabling direct visualization of synaptic vesicle dynamics at unprecedented resolution, this innovation will fundamentally advance our understanding of neuronal communication. Moreover, this breakthrough in imaging technology has the potential to catalyze the development of novel therapeutic strategies for neurological disorders associated with synaptic dysfunction.

NIH Research Projects · FY 2026 · 2026-05

Aaron Murnan, PhD, LMFT is a tenure-track Associate Professor at the Martha S. Pitzer Center in the College of Nursing at The Ohio State University. His career goal is to become an independent clinician-scientist with expertise in co-designing novel interventions to address salient treatment needs among families disproportionately impacted by opioid use and justice-involvement, specifically women in the sex trade. The term ‘co-design’ refers to a community-based participatory research methodology in which individuals with lived experience are engaged as study team members to lend their hands-on expertise and thoroughly participate in the intervention development process. Given well-documented risks associated with the sex trade and opioid use, as well as a lack of interventions prospectively designed to address challenges experienced by this population, the current study represents a critical next step towards promoting the health and well-being of women in the sex trade. The proposed five-year Mentored Patient-Oriented Research Career Development Award (K23) will provide Dr. Murnan a perfect opportunity to achieve his career goals through training in: 1) Community-Based Participatory Research; 2) ADAPT-ITT opioid use intervention adaptation methodologies; and 3) conducting research within the criminal justice system. Dr. Murnan’s proposed study aims to evaluate the feasibility and acceptability of co-designed family-based intervention offered as an adjunct to treatment diversion programming (offered through the criminal justice system) using a pilot RCT design. In Aim 1, participant family dyads (n=15 women paired with one supportive family member) will be recruited to explore intervention needs, motivations, and barriers related to participating in a family-based intervention, as well as delivery preferences. This information will be used by the co-design team (individuals with lived experience, the PI, and other research personnel) to adapt a family-based intervention manual. For Aim 2, additional family dyads (n=60) will be recruited to participate in a randomized controlled trial in which they will be randomly assigned to one of two treatment arms: 1) co-designed family-based intervention + diversion treatment programming or 2) diversion treatment programming only. Feasibility, acceptability, and preliminary efficacy of both interventions will be evaluated (Aim 2). Dr. Melinda Butsch Kovacic, a leading expert in community-based participatory research, will serve as Dr. Murnan’s primary mentor. Dr. Butsch Kovacic in conjunction with his co-mentors (Drs. Jennifer Brown and Sarah Manchak) will provide him with necessary training in areas of community-based participatory research, substance use/opioid use intervention adaptation methodologies, and conducting research within the criminal justice system, that will launch his independent research career as a clinician scientist with the requisite skills to conduct high quality intervention research to promote best outcomes among families disproportionately impacted by opioid use and criminal justice involvement.

NIH Research Projects · FY 2026 · 2026-05

Abstract The Ohio State University Comprehensive Cancer Center (OSUCCC) Genomics Shared Resource (GSR) is requesting funds to purchase an Oxford Nanopore Technologies (ONT) PromethION 24 long-read nucleic acid (DNA and RNA) sequencer. There are several innovative and exciting features of the PromethION 24 which is the “third generation” sequencing instrument that best meets the needs of OSU researchers. Features include 1: the ability to assess native DNA and RNA modifications; 2: the ability to generate long-reads of a variety of lengths from a few kb to over 2 Mb of continuous sequence; 3: the ability to perform long-read single cell sequencing from 10X Genomics single-cell libraries enabling evaluation of expression of mRNA isoforms, variants and mutations from single-cells; 4: adaptive sampling which is on-instrument targeted sequencing strategy in which nucleic acids not of interest are rejected from the pores; 5: the ability to run up to 24 flow cells synchronously or asynchronously which adds flexibility in project management and ability to run multiple types of projects at the same time as well as being able to easily accommodate larger projects. The PromethION has advantages over long-read instruments from other companies because of it being able to sequence longer (>25 kb) of contiguous sequence and the ability to directly sequence RNA and interrogate multiple different RNA modifications. Furthermore, ONT offers a variety of on-instrument and cloud-based open-source software (free to users) that range from straight-forward “point and click” user-friendly tools to more sophisticated analytical programs. There are also links to community-based GIT-hub software on the ONT website. Seven major and six minor users have current NIH-funded projects that could benefit from having this technology on site. Examples of types of projects that would benefit from access to a PromethION 24 on site include: single-cell RNA- sequencing to identify allele-specific expression and escape from X-chromosome inactivation, single-cell isoform expression, long-read sequencing of single-cell spatial (10X Visium) B-cell receptors (BCR)/T-cell receptors (TCR), long-read telomere sequencing (Telo-Seq), long-read ribosomal RNA sequencing to understand ribosomal RNA processing, long-read DNA sequencing to understand genomic complexity at the Spinal Motor Neuron (SMN) locus and its impact on phenotype, and long-read direct RNA sequencing for transcript and isoform analysis in T-cells during parasitic infection. The Promethion 24 will be housed in the GSR with administration and financial support from the OSUCCC. The instrument will be available to all OSU investigators as well as outside investigators. Oversight of the instrument will be supported by the GSR technical director who has ONT experience as well as 3+ staff with NGS library expertise. There are currently no long-read PromethION 24 sequencers available in any shared resource at OSU so this instrument fills a critical need and will support cutting-edge applications and science.

NIH Research Projects · FY 2026 · 2026-04

PROJECT SUMMARY--------------------------------------------------------------------------------------------------------------------- The timely and accurate genetic diagnosis of Mendelian diseases enables interventions to prevent catastrophic outcomes such as progressive heart failure and metastatic cancer. These diagnoses depend on the ability to distinguish pathogenic variants from benign. However, the vast majority of variants that have been identified in disease genes are currently classified as variants of uncertain significance (VUS), which are not clinically actionable. This “VUS problem” represents a major barrier to the implementation of Genomic Medicine. This proposal aims to address the VUS problem by employing a novel, integrative approach to prospectively assess variant pathogenicity. This approach will leverage the enormous genotypic and phenotypic variation present in large, DNA-linked biobanks together with the results of high-throughput, in vitro functional assays with the goal of generating sufficient evidence to reclassify VUS as either pathogenic or benign, at scale. To capitalize on the PI’s expertise in computational electrophysiology and Cardiovascular (CV) Medicine, the proposed research will focus on CV disease-associated genes; however, the methods developed in this proposal will be designed to generalize to Mendelian disease in any organ systems. The overall aim of this proposal is to accelerate the implementation of Genomic Medicine through the prospective reclassification of disease-gene VUS. The first aim of this proposal is to develop and validate CV gene-specific phenotype risk scores (PheRS) in the large, DNA-linked biobanks All of Us, the UK Biobank, and Vanderbilt’s BioVU, which, collectively, will have whole genome sequences for over 1 million participants linked to phenotypic data. These PheRS will be developed to capture a spectrum of phenotypes detectable in the biobank records with the purpose of amplifying the often- faint phenotypic signal associated with rare pathogenic variants in population studies. The second aim of this proposal will generate evidence to reclassify CV disease-gene VUS in the biobanks by deploying the biobank- calibrated PheRS in conjunction with Multiplexed Assays of Variant Effects (MAVEs), high-throughput, in vitro assays which generate functional data for nearly all possible protein-coding variants in a gene of interest. This proposed research will be conducted at Vanderbilt University Medical Center (VUMC), under the guidance of an exemplary mentorship team and in parallel with a thoughtful career development plan. VUMC is a world leader in Genomic Medicine and Biomedical Informatics, and thus an ideal environment for the PI’s continued training. The career development plan has been tailored to develop further expertise in human genetics, biomedical informatics, and Genomic Medicine. Overall, the research, environment, mentorship, and training in this proposal will give the PI with the necessary experience to successfully transition into an independently- funded physician-scientist.

NIH Research Projects · FY 2026 · 2026-04

PROJECT SUMMARY Human skin scarring after trauma or surgery remains a major medical problem. The transformation of fibrotic healing processes into regenerative ones, in which original tissues are restored, would constitute an enormous advancement in human health. Unfortunately, effective treatments to reduce scarring are still lacking. Addressing the complex post-injury immune responses is critical to devising effective therapies for skin wound scarring. Our overarching goal is to elucidate a novel wound-healing mechanism that could achieve “scar-free regeneration” in humans. Mast cells (MCs) are the first responders to local skin injury. Recent studies unraveled a unique property of MCs: their IgE-independent activation through the Mas-related G-protein-coupled receptor (Mrgpr)- b2. Mrgprb2 has been previously shown to mediate neurogenic inflammation and post-operative pain in injured skin. Our preliminary data further suggest a role for this novel non-canonical MC signaling (Mrgprb2 and its human homolog MRGPRX2) in wound healing and scar formation. This research will investigate three Specific Aims to test our central hypothesis that Mrgprb2 signaling in connective tissue MCs is a key contributor to post- injury scar formation through its immunoregulatory activities. In Aim 1 (in vivo studies), using Mrgprb2 gene- modified mouse lines, we will test the effects of manipulating Mrgprb2 signaling on skin scarring and regeneration. Mechanistically, we will dissect the pro-inflammatory and pro-fibrotic properties of Mrgprb2 signaling in wound healing animal models. In Aim 2 (in vitro studies), using skin wound-related cells derived from both mice and humans, we will test the hypothesis that Mrgprb2/X2-activated MCs drive immune cell polarization, endothelial- to-mesenchymal transition, and fibroblast proliferation and migration. We aim to decipher human MRGPRX2- regulated cellular interactions as key mechanisms of MC-driven fibrosis. In Aim 3 (human tissue studies), we will ultimately determine effects of MRGPRX2 signaling in human scar tissues using samples collected in the dermatology and plastic surgery clinics. We will also employ ex vivo human skin wound models to test MRGPRX2 signaling in early healing processes. This proposal is innovative because it will unravel a unique mechanistic link between inflammation and fibrosis by elucidating the essential roles of Mrgprb2/X2 signaling in tissue remodeling. The proposed research is significant because it may help develop novel immunological therapies based on using MC-specific GPCR as a target for tissue regeneration in fibrotic diseases of skin.

NIH Research Projects · FY 2026 · 2026-04

An emerging infectious disease, human granulocytic anaplasmosis (HGA) caused by infection with Anaplasma phagocytophilum (Aph), is first in mortality among tickborne diseases, and second in prevalence only to Lyme disease. Aph causes systemic infection by effectively replicating within membrane-bound inclusions/vacuoles of granulocytes and endothelial cells by subverting innate immune mechanisms and exploiting host nutrients. Therefore, understanding Aph machinery and mechanisms essential for this process in the context of the host response will inform the design of potential therapeutic/prophylactic targets. Aph have a Type IV secretion system (T4SS), similar to the virB/virD system, which allows pathogens to manipulate host cells viaT4SS effectors. We identified a new T4SS effector, endoplasmic reticulum (ER)-Golgi exit site (ERES) protein of Anaplasma (EgeA), and two host cell receptors of EgeA that are ERES proteins: Transport and Golgi organization protein 1 (TANGO1) and Sec1 family domain-containing protein 1 (SCFD1). EgeA, TANGO1, and SCFD1 are all required for Aph infection of human cells and appear to localize to Aph inclusions. On the peripheral ER, ERES exists as specialized domains in a dispersed pattern. At ERESs, vesicles bud off and fuse with cis-Golgi. TANGO1 proteins organize the early secretory pathway from the ER to cis-Golgi to sort cargoes that are too bulky or secreted in volumes too great to use generic mechanisms. Thus, our first hypothesis is EgeA facilitates ER nutrient delivery to Aph inclusions by redirecting TANGO1 and SCFD1 protein transport/ER exit machinery to Aph inclusions. TANGO1 functions to relieve ER traffic jams due to bulky cargo retention at ERES. Thus, our second hypothesis is EgeA alleviates ER stress via TANGO1-mediated ER protein exit. In this proposal, we will test these hypotheses in three major stages: In Aim 1 we will elucidate how EgeA hijacks TANGO1- and SCFD1-mediated protein transport machinery at ERES to deliver ER nutrients to Aph inclusions by assessing EgeA domains interacting with host molecules at ERES and inclusions and binding affinities of host molecules and spatial topology between ERES and Aph inclusions, and EgeA-dependency of delivery. In Aim 2 we will analyze how EgeA interacts with host protein components to modulate ER stress in Aph-infected cells and the unfolded protein response (UPR) signaling pathway. In Aim 3, we will test if EgeA functions and Aph infection can be blocked by intracellular delivery of lipid nanoparticle-encapsulated anti-EgeA nanobody mRNA in cell culture and in mice. The proposed studies will be crucial to understanding biomolecular mechanisms of EgeA functions in Aph infection. Impact: By ascertaining how intracellular Aph acquires ER nutrients and modulates ER stress and the UPR, infection may be pharmacologically or immunologically inhibitable, yielding new and innovative approaches to reduce human risk of acquiring or developing severe HGA. Further, the results will reveal a mechanism of TANGO1 and SCFD1 modulation in ERES transport and ER stress with an exogenous molecule, benefiting the broader fields of infectious diseases and ER homeostasis.

NIH Research Projects · FY 2026 · 2026-04

PROJECT SUMMARY Microorganisms residing in the intestinal lumen have a significant impact on brain function and behavior. Perturbation of microbe-gut-brain communication is thought to underlie the increased visceral sensitivity and anxiety that occur in disorders of gut-brain interaction (DGBI) like irritable bowel syndrome (IBS). Recent studies suggest that enterochromaffin cells (ECs), a group of enteroendocrine cells in the intestinal epithelium that secrete serotonin, sense microbial irritants. EC activation drives visceral pain and anxiety, suggesting a critical role of EC in mediating the pathophysiological development of IBS. However, the cellular signaling and neuronal circuitry by which EC activation drives anxiety remain largely unknown. Whether and how gut microbiota modulates ECs to promote anxiety also remains unclear. Using in vivo real-time measurements of cell activity in zebrafish, our recent research revealed that specific gut bacteria secrete tryptophan metabolites indole and indole-3-aldehyde (I3A) to directly activate ECs through the receptor transient receptor potential ankyrin A1 (Trpa1). Using optogenetics, gut-brain in vivo imaging, and behavioral analysis, new preliminary data from our laboratory demonstrated that the Trpa1-expressing ECs directly connect and communicate with vagal sensory fibers. Stimulating Trpa1-expressing ECs activate a subset of vagal sensory neurons that express Parvalbumin 7 (Pvalb7). Stimulating Trpa1-expressing ECs with microbial irritants promotes anxiety. This proposal will test the central hypothesis that indole and I3A secretion by enteric irritant bacteria activates the Trpa1+EC- Pvalb7+vagal sensory neuron pathway to promote anxiety. Three Aims were proposed. Aim 1 will use zebrafish and bacterial genetic manipulation to determine molecular mechanisms that promote the formation of EC-vagal synaptic connections and test the hypothesis that microbial irritant signals promote Trpa1+ECs to form synaptic connections with Pvalb7+vagal sensory neurons. Aim 2 will use optogenetics, vagal calcium imaging, and novel genetic reporter lines to image real-time EC-vagal sensory neuron communication in vivo to determine the precise molecular and signaling mechanisms that mediate Trpa1+EC-Pvalb7+vagal sensory neuron communication. Aim 3 will use genetics, optogenetics, brain calcium imaging, and behavioral analysis to determine the key brain region that is activated by the Trpa1+EC-Pvalb7+vagal sensory pathway and underlies microbial irritants-induced anxiety. The proposed project is expected to provide a mechanistic understanding of how microbial irritants perturb EC-vagal communication to alter brain activity and promote anxiety in a microbial, molecular, and cellular-specific manner. The results are expected to provide evidence for new therapeutic approaches that target EC-vagal circuitry to treat DGBIs associated with microbial dysbiosis.

NIH Research Projects · FY 2026 · 2026-04

Project Summary/Abstract Mosquito population control is a primary means to prevent mosquito-borne disease worldwide, and many novel approaches to population control involve traditional or modified versions of Sterile Insect Technique (SIT). One thing these methods all have in common is the need to rear healthy adult male mosquitoes en masse. These males are then released to mate with wild females, which they sterilize and prevent from producing offspring, thus causing a decline in mosquito population size. Males used for SIT must be able to compete for mates in a field setting for the method to be successful. Moreover, modifications to rearing that improve mass production of males, increase their competitive ability, or increase the duration of their field efficacy, will improve the success and reduce the cost of these programs. The microbes found in larval development water and within the mosquito body have been shown to impact multiple traits that are relevant to SIT and mass rearing, including pupation time and body size. Here, we propose to comprehensively determine the impact of the microbiota on adult male life history traits relevant to the success of SIT. We will experimentally colonize mosquitoes with multiple candidate bacterial communities and determine the impact of different communities on male longevity, flight capacity, mating success, sperm quantity, and ability to induce refractoriness to re-mating in female mosquitoes. Additionally, we will assess the impacts of these colonies on male traits in semi-field conditions We will also assess the impact of the microbiota on male life history traits across a genetically differentiated group of mosquito strains. Finally, we will assess the role of B vitamins as a mechanistic determinant of male mosquito-microbe interactions. This work will reveal, for the first time, the comprehensive impact of the microbiota on traits relevant to the success of SIT. Additionally, it has the potential to reveal specific bacterial treatments that could be used to improve male quality in mass rearing operations and the extent to which effects of the microbiota may differ depending on mosquito genetic background. Each component of the proposal is critical to understanding mosquito-microbe interactions in male mosquitoes and will provide key foundational knowledge as well as potentially actionable findings to improve the success of SIT.

NIH Research Projects · FY 2026 · 2026-04

7. Project Abstract Approximately 3.6 million concussions occur every year in the United States. The largest affected group is 11- to 18-year-olds, and the diagnosis of concussion in that age group is increasing. Children and adolescents are particularly vulnerable to the harmful effects of concussion because their higher-level cognitive functions are still developing. Although many adolescents fully recover from their concussion within four weeks, a significant number of adolescents have a prolonged recovery that may last weeks or even months. Common symptoms reported during the recovery from concussion are difficulty concentrating, blurred and/or double vision, losing one’s place when reading, and headaches. At least one vision disorder is reported to be present in 40-70% of adolescents with concussion. Persistent vision disorders during the recovery period are associated with decreased cognitive function and may be associated with prolonged concussion recovery. Despite the reported frequency of vision disorders in adolescents with concussion and the clear overlap in non-visual symptoms for concussion and vision disorders, it is still unclear what role vision disorders may play in the recovery process and their longer-term impact. Through three specific aims we will: 1) Evaluate visual function among adolescents with concussion at <28 days from injury and 90 days post injury, 2) Define the relationship between the cognitive and neurological performance, concussion symptoms, and vision disorders and symptoms in the first 28 days post-injury, and 3) Establish which vision findings observed in the first 28 days post- concussion predict delayed vision recovery as defined by both symptoms and visual dysfunction. These results will provide guidance to concussion doctors about what signs and symptoms would indicate the need for a comprehensive vision exam, as well as what to look for as predictors of persistent symptoms.

NIH Research Projects · FY 2026 · 2026-04

Project Summary (Abstract): Glioblastoma (GB) remains one of the most lethal cancers, characterized by a profoundly immunosuppressive tumor microenvironment (TME) that limits the effectiveness of current immunotherapies. While research has heavily focused on traditional regulators of immune cells, nociceptors—pain-sensing sensory neurons known to regulate immune responses in the periphery—have not been studied in GB, creating a significant gap in our understanding of immune regulation in the disease. Of note, in the cranial region, nociceptors are densely concentrated in the dural layer of the meninges but are absent from the brain parenchyma. This anatomical separation from GB tumors has likely contributed to their historical neglect in GB research, overlooking their potential as critical regulators of anti-tumor immunity. Our preliminary data provide compelling evidence that nociceptors play an active role in GB pathogenesis. In syngeneic orthotopic GB mouse models, we observed heightened activation of dural nociceptors in the presence of tumors, marked by increased production of calcitonin gene-related peptide (CGRP), a neuropeptide with known immunomodulatory functions. Furthermore, cerebrospinal fluid (CSF) from GB-bearing mice promotes pronounced axonal elongation in cultured primary trigeminal nociceptors, indicating that tumor-derived factors can directly modulate these neurons. Strikingly, nociceptor ablation in GB-bearing mice leads to transformative changes: prolonged survival, a shift in the TME from an immune-suppressive ‘cold’ state to an immune-activating ‘hot’ state and enhanced responsiveness to immune checkpoint blockade (ICB) therapy. These findings demonstrate that nociceptors, despite their physical separation from the tumor, can remotely regulate GB progression by modulating the immune landscape. To elucidate the mechanisms underlying nociceptor-mediated immune regulation in GB, we are employing methodologies including ELISAs, in vitro neuronal culture assays, single-nucleus and single-cell RNA sequencing (snRNA-seq, scRNA-seq), multi-dimensional flow cytometry, and survival studies under conditions of immune perturbation. These approaches will elucidate the tumor-derived factors that modulate nociceptors and define the bidirectional interactions between nociceptors and immune cells within the TME, revealing the mechanisms by which these neurons regulate anti-tumor immunity. Beyond advancing fundamental understanding, this work holds significant therapeutic potential. By targeting nociceptor-driven immune regulation, we aim to develop strategies to reverse immune suppression in GB, including repurposing existing nociceptor-targeting therapies to enhance efficacy of immunotherapies. Ultimately, our goal is to uncover new therapeutic avenues for improving survival outcomes in patients with this devastating disease.

NIH Research Projects · FY 2026 · 2026-04

SUMMARY Pseudomonas aeruginosa is a ubiquitous opportunistic pathogen that causes acute and chronic pneumonia, sepsis, urinary tract infections, and surgical-site infections. Its success as a human pathogen is due in large part to its ability to form biofilms, which confer resistance to the host immune system, antibiotics, and other stresses. Biofilms are structured communities held together by an extracellular matrix composed primarily of exopolysaccharides (EPS) encoded in long operons. As all RNA polymerases are prone to premature termination, the expression of long operons commonly relies on dedicated antitermination factors that belong to the universally conserved family of NusG transcription regulators. NusG paralogs bind to the elongating RNA polymerase and enable uninterrupted synthesis of long RNAs in many bacteria, including Bacillus velezensis, Bacteroidetes fragilis, and Escherichia coli. Surprisingly, P. aeruginosa does not encode NusG paralogs, and the scarce available data suggest that its regulation of transcription termination and antitermination is very different from that of E. coli. We hypothesize that many extended P. aeruginosa operons are subject to premature termination by Rho, possibly with the assistance of NusG and histone-like Mva proteins, and that they use unique antitermination mechanisms to ensure complete synthesis of their multi-cistronic RNAs. We propose to investigate the regulation of P. aeruginosa RNA chain elongation, focusing on two EPS operons, pel and psl. This choice is dictated by outsized contributions of these gene clusters to P. aeruginosa pathogenesis and their expected dependence on antitermination to complete the synthesis of 10-19 kb RNA chains. In Aim 1, we will determine if Rho, NusG, and Mva proteins induce premature termination during transcription of the pel and psl operons using in vivo RNA probing and in vitro assays with purified P. aeruginosa RNA polymerase and transcription factors. In Aim 2, we will identify accessory factors that enable the processive synthesis of long EPS RNAs. Together, these experiments will provide first insights into the poorly understood post- initiation control of transcription in P. aeruginosa.

NIH Research Projects · FY 2026 · 2026-04

Adipose tissue (AT) is an active metabolic organ that contains a network of immune cells whose crosstalk regulates adipose tissue homeostasis. In response to obesity, these immune cells can become activated, expand, and secrete adipokines and pro-inflammatory cytokines that modify local and systemic insulin sensitivity and contribute to the development of metabolic dysfunction and type 2 diabetes. Promotion and maintenance of adipose tissue inflammation in the obese state has been shown to involve the activation of adipose tissue T cells (ATT), which potentiate the activity of pro-inflammatory adipose tissue macrophages (ATMs) in mouse models of obesity. There is a fundamental gap in our understanding of the mechanisms by which ATTs are maintained, their diversity relative to other T cell subsets, and the signals to which ATTs respond during adipose tissue remodeling and expansion. Our preliminary data assessing early ATT kinetics demonstrated that ATTs proliferate in response to the rapid AT expansion induced by short-term high-fat diet (HFD) feeding. Timepoints assessed in these experiments precede the substantial infiltration of pro- inflammatory ATMs associated with the obese state, suggesting ATTs play vital roles in the initiation of AT inflammation observed in the obese state. This proposal seeks to test the hypothesis that ATT proliferation in response to short-term HFD feeding requires signals from antigen presenting cells (APCs). Further, the proposed research will interrogate the establishment of memory ATT and the determinant which drive the priming of their responses to AT changes in the obese state. Lastly, the PI’s prior research uncovered a unique population of TCRαβ CD3+ CD4- CD8- double negative (DN) ATTs and will test the hypothesis that DN ATTs possess an immunoregulatory phenotype and function in maintaining AT homeostasis and controlling effector ATTs during development of obesity. To test these hypotheses, the proposed project will pursue the following Aims during the K99 phase of this award: 1) To determine driver(s) of ATT proliferation during short-term HFD feeding and 2) to identify phenotype and function of DN ATTs in the lean and obese state. During this time, the PI will receive research training in nutrient metabolism in mice, bioinformatics and sequencing analysis, and translational research in obesity and diabetes. During the independent R00 phase, the PI will continue these Aims and seek to 3) determine establishment of resident memory ATTs and signals required for their induction and maintenance. Completion of this work will have significant impact on identification of the regulators of ATT functions in lean and obese states and elucidate novel targets to suppress obesity-associated AT inflammation mediated by ATT responses. Research outlined in the R00 phase will build the foundation of the PI’s independent research program centering on obesity-associated inflammation and metabolic dysfunction, and how ATTs contribute to the initiation, orchestration, and maintenance of AT inflammation and subsequent insulin resistance.

NIH Research Projects · FY 2026 · 2026-04

A sustained decline in practicing surgeon–scientists has been identified as a major contributor to the bench-to-bedside translation gap in otolaryngology. This R25 program directly addresses these barriers by implementing an interdisciplinary, structured, and holistic training program designed to launch successful, self-sustaining careers in a cohort of next-generation otolaryngology surgeon–scientists. The program will achieve this goal through three integrated objectives: (1) recruitment of a cohort of medical students and otolaryngology residents with strong potential for academic research careers; (2) delivery of a comprehensive training curriculum encompassing research methods, outcomes research, statistics, team science, scientific communication, grant writing, wellness, and professional development; and (3) creation of an interdisciplinary and collaborative research environment that supports mentorship, team-science, and long-term career sustainability. By leveraging Ohio State University’s extensive translational research infrastructure, interdisciplinary faculty expertise, and trainee wellbeing, this program aims to strengthen the otolaryngology surgeon–scientist pipeline and advance NIDCD-relevant research across the lifespan.

NSF Awards · FY 2026 · 2026-04

This Research Experiences for Undergraduates (REU) site award to The Ohio State University located in Columbus, Ohio, supports the training of 10 students for 10 weeks during the summers of 2026–2028. In this program, funded by the Chemistry section, students conduct research under a faculty mentor and participate in weekly meetings that provide professional development training for future postgraduate degrees in STEM fields. The program supports participants from institutions that lack specialized instrumentation for research, especially in underserved Rust Belt cities and Appalachian counties in Ohio and neighboring states. The REU program, augmented by long-term outcome tracking and assessments of program effectiveness, provides these students with mentoring and networking opportunities that will enhance their career opportunities in science and technology. The research training in this program is focused on how to use laboratory instruments and computers to learn about molecules and materials using electromagnetic radiation at wavelengths spanning the spectrum from radio frequencies to visible and ultraviolet light to x-rays, for a broad range of applications. The spectroscopy research theme overlaps foundational ideas taught in initial chemistry courses, ensuring that even beginning students have some existing subject-matter knowledge upon which they can build. Training in the interactions between light and matter (spectroscopy) is critical for creating a workforce prepared for 21st century jobs in fields such as biomedical imaging, materials characterization, and quantum information science. In addition, the program features a weekly workshop on math and coding that will include elements of artificial intelligence (AI) fluency and machine learning (ML) as applied to laboratory experiments in spectroscopy. These activities will prepare students to enter a cyber- and AI-enabled workforce where coding skills are often necessary even for experimental spectroscopists. 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.

NSF Awards · FY 2026 · 2026-03

Researchers in the social sciences increasingly utilize event data sets when studying crime, protests, and terrorism. These data sets provide information on each incident, including where it occurred, who was involved, what the consequences were, etc. Unfortunately, the recorded location of incidents in these data sets are often inaccurate, due to limitations in the available information from which they are drawn (ex. incomplete media reports). Left unaddressed, these geolocation errors impair one’s ability to effectively learn about the underlying process of interest from these data. For example, geolocation errors may cause researchers to infer spatial patterns from these data that would not be found with the correct locations. In this research, investigators will develop statistical methods to better account for geolocation errors in these kinds of data. The statistical methods developed will then be applied to data on political violence, demonstrating their importance for improved understanding of real-world problems. The multidisciplinary project will also provide training for the next generation of researchers at the intersection of statistics and the social sciences. This collaborative project includes support and mentorship for graduate students. Spatial point processes are a natural approach for modeling event data. However, geolocation errors produce two distinct, but related, problems for these methods: i) duplicate event locations, and ii) inaccurate spatial coordinate information. In this project, investigators will address both issues, developing a computationally efficient statistical inference method to account for geolocation error in spatial point pattern data within the Log-Gaussian Cox Process framework. Various geolocation error structures will be considered, including nonstationary errors, to better reflect complex real-world applications. The project will include research on both the finite-sample performance and asymptotic behavior of the estimators from the developed inference methods. These methods will be used to analyze real-world political violence data from various sources. 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 2026 · 2026-03

PROJECT SUMMARY/ABSTRACT This career development award will provide the didactic coursework, experiential training, and mentorship necessary for the candidate to become an independent clinician-scientist capable of developing and evaluating emergency medical services (EMS)-based interventions for substance use disorders (SUD). The research plan focuses on the opioid epidemic and emerging EMS interventions for harm reduction, treatment linkage, and recovery support for individuals following nonfatal overdose. Despite a rapid expansion of EMS interventions for SUD, data to guide the most effective approaches remain limited. There is an urgent and critical need to develop and evaluate strategies that reach and engage at-risk individuals earlier and more intensively in harm reduction, treatment linkage, and recovery support. EMS has traditionally focused on emergency stabilization and transport, but its emerging public health arm – mobile integrated healthcare & community paramedicine (MIH-CP) – expands services to include community outreach, prevention, and longitudinal care. A common MIH-CP example is the post-overdose quick response team (QRT), which delivers EMS-based outreach, harm reduction materials, and treatment linkages after overdose. Though increasingly popular, demand for QRT has outpaced evidence, leaving its optimal design, services, and outcomes unclear. QRT has focused almost exclusively on opioid use despite the broader burden of polysubstance use and remains largely reactive, engaging individuals only post-overdose. Expanding QRT to proactively reach at-risk individuals (i.e., pre-overdose) and broadening its consideration of co-occurring SUD represents a compelling yet untested approach to enhance early intervention and improve outcomes. The objective of this study is to rigorously observe and evaluate an exemplar QRT in Columbus, Ohio to characterize processes and key components associated with improved outcomes. Our aims are to: 1) evaluate the association of proactive (pre-overdose) and reactive (post- overdose) outreach on participant engagement in QRT and 2) characterize predictors of change in substance use and health outcomes three months after QRT engagement. The contribution of this research will be to characterize the real-world outcomes of QRT intervention and identify critical components that improve individual and public health. With mentorship from a qualified and multidisciplinary team, the candidate will generate a preliminary dataset evaluating QRT approaches, guiding the next generation of EMS-based SUD interventions by supporting NIDA’s strategic goal to integrate harm reduction and treatment into real-world settings. The research plan complements and supports the candidate’s development as an independent clinician-scientist focused on EMS-based interventions for SUD by providing expertise in field-based longitudinal research, SUD outcomes assessment, advanced statistical analysis (including causal inference methods), and mixed-methods approaches to evaluation.

NIH Research Projects · FY 2026 · 2026-03

PROJECT SUMMARY Microbial carbohydrate metabolism is typically assayed by measuring biomass accumulation in pure cultures on a defined carbon source. This approach may obscure what occurs in natural microbial communities where cooperation or competition with other species may exist, where metabolic intermediate are generated that other species may utilize, and it ignores the physical features of the environment where that community resides. Humans host a taxonomically and functionally diverse microbial community in their gastrointestinal tracts that is influenced by multiple intrinsic and extrinsic factors that remain ill-defined. Extrinsic carbohydrate nutrients (diet- or host-derived) can support the growth of microbes that can access and utilize those nutrients. How bacteria intrinsically alter the nutrient landscape of their environment and how/whether active bacterial processes promote competition or cooperation for carbohydrate nutrients remains unclear. We have identified oligosaccharide intermediates produced during dietary plant polysaccharide (fiber) degradation that are structurally modified by bacteria before being released into the environment. We hypothesize that select human gut bacteria generate modified oligosaccharides to create extracellular nutrient reservoirs, enhancing their fitness and that of their kin in competitive environments. Over the next five years, we will first define the structures of modified oligosaccharides produced by gut bacteria to determine how human gut microbes synergize anabolism and catabolism to alter the pool of available carbohydrate nutrients. Next, we will establish how oligosaccharide structures influence the ability of bacteria to acquire, degrade, and utilize nutrients by employing forward- and reverse genetics approaches and synthetic microbial communities. Lastly, we will establish how microbial community composition and host diet shape oligosaccharide nutrient availability along the length of the gastrointestinal tract using mass spectrometry imaging. Using the mammalian gut microbiota as a model microbial community, we will define an intrinsic role for bacteria in shaping carbon nutrient accessibility. Our study will reveal an active biochemical mechanism that bacteria employ in nutrient- competitive environments to obtain carbohydrate nutrients and compete against other species.

NIH Research Projects · FY 2026 · 2026-03

PROJECT SUMMARY/ABSTRACT Deep tooth injuries and inflammation affect millions of Americans, often progressing into pulp necrosis and treated by removing, i.e. devitalizing, and replacing the dental pulp tissue with artificial materials. Vital pulp therapies (VPTs) have the potential to promote cell signaling that stops the infection and restores pulp health, but their effectiveness is limited once pulpitis is established. This is largely due to the large gap in knowledge of what signaling cascades regulate the immune response to defend the pulp versus promote tertiary dentino- genesis to repair the damage. Studies have demonstrated that sensory axons sprout during infection and injury and communicate with immune and stem cell populations during tissue healing. Insights into how neurons pro- tect and enable dentin repair can therefore create therapeutic strategies that harness this axis to heal, rather than remove, the dental pulp tissue. The long-term goal of this project is to understand the cellular mecha- nisms that protect teeth via immune and mineralizing cell responses to inflammation and damage. The applica- tion objective is to determine the role of calcitonin gene-related peptide (CGRP) in activating immune and min- eralizing cells during tooth healing, i.e. reparative dentinogenesis. Our central hypothesis is that neuronal-de- rived CGRP regulates the function of neutrophils & monocytes, surviving odontoblasts and odontoblast progen- itors via its receptor, receptor activity modifying protein 1 (Ramp1), during reparative dentinogenesis. The ra- tionale for this project is that a detailed scientific framework of cellular crosstalk during reparative dentinogene- sis is likely to identify cell-specific mechanisms that sustain and enhance tooth health. The central hypothesis will be tested with the following specific aims: 1) Determine the cell-specific effects of CGRP signaling after deep dentin trauma; 2) Identify the role of CGRP signaling in regulating inflammation and subsequent tertiary dentinogenesis after direct pulp capping procedures; and 3) Evaluate the effect of increased RAMP1 expres- sion and/or exogenous CGRP on promoting pulp healing and homogeneous mineralized tissue barrier for- mation after direct pulp capping. In aim 1, direct pulp capping will be performed on mice with conditional dele- tions of the CGRP receptor, RAMP1, in immune or mineralizing cells. The dentin pulp complex will be analyzed throughout the healing period of 1-56 days for inflammatory and mineralization responses. For aim 2, a model of pulpal inflammation will be created with an initial shallow injury on molars with lipopolysaccharide applied. This will be drilled out and restored 24 hours later, followed by a similar timeline of investigations into pulpal responses. The last aim proposes to enhance RAMP1 and therefore the ability to respond to CGRP released during injury and inflammation to establish whether this can promote pulp healing and tissue barrier formation. The research proposed in this application is innovative because it provides a clinically relevant model of deep dentin damage and repair with which to study cell-specific repair mechanisms. The proposal is significant be- cause it is expected to identify cellular mechanisms that sustain tooth vitality to advance the development of next-generation VPTs.

NIH Research Projects · FY 2026 · 2026-03

PROJECT SUMMARY The long-term goals of our research program are to discover new microbial natural products (NPs) of chemotherapeutic value, establish a mechanistic understanding of their underlying their biosynthesis and bioactivities, and to leverage these molecules, enzymes, and metabolic pathways for biomedical and biotechnological advancement. Evolved over millennia, NPs are chemically diverse small molecules produced from living organisms that exhibit with a wide range of useful inhibitory activities. NPs are a critical source of new drugs and often the initial chemical catalysts driving drug development. Phosphonic acids (Pn) are a privileged class of NPs with a prolific history of development and commercialization as antibiotics, antivirals, antimalarial, anticancer, and herbicidal agents. Universally defined by a carbon-phosphorous (C-P) bond, this moiety enables unique targeting of essential pathways in cellular metabolism thru chemical mimicry of enzyme substrates, products, and transition-state intermediates. These traits, combined with a near boundless collection of uncharacterized biosynthetic gene clusters for Pn encoded in microbial genomes and metagenomes, suggest that a significant reservoir of novel compounds remain for this important NP class. Genome mining is a proven method to accelerate discovery of NP leads through prediction of their pathways, molecules and activities from biosynthetic gene cluster sequence. However, fundamental gaps in our biosynthetic and chemical understanding of Pn NPs significantly limits the predictive power for this compound class. Thus, universal genetic, biochemical, and bioactivity principles of Pn NPs must be established to fully exploit their encoded genomic potential. In this proposal, we propose two research directions to address these goals. Direction 1 seeks to answer challenging, foundational questions central to Pn biosynthesis that will generate conceptual innovations necessary to improve their discovery. Using our expertise in bioinformatics, molecular microbiology, biochemistry, and chemical analysis, we will define essential characteristics of C-P bond enzymes, discover of new mechanisms C-P bond formation, and elucidate pathways for new Pn NPs. In Direction 2 our expertise in genome mining, NP isolation, and structure elucidation will be combined to isolate new inhibitory Pn NPs from diverse biosynthetic gene clusters of marine, soil dwelling, and plant associated microbes predicted to encode novel chemistries. Both directions afford natural integration throughout the project, as biosynthetic findings will support genome mining of new NPs, and newly uncovered molecules will drive studies that illuminate previously unknown paradigms in biosynthesis. Outcomes of this research program are expected to significantly advance Pn drug discovery thru refinements of their biosynthetic landscape and with the discovery of new NPs with potential therapeutic value.

NIH Research Projects · FY 2026 · 2026-02

ABSTRACT: Our laboratory discovered and confirmed STING (Stimulator of Interferon Genes) as an essential component of the cytosolic DNA-mediated innate immune response pathway. We subsequently demonstrated that STING signaling is commonly suppressed in microbial infected cells or human cancers, which enable such cells to escape the host immunosurveillance system. Our new data, enclosed herein, indicates that intrinsic innate immune STING signaling stimulates the production of essential innate immune proteins that in combination with microbial or self-nucleic acid, renders such cells highly immunogenic. Here, we intend to clarify the mechanisms of how intrinsic STING signaling facilitates the trans-activation of phagocytes and the cross-presentation of microbial or tumor antigen. Aim I: We aim to evaluate the mechanisms by which innate immune STING signaling renders microbial infected cells or DNA-damaged cells immunogenic, compared to normal apoptotic cells (which are non-inflammatory). Our data indicates that cytosolic DNA triggered STING-inducible genes combine with and protect, cytosolic DNA species to enable them to escape DNase-mediated degradation and activate STING signaling, in trans, in engulfing phagocytes. We have now identified these genes and will further characterize their novel mechanisms of action. Aim II. Our data indicates that innate immune STING signaling is suppressed in aged cells by epigenetic silencing. Such cells, for example, STING -/- MEFs or transformed cells lacking cGAS or STING expression, are non- immunogenic following DNA-damaging events, even though they produce cytoplasmic micronuclei. We hypothesize that reconstituting intrinsic STING signaling in vitro and, in vivo, will render cells immunogenic and trigger the innate activation of engulfing phagocytes. We believe that these strategies could shed insight into mechanisms of resistance to cancer therapies, (many of which invoke micronuclei formation) as well as lead to new strategies to help improve the treatment of a wide array of inflammatory and malignant disease.

NIH Research Projects · FY 2026 · 2026-02

SUMMARY Systemic lupus erythematosus (SLE) is a chronic autoimmune disease characterized by systemic immune dysregulation, with lupus nephritis (LN) being one of its most severe manifestations. While systemic immune abnormalities in lupus are well recognized, the immune landscape within lupus-affected tissues remains poorly characterized, and the role of immune checkpoint-mediated regulation in autoimmune microenvironments is a critical yet largely unexamined frontier. PD-1H (Programmed Death-1 Homolog, also known as VISTA) is a unique immune checkpoint expressed on both T cells and myeloid cells, functioning as a key regulator of immune suppression. Our preliminary studies reveal that PD-1H deficiency leads to spontaneous autoimmunity resembling lupus, and PD-1H is upregulated in lupus-affected tissues, suggesting its critical role in local immune regulation. These findings set the stage for investigating how PD-1H regulates immune pathogenesis in lupus and evaluating PD-1H-targeted immune checkpoint activators (ICAs) as a novel therapeutic approach. Aim 1 will determine the role and mechanisms of PD-1H signaling in the regulation of pathogenesis in the lupus affected tissue microenvironment. We hypothesize that PD-1H functions as a checkpoint receptor on pathogenic T cells and anti-inflammatory macrophages, mitigating inflammation and reducing tissue damage. To test this, we will (1) map the spatial distribution of PD-1H in kidney and skin biopsies from lupus patients using multiplex immunofluorescence and spatial transcriptomics and (2) determine how PD-1H regulates immune infiltration and modulates pathogenic immune activity using induced lupus models in tissue-specific PD-1H- deficient mice. Furthermore, our preliminary studies reveal that PD-1H interacts with key kinases in T cell receptor (TCR) signaling. We hypothesize that this interaction mediates PD-1H’s inhibitory effects on T cell activation, which we will test using structural mutants, co-immunoprecipitation assays, and phosphor-proteomics- based analyses to map PD-1H-regulated signaling networks in lupus pathogenesis. Aim 2 will Develop Novel Therapeutic Strategies for Lupus Using PD-1H Immune Checkpoint Activator (ICA). We will assess the efficacy of mouse PD-1H ICA in lupus and LN models, performing functional assays to determine its impact on T cell suppression and macrophage modulation. To advance these findings toward clinical translation, we will evaluate the effects of a newly developed human PD-1H ICA in a lupus patient-derived xenograft (PDX) model. These studies will establish PD-1H ICA as a novel strategy to modulate autoimmune pathogenesis while preserving immune homeostasis. This project integrates mechanistic studies with innovative therapeutic strategies to establish PD-1H as an essential regulator of lupus pathogenesis. These findings will not only reshape our understanding of immune regulation in lupus but also provide a foundation for developing next-generation immune checkpoint-targeted therapies for autoimmune diseases.