University Of California At Davis

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY (ABSTRACT) Enteric fever is caused by the bacteria Salmonella enterica serovars Typhi and Paratyphi (S. Typhi and S. Paratyphi). It remains a significant public health challenge in South Asia, Southeast Asia, and sub-Saharan Africa, particularly affecting children under five years old. In 2017, an estimated 14.3 million cases resulted in approximately 135,900 deaths, imposing substantial healthcare costs and economic burdens on lower- and middle-income countries. Clinical manifestations of enteric fever include prolonged fever, abdominal pain, headache, and malaise, with potential complications such as intestinal perforation, septic shock, and death. Diagnosis remains challenging due to non-specific symptoms and the limitations of current diagnostic methods, which are often expensive, inaccessible, and slow, leading to underreporting and inadequate surveillance. The emergence of antibiotic-resistant strains of S. Typhi further complicates treatment strategies, underscoring the urgent need for effective interventions. Vaccination with the typhoid conjugate vaccine (TCV), which offers over 70% efficacy and up to seven years of protection, presents a viable preventive measure. However, widespread vaccine implementation is hindered by a lack of reliable epidemiological data to inform policy decisions. Seroepidemiology, the collection and analysis of data on the presence and quantity of antibodies in blood to study the distribution and determinants of infection in populations, can provide cost-effective tools to estimate seroconversion rates and better understand infection dynamics, facilitating targeted vaccine deployment. New methods are available that using quantitative antibody responses to HlyE and LPS antigens to characterize seroconversion rates in population-based samples. However, several important limitations remain to scale these methods to inform and evaluate public health interventions. This project aims to address critical gaps in enteric fever surveillance by advancing seroepidemiological methods to support disease detection and prevention. Aim 1 will develop a method to accurately characterize enteric fever seroconversion rates in settings with high disease burden and frequent re-exposures. Aim 2 will extend these methods geospatially identify populations with the highest enteric fever seroconversion rates. Aim 3 will develop a novel method to distinguish S. Typhi from S. Paratyphi based on quantitative antibody responses and modeled antibody decay curves. This pre-doctoral research will enhance the accuracy of seroepidemiological methods for enteric fever, informing vaccine introduction and other disease prevention efforts. The accompanying training plan will to provide the trainee in with the skills and professional expertise necessary for a successful career as an independent academic researcher.

NIH Research Projects · FY 2025 · 2025-09

Project Summary/Abstract All cells in the body are covered with a sugar- and protein-rich material called the glycocalyx. Specific changes to the composition and organization of the glycocalyx are linked to lethal cancers and other debilitating diseases, but the mechanisms are not fully understood. Our group’s mission is to uncover the physical principles that define glycocalyx function with the hope that we can reprogram cells back to normal, healthy states or other desired phenotypes through rational manipulation of the glycocalyx. An interwoven mission is to develop the infrastructure—custom-tailored experimental tools and computational models—necessary to propel the early stages of biophysical inquiry in glycobiology. Our proposal will develop imaging tools intended to resolve the nanoscale architecture of the glycocalyx and probe its physical properties. We will continue to advance a systematic approach for engineering the biochemical and biophysical states of the glycocalyx through genetic methods. Theoretical models will be constructed to understand structure-function relationships for the glycocalyx. The tools and models that we develop for glycocalyx research will be applied to understand how the glycocalyx physically regulates various modes of intercellular communication, including through cell-surface receptors, highly curved membrane structures, and extracellular vesicles. Cell surface receptors reside and operate within the densely crowded glycocalyx. We will test the specific hypothesis that entropic forces arising from macromolecular crowding in the glycocalyx can strongly influence receptor assembly and activation. We will initially consider the effects of glycocalyx crowding on the dynamics and activation of growth factor receptors, including the epidermal growth factor receptor family. Works from our group have implicated transmembrane mucins and other membrane-anchored polymers in the glycocalyx as membrane curvature generating machines. Here, we will investigate how membrane-bending by the glycocalyx can support the projection of microvilli and the secretion of extracellular vesicles that bud from the plasma membrane and tips of microvilli. We will consider whether the residual glycocalyx on vesicles controls vesicle stability, fusogenic properties, and communication, including through the glycan epitopes that the vesicles carry. Our overarching hypothesis is that the biophysical properties of the glycocalyx play key roles in the transmission and receipt of biological information to and from cells. Emerging from our hypotheses is a vision that glycocalyx engineering and pharmacological targeting of the glycocalyx represent viable strategies to harness control over cellular interactions in tissue engineering, regenerative medicine, and cell-based therapeutics. Insights from our work are intended to inform the development of therapeutic strategies that can correct the broken channels of intercellular communication that are ubiquitous in diseases such as cancer.

NIH Research Projects · FY 2025 · 2025-09

Alzheimer’s disease and related dementias (ADRD) are the most significant health concerns of our time. More than 6 million older Americans have ADRD, and by 2050 there will be 12.7 million cases of ADRD costing over $1 trillion. Environmental risk factors, including extreme temperatures and air pollution, and environmental resilience factors, including greenspaces and walkable built environments, may be important drivers of brain health on both the short- and long-term. Although these exposures are ubiquitous and modifiable, little is known about how these exposures influence cognitive function on a day-to-day basis, or whether chronic exposure to these factors over time translates into long-term cognitive health, dementia risk, and neuropathologic burden. Exposure science advances now enable measurement of environmental factors that may drive brain health with unprecedented precision and specificity. Using data from participants in the nationwide Nurses’ Health Study II (NHSII) (ages 59-76 years old), we aim to estimate how smartphone global positioning system-based exposure to temperature, greenspace, and the built environment is linked to repeated monthly smartphone-based cognitive tests in 1,000 participants followed continuously over a year. In a subset of NHSII participants who underwent repeated web-based cognitive tests every 6-12 months starting in 2014 (N=20,532, mean age 69 at baseline) and in the Rush University Cohorts (the Religious Orders Study, Rush Memory and Aging Project, the Minority Aging Research Study, the Clinical Core Study, and the Latino Core Study (combined N=5,308, mean age 61 at baseline)), we will estimate how long-term residence-based exposures to these same environmental factors are associated with cognitive decline. Using data from the Rush University Cohorts, we aim to estimate how long-term residence-based exposure to environmental factors influence incidence of dementia and neuropathologic measures upon autopsy. These data combined will provide a comprehensive picture of how environmental exposures may impact both short-term cognitive function, long-term cognitive decline, and the development of Alzheimer’s disease and related dementias including their underlying neuropathologies. The proposed analyses will use cutting-edge exposure science methods to quantify relationships between environmental exposures and brain health across multiple time horizons, shedding light on which environmental factors influence cognitive function, long-term declines in cognition, dementia incidence, and ADRD neuropathologic burden. These results can be used to identify intervention targets in mitigation and adaptation policies to optimize brain health.

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT: Myocardial infarction (MI) is a leading risk factor for ventricular arrhythmias. Due to significant MI- induced remodeling within the sympathetic nervous system and cardiac adrenergic signaling cascades, blocking cardiac b-adrenergic receptors (b-ARs) and downstream cyclic adenosine monophosphate (cAMP) signaling remains one of the most effective anti-arrhythmic strategies post-MI. In cardiomyocytes, cAMP is tightly controlled within a complex at the b-AR that includes phosphodiesterase 4D (PDE4D, cAMP degrading enzyme), and this complex may be impacted by sympathetic innervation. During the previous funding period, we developed a novel multi-parametric whole-heart imaging system and two transgenic mice expressing biosensors for cAMP and norepinephrine (NE) to assess cAMP and NE signaling in healthy and post-MI hearts. Using this approach, we found that cellular cAMP decay is significantly slowed in the post-MI heart following sympathetic stimulation, potentially implicating reduced PDE activity as an important – and previously unrecognized – contributor to arrhythmias. Preliminary data with the NE biosensor further suggest that reduced NE reuptake at the neuro- cardiac junction may also play an important role in slowing cAMP break-down. Finally, we found that restoring sympathetic innervation pharmacologically restored normal cAMP break-down and prevented arrhythmias during sympathetic stimulation. Therefore, this renewal application aims to test the hypotheses that: 1) the regulation and termination of cAMP signaling in the normal heart is governed by clearance of catecholamines from the synaptic cleft by the NE transporter (NET) and by PDE4D activity in the b-AR complex; 2) regional hypo-innervation post-MI leads to reduced NE reuptake and regional disruption of b-AR-cAMP signaling to promote arrhythmias; and 3) restoring sympathetic innervation to the infarct restores cAMP break-down to prevent arrhythmias. We will combine our multi-parametric whole-heart imaging system with cellular and biochemical analyses and functional optical mapping of transmembrane potential (Vm) and intracellular Ca2+. Aim 1 will characterize regional and sex differences in NE reuptake and PDE activity in modulating cAMP signaling and functional responses in the normal heart. In Aim 2, we will determine the role of NE reuptake vs. PDE activity in the termination of cAMP signaling in the post-MI heart and how this contributes to ventricular arrhythmias. Aim 3 will determine the mechanisms by which pharmacological sympathetic re-innervation post-MI accelerates and restores cAMP break-down to prevent arrhythmias. These studies will provide unprecedented insight into how changes to cAMP break-down contribute to arrhythmias from cellular signaling up to macro-scale multi-organ interactions. We will also determine detailed mechanisms of action of novel neuromodulatory drugs that may have therapeutic potential beyond the post-MI environment.

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT Dup15q syndrome is a neurodevelopmental disorder marked by severe intellectual disability, autism spectrum disorder (ASD), epilepsy, and motor delays, resulting from duplications of the long arm (q) of maternal chromosome 15. It affects approximately 1 in 5,000 individuals and is 10 times more common in people with ASD or intellectual disability. More than half of patients with Dup15q experience epilepsy, which increases the risk of sudden unexpected death in epilepsy (SUDEP). The primary genetic cause is maternal duplications or triplications of the 15q11.2-q13 region, leading to overexpression of the UBE3A gene. The high medical costs associated with caring for these chronically ill children place a significant burden on families, particularly those with limited resources. Currently, no curative treatments exist, making the development of Cas9 mRNA therapy delivered via lipid nanoparticles (LNPs) a promising approach. This project aims to develop a non-viral gene- editing therapy using LNP/mRNA complexes to correct UBE3A overexpression in neural stem progenitor cells (NSPCs) during fetal brain development. By intervening early, this therapy has the potential to prevent or alleviate neurodevelopmental deficits, seizures, and cognitive impairments characteristic of Dup15q syndrome. Delivering Cas9 mRNA via LNPs offers a less immunogenic, non-viral alternative to traditional viral-based gene therapy approaches. Early removal of the duplicated 15q region may prevent or rescue Dup15q phenotypes. Administering this therapy in utero creates an opportunity to correct the genetic defect before symptom onset, potentially preventing neurodevelopmental damage. This approach could revolutionize treatment for genetic disorders by addressing the root cause during critical prenatal development. In utero gene editing offers unique advantages: 1). Stem and progenitor cells in the fetal brain are abundant and actively proliferating, which enhances gene-editing efficiency and functional outcomes. 2). The fetal immune system is more tolerant, reducing the risk of immune rejection of foreign gene-editing enzymes. 3). The smaller fetal size allows for optimal dosing, improving cost efficiency, a critical factor given the financial constraints of gene editing. 4). In utero treatment is the earliest possible intervention and is unlikely to cause germline editing after the seventh week of gestation. Early treatment of prenatally diagnosable conditions like Dup15q syndrome could significantly alleviate symptoms before disease onset. Achieving the goals of this project will advance this regenerative therapy toward clinical use, providing both healthcare and financial benefits. The development of non-viral, in utero gene editing has the potential to functionally treat Dup15q syndrome and reduce the need for intensive medical care. While this project focuses on Dup15q syndrome, the platform technology could be applied to other genetic disorders, making it valuable for addressing a range of conditions affecting diverse populations.

NIH Research Projects · FY 2025 · 2025-08

Project summary Opportunistic Clostridioides difficile (previously Clostridium difficile) infection is a significant public health threat, primarily affecting hospitalized patients and leading to severe, life-threatening diarrhea. The CDC classifies C. difficile infection as an urgent health threat due to its high incidence, morbidity, and mortality rates. The infection typically follows antibiotic therapy (i.e., treatment with 3rd generation cephalosporins, fluoroquinolones, and clindamycin), which disrupts colonization resistance mediated by the normal colonic microbiota, allowing C. difficile to colonize the gastrointestinal tract and begin producing toxins to trigger antibiotic-associated colitis. Our long-term research goal is to understand how colonization resistance can be strengthened to prevent C. difficile infection. Our central hypothesis is that restoring epithelial hypoxia after antibiotic treatment can accelerate microbiota recovery to restore colonization resistance against C. difficile. We will test our hypothesis using the following specific aims: (i) determine whether treatment with a second-generation probiotic, Anaerostipes caccae, can accelerate microbiota recovery after antibiotic treatment to restore colonization resistance against C. difficile, and (ii) determine whether pharmacological activation of epithelial peroxisome proliferator–activated receptor gamma signaling can accelerate microbiota recovery to restore colonization resistance against C. difficile. The proposed experiments provide innovation by establishing a proof of concept for a completely new and original starting point for preventing C. difficile infection. This outcome will be significant by providing mechanistic insights into colonization resistance against an important opportunistic pathogen, which will be of wide appeal among researchers interested in antimicrobial-resistant infections, microbiota research, and intestinal biology.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT Dysbiosis in the female reproductive tract (FRT) occurs when unknown ecological factors in the vaginal microenvironment drive the expansion of suboptimal bacterial communities that contribute to disease. Our understanding of the host and microbial-derived factors that contribute to FRT dysbiosis is limited, and as such we lack effective strategies for restoring vaginal homeostasis in at-risk women. Aerobic vaginitis occurs when enteric opportunists such as Escherichia coli subvert the dominance of glycogen fermenting commensals to expand in the FRT, causing severe inflammation and tissue atrophy. The host-derived factors that drive the bloom of facultative anaerobes in the FRT during aerobic vaginitis are unclear, but risk factors include hormonal changes during pregnancy, menopause, etc. We and others have observed that mice in specific reproductive phases (estrus and metestrus) are highly susceptible to vaginal E. coli colonization suggesting hormonal-driven changes in the FRT may influence pathogen susceptibility in mice. Our preliminary findings show that estrogen alters vaginal epithelial cell metabolism, reduces epithelial hypoxia, and drives a respiration-dependent bloom of E. coli that induces subsequent inflammation in the FRT. From these findings, I hypothesize that hormone-induced metabolic reprogramming of the host epithelium towards aerobic glycolysis increases luminal oxygenation of the vaginal tract, allowing E. coli to outcompete fermenting anaerobes and induce an inflammatory response that perpetuates chronic aerobic vaginitis. Here, I propose to use genetically tractable uropathogenic E. coli (UPEC) in combination with specialized animal and tissue culture models to identify the specific host and bacterial metabolic pathways that contribute to aerobic vaginitis. In AIM1, we will determine how host cell metabolism controls vaginal community composition during homeostasis. In AIM2, we will use UPEC to identify host and microbial metabolic pathways that facilitate the bloom of facultative anaerobes during vaginal dysbiosis. In AIM3, we will explore how UPEC ecosystem engineering facilitates persistent colonization of the female reproductive tract during aerobic vaginitis. Together, these studies will provide much-needed insight into the factors that influence vaginal health and disease and reveal new therapeutic targets for supporting women’s reproductive health. The support of this K99/R00 Pathway to Independence award will facilitate the training necessary to achieve these aims, including training in advanced microbiome techniques, specialized conventional and gnotobiotic mouse models, and further career development. With the combined support of an outstanding mentorship team and the world-class technical and intellectual resources available at UC Davis, the proposed scientific aims and training objectives will form the foundation for an independent research program aimed at uncovering the ecological drivers of dysbiosis in the FRT.

NIH Research Projects · FY 2026 · 2025-08

Project Summary The goal of the Mentoring and Skills Development Training Program in One Environmental Health Toxicology (MSDT-OEHT) dovetails with NIGMS R25 IPERT objectives to enhance workforce training in an area of unmet biomedical need. Environmental exposures have a negative impact on human and animal health and One Environmental Health Toxicology (OEHT) is an emerging field of converging biomedical research at the interface of Toxicology and One Health (OH). The premise of OH is that common mechanisms can impact the health of humans and animals. OH research is dominated by infectious disease experts working on zoonotic and emerging pathogens but toxicology is an underrepresented discipline in OH research. There is a need to bridge clinical skills with toxicology knowledge and practice especially in relation to impacts of environmental exposures on affected communities. A critical gap in traditional toxicology training programs is a lack a focus on OH, producing toxicologists who work in silos on narrow topics. An unmet need is toxicologists and clinicians equipped with knowledge and research skill sets suitable for transdisciplinary multisectoral transformative OEHT research in human and animal populations. Yet, the environment has a major impact on both human and health and disease. The long-term objective of MSDT-OEHT is to produce a cadre of biomedical and clinical scientists trained and engaged in OEHT research. MSDT-OEHT is a yr long national mentoring and skills development training program targeting 2 critical career stages 1) STEM undergraduates and 2) early career researchers. At these stages decisions about career path are made, and often OEHT is not considered. The MSDT-OEHT program is delivered by a consortium of institutions contributing to national prominence. The program will achieve its goals through a unique curriculum and by bringing together biomedical scientists from different disciplines and career trajectories. Funding through this program will be used to generate e-modules and case studies on the fundamentals of OEHT to be delivered through an already operable web based system. We will also include a yr long mentoring program for 100 STEM undergraduate students interested in OEHT during the life of the grant, and to develop and deliver a professional skills development and mentoring program for early career scientists with interest in OEHT. The goal for the early career researchers is to increase their competitiveness for funding and skills in OEHT, including community engaged research. The program is nationwide and open to all, but participants from marginalized and vulnerable communities, most impacted by environmental toxicants, will be prioritized. MSDT-OEHT content will be freely accessible globally to anyone with a handheld device or computer connected to the internet, expanding the impact of the program. The goal is to increase the number of biomedical research scientists with toxicology knowledge to advance transformative transdisciplinary research on the impact of the environment on human and animal health.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY The UC Davis (UCD) Stimulating Access to Research in Residency (StARR) Training Program in human and veterinary medicine will recruit outstanding residents with diverse backgrounds who have demonstrated keen interest and potential in pursuing a career as clinician-scientists. The program will include two postdoctoral Internal Medicine MD/DO residents and two postdoctoral DVM residents and will offer a formal Master’s in Clinical Research Training Program (MCRTP) supported by matching funds from the deans of the UCD School of Medicine and School of Veterinary Medicine. This is a special arrangement as there are currently no other StARR programs that cross-train medical and veterinary residents. UCD attracts an outstanding resident pool from both disciplines as potential candidates for the StARR program. Aim 1 will select the most suitable trainees through an extensive recruitment and competitive application process drawing applicants from both professional schools while promoting diversity and inclusion in research training in an intentional manner. Personalized research mentoring committees will be established to help achieve program goals, evaluate progress, and track scholar’s Individual Development Plans. We will train ten scholars during the first grant cycle. Aim 2 will perform a rigorous training that will propel trainees to successful research-intensive fellowships and subsequent independence as clinician-scientists. Residents will complete a mentored research project in a specific area supported by their mentoring committees with emphasis on the concepts of comparative medicine and One Health, basic and translational cardiopulmonary research methods, research communication, grant and manuscript writing, rigor and reproducibility in science, and team science. Faculty preceptors will direct research training in three primary areas: 1) Cardiopulmonary biology and molecular medicine, 2) Emerging infectious diseases and host responses, and 3) Data Sciences and Engineering in Translational Research. Aim 3 will enhance training and individual career development opportunities through advanced training in a Clinical Research Certificate program or an optional Master of Advanced Studies in Clinical Research program already attended by SOM and SVM junior faculty. Graduated StARR alumni will continue their engagement with the program as mentors to promote a sustained culture of resident research on their path to becoming clinician- scientists and academic leaders. Residents will be evaluated by competency-based milestones. Program metrics of success will include a) Scholarly productivity (publications, presentations), b) Diversity of trainees, c) Acceptance of residents into research-intensive fellowships or academic faculty positions, and d) Future T32, K and R grants. The existing highly collaborative environment between the Schools of Medicine and Veterinary Medicine will provide the UCD StARR resident investigators with a creative and lively training and scientific milieu to help them become members of high-level translational science teams and successful clinician-scientist.

NIH Research Projects · FY 2025 · 2025-08

Project Abstract The Translating Engineering Advances to Medicine (TEAM) Lab, a core facility within the Biomedical Engineering Department at the University of California, Davis, seeks support to acquire the Stratasys J5 Digital Anatomy 3D Printer to modernize our shared-use biomedical research facility and overcome current limitations in the university's 3D printing technologies. The J5 Digital Anatomy Printer offers unique capabilities, including the ability to incorporate multiple materials into a single print to replicate the biomechanical properties and radiopacity of human tissue. Its advanced software comes pre-programmed with clinically validated presets that mimic a range of human tissues, while also allowing precise control over material composition and microstructures to replicate various tissue pathologies. By enhancing our ability to produce realistic anatomical models, the J5 printer will significantly elevate our research capabilities across multiple disciplines. It will facilitate groundbreaking studies in medical device development, surgical planning for rare and difficult-to-treat conditions, optimization of radiation dose delivery treatments, and more. Moreover, the ability to 3D print identical replicas of pathologies allows for more precise experimental testing and controls, reducing reliance on cadaver and animal models, which are costly, difficult to source, and generate biological waste. Acquiring the Stratasys J5 will enable the TEAM Lab to support new research initiatives currently constrained by technological limitations. By providing advanced simulations and realistic models, the printer will drive the development of new treatments for challenging disease states, leading to better patient care and outcomes. This enhancement aligns with UC Davis's mission to advance public health through pioneering research and innovation, ultimately fostering improved healthcare solutions and societal well-being. We plan to integrate the equipment into the TEAM Lab expansion, located at the UC Davis Sacramento Campus, is fully prepared for the new printer without requiring any site modifications. This expansion demonstrates the department’s and university's continued support for the TEAM Lab, operational since 2010 under the proposed PI’s management.

NIH Research Projects · FY 2025 · 2025-08

Project Summary: Trauma-induced Heterotopic Ossification (tHO) is an extremely painful condition, caused by aberrant bone formation in severely injured soft tissues, with no effectual prophylaxis or treatment. The recent observation that tHO initiation is linked to nociception induced neuroinflammation (NINI), offers a potential therapeutic target. This proposal presents a four-year research career development plan focused on modulating NINI to mitigate tHO formation. The candidate is a board-certified Plastic Surgeon with fellowship training in upper extremity and peripheral nerve surgery, who decided to pivot to a research-oriented academic tract, to study this devastating pathology, given the high prevalence of tHO in high-impact injuries in the candidate’s practice at a Level I trauma center. This proposal builds on his clinical expertise, prior research on osteogenesis, and merges them with two new domains of expertise represented by his mentors Dr. Sean Adams (Vice Chair of Research, UC Davis) – inflammation; and Dr. Benjamin Levi (Chair of Burn Surgery, UTSW) – tHO. The proposed work, didactics, mentorship, and unencumbered research time, will raise the level of the candidate’s science, establish credibility in the tHO research space, and ensure his successful transition to an impactful researcher. Trauma-induced nociception results in sensory nerve over-expression of neuroinflammatory peptide Calcitonin Gene-Related Peptide (CGRP), which activates the Bone Morphogenic Protein (BMP) pathway, causing osteogenic differentiation of resident progenitor cells. Targeted Muscle Reinnervation (TMR) is a peripheral nerve transfer procedure that connects amputated nerve stumps to motor branches in the residual limb of amputees, and significantly reduces pain and neuroinflammation. It is only indicated in cases with intractable pain or phantom pain. Fremanezumab is a CGRP inhibitor currently used in migraine but has not been considered for tHO. The central hypothesis of this proposal is that reducing NINI surgically (TMR) or medically (Fremanezumab) will diminish sensory nerve CGRP expression and ultimately tHO formation. This will be investigated using an in vivo rodent model to test the efficacy of TMR; and a neuroinflammatory in vitro model to test the efficacy of Fremanezumab. Utilizing existing surgical and medical modalities to inhibit NINI in tHO is innovative not only because of their translational potential, but also because of the novel mechanistic insight they provide into tHO initiation and the role of sensory nerve-stem cell communication in it.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Colorectal cancer (CRC) is the second leading cause of cancer-related mortality globally, with current screening techniques, such as colonoscopy, demonstrating limitations in accurately detecting subtle structural abnormalities and biochemical changes in colorectal lesions. These limitations contribute to missed diagnoses and incomplete resections, which account for over 85% of post-colonoscopy CRC cases. This project aims to address these challenges by integrating label-free fluorescence lifetime imaging (FLIm) into standard colonoscopy procedures to enhance the detection and characterization of colorectal lesions in real-time. FLIm offers a novel approach to providing detailed biochemical information by capturing autofluorescence signals that reflect changes in tissue composition, metabolism, and structure. We hypothesize that FLIm can capture biochemical changes associated with inflammation and carcinogenesis, facilitating better staging of colorectal lesions. The proposed research is based on three specific aims and will first build a comprehensive database of in vivo FLIm signatures from colorectal lesions in 200 patients undergoing routine colonoscopy, correlating these signatures with histopathological findings to establish robust markers for lesion detection and classification (Aim 1). We will then develop advanced, user-friendly algorithms to enable rapid, real-time analysis and visualization of FLIm data during colonoscopy, making the system practical for clinical use (Aim 2). Finally, we will evaluate FLIm's sensitivity and specificity in identifying and staging a range of colorectal lesions, from benign polyps to malignant tumors, using machine learning techniques (Aim 3). By integrating FLIm into colonoscopy, this project seeks to overcome the current limitations of endoscopic imaging, providing clinicians with real-time biochemical insights that improve lesion detection, diagnosis, and decision-making. The expected outcomes include a fully developed FLIm-colonoscopy system that can significantly enhance CRC screening accuracy and early-stage diagnosis. The long-term impact of this work will be the reduction of CRC incidence and mortality through improved screening technologies and better clinical management of colorectal diseases.

NIH Research Projects · FY 2025 · 2025-08

Project Summary/Abstract Many survivors of premature birth and perinatal brain injury suffer from long-term neurological sequelae including seizures, motor and cognitive deficits, elevated rates of hyperactivity, and autism. These newborns urgently need early effective and safe interventions for neuroprotection, a medical goal that is currently unmet. Oxygenated lipid mediators (oxylipins) derived from polyunsaturated fatty acids (PUFAs) are potent signaling molecules that regulate a multitude of cellular and systemic responses, including inflammation. Epoxy fatty acids (EpFAs) are cytochrome P450 (CYP)-dependent derivatives of PUFAs. They are a group of lipid mediators with potent anti-inflammatory and pro-resolving properties. However, their activities are extremely short-lived as soluble epoxide hydrolase (sEH) quickly converts EpFAs to pro-inflammatory diols. Our therapeutic hypothesis is that inhibition of sEH will increase and prolong the anti-inflammatory and pro- resolving actions of chemically stable EpFAs to exert neuroprotection. This proposal, built upon our preliminary data, will examine this therapeutic hypothesis in perinatal brain injury (PNBI). We employed a well-established mouse model of neonatal lipopolysaccharides-sensitized hypoxia-ischemic injury (LPS-HI) to replicate a major form of PNBI in which perinatal infection/inflammation sensitizes the brain to subsequent HI insult and augments brain injury. In Aim 1, we will use both genetic knockout of sEH and pharmacological inhibition of sEH by an inhibitor called TPPU to determine if sEH inhibition protects neonatal mice from LPS-HI injury. Because our preliminary data suggests that liver (hepatocyte)-targeted knockout of sEH may also protect mice from lPS-HI, we propose a sEH-regulated liver-brain mechanism in PNBI via which the liver protects the brain in a long-range fashion via regulating the lipidomic structure of the blood transporting EpFAs. To examine this hypothesis, we will conduct lipidomic profiling of liver, plasma, and brain samples to construct a basic framework for the liver → blood → brain sEH-regulated mechanism. Together, the two aims will provide proof- of-concept that sEH is a novel therapeutic target for PNBI and transformative insights into how liver may play a critical role in regulating or maintaining brain integrity during perinatal period. Completion of this project will provide a potential new avenue for managing perinatal brain injury.

NIH Research Projects · FY 2025 · 2025-08

Abstract . Intravascular pressure is a key stimulus controlling arterial tone and blood flow delivery. Pressure-induced constriction (i.e. myogenic response) is recognized as foundational to vascular hemodynamic control in health and disease. A unique feature of this response is its near complete dependence on vascular smooth muscle (VSM) L-type CaV1.2 channels. While the current dogma assumes that membrane depolarization is the sole factor driving CaV1.2 activation in the myogenic response, pressure itself, independent of electrical coupling, may also directly influence VSM CaV1.2 function. Yet, whether pressure modulates CaV1.2, the underlying mechanisms, and potential (patho)physiological impact are key knowledge gaps that this application aims to address. By utilizing a sophisticated toolkit, exciting and rigorous preliminary data support the central hypothesis that pressure dynamically regulates CaV1.2 spatiotemporal properties via the engagement of PKCα and pS1928 to control VSM contractility and vascular reactivity in health and disease. This hypothesis will be tested in three specific aims. Aim 1 is to test the hypothesis that pressure regulates CaV1.2 spatiotemporal properties in VSM. Aim 2 is to test the hypothesis that pressure regulation of CaV1.2 requires kinase activity and pS1928. Finally, Aim 3 is to test the hypothesis that pressure regulation of CaV1.2 spatiotemporal properties is altered in disease states. The proposal has high translational impact as it will advance understanding of VSM Ca2+ dynamics and its role in setting the myogenic response and nutritive blood flow in health and disease, and it may open new avenues for therapeutic manipulation of CaV1.2 in disease states.

NIH Research Projects · FY 2025 · 2025-08

PROJECT ABSTRACT This proposal will lay the foundation for developing a molecular therapy for ADNP Syndrome. Also known as Helsmoortel-Van Der Aa Syndrome, ADNP Syndrome is a neurodevelopmental disorder characterized by severe intellectual disability, a myriad of motor dysfunctions, autism spectrum disorder, impaired sensory processing, seizures and sleep disturbances. ADNP Syndrome has an incidence of ~1:10,000 live births but accounts for ~0.2% of autism diagnoses, labeling ADNP as one of the highest causal risk genes for autism spectrum disorder. ADNP Syndrome is caused by mutations in Activity Dependent Neuroprotective Protein (ADNP), a chromatin modifier that affects over 400 genes during embryonic development and thousands postnatally. The mutations occur de novo, many of which are heterozygous haploinsufficient loss-of-function (LoF) mutations. This suggests that one approach to therapy could be to rescue the haploinsufficiency by increasing the amount of ADNP protein. Through private pilot funding, we have created antisense oligonucleotides (ASOs) that are able to increase ADNP protein expression in iPSC-derived human neurons. The presumed mechanism of action, blocking unproductive upstream open reading frames (uORFs), will be tested in Aim 1 of this proposal. Also, because mice do not have the same uORF structure as in human, the ASO cannot be tested in a mouse model. We have therefore designed a fully humanized ADNP mutant mouse model. An initial characterization of the molecular features of this model will be investigated in Aim 2 of this proposal to demonstrate its utility for ADNP therapeutic development. The results of the proposed study should provide foundational preliminary data to support more substantial subsequent studies to create the first ever ASO therapy for ADNP syndrome. The results from this study will further advance the translational potential for precision therapeutics for children suffering from ADNP and other genetic haploinsufficient disorders.

NIH Research Projects · FY 2025 · 2025-08

Abstract We have developed a transformable nanoparticle (TNP) platform capable of transforming its morphology from nanoparticles to nanofibrils when interact with receptors (e.g., EGFR or α3β1 integrin) over-expressed on the cell surface of many tumor types, including non-small cell lung cancer (NSCLC). TNPs displaying peptide ligands against these receptors, will transform into nanofibrillar networks surrounding the tumor cells, in vitro or in vivo. Through the above work, we found that the nanofibrillar network generated was able to retain at the tumor microenvironment (TME) for a week, whereas nanostructures retained at normal organs such as liver and lung disappeared after 2 days. Here, we want to exploit this unique tumor retention property of TNP by utilizing the highly specific bioorthogonal chemistry to introduce, on- demand, desired therapeutic payloads, to modulate the tumor microenvironment (TME), while sparing normal tissues. This novel two-component two-step (TCTS) nanotherapeutic strategy is based on the use of (i) TNP for pre-targeting, and (ii) the rapid and highly specific bioorthogonal click reaction for on- demand targeted delivery of peptides, small molecules and proteins that can modulate the TME. Our hypothesis is that this highly versatile TNP/TCTS therapeutic platform will allow effective on- demand sequential or combination delivery to the TME orthogonal immunomodulatory molecules such as immune cell capturing ligands (e.g., LLP2A), small molecule immunostimulants (e.g., resiquimod), and sialidase (to shave the immunoinhibitory oligo-siliac acids off the tumor cell membrane). As a consequence, a robust anti-tumor immune response will be elicited, resulting in durable regression of the cancer. The 3 specific aims of this project are: Aim 1. To design, synthesize and characterize EGFR and α3β1 integrin targeting transformable nanoparticle (TNP) constructs. Aim 2. To use in vivo and ex vivo optical imaging to optimize TNP/TCTS platform for cancer immunotherapy. Aim 3. To determine the in vivo toxicity and efficacy of the novel TNP/TCTS immunotherapeutic approach using syngeneic murine Lewis lung cancer model.

NIH Research Projects · FY 2025 · 2025-08

The importance of differences in islet architecture between rodent and human islets is much debated and remains understudied, highlighting a need to further understand the relationship between islet architecture and function. A large-scale comparative approach to characterize species differences at the morphological and functional levels can provide important context to variations observed between rodent and human islet physiology. Although evolutionary changes may contribute to broader differences in islet architecture across distantly related species, they do not explain differences in pancreas architecture between closely related species. These variations may instead be attributed to differences in diet and metabolism. This is supported by evidence that even individual increases in metabolic demand are associated with architectural changes within the islet. This has informed the hypothesis that species-specific variations in metabolism and dietary composition influence pancreas architecture and function in addition to phylogenetic relatedness. Reports of islet architecture have been described for some species, but these data are based on the assessment of a limited number of islets, often from a single animal for a relatively limited number of species. This gap will be addressed by utilizing a systematic, high-throughput comparative approach to quantify pancreas architecture differences across species and correlating these changes to functional differences at the cellular level in response to variations in macronutrient stimulation. The study of species with alterations in their reliance of various macronutrients, namely glucose, provides a unique opportunity to explore the role of α and β cells as nutrient sensors of the islet. Aim 1 will systematically quantify variations in morphology and paracrine factors using high-throughput image analysis across carefully selected samples from unique collection of 11,000 pancreas samples representing all major vertebrate taxa banked at the UC Davis School of Veterinary Medicine. Aim 2 will compare rodent and human Ca2+ and cAMP dynamics to those of live pancreatic slices from carnivores and ruminants, whose unique dietary and metabolic adaptations favor a constant gluconeogenic state. These approaches are innovative as they employ a systematic, quantitative approach to understand islet architecture and function through a comparative lens. These proposed aims are significant as they will provide an understanding of how variations in islet architecture correlate with the macronutrient sensitivity of α and β cells within the islet. This foundational knowledge may prove essential to understanding the consequences of islet architectural defects on dysregulated hormone secretion in human diabetes.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Homologous recombination is a critical pathway for repair of DNA lesions, including double-stranded breaks, ssDNA gaps, and stalled or collapsed replication forks. Of particular importance is the selection of the duplex template for repair. Recombination between non-allelic repeats instead of the sister chromatid can lead to genome rearrangements and loss of heterozygosity, both of which have demonstrated significance to an array of human diseases, particularly cancer. The mismatch repair proteins MSH2 and MSH6 form a heterodimer that detects base-base mismatches and small insertions/deletions. Not only does it function in postreplicative mismatch repair, MSH2-MSH6 has also evolved to prevent recombination between mismatched sequences. Despite the importance of this activity to genome stability, the mechanisms involved remain poorly characterized. In this proposal, I will use a combination of genetic and genomic approaches to delineate the mechanism(s) through which MSH2-MSH6 rejects mismatched recombination intermediates and the define the clinical significance of loss of this important activity in MSH2/6-deficient patient tumors. In Aim 1A (K99 phase), I will use novel proximity ligation-based assays in the model organism Saccharomyces cerevisiae in combination with genetic analysis to delineate the mechanism of MSH2-MSH6 rejection of mismatched recombination intermediates. I will examine recombination between an inducible double-stranded break and each of two templates, one that is perfectly matched, and one that contains ~5% mismatches. I will determine how MSH2-MSH6 biases recombination towards the perfectly matched template and identify the effectors that it recruits to achieve this outcome. In Aim 1B (R00 phase), I will transfer the proximity ligation assays from yeast to human cells and characterize MSH2-MSH6-dependent regulation of mismatched recombination intermediates in this system. In Aim 2, I will use two different approaches to systematically characterize the mutational signatures associated with loss of regulation of recombination by MSH2-MSH6 in MSH2/6-deficient patient tumors as compared to matched healthy tissue controls. In Aim 2A (K99 phase), I will use unbiased long-read whole-genome sequencing and a genome-wide bioinformatic approach to identify de novo structural variants in the patient tumor samples. In Aim 2B (R00 phase), I will use targeted sequencing of long reads containing human lineage-specific LINE-1 elements, which are expected to be disproportionately involved in recombination-mediated rearrangements as a result of their length and the high sequence identity of elements within relatively young LINE-1 subfamilies to one another. This work will provide important insight into the processes that prevent recombination between ectopic repeats and will serve as the foundation of my research program as I transition to an independent faculty career.

NIH Research Projects · FY 2025 · 2025-08

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

NIH Research Projects · FY 2025 · 2025-08

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

NIH Research Projects · FY 2025 · 2025-08

The impact of targeted radiopharmaceuticals on oncology spans from diagnosis to therapy. Targeted diagnostic radiopharmaceuticals are molecular imaging agents labeled with a diagnostic radioisotope, allowing non-invasive characterization and assessment of the extent of disease, patient stratification, and monitoring response to treatment through tomographic imaging. Radiotherapeutics are targeted agents, labeled with a therapeutic radioisotope, that enable tumor-specific targeted radionuclide therapy (endoradiotherapy). Diagnostic and therapeutic radiopharmaceuticals based on the same molecule allow for a combined targeted theranostic approach to imaging, therapy, and response monitoring. That is we treat what we see and we see what we treat. Challenges include the complex design, synthesis and optimization of the radiopharmaceutical, and the establishment of robust syntheses for deployment to the clinic. There is a clear need for exceptional laboratory-based scientists with extensive research experience encompassing basic compound identification and optimization; preclinical evaluation; and clinical translation. I have dedicated my scientific post-graduate career for over 20 years in Dr. Sutcliffe’s laboratory (my Unit director) to the initial design-stages of agents targeting cancer-associated cell-surface receptors for positron emission tomography (PET) imaging and endoradiotherapy, including extensive chemistry and radiochemistry research; wide-ranging pre-clinical in vivo testing; through to the development of clinical grade compounds including the cGMP-synthesis of a range of radiopharmaceuticals currently under evaluation in several clinical trials under the leadership of Dr. Sutcliffe. My work was and remains instrumental for the success of Dr. Sutcliffe’s NCI and other grants, as highlighted by the 23 research publications jointly authored with her, our 2023 Society of Nuclear Medicine and Molecular Imaging Image of the Year Award - chosen from over 1500 submitted abstracts - because it “best exemplifies the most promising advances in the field of nuclear medicine and molecular imaging”, and my development and preclinical work directly leading to the 3 novel radiopharmaceuticals used in the 6 clinical imaging and therapy trials led by Dr. Sutcliffe. In this application, in close collaboration with Dr. Sutcliffe, my goals are to 1) further develop radiotheranostics with improved pharmacokinetics, 2) develop, refine and simplify radiolabeling protocols for manufacturing scale-up and wide-spread application, 3) collaboratively with other Sutcliffe lab members advance and support work on targeted theranostic peptide-conjugates to investigate their ability for cancer- specific therapy, and 4) continue to support of the current diagnostic/theranostic clinical trials targeting the cancer-associated integrin αvβ6. This award is an excellent opportunity for me to contribute to basic, translational, and clinical science. I am highly motivated by our encouraging results obtained to date and look forward to more innovative research contributions, and to improving the lives of patients with cancer.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Adeno-associated viruses (AAVs) are widely employed for retinal gene therapies, but conventional routes of administration have several challenges. Subretinal AAV delivery enables only focal transduction and risks surgical complications from vitrectomy. Intravitreal AAV injections are easier to perform in the office, but do not efficiently transduce photoreceptors and induce greater intraocular inflammation. We recently demonstrated the successful use of transscleral microneedles to deliver AAV8 vectors into the suprachroidal space of nonhuman primates (NHPs). Suprachoroidal AAV injections are more easily performed in the office and less invasive than subretinal injections, and enable broad retinal transduction that cannot be achieved with intravitreal delivery. Although suprachoroidal gene therapy has begun entering human clinical trials, our recent studies showed that suprachoroidal AAV8 delivery has (1) restricted biodistribution to the peripheral retina, (2) limited cellular tropism for mostly retinal pigment epithelium (RPE), and (3) short durability of transgene expression that may be attributed to host immunity to viral transduction outside the blood-retinal barrier. We hypothesize that suprachoroidal AAV delivery can overcome the limitations of conventional subretinal or intravitreal AAV administration, if biodistribution, cellular tropism, durability, and safety can be properly optimized. In this study, we will evaluate the use of microcatheterization, as well as the impact of increasing injection volume or viscosity, to enhance posterior biodistribution of suprachoroidal AAV delivery to the macula. Next, we will screen a barcoded library of naturally-occurring and engineered AAV capsids designed to penetrate the blood-brain barrier in NHPs, with the goal of identifying and validating AAVs that can cross the blood-retinal barrier from the suprachoroidal space to transduce photoreceptors. Finally, we will characterize host local and systemic immune responses to capsids and transgenes after suprachoroidal AAV delivery by measuring aqueous and plasma cytokines, neutralizing antibodies (NAbs), leukocyte responses, and cellular-level transcriptional profiling, with and without systemic prophylaxis with corticosteroids or the sphingosine 1-phosphate receptor modulator fingolimod. Together, our studies will address critical barriers to optimizing suprachoroidal injections as a novel route of AAV administration for effective, safe, and durable gene therapy strategies to treat patients with retinal diseases.

NIH Research Projects · FY 2025 · 2025-08

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

NIH Research Projects · FY 2025 · 2025-08

Organophosphate cholinesterase inhibitors (OPs) were developed as agricultural insecticides and adapted to function as chemical weapons. OPs inhibit the acetylcholinesterase enzyme and acute intoxication can lead to a cholinergic crisis, status epilepticus (SE) and, for individuals who survive, the development of spontaneous recurrent seizures (SRS) and cognitive deficits. The current CounterACT PAR-24-030, supports research related to the 1) evaluation of the natural history to 2) elucidate mechanisms related to long-term effects of acute- intoxication, and in particular the development of SRS and cognitive dysfunction, that will 3) identify relevant biomarkers to predict long-term outcomes and 4) identify new targets for therapeutic intervention. To date, little is known regarding the acute- or long-term effects of acute-OP intoxication on sleep architecture. In the field of epilepsy in general, the link between seizures and sleep disruption is complex, but well documented. Epilepsy and vigilance stages show a bi-directional relationship, with epilepsy disrupting sleep, and sleep disruption promoting seizures. Vigilance stages and seizures are each characterized by signature cortical electrical activity. Moreover, it is suggested that sleep-wake deficits are, in part, responsible for cognitive deficits observed in patients with epilepsy disorders. In fact, sleep enhancement is being pursued as an interventional strategy to reduce the burden of the epileptic syndrome. We hypothesize that sleep disruptions in the days-to-months following acute intoxication with the OP diisopropylfluourophosphate (DFP) play a key role in the development and maintenance of lasting pathophysiology including sleep disorders, seizures and cognitive dysfunction. We further hypothesize that restoring a normal sleep-wake cycle will be protective, preventing the development of the worst outcomes when provided acutely, and, in the absence of early intervention, counteracting symptoms when delivered chronically. In testing our hypotheses, we will necessarily assess each of the four topic areas related to the CounterACT RFA as described above. Specifically, we will determine whether acute intoxication with DFP leads to dynamic changes in sleep architecture in the days-to-months post-intoxication (natural history; goal 1) and predict that the animals with greater sleep disturbance experiencing prolonged cognitive and seizure disorders (sleep as a mechanism; goal 2). We further hypothesize that changes in sleep-wake phenotypes observed in the initial hours-to-days following acute intoxication will predict the extent of cognitive and seizure outcomes (sleep as a biomarker; goal 3). Finally, we propose that early intervention with the FDA-approved dual- orexin receptor antagonist (DORA) Lemborexant will improve sleep phenotypes and protect against the development of the worst seizure and cognitive disorders while delayed intervention will reduce seizure frequency and duration and improve cognitive outcomes (sleep as a new target for intervention; goal 4). This collaborative proposal, therefore, fills a critical unmet need in the field of countermeasures research, while evaluating a potential therapeutic target and FDA-approved candidate therapy.

NIH Research Projects · FY 2025 · 2025-08

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