University Of Colorado Denver

NIH Research Projects · FY 2026 · 2025-01

Inhibiting Free Fatty Acid Transport to Improve CAR-T Cell Therapy of Relapsed B-cell Acute Lymphoblastic Leukemia Abstract Chemotherapy-resistant B-cell acute lymphoblastic leukemia (B-ALL) remains a leading cause of cancer-related death for children and young adults. While CD19-directed chimeric antigen receptor T cell (CAR-T) therapy offers promise for relapsed/refractory cases, high long-term failure rates necessitate a deeper understanding of resistance mechanisms. Our preliminary studies highlight free fatty acid (FFA) uptake inhibition as a potential metabolic vulnerability in human B-ALL to enhance CAR-T efficacy, particularly in cases harboring TP53 mutations associated with relapse and chemotherapy resistance. Using genome-wide CRISPR/Cas9 screening on TP53-wildtype and TP53-mutated CD19+ B-ALL cell lines exposed to CD19-CAR-T treatment, we identified the targeting of fatty acid transporter FATP2 (encoded by SLC27A2) as a candidate for enhancing CAR-T efficacy. Subsequent pediatric B-ALL patient cohort analyses correlated high SLC27A2 mRNA expression with worse overall survival. FATP2-expressing B-ALL lines exhibited increased long-chain FFA uptake, with in vitro studies validating a leukemia-intrinsic role for FATP2 in promoting CD19 CAR-T resistance, with reversible effects upon depleting exogenous FFA. Stable isotope FFA uptake tracing revealed a role for FATP2 in promoting succinate accumulation in TP53-mutated B-ALL cells, which may impact leukemic gene expression through α-ketoglutarate inhibition. This proposal seeks to evaluate exogenous FFA uptake inhibition via genetic and pharmacological FATP2 inhibition to overcome CAR-T resistance in human B-ALL cell lines in vivo and primary B-ALL patient biospecimens ex vivo. Through metabolomic analyses and stable isotope-labeled linoleic acid tracing, we will then elucidate the precise metabolic pathways impacted by FATP2-dependent FFA utilization in primary human B-ALL, including investigating the mechanistic impact of succinate accumulation on TP53-mutated B-ALL gene expression and its role in CAR-T responsiveness. This interdisciplinary research seeks to enhance CD19 CAR-T therapy for relapsed/refractory B-ALL by targeting FFA uptake and uncovering gene-regulatory changes. The study aims to determine whether succinate or related metabolic intermediaries mediate B-ALL cell survival during CAR-T therapy. The comprehensive approach leverages the expertise of a diverse research team, ensuring a nuanced understanding of B-ALL biology and potential avenues for improving CAR-T-treated B-ALL patient outcomes.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY Regulation of genome access underlies development, differentiation, and response to stimuli. Genome access in turn is determined by nucleosome packaging and modification, the “chromatin landscape”. Static snapshots of the chromatin landscape cannot explain how stable cellular states arise despite constant chromatin dynamics. By developing new approaches in time-resolved structural epigenomics, we will uncover chromatin states that are invisible with current methods, to identify the molecular determinants of cellular identity. A key pathway that determines cellular identity during development is the polycomb pathway, an epigenetic mechanism of maintaining cell states through gene repression. By using our novel tools to track temporal dynamics of heterochromatin in unperturbed cells, we can map the recovery of heterochromatin after its dilution due to replication. We will identify the factors that determine the locations where polycomb enzymes are first recruited after replication. By tracking transcription and chromatin states across the cell cycle, we will identify bistable states of genes where they are expressed in one phase of the cell cycle, and they are packaged as heterochromatin in another phase of the cell cycle. Overall, our methods will resolve how the cell cycle influences chromatin states and how chromatin states in turn regulate the cell cycle. In summary, our approach of time-resolved- structural genomics will uncover not only the determinants of cellular memory in fundamental systems of mammalian development, but also provide the means to understand gene regulation across the cell cycle.

NIH Research Projects · FY 2025 · 2025-01

PROJECT SUMMARY From Dr. Goldberg’s (principal investigator’s) NIA GEMSSTAR R03 and BEESON K76 research, there is promising evidence that we can prevent fall-related emergency department (ED) visits among older adults with the Geriatric Acute and Post-Acute Fall Prevention (GAPcare) intervention. The GAPcare randomized, controlled trial (RCT) (n=110, two sites) had high patient and caregiver satisfaction, did not prolong ED length of stay, and resulted in 66% fewer 6-month fall-related ED visits, as compared to usual ED care. However, GAPcare requires in-person pharmacists and physical therapists (PTs), which limits scalability to large EDs, which account for only 43.8% of all ED visits. If we fail to scale this intervention, valuable prevention opportunities will be missed for the 3 Million older Americans who seek care for falls in EDs each year. Our over-arching goal is to expand the reach of GAPcare by testing a video telehealth GAPcare (e-GAPcare) intervention in a community ED without existing pharmacy and PT services. The rationale for this research is that telehealth for specialist evaluations have already been successfully implemented in EDs, suggesting that GAPcare could be adapted for patients with falls who would benefit from pharmacy and PT consultation, but cannot receive them due to staffing limitations in rural or smaller community EDs. Our long-term goal is to prevent fall-related ED visits. The objective of this R21 is to evaluate feasibility outcomes, assess acceptability, and to obtain estimates of effect on the outcome measures. Our central hypothesis is that e-GAPcare will help ED clinicians identify fall risk factors, such as gait, balance, and fall-risk increasing medication, and could ameliorate these risk factors through action plans co-created with patients preventing the next fall. GAPcare’s specific aims are to: (1) qualitatively elicit perspectives from content experts in telehealth (n=18- 24) to gain insights about the practical considerations of implementing telehealth consultation in the ED; (2) Adapt the GAPcare intervention and study protocol based on interviews with experts and workgroup feedback from ED staff, patient and caregivers (n=20); and (3) determine feasibility of e-GAPcare by conducting a single arm feasibility study with older ED patients (n=40) with a recent fall. Feasibility measures include our ability to recruit and retain participants, complete telehealth consults, and participants’ ability to initiate and perform necessary actions (with or without staff assistance). We will also assess acceptability using the Net Promoter Score and uptake of action plans. Using phone calls at 1, 3, 6 months and the electronic health record we will measure subsequent 6-month ED visits for falls, to inform initial efficacy for our subsequent large-scale RCT. Upon successful completion of this research, we will have a fully piloted telehealth intervention tailored to the unique health needs of older adults. This contribution is expected to be significant because it will provide a mechanism for patients in smaller or rural EDs to access geriatric-trained professionals and achieve health outcomes – reduced ED visits for falls – like patients in higher resourced larger academic medical centers.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY ABSTRACT Patients with terminal illnesses face many complex decisions regarding their care at the end of life. Options range from aggressive life-prolonging care to those that aim to minimize suffering and include palliative care (which can occur simultaneously with life-prolonging care) and interdisciplinary hospice care. Enter into this already complicated decisional context a legal option for patients to voluntarily end their lives before their illness advances further: Medical Aid in Dying (MAiD). In 1997, the “Death with Dignity Act” was passed, legalizing MAiD in Oregon. Since that time, 10 states and the District of Columbia have passed MAiD laws, and 8-9 states are considering similar legislation as of 2021-22. Yet, despite growing widespread access to MAiD, there is a paucity of research about physicians' participation in MAiD. There are no national data to understand the epidemiology of MAiD from the physician perspective. Further, there are no data on the role that ethical and patients' quality-of-life (QOL) considerations do or don't not play in physicians' participation in MAiD activities. To address these large knowledge gaps, we propose to conduct the first ever national study of physician participation in MAiD. The specific aims are: Aim 1: to create a claims-based MAiD-targeted sample of doctors practicing in states where MAiD is legal; Aim 2: To conduct a national survey of doctors practicing in states where MAiD is legal regarding the nature, extent, and consequences of their participation in MAiD; and Aim 3: To explore the unique role that ethical principles and physician assessments of the quality of life patients have on physicians' attitudes towards and participation in MAiD activities. Given MAiD appears to be expanding across the US, the data provided by this study will be critical to inform MAiD discussions and evidence-based policies and practices of physicians in the US.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY Cancer health disparities in the US persist, particularly between racial/ethnic groups and urban/rural residency, despite nearly 92% of Americans having health insurance coverage. In 2021, Medicaid covered ~19% of people; 54.2% were covered by employer-based insurance; Medicare covered another 18.4%; 10.2% were covered by direct purchase plans, and the remaining 3.5% were covered by military plans. Previous research has focused on patients without insurance, but factors associated with coverage, such as cost sharing, out-of-pocket limits, provider networks, and drug formularies can vary widely, potentially contributing to substantial disparities in access to and costs of care. Progress toward ameliorating disparities is hindered by the absence of multiple-payer data that could inform policies. Population-based registries do not adequately capture health insurance coverage or the full cancer care experience. Moreover, few data sources capture OOP costs that can both influence treatment decisions and lead to financial hardship. We address this gap by building on an established linkage between the Colorado Central Cancer Registry (CCCR) and the Colorado All-Payer Claims Data (APCD). APCDs are an emerging source for comprehensive data on healthcare utilization within a state. We supplement these data with plan-level information and census and health services files to better understand the social determinants of health context in which patients reside. The resulting data will contain person, treatment, provider/facility, plan, and geographic data for the years 2012-22 that allows longer-term survivorship assessments. Our specific aims are to: Aim 1: Compare differences in outcomes by race/ethnicity and urban/rural residency, controlling for plan type, patient demographics, market and provider characteristics, and social determinants of health. Aim 2. Decompose the relative contribution of plan type, patient demographics, market and provider characteristics, and social determinants of health in explaining observed outcome differences. Aim 3: Estimate the burden of OOP expenditures after a cancer diagnosis by plan type and by race/ethnicity and urban-rural residency.

NIH Research Projects · FY 2025 · 2025-01

PROJECT SUMMARY End-stage heart failure (HF) due to dilated cardiomyopathy (DCM) is the most common indication for cardiac transplantations in children over the age of 1 year. Though DCM affects both adults and children the causes can be different, with ischemic heart disease being the most frequent cause of HF in adults whereas in children the most common cause of DCM is idiopathic. However, the therapeutic strategies in children with heart failure are based on adult clinical studies and are less effective in managing HF in children. This suggests that understanding the mechanisms of heart failure in children is critical to identify novel age-specific therapeutic targets in this population and improve the translational barrier roadblock in pediatric DCM patients. My preliminary data suggests that miR-301b is highly expressed in pediatric DCM hearts and its predicted target genes are implicated in autophagy. In addition, primary cardiomyocytes over-expressing miR-301b demonstrate inhibited phagosome formation associated with reduced RAB5A protein expression. RAB5A is known to be highly expressed in the heart and is a miR-301b predicted target gene. In this application, I propose to determine the expression level of autophagy-related proteins in the heart tissue of pediatric DCM patients (Aim1), establish the effect of miR-301b over-expression on autophagy and apoptosis in primary cardiomyocytes (Aim2) and Determine in vivo regulation of cardiac autophagy by miR-301b and its consequence on cardiac function in neonatal mouse model. These studies will confirm the negative effect of miR-301b on autophagy, apoptosis and most importantly on cardiac function by directly targeting Rab5A and contribute to the development of novel age-specific therapies in children with heart failure addressing the translational barrier roadblock in this population.

NIH Research Projects · FY 2025 · 2025-01

Project Summary/Abstract Obesity is an urgent global epidemic demanding attention. A formidable obstacle in tackling obesity is the tendency of weight losers to rebound. While multiple factors contribute to weight regain, we hypothesize that heightened energy intake during meal events emerges as a primary driver of positive energy balance. Understanding the regulation of meal energy intake is imperative in addressing this critical issue. This mentored F32 proposal aims to investigate the physiological regulation of energy intake at meals following weight loss. This research will involve analyzing energy balance and gut hormone data from the ARROW study (R01DK114272), which aims to identify predictors of long-term weight loss maintenance in subjects who lose weight through dieting with or without exercise compared to a control group with obesity. Additionally, participants from the Time2Eat study (R01DK132372) will be leveraged to investigate gut hormones, metabolites, and subjective measures of appetite before and after breakfast and dinner test meals in subjects engaging in early time-restricted eating versus late time-restricted eating. Dr. Ezpeleta's background in intermittent fasting and experience in analyzing energy balance physiology in his current T32 program uniquely positions him to investigate these factors. Dr. Ezpeleta has assembled a team of mentors who have developed a comprehensive training plan for his success. The plan includes developing expertise in energy balance, gut hormones, and measures of appetite, designing and conducting clinical trials to analyze appetite in various weight statuses and conditions, gaining proficiency in using quantitative questionnaires to assess eating behaviors, psychosocial measures, appetite, and other ingestive behaviors, and enhancing his proficiency in clinical research methods while receiving additional training in manuscript development, grant applications, and presenting research findings. The research and training will take place at the University of Colorado Anschutz Medical Campus (CU- AMC), which offers state-of-the-art resources, career development opportunities, and pilot grant funding. Successful completion of this proposal will lead to five first-authored publications and generate thrilling preliminary data to support a K-award application. In summary, this F32 proposal will expand Dr. Ezpeleta's expertise in energy balance and appetite factors, contribute profoundly to the understanding of physiological regulations behind meal energy intake, and assist in establishing Dr. Ezpeleta as an independent investigator.

NIH Research Projects · FY 2026 · 2024-12

SUMMARY Each cell cycle, the genome must be faithfully duplicated and then divided appropriately into two progeny cells. Most of the genome is replicated in S phase of the cell cycle, and the mechanisms of DNA replication are well-characterized. However, cells sometimes have not finished replicating the entire genome as they enter mitosis, the phase of the cell cycle when chromosomes are segregated eventually leading to cell division. Instead, these remaining regions are replicated in mitosis in a likely “last-ditch effort” to produce separable chromosomes: cells without it face aneuploidy or mitotic catastrophe. Despite the importance of mitotic DNA replication, the mechanisms governing it are poorly understood. To uncover the molecular mechanisms of mitotic DNA replication, our research program will use the biochemical reconstitution of DNA replication using purified budding yeast proteins, which is ideally suited for addressing direct mechanistic questions. We will complement this with yeast genetics approaches, known for their significant contributions to our understanding of the cell cycle and DNA replication. Additionally, we have developed innovative strategies that combine reconstituted replication forks with yeast cell extracts to discover novel factors and explore the regulated timing of mitotic DNA replication. Our model of mitotic DNA replication is a multi-step process starting with the regulated inhibition and dismantling of the S phase replication machinery present at the replication fork to present a different structure and make space for mitotic proteins. This dismantling is followed by regulated unwinding and cleavage events, ultimately leading to a break-induced replication mechanism of strand-invasion, synthesis, and resolution. Our novel and innovative approaches will test this complex model and determine the coordination of these steps with other mitotic events occurring in distinct mitotic phases. We will also explore how and when mitotic DNA replication particularly occurs at hard-to-replicate regions of the genome like chromosome fragile sites and telomeres. S phase replication and the proteins so far implicated in mitotic DNA replication are well-conserved, so the mechanisms we uncover using the yeast system will provide a mechanistic framework that we will explore in other contexts in the future. We are interested in how genomic factors such as chromatin state and the presence of DNA damage or transcriptional conflicts play a role in mitotic DNA replication. Additionally, we seek to understand how diverse cellular contexts like developmental stage or disease state result in variable reliance on mitotic DNA replication, ultimately contributing to genome stability and cell viability.

NIH Research Projects · FY 2025 · 2024-12

Project Summary Navajo people (the Diné) experience more health disparities compared to the general population and have higher prevalence of certain genetic diseases and disorders. There is limited research on disease, treatment, and prevention, and underrepresentation in genetic and genomic research studies remains a concern. The Navajo Nation’s Health and Human Services Committee (HHSC), representing the largest tribe in the southwestern U.S. with over 400,000 enrolled citizens, approved a moratorium in April 2002 “on genetic research studies conducted within the jurisdiction of the Navajo Nation until such time that a Navajo Nation Human Research Code has been amended by the Navajo Nation Council” after long consultations with tribal leaders, traditional healers, and Navajo people with medical and scientific training. An underlying concern was that the Navajo Nation lacked policies or guidelines for modern genetic research protocols that would prevent research harms. In recent years, the Navajo Nation has begun to reexamine the moratorium and several public discussions have taken place with Navajo people about emerging genetic and genomic technologies and its associated ethical dimensions, yet there has not been an opportunity to convene the wider Navajo public and other tribal nations to discuss the benefits and risks of genomic research (including emerging initiatives such as precision medicine). The proposed Navajo Genetics conference to be held in April 2025 will be timely as the Navajo Nation reconsiders the moratorium. Our main goal is to improve genetic literacy and knowledge of genetics and genomics on the Navajo Nation and within the larger Indigenous community, which is valuable and important for making future health decisions and advancing health research. We also strive to support Navajo tribal sovereignty by supporting ways that discussion and decision- making can be shared as a robust genetic research policy is created by the Navajo tribal leaders.

NIH Research Projects · FY 2026 · 2024-12

PROJECT SUMMARY/ABSTRACT Alzheimer's disease is a leading cause of death in the U.S. and there is an urgent and unmet need to develop digital technology for early detection and monitoring of Alzheimer's disease and related dementias. Single- channel sleep electroencephalography (EEG) is inexpensive, easily scalable, and can be recorded in a home setting from currently available “wearable” headband devices. Within sleep EEG, oscillatory events that reflect activity in memory circuits have been associated with aging, cognition, and markers of preclinical Alzheimer's disease. Thus, single-channel sleep EEG provides the framework of practical digital technology that can be used to build a powerful tool for monitoring brain health and for detection of preclinical neurodegenerative disease. Real world application of this technology will require significant work to better understand how oscillatory events in sleep EEG reflect brain aging and neurodegenerative processes, as current knowledge of this neurophysiology remains insufficient to build a robust digital biomarker. Moving to bridge this critical gap, our team has pioneered advanced signal processing methods and generated compelling preliminary data from machine learning approaches that demonstrate convincing predictions in aging, cognitive measures, and biomarkers of Alzheimer's disease pathology. Here we propose to substantially advance this approach by applying a robust machine learning approach that will build the foundational algorithms of a digital biomarker for detecting pathological aging and early stages of neurodegeneration. Our innovative approach will incorporate our team's domain knowledge of sleep's oscillatory events with the power of large-scale machine learning. Utilizing more than 15,000 sleep recordings from nine existing cohorts, we will train and independently validate predictive models as the basis for novel digital biomarkers. Our overall goal is to create digital biomarkers that are sufficiently accurate to monitor personalized brain health via a single channel of sleep EEG. The overarching hypothesis of our proposal is that metrics of sleep EEG capturing the health and consistency of memory processing circuits can be incorporated into machine learning models to provide robust predictions of 1) pathological brain aging, 2) cognitive decline, and 3) neuroimaging and molecular biomarker changes that occur early in Alzheimer's disease pathogenesis. We will interrogate this hypothesis with the following Specific Aims: (Aim 1) Elucidate the oscillatory event features of sleep EEG that best predict brain age and determine the performance of these features in assessing whether an individual is experiencing more “youthful” or “accelerated” brain aging, (Aim 2) Examine the relationship between sleep's memory-processing oscillatory circuit integrity and cognitive decline, (Aim 3) Delineate the relationships between memory-processing oscillatory circuitry integrity and Alzheimer's disease-related neuroimaging and molecular biomarkers. Our proposal is significant because it will seek to build the foundation for a digital biomarker that can monitor brain health and identify pathological aging and early neurodegenerative changes using practical and accessible digital technology.

NIH Research Projects · FY 2026 · 2024-12

PROJECT SUMMARY Group B coxsackieviruses (CVB) are in the genus Enterovirus and Family Picornaviridae. CVBs are spread by fecal-oral transmission and can cause diseases including intestinal inflammation, pancreatitis, myocarditis and central nervous system diseases. Sensing of CVB3 RNA by melanoma differentiation-associated gene (MDA5) induces a Type 1 Interferon (T1IFN) response that is critical for reducing viral replication and dissemination within cells and the host. The pattern recognition receptor (PRR) Nucleotide Oligomerization Domain protein 2 (NOD2), which is critical for intestinal immunity, is classically known to detect bacterial peptidoglycan. Once stimulated, NOD2 requires RIP2 kinase (RIP2K) to activate NF-kB leading to a pro-inflammatory response. More recent data has demonstrated that NOD2 can also function as a cytoplasmic viral PRR by activating the production of T1IFNs in response to Influenza A virus. Interestingly, NOD2 has been implicated in a proviral role in CVB3-induced myocarditis, however, the underlying mechanism is not well understood. Because NOD2 is especially important in regulating the intestinal microbiota and maintaining intestinal immune homeostasis and CVB3 is an enteric virus that initially infects the intestinal tract leading to virus dissemination to other tissues, we sought to determine whether NOD2 expressed intestinal epithelial cells (IECs) would impact CVB3 infection. We observed that IEC NOD2 exacerbates CVB3 pathogenesis and fecal shedding in mice that were infected with CVB3. Additionally, we observed that the intestinal microbiota is required for CVB3 pathogenesis, suggesting that the microbiota may influence CVB3 infection by modulating host immunity through IEC NOD2. Furthermore, we demonstrate that IECs deficient for NOD2 have increased T1IFNs and Interferon Stimulated Genes (ISG) expression that limits CVB3 replication. CVB3 replication and ISG expression was rescued when IFIH1 (MDA5) expression was reduced with small interfering RNAs (siRNAs). In addition, NOD2-/- IECs have impaired mitochondrial health measured by increased ROS production and loss of mitochondrial membrane potential. In this proposal we will test our hypothesis that microbiota activation of IEC NOD2 induces mitophagy which blunts T1IFN responses and MDA5 expression resulting in exacerbated CVB3-induced pathogenesis. We will test key aspects of our hypothesis using the logical and innovative approach outlined in the following specific aims. Aim 1. Examine if CVB3 infection alters the microbiome causing dysbiosis that activates IEC NOD2 to contribute to CVB3-induced pathogenesis. Here we will define whether IEC NOD2 and the microbiota are initiators of CVB3-induced pathogenesis. Aim 2. Determine if NOD2/RIP2K expressed in IECs induces mitophagy to inhibit MDA5 activation and T1IFN responses. We will test whether NOD2/RIP2K signaling contributes to effective mitophagy leading to decreased MDA5-mediated antiviral responses and susceptibility to CVB3 infection. Characterizing and understanding the mechanisms of NOD2-dependent proviral responses are innovative new concepts that would markedly influence the current beliefs of NOD2 biology.

NIH Research Projects · FY 2026 · 2024-12

Project Summary Lung cancer is the leading cancer killer. Preventing lung cancer is vital to reducing the global cancer burden. Pharmacological prevention is an appealing strategy because it reduces the burden of treating tumors and pro- vides options for former smokers or lung cancer survivors who are highly motivated to minimize their lung can- cer risk. Prevention agents can be targeted to high-risk patients with premalignant lesions (PMLs), but lung PMLs often spontaneously progress and regress, making it difficult to avoid overtreatment. Recently, we identi- fied molecular patterns that separate lung PMLs likely to persist to carcinoma from those likely to regress. However, there is an urgent need to identify interception targets in these persistent PMLs. Identifying key mechanisms in PML persistence targeted by prevention agents will empower a precision approach focused on intercepting PMLs, which could significantly reduce tumor development high-risk patients. Our long-term goal is to use targeted prevention agents to intercept lung tumor development in high-risk populations. The main objective of this proposal is to identify interception targets in persistent lung PMLs. We identified a profile that differentiates lung dysplasias likely to persist from those likely to regress. This profile implicates loss of cell cy- cle control, altered adherens junction components, decreased M1 macrophages, and increased regulatory T- cells in dysplasia persistence and includes a gene expression panel for persistence. We chose Iloprost and PD-1 inhibition as valuable tools for investigating persistence pathways in lung PMLs, based on their efficacy in clinical trials and known targeting of epithelial or immune cells, both components of the persistence profile. We hypothesize that persistence pathways can be targeted to intercept the lung PMLs most likely to progress to carcinomas, and that changes in the persistence profile correlate with response to prevention agents. We will use an innovative and human-relevant reverse translational approach to test this hypothesis in the following specific aims: 1) Identify interception targets in the persistence profile in clinical trial biopsies, 2) Determine ef- fects of prevention agents on persistence pathways in vivo, 3) Identify the mechanistic targets in persistence pathways for prevention agent activity. After successful completion of this work, the expected outcome is demonstration that prevention agents targets persistence mechanisms in lung PMLs at risk of progressing to carcinoma. This project is innovative because it uses state-of-the-art assays and shifts the prevention research paradigm away from bench to beside translation, instead using clinical data to inform human-relevant mecha- nistic studies at the bench and cage. Our results will provide a strong basis for further development of precision application approaches for medical prevention, which would significantly impact populations at high risk of lung cancer by targeting the patients most at risk of progressing to invasive carcinoma. This project will advance lung cancer prevention research and address a critical need of the NCI Division of Cancer Prevention.

NIH Research Projects · FY 2025 · 2024-12

PROJECT SUMMARY This proposal aims to elucidate the mechanisms that drive the development of chronic chikungunya virus (CHIKV) arthritis (CCA). CHIKV is a mosquito-borne RNA alphavirus of global health concern. The acute phase lasts for 4-8 days and involves severe joint and muscle pain. In many patients, acute disease resolves without lasting symptoms; however, up to 50% of patients develop CCA characterized by incapacitating inflammatory musculoskeletal disease that persists for months to years. There are no approved therapeutics to treat CCA and the pathogenic mechanisms remain poorly understood. In preliminary studies using a mouse model of chronic CHIKV disease, I identified an increase in pro-inflammatory macrophages defined by elevated expression of NLRP3, TNF, and IL-1β, implicating macrophages as pathogenic effectors of chronic CHIKV disease. In addition, using scRNAseq and cell sorting approaches, I found that the persistent CHIKV RNA detected in joint-associated tissues is almost exclusively harbored in macrophages. Given that the NLRP3 inflammasome can be activated by viral RNA, these data offer a mechanism by which persistent viral RNA in macrophages promotes chronic inflammation. I also discovered that joint tissue CD4+ T cells during chronic CHIKV infection express elevated IFN-γ, suggesting a role for CD4+ T cells in the activation of macrophages. Finally, flow cytometric analysis revealed an increase in SLAMF7 on macrophages and CD4+ T cells during chronic disease. SLAMF7 contributes to TNF and IL-1β expression in macrophages and this is regulated by IFN-γ. Notably, in rheumatoid arthritis, macrophages are pathogenic due to elevated expression of SLAMF7 and inflammatory genes including IL-1β, TNF, and IL-6 that promote cartilage and bone destruction. These findings suggest macrophages share a similar pathogenic role in CCA. I hypothesize that CHIKV RNA persists in macrophages and induces inflammasome- mediated pro-inflammatory programming. In addition, SLAMF7 and CD4+ T cell interactions contribute to macrophage mediated CCA. My proposed studies will (i) define a viral reservoir in macrophages in joint associated tissue, (ii) assess mechanisms that drive pro-inflammatory programming in macrophages, (iii) elucidate the role of CD4+ T cells in regulating pro-inflammatory macrophages, (iv) identify the contribution of SLAMF7 in CHIKV infection, and (v) provide insight on the consequence of pro-inflammatory macrophages in CCA. The knowledge gained from these studies will help identify potential therapeutic targets to mitigate chronic CHIKV disease.

NIH Research Projects · FY 2026 · 2024-12

PROJECT SUMMARY/ABSTRACT Each year, 12-18 million infants worldwide, representing ~10% of all births, are born preterm (before 37 weeks of gestation). The mechanisms underpinning preterm birth are poorly understood and, with the possible exception of progesterone in a limited number of women, no interventions are currently available to prevent preterm labor. The management of the preterm infant has improved over the last 30 years, however despite this progress, prematurity remains the second most common direct cause of death among children under 5 years of age. In addition, infants born preterm are at risk of neonatal morbidity (e.g., intraventricular hemorrhage, bronchopulmonary dysplasia and necrotizing enterocolitis) and long-term sequelae including chronic lung disease, retinopathy and poor neurodevelopmental outcomes. Emerging evidence in animal studies show that factors secreted by the placenta are critical for normal fetal organ development. One of the most fundamental differences between fetal and postnatal life is the instantaneous discontinuation of the umbilical circulation at delivery, depriving the premature infant of placental factors, such as proteins, critical for fetal organ development. We reported that 434 proteins are secreted by the placenta into the fetal circulation before 32 weeks of gestation. Remarkably, a substantial number of these proteins could be linked to processes such as angiogenesis, pulmonary development and neurogenesis, suggesting that a subset of these proteins are critical for the normal development of fetal organs that are often injured in premature infants. Inresponse to PAR-23-130 Translational Research in Maternal and Pediatric Pharmacology and Therapeutics, we will test the central hypothesis that supplementation with candidate human placental proteins, i.e., that are secreted into the fetal circulation and associated with the development of the brain, lung, retina, intestine and cardiovascular system, improve outcomes in premature guinea pigs. Our approach will be to induce preterm delivery in guinea pigs, provide them with 24/7 support in a guinea pig neonatal intensive care unit. Pups will either receive a single candidate human placental protein or vehicle by a subcutaneous mini-osmotic pump for 7 days after preterm delivery. We propose three aims: Aim 1: Determine the effect of candidate human placental proteins on mortality and short-term respiratory outcomes in premature guinea pigs. Aim 2: Establish the effect of candidate human placental proteins on short-term neurodevelopmental outcomes in premature guinea pigs. Aim 3: Determine the effect of candidate human placental proteins on long-term lung and neurodevelopmental outcomes in premature guinea pigs. Our proposal is conceptually highly novel because it represents a shift in the paradigm of how to approach prematurity by supplementing with placental proteins supporting organ development. This work may lead to a specific novel intervention that will alleviate perinatal mortality and morbidity and long-term sequelae in extremely premature babies and that is scalable and universally applicable.

NIH Research Projects · FY 2026 · 2024-12

PROJECT SUMMARY / ABSTRACT: Rates of obesity continue to rise across the world, hastening the need for new and improved treatment interventions. Currently, bariatric surgery is the most effective strategy to induce and sustain significant weight loss for individuals with obesity. However, this treatment option is often accompanied with metabolic complications, most notably postbariatric hypoglycemia (PBH). Latest estimates suggest that PBH occurs in ~30% of patients, after both Roux-en-Y gastric bypass (RYGB) and vertical sleeve gastrectomy (VSG). Despite the high prevalence of PBH, many patients do not experience the symptoms that should normally alert them of hypoglycemia. If untreated, severe hypoglycemia can lead to accidents, brain damage, and even sudden death. Thus, PBH is a serious issue that often goes unnoticed and therefore cannot be easily treated until we increase our understanding of the causes and mechanisms. The dramatic changes in feeding behavior after bariatric surgery and the lack of neurological symptoms in response to hypoglycemia clearly implicate altered brain function, but very little is known about how bariatric surgery alters the central circuits regulating feeding and glucose homeostasis. Recent work suggests that bariatric surgery reduces counterregulatory responses (CRR) and that dysregulated hypothalamic response to hypoglycemia may be involved. However, there are currently no effective treatment strategies for PBH, and more preclinical work is needed to identify therapeutic targets. In our lab, we have found that mouse models of VSG recapitulate key features of PBH, including postprandial and prolonged insulin-induced hypoglycemia. Overall, our data suggest that CRR signaling is dysregulated after VSG, and we will investigate neuronal mechanisms which may be underlying these deficits. The current proposal aims to study whether the glycemic threshold for CRR hormone release, glucoprivic feeding, and brain activation is shifted lower after VSG, and to identify the hypothalamic circuits and molecular mechanisms that may contribute. To accomplish this research objective, we will utilize glucose clamp strategies to characterize the glycemic threshold for CRR hormone release, and we will perform dose-response studies to investigate dynamics of glucoprivic hunger and hypothalamic activation (Aim 1). In Aim 2 we will perform stereotaxic surgeries to manipulate ventromedial hypothalamus signaling to determine the mechanisms contributing to PBH. By investigating how VSG dysregulates central glucoregulatory mechanisms, the current project will identify new therapeutic targets that could help mitigate or prevent PBH, thus making bariatric surgery safer for a broader range of patients.

NIH Research Projects · FY 2026 · 2024-12

Abstract Head and Neck Squamous Cell Carcinoma (HNSCC) is a deadly disease with proclivity for regional spread to the lymphatic chains following definitive therapy (regional recurrence). This recurrent disease in the lymph nodes remains a clinical burden that severely dampens overall survival, yet the biological underpinnings of this phenomenon remain elusive. Our lab has shown in murine models of HNSCC that targeting immunosuppressive T-regulatory cells (Tregs), prior to tumor inoculation rather than following, ablates regional recurrence altogether. Mechanistically, we find that Tregs interfere with High Endothelial Venules (HEVs), specialized postcapillary venules in the lymph node that allow for lymphocyte transmigration, early during tumor development. This interference of HEVs leads to the engorgement and de-differentiation of HEV structure, associated decreased lymphocyte trafficking and promotion of cancer cell quiescence. Indeed, we find that quiescent cancer cells, measured by p27+ expression, uniquely occupy the perivascular niche (PVN) surrounding the HEVs early during tumor progression suggesting that de-differentiated HEVs interact directly with the cancer cells to promote regional recurrence. These data led us to believe that targeting HEV function could prove to mitigate regional recurrence. We therefore sought to target HEV functioning during early tumor formation by using an agonist for Lymphotoxin Beta Receptor (LtBR), which has been shown to restore function of HEVs and increase lymphocyte presence within tumors. Upon addition of a systemic LtBR agonist to therapeutic radiation and Treg depletion, we observed a striking reduction in regional recurrence and a significant increase in lymphocyte presence within the tumor draining lymph node (tDLN), the site of regional recurrence. These preliminary findings lead us to hypothesize that restoring HEV function through LtBR agonism provides a rational approach towards treated regional recurrence. Aim 1 will focus on the mechanisms of HEV de-differentiation by Tregs in murine models of regional recurrence. In Aim 2, we will explore the impact of HEV de-differentiation on cancer cell seeding and quiescence. We expect that this proposal will provide a mechanistic understanding of how regional recurrence is established and will identify druggable axes to mitigate this phenomenon.

NIH Research Projects · FY 2026 · 2024-12

PROJECT SUMMARY: Despite the advent of immune-checkpoint blockade (ICB), melanoma remains a deadly skin cancer. Notably, the indication for ICB therapies continues to expand for melanoma, and they are now used in the adjuvant, neoadjuvant and first-line treatment settings. However, these agents can have deadly side effects and the field remains unable to predict response to these agents. Therefore, refining our ability to predict (A) patients with early-stage disease at high risk of disease progression who would benefit from ICB adjuvant therapy and (B) patients who will not respond to ICB treatment remains a top priority in melanoma, and cancer research more broadly. To address these clinical needs, in this proposal we will study primary tumor invasive capacity to predict risk of progression and tumor immune evasion mechanisms to predict response to ICB therapy. Interestingly, placentation (the formation of the placenta) is a naturally occurring model in which fetal tissue invades into the maternal uterine line (decidua) and evades maternal immune cell destruction. Indeed, there is strong evidence that cancer cells can co-opt mechanisms that evolved for placentation to increase their own malignant potential. The cell type that regulates placentation is called a decidual Natural Killer cell (dNK cell) that, unlike circulating NK cells, is proangiogenic and immunoregulatory. NK cell function is largely regulated by Killer-immunoglobulin like receptors (KIRs) and dNK cells express very high levels of KIRs that bind to HLA-C expressed on invading fetal cells. This interaction promotes tissue invasion and immune evasion during placentation. Importantly, the KIR/HLA-C interaction is highly polymorphic, leading to varied NK cell effector functions across individuals and specific KIR/HLA genotypes are associated with disorders of placentation. Therefore, KIR/HLA genotypes impacting NK cell function within melanoma patient could potentially explain differing therapy responses. Notably, decidual-like NK cells (dl-NK cells) with similar immunosuppressive and proangiogenic functions have been reported in numerous other solid cancers and we have exciting preliminary data showing that these cells are also present within melanoma tumors and express high levels of KIR. Therefore, we hypothesize that the KIR/HLA axis acting on dl-NK cells facilitates both tissue invasion (Aim 1) and immune evasion (Aim 2) in melanoma. To test this hypothesis, we will use state-of-the-art techniques including multiplexed immunohistochemistry, single-cell spatial transcriptomics, patient derived organoid models and high-resolution allelic level sequencing of the KIR/HLA loci, to evaluate these mechanisms. Our primary objective with this proposal is to identify biomarkers that will predict risk of disease progression and response to ICB therapy. Our secondary objective is to define new therapeutic targets to either limit disease progression or improve response to ICB therapy.

NIH Research Projects · FY 2026 · 2024-12

PROJECT SUMMARY Genetic alterations in cancer cells can cause activation of regulatory factors that bind chromatin at target gene loci to turn on transcription programs that drive tumor initiation and progression. However, despite identification of these oncogenic regulatory factors in multiple tumor types, a gap in this rapidly evolving field is the delineation of molecular mechanisms that open opportunities to target those regulators. Thus, my long-term goal is to define interaction surfaces within chromatin-bound regulators critical for their oncogenic functions. In the F99 phase of this proposal I will focus on the transcription regulator KAP1 and its previously unprecedented role in activating oncogenic WNT signaling in colorectal cancer (CRC). Despite the well- established knowledge that WNT signaling drives CRC transformation and promotes tumor progression, a therapeutic arm that successfully inactivates WNT in CRC has yet to be employed. Potentially addressing this gap in knowledge, my studies have discovered that KAP1 is required for expression of WNT target genes in response to oncogenic WNT stimulation. Importantly, compelling preliminary data support two non-mutually exclusive mechanisms explaining how KAP1 may regulate WNT signaling. First, KAP1 could directly activate WNT by scaffolding key transcriptional machinery to WNT target gene promoters using its chromatin reader module. Second, KAP1 could interact with and regulate β-Catenin stability (the WNT transcription effector). I will test these two models and then evaluate whether perturbing the KAP1–chromatin and/or KAP1–β-Catenin interactions will block WNT-induced CRC phenotypes. In the K00 phase, I will shift focus to another biomedically relevant chromatin regulatory complex (SWI/SNF). SWI/SNF normally remodels nucleosomes on chromatin to activate target gene expression, but its dysregulation in cancer can cause aberrant activation of oncogenic programs. Likewise, in the soft-tissue malignancy Synovial Sarcoma (SS), the transforming genetic alteration is translocation of SS18, a member of SWI/SNF, to the SSX transcription factor. Because SSX, but not SSX18, normally binds modified nucleosomes, SSX abnormally redirects SS18 to a cancer-specific set of genomic sites, causing upregulation of genes that promote tumorigenesis. Despite this mechanistic understanding, the molecular and structural basis of the SS18-SSX– nucleosome interaction remains unclear, and a strategy to target SS18-SSX is currently undefined. To achieve these unmet needs, I will biophysically characterize the SS18-SSX-nucleosome interaction and then identify small-molecule inhibitors that disrupt the SS18-SSX-nucleosome interaction. Finally, I will test lead inhibitor candidates in hallmarks of cancer assays (proliferation and invasion). The training obtained under both phases will fulfill my long-term goal of running my own lab with an emphasis on molecular mechanisms of gene regulation, allowing me to have a positive impact on cancer patients.

NIH Research Projects · FY 2025 · 2024-12

PROJECT SUMMARY/ABSTRACT Autism spectrum disorder (ASD) has increased in prevalence considerably over the past several years and is marked by social cognitive challenges, such as perspective taking. Such challenges can have significant influence on one’s quality of life, with adolescence representing a particularly difficult phase due to the increase in social demands that occur during this developmental period. ASD is associated with aberrant activity in brain regions involved in the default mode network (DMN) as well as low-level systemic inflammation (i.e., increased proinflammatory cytokine levels), both of which may contribute to the impaired social cognition observed in this population. Exercise (EX) interventions have been shown to alter DMN function and reduce inflammation in non-autistic samples, and preliminary studies suggest EX may enhance social functioning in autistic individuals. Therefore, EX may be a promising, low-cost intervention approach for supporting social cognition in autistic adolescents, and the proposed project is designed to test this hypothesis. Specifically, we aim to determine if a 10-week EX intervention, relative to a social gaming control condition, can 1) enhance brain response in DMN regions during a theory-of-mind (ToM) task; 2) improve social-cognitive functioning; and 3) reduce levels of circulating proinflammatory cytokines in autistic adolescents. The project will deepen current understanding of the neurobiological and immune mechanisms underlying EX interventions for autistic adolescents, which is a population with high risk for sedentary behavior and few accessible, empirically supported treatment options. Using functional magnetic resonance imaging (fMRI) and systemic inflammation as biomarkers adds an innovative dimension to the project and may allow for more sensitive measures of intervention-related changes than traditional behavioral markers. Data for the project will be derived from an ongoing parent study led by Dr. Legget (Sponsor), who will primarily oversee Dr. Cosgrove’s (PI) research and training during the fellowship period. Co-Sponsor Dr. Tregellas and a strong team of consultants will provide additional, complementary expertise to Dr. Cosgrove’s training in the areas of neuroimaging, intervention research, neurodevelopmental disorders, and immunology. Dr. Cosgrove has access to state-of-the-art equipment and resources at the University of Colorado Anschutz Medical Campus that will allow her to complete the proposed research and training. Altogether, the research and training goals planned for the two- year fellowship period will support Dr. Cosgrove in developing as an independent researcher with expertise in the application of clinical neuroscience methods to inform development of novel intervention approaches for youth with ASD and related neurodevelopmental disorders, such as intellectual disability.

NIH Research Projects · FY 2026 · 2024-12

PROJECT SUMMARY Group B Streptococcus (Streptococcus agalactiae, GBS) is a Gram-positive, β-hemolytic bacterium and a leading etiologic agent of neonatal bacterial meningitis and major opportunistic pathogen in neonatal, pregnant, elderly, and diabetic populations. During pregnancy, GBS asymptomatically colonizes the vaginal tract of 30% of healthy women but can result in adverse pregnancy outcomes and severe neonatal disease. GBS is also increasingly isolated from adult diabetic wound infections. Both the vaginal tract and diabetic wound are complex polymicrobial environments. However, despite the public health burden of GBS, little is known about how GBS competes with these diverse populations to establish a niche and persist in either host environment. The Type VII Secretion System (T7SS) in Firmicutes secretes diverse toxic effectors that play an important role in interbacterial competition and virulence. Prior work from our laboratory has shown that the GBS T7SS contributes to cytotoxicity and virulence during meningitis as well as persistence in the murine vaginal tract during colonization. However, the specific mechanisms by which the GBS T7SS contributes to colonization as well as its role in other host environments remain unknown. My preliminary data indicate that the GBS T7SS inhibits growth of the important vaginal pathobiont and diabetic wound pathogen Enterococcus faecalis in vitro and in vivo in the murine vaginal tract. Additionally, I have found that a T7SS-associated LXG protein is a putative intracellular toxin that inhibits growth of E. coli and is important for persistence in the murine vaginal tract and diabetic wounds. Based on this, I hypothesize that the GBS T7SS mediates competition with neighboring bacteria in the vaginal tract and diabetic wound and may be mediated by a secreted LXG toxin with nuclease activity. This proposal seeks to evaluate the interaction of T7SS with the native vaginal microbiota in GBS colonization of the vaginal tract, interrogate the role of a T7SS-associated toxin in interbacterial competition, and elucidate the importance of GBS T7SS to diabetic wound infection. These goals will be addressed in vitro and in in vivo murine models of vaginal colonization and diabetic wound infection in the following specific aims: Aim 1: Characterize the importance of T7SS to GBS interbacterial competition and vaginal colonization. Aim 2: Evaluate function of a GBS LXG toxin and its contribution to T7SS interbacterial competition. Aim 3: Investigate the role of GBS T7SS in diabetic wound persistence.

NIH Research Projects · FY 2026 · 2024-11

PROJECT SUMMARY Outer membrane vesicles (OMVs) released by extracellular Gram-negative bacteria can deliver lipopolysaccharide (LPS) and other bacterial molecules to the cytosol of macrophages. OMVs enter macrophages through endocytosis and reach the endosome of macrophages. Subsequently, components of OMVs including LPS cross the endosomal membrane and reach the cytosol to trigger pyroptosis, an inflammatory form of cell death. While the downstream pyroptotic signaling steps were previously characterized, the upstream stage of OMV entry into macrophages remains largely unknown. The goal of this project is to bridge this major knowledge gap. In our preliminary studies, we established assays to measure OMV-induced pyroptosis in RAW 264.7 macrophages. Using these assays, we performed an unbiased genome-wide CRISPR genetic screen to dissect OMV-induced pyroptosis. The screen isolated known mediators of pyroptosis and a large number of candidate genes not previously linked to pyroptosis or OMV function. In this proposed research, we will validate and characterize the candidate genes isolated in the CRISPR screen to identify new regulators of OMV entry into macrophages. These experiments are expected to establish mediators of OMV-macrophage interaction, OMV endocytosis, and translocation of OMV components across the endosomal membrane. These exploratory studies will shed light on the molecular basis of OMV entry into macrophages and will broaden our knowledge of inflammatory responses. Insights gleaned from this research will facilitate the development of safer and more effective strategies to combat bacterial infection and inflammatory diseases.

NIH Research Projects · FY 2026 · 2024-11

PROJECT SUMMARY Streptococcus agalactiae microbiota However, disease factor 30%. or Group B streptococcus (GBS) is a pathobiont that is often part of the normal found in the gastrointestinal (GI) tract and reproductive tract of healthy people. it onset ( LOD), which presents as meningitis. Maternal GBS colonization represents the most important r isk for adverse pregnancy outcomes and neonatal infections, worldwide range from 10- F female (FRT) is a major cause of serious invasive disease in neonates, including pneumonia, sepsis and late and carriage rates etal and neonatal infection can occur following i) ascending infection of the placental membranes in utero, ii) passage through the birth canal, by aspiration of infected vaginal fluids, or iii) colonization of the neonatal GI tract, which can occur during childbirth, or ingestion of infected breast milk postnatally. The fungus Candida albicans (Ca) is also a frequent inhabitant of the GI tract and FRT, with asymptomatic carriage rates of approximately 30% in women, and a higher prevalence often reported with age and in pregnancy. A growing body of evidence has identified C. albicans as an independent risk factor for vaginal colonization by GBS. However, surprisingly little is known about the cross-kingdom interactions that govern these bacterial- fungal associations within the GI and genital tract mucosa. We now have data using our mouse model of vaginal colonization that indicates co-colonization with Ca increases GBS vaginal persistence in vivo; however, the specific mechanism(s) for this synergy is unknown. We also show that Ca significantly enhances GBS attachment to vaginal mucosa using a newly developed multi-layered, highly differentiated human Vaginal Epithelial Cell-Air Liquid Interface (hVEC-ALI) model. Using triple RNA-sequencing during co-culture we have observed that Ca reprograms GBS transcription to induce adhesins and  -hemolysin/cytolysin (  -h/c) that are requiredfor GBS colonization and virulence, and that Ca responds to GBS by upregulating genes involved in arginine biosynthesis and uptake. We hypothesize that a GBS-Ca direct physical interaction promotes this transcriptional remodeling to promote GBS host cell attachment, vaginal persistence, and ascending infection. Further, that Ca arginine biosynthesis promotes GBS survival and virulence potential. We will also explore the hypothesize that Ca will similarly promote GBS neonatal intestinal colonization which could impact LOD, and that GBS mucosal colonization can be Ca Als3-based vaccine. We will examine these hypotheses in the following aims: 1) Determine the consequence of Ca-induced transcriptional reprogramming to GBS vaginal colonization and disease, 2) Elucidate the role of Ca arginine biosynthesis to GBS-Ca synergy and vaginal persistence, 3) Examine the effect of Ca on GBS intestinal targeted by therapeutic intervention with a colonizationand LOD, and the potential for maternal vaccination in preventing cross-kingdom synergy.

NIH Research Projects · FY 2026 · 2024-11

PROJECT SUMMARY Proinflammatory cytokines are key players in innate immune responses. Dysregulation of proinflammatory cytokines is associated with a number of human diseases such as arthritis, cardiovascular diseases, and metabolic disorders. In particular, the proinflammatory cytokines interleukin 1β (IL-1β) and tumor necrosis factor alpha (TNFα) interfere with important cellular signaling pathways such as AKT signaling and disrupt physiological processes dependent on the signaling pathways. It remains poorly understood how IL-1β and TNFα compromise AKT signaling at the molecular level. In this exploratory project, we will address this key question by identifying mediators of cytokine-induced inhibition of AKT signaling, which regulates a range of physiological responses. In our preliminary studies, we developed assays to measure the inhibitory effects of IL-1β and TNFα on AKT signaling. Moreover, using RNA-Seq, we identified genes upregulated or downregulated in cytokine-treated cells. We also developed CRISPR screening platforms to genetically dissect complex biological pathways at the genomic scale. In this project, we will capitalize on these preliminary data to perform genome-wide genetic screens to identify factors mediating cytokine-induced inhibition of AKT signaling. Next, we will validate candidate genes identified in the screens using pooled secondary screens and individual gene validation. Finally, we will gain mechanistic insights into the molecular functions of selected factors identified in the screens. If successfully accomplished, this exploratory project will provide key insights into cytokine-induced inhibition of cellular signaling and will facilitate the development of new therapeutics for cytokine-linked diseases.

NIH Research Projects · FY 2026 · 2024-11

PROJECT SUMMARY Group B Streptococcus (GBS) is a Gram-positive pathobiont that asymptomatically colonizes the female genital tract (FGT) of up to 30% of healthy women. However, during pregnancy it is associated with adverse pregnancy outcomes, including premature rupture of membranes (PROM), chorioamnionitis, stillbirth, and preterm birth; notably 10% of preterm births are caused by GBS. Further, GBS can be transmitted to the fetus in utero, or the newborn during vaginal birth, resulting in invasive neonatal disease. GBS is the leading cause of meningitis and bloodstream infections in newborns, leaving them at risk for long-term problems. During pregnancy, GBS colonization of the FGT is intermittent and can be transient. However, the bacterial and host determinants that promote GBS vaginal colonization and ascending infection, as well as the role of native microbiota are largely unknown and remain important questions to be studied. Previous microbiome studies have demonstrated that GBS colonization impacts vaginal microbial diversity. Using a mouse model of GBS vaginal colonization, we found that GBS persistence is driven by computationally predictable changes in certain taxa, including Akkermansia muciniphila, a Gram-negative obligate anaerobe that degrades mucin. We have further shown that co-colonization with A. muciniphila increases GBS vaginal persistence, however, the specific mechanism(s) for this synergy is unknown. To date, we demonstrate that GBS adherence to human vaginal and endocervical epithelial cells increases in the presence of A. muciniphila. To further explore this synergism, we performed dual and triple RNA sequencing with human vaginal epithelial cells (hVECs) in order to observe the effect of each organism during infection. Upon analysis, we identified 225 unique GBS genes that were differentially expressed in the presence of hVECs and A. muciniphila. Interestingly, our data showed significant increase in expression of genes encoding for GBS pili, and preliminary data suggest that the observed increase in adherence to hVECs is occurring in a pili-dependent manner. Additionally, the host transcriptome demonstrated major changes in numerous cellular processes in the presence of GBS and A. muciniphila, including innate immune responses. We hypothesize that the observed increase in GBS burden during co-culture with A. muciniphila may be due to: 1) induction of GBS pili production; 2) alteration of mucin abundance; 3) modulation of host immune responses. In my proposed studies, I will focus on identifying the specific mechanisms by which A. muciniphila promotes GBS host cell interaction, as prolonged GBS vaginal persistence may potentiate the risk of invasive infection in utero and in the newborn.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY/ABSTRACT The way antibiotics are delivered to hospitalized children matters. When intravenous (IV) and oral antibiotics are equally effective, the use of oral antibiotics may avoid IV-related harms, improve patient experience, and decrease resource utilization, hospital length of stay, and cost. Pneumonia is the ideal infection to study the overuse of IV antibiotics as it is one of the most common reasons for hospitalization and there is emerging evidence that initial oral antibiotics are likely as effective as IV antibiotics. Despite existing evidence, most children hospitalized with pneumonia still receive IV therapy. This gap in translating evidence into practice highlights the critical need for strategies to increase oral (and reduce IV) antibiotic use. The overall hypothesis is that clinician-focused strategies that target initial antibiotic route in the Emergency Department are feasible, increase oral antibiotic use, and improve patient outcomes. The PI will evaluate these strategies in a pilot hybrid trial that will evaluate both effectiveness and implementation outcomes. Through this, the PI will simultaneously gather data on the effectiveness of IV vs. oral therapy and investigate ways to translate evidence into practice. Specifically, the PI, Jillian Cotter, MD, MSCS, in collaboration with her mentoring team will pursue the following aims: (1) Identify barriers and facilitators to increasing initial oral antibiotics for children hospitalized with pneumonia; (2) Develop a set of implementation strategies to increase initial oral antibiotics; and (3) Conduct a pilot hybrid trial to determine the feasibility and acceptability of the implementation strategies. This will prepare her to submit an R01 grant for a multisite hybrid trial to definitively evaluate implementation and effectiveness outcomes. This proposal focuses on children, an AHRQ priority population, and aligns with AHRQ’s goals to implement studies to accelerate the spread of evidence-based practices. As a pediatric hospital medicine physician and clinical researcher with growing expertise in pneumonia and antibiotic stewardship, Dr. Cotter is uniquely positioned to accomplish the proposed research and training aims. Her career goal is to become an independently funded investigator and national leader in the implementation of antibiotic stewardship practices. Dr. Cotter has developed a detailed career development plan that builds on her research skillset and fills key knowledge gaps that are critical to advancing her career. Through coursework, experiential learning, and guided mentorship, she will gain experience in the: (1) design and conduct of qualitative research, (2) development of evidence-based interventions using implementation science, and (3) conduct and evaluation of pragmatic trials. Dr. Cotter has assembled a strong multidisciplinary mentorship team with expertise in pneumonia, qualitative methods, implementation science, and pragmatic trials, which will ensure her success in achieving her stated aims and training goals. This line of inquiry will prepare Dr. Cotter for future R01 funding and her transition to independence.