Colorado State University

NIH Research Projects · FY 2026 · 2024-08

Project Summary: Maternal obesity with increased circulating inflammatory markers can lead to adverse pregnancy outcomes, such as preeclampsia (PE). The clinical signs of PE include maternal hypertension and proteinuria during the second half of gestation. While PE presents later in pregnancy, its origins are thought to begin early in pregnancy or even before conception. Importantly, maternal hypertension only resolves after delivery of the placenta; therefore, it is widely accepted that abnormal placentation plays a causal role in PE pathogenesis, though the etiology of this is unknown. It is our hypothesis that maternal adiposity contributes to heightened inflammation and subsequent abnormal placental vascular development. The overarching goal of these proposed studies is to test the hypothesis that pro-inflammatory mediators produced by maternal adipose tissue (leptin) reduce pro- angiogenic immune cells and factors at the maternal-fetal interface. We will test our hypothesis in the BPH/5 mouse model of PE combining novel in vivo and ex vivo experiments. In Aim 1, we will determine if hyperleptinemia promotes decidual inflammation at the maternal-fetal interface in early pregnancy. Leptin- STAT3 inflammatory mediators will be measured in control+leptin mice and BPH/5 mice that have hyperleptinemia, which is attenuated with weight loss. In Aim 2, we will use single cell sequencing to isolate the source of decidual inflammation in these pregnancies as well as placental angiogenic deficiency. In Aim 3, offspring outcomes will be assessed to determine whether reversal of maternal obesity via pair-feeding prevents aberrant fetal cardiac angiogenesis and adult onset cardiometabolic dysfunction. These experiments will combine the gold standard radiotelemetric blood pressure recording with metabolic cages to ascertain cardiometabolic fitness in offspring when maternal hyperleptinemia is blunted and PE is prevented. These findings will provide groundbreaking prenatal mechanisms to prevent adult-onset cardiometabolic disease, hypertension, in PE born offspring.

NIH Research Projects · FY 2026 · 2024-08

PROJECT SUMMARY Molecular tools that can precisely modulate gene expression provide unique opportunities to both study the relationships between genotype and phenotype, as well as correct the pathologies caused by dysregulated gene expression. However, restricting the activity of these tools to the appropriate times or cell types remains a major hurdle to their effectiveness at interrogating biology and their safety in therapeutic settings. Our research program’s main goal is to elucidate the design principles for synthetic signaling systems that enable precision control of gene expression and, in doing so, to create fundamental insights into the mechanistic basis of natural signal transduction. Our work spans 3 methods of modulating expression and in each case seeks to overcome a major engineering challenge by generating novel fundamental insights into the transduction mechanism through a combination of high-throughput screening and machine learning. The first are Cas9-based synthetic transcription factors, which enable targeted changes to the transcription of a gene. We plan to restrict their activity via fusion to nuclear receptors that will make their nuclear localization, and hence regulation, conditional on a chemical inducer. We aim to elucidate how the structure of nuclear receptors encodes their nuclear trafficking kinetics and dynamic range, and then use these insights to design controls systems that can rapidly implement strong regulation in response to a non-toxic chemical cue. The second are ribozyme-based tools that regulate expression at the RNA level through splicing or trans- cleavage. We plan to make their activity contingent on the presence of either native mRNAs, through template dependent splicing, or chemicals, using aptazymes. We aim to understand how changes to the sequence, and resulting structure, of these RNA devices alter their capacity to transduce their triggers into catalysis. The resulting insights will be used to identify a combination of mutations that can overcome the low catalytic efficiencies often associated with these tools. The third are chemicals that activate human G-protein coupled receptors (GPCRs) to modulate expression of the genes they regulate. We plan to identify plant metabolites that act as selective agonists by developing a high-throughput screen that enables massively parallel characterization of GPCR-ligand interactions. We aim to elucidate the design principles for functional expression of human GPCRs in yeast and use the resulting biosensors to reveal the ligand features necessary for selective activation of GPCRs. Overall, this research will create novel tools to enable precision control of gene expression and generate fundamental insights into how molecular architecture, structure, and ligand specificity impact signal transduction. Thus, it both aligns with the NIGMS mission and fills the need for “transcriptional control tools” recently identified in the NIGMS NOSI: Synthetic Biology for Biomedical Applications (NOT-EB-23-002).

NSF Awards · FY 2024 · 2024-07

This award supports research in relativity and relativistic astrophysics, and it addresses the priority areas of NSF's "Windows on the Universe" Big Idea. Since 2015, when the twin NSF's LIGO gravitational wave detectors (GWD) in the United States detected the first signals originating from the coalescence of two massive black holes, there have been multiple detections at a pace of about one a week. Presently in the LIGO-Virgo-KAGRA fourth observation campaign, detections are revealing new mergers with unexpected results. Key to these remarkable achievements has been the improvements in the sensitivity of GWD, among which the reduction in the coating thermal noise (CTN) of the test masses of the interferometer has had a large impact. The projected 2x reduction in CTN with respect to the current LIGO coatings and beyond for third-generation (3G) GWD is pushing the scientific and engineering boundaries in dielectric coatings to understand and control the fundamental mechanisms in amorphous oxides that contribute to mechanical loss. The project aims to demonstrate next-generation multilayer dielectric coatings that reduce the thermal noise by a factor of two and pave the way to identify strategies to further reduce CTN for 3G GWDs. Toward this goal, new amorphous oxide mixtures will be investigated for their potential to reduce CTN when incorporated into mirror coating stacks. The proposed research on amorphous thin films by ion beam sputtering supports the projected development of GWDs. The fundamental materials and coating design aspects of the project will have a broad impact as near-infrared dielectric coatings are ubiquitous in ultra-stable optical cavities and other laser systems. The know-how to be developed is translational, as the deposition method and equipment we will use are standard in the coatings industry. The proposed research will offer a diverse group of students at all levels multiple opportunities to gain an in-depth understanding of the physical mechanisms that affect internal friction in amorphous materials that are the backbone of interference coatings for ultra-high finesse optical cavities and at the same time gain valuable expertise in optical sciences. This interdisciplinary research project will train students in STEM areas, to contribute to advancing science and technology in academic, national laboratory, and industrial settings. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-07

Alzheimer’s disease (AD) is a progressive neurodegenerative disease with a growing humanitarian and economic burden. Advanced age is the greatest risk factor for AD so the long- term goal of this research is to better understand how advancing age contributes to AD occurrence and progression. Aging and AD pathology are characterized by a decline in the cell’s ability to eliminate damaged and pathological proteins through endosome trafficking (NOT- 21-034) which contributes to the accumulation of protein like tau within neurons and other cells of the brain. We have identified a key protein involved in the formation of endosomes, the vehicle for endosome trafficking, that is decreased in neurons across the lifespan and further decreased in neurons from donors with AD. We have shown that we can restore expression of this protein in patient-derived human neurons using adeno-associated virus (AAV) and that this decreases intracellular tau. Based on these findings, the Specific Aims of this proposal are to: 1) determine how increasing endosome formation decreases tau in neurons and 2) define how increasing endosome formation influences cognition and AD pathology in transgenic and aged wild type mice. If successful, these studies will establish decreased endosome formation as a cause of tau pathology in AD and support the development of therapeutic strategies to enhance endosome formation for the treatment of AD and other proteinopathies (i.e., frontotemporal dementia, Parkinson’s disease and others).

NIH Research Projects · FY 2025 · 2024-07

Abstract Down syndrome (DS) is associated with executive function (EF) challenges throughout the lifespan. Although early intervention has the potential for positive downstream effects on adaptation in DS, to date, intervention approaches have not proactively aimed to strengthen EF foundations in ways that capitalize on early neuroplasticity in this population. This R61/33 proposal will refine and test the effects of EXPO (EXecutive Function Play Opportunities), a 12-week caregiver-mediated intervention designed to strengthen EF in young children with DS. EXPO is tailored to the behavioral strengths and challenges associated with DS, thus reducing cognitive and linguistic barriers to EF intervention participation. A preliminary implementation of EXPO during Spring/Summer 2023 has demonstrated promising feasibility and acceptability trends, as well as preliminary efficacy trends at the overall group level. Additionally, heterogeneity in child intervention response was also observed, which likely corresponds to varying degrees of delay in developmental skill acquisition observed in this population during childhood. In line with the growing recognition of the need for personalized and precision approaches to intervention, in this project, we will transform EXPO into an adaptive intervention to better meet the needs of children who are slower to respond to the current intervention design. During the R61 phase, we will incorporate usability and feasibility caregiver feedback and the preliminary efficacy data from the Spring/Summer 2023 trial to refine the structure, organization, and materials of EXPO. We anticipate that we will develop two adaptive EXPO pathways to be subsequently tested and compared: (1) lengthening the duration of EXPO to allow for more practice of each skill, and (2) augmenting EXPO with activities that strengthen play foundations necessary for EXPO engagement (e.g., establishing social routines). During the R33 phase, we will test the EXPO adaptive intervention conditions via a two-arm, Singly Randomized Trial. Findings from this R61/33 project will yield a novel adaptive EF intervention designed to meet a wide range of needs within the population of young children with DS, in preparation for a large Randomized Controlled Trial.

NIH Research Projects · FY 2026 · 2024-07

Project Summary/Abstract Precision environmental health focuses on individual risk assessment to inform targeted disease prevention strate­ gies. Identifying individuals with increased sensitivity to environmental exposures is especially challenging with mixture exposures. The health effects of exposure to mixtures are likely to depend on the composition of the mixture, characteristics specific to the espoused individual including individual­ and neighborhood­level factors, and the developmental stage at which an individual is exposed. We propose to develop statistical methods for precision environmental health with mixture exposures. The proposed methods will estimate mixture­exposure­ response relationships that are individualized based on multiple candidate modifying factors. The framework we develop will allow for data­driven discovery of novel combinations of individual­ and neighborhood­level factors that define susceptible subgroups. We will address three specific data settings. In Aim 1 we propose a general framework for effect heterogeneity using established mixture methods including Bayesian multiple index models. This will include heterogeneous versions of Bayesian kernel machine regression and linear index models. In Aim 2 we develop methods to identify critical windows of susceptibility to mixtures that are assessed longitudinally. The methods will allow for identification of individualized windows of susceptibility to a mixture and estimation of individualized mixture­exposure­time­response functions. In Aim 3 we develop heterogeneous mixture methods for multiple outcomes. The multiple outcome methods will apply to trajectories defined by repeated measures of common endpoint or pathway as well as shared information across multiple related endpoints, such as multiple measures of a common pathway. In Aim 4 we will develop software to implement the methods, along with vignettes and tutorials. We will use the methods developed to analyze air pollution mixtures in a large administrative birth cohort and in a Northeastern United States longitudinal perinatal cohort drawing from multiple source populations. We will estimate individualized mixture­exposure­response functions for birth weight and multiple neurodevelop­ mental endpoints assessed at multiple times. The methods we develop will allow for new avenues of precision environmental health to better identify individuals at increased risk of adverse effects of the environment, which will better inform targeted disease prevention strategies.

NIH Research Projects · FY 2025 · 2024-07

To cure and control Tuberculosis (TB) disease we need shorter, simpler and safer therapies. The goal is to develop therapies of low pill burden; of shorter duration (ideally 2-3 months); with 3-4 drug regimens that avoid resistance and of limited toxicity. Today test regimens are identified in preclinical in vitro and animal models and the most promising multi-drug regimens are subsequently evaluated as a unit in clinical trials. This strategy was used to develop one of the most successful TB regimens so far; the 6moBPaL regimen (a 6-month all oral drug regimen consisting of bedaquiline (B), pretomanid (Pa) and linezolid (L)) Unfortunately, a high rate of adverse events associated with long-term administration of linezolid (a protein synthesis inhibitor) was also reported with this regimen. An option to improve the BPaL regimen is to replace L with spectinamides, another protein synthesis inhibitor without oral bioavailability but an excellent safety profile and with potent efficacy against drug sensitive (DS) and multidrug or extensively drug resistant (MDR-XDR) Mtb strains. The preclinical lead compound 1810 (backup 1599) administered in vivo as injectable, or directly into the lungs via aerosol, demonstrated potent synergistic effects with other oral TB drugs. Moving forward, we propose to develop new 3-4 drug regimens consisting of spectinamide (S) administered via aerosol and combined with BPa (aka: BPaS regimen) and BPaS in combination with pyrazinamide (Z) (aka: BPaSZ regimen). We have shown that 4-weeks of BPaS treatment resulted in similar bactericidal effect to that of the BPaL regimen in BALB/c and C3HeB/FeJ mice. Moreover, the BPaS regimen avoided the myelosuppression and anemia observed in mice treated with BPaL. Our preliminary data also suggests that sterility with BPaS is possible. The studies proposed here test inhalational therapies with spectinamides within new regimens of TB therapy for treatment of DS-TB and MDR-XDR TB. (Aim 1) We had produced small quantities of dry powder 1810 (DP1810) for inhalation and here we will manufacture and physico-chemically characterize larger scale of DP1810 and evaluate their aerodynamic performance for human or animal use. Aim 2 will use pharmacokinetics studies to determine the optimal dose level and dosing frequency for regimens BPaS and BPaSZ. Aim 3, using BALB/c and C3HeB/FeJ TB models will assess the efficacy and sterilization potential of BPaS and BPaSZ regimens. Aim 4 will develop PK/PD simulation-based allometric scaling to aid in human dose projections. Finally, in preparation for the IND preclinical package, Aim 5 will develop non- GLP toxicologic dose range finding studies in rats after inhaled DP1810 administration. Our Multi-PI team led by Dr. Gonzalez-Juarrero at CSU, Dr. Hickey from the RTI International, and Dr. Meibohm at the University of Tennessee will work together with experts in TB drug modeling, TB drug combinations and clinical pathology experts (Drs. Lyons, Robertson and McNeil at CSU respectively), medicinal chemistry (Dr. Lee at St. Jude Children’s Research Hospital) and a world expert physician in TB therapy (Dr. Daley at National Jewish Health) and will receive advice from our partner company for drug commercialization of spectinamides (Microbiotix Inc.).

NIH Research Projects · FY 2025 · 2024-07

Project Summary Culture-based diagnostics for the leprosy-causing agent Mycobacterium leprae are challenged by our inability to culture the pathogen in vitro. Moreover, obtaining a sufficient quantity of bacilli relies on in vivo models such as mice or armadillos in a process that requires significant resources and time because of the slow growth of the pathogen. This extended timeframe, coupled with our inability to engineer recombinant M. leprae strains in vitro, presents significant obstacles to investigating the mechanisms behind its pathogenicity and drug resistance. Furthermore, to thoroughly study M. leprae's epidemiology, transmission, and evolution, it is imperative to sequence M. leprae stains from all clinical and field origins. However, obtaining sufficient bacterial DNA from human and animal samples is challenging. Over the past decade, extensive efforts have been dedicated to enhancing our ability to sequence M. leprae genomes. Currently, two methods are employed for this purpose, but they are limited in either their sensitivity, applicability to different tissue samples, or high cost. As a result, these limitations are currently impeding the widespread use of genome sequencing in leprosy research. We propose to develop a new method based on the selective whole genome amplification (SWGA) of a minuscule quantity of M. leprae directly from complex samples. Our preliminary data on DNA collected from laboratory-infected armadillos shows that SWGA could combine sensitive, accurate, easy-to-implement, and cost-effective characteristics to successfully sequence M. leprae from complex samples. This exploratory project aims to develop the SWGA method and compare it with the gold standard bait capture to validate its use to sequence the genome of M. leprae from infected humans and animals. Aim 1 will evaluate the sensitivity and accuracy of SWGA combined with short-read sequencing and its applicability to long-read sequencing on M. leprae DNA extracted from laboratory and naturally infected armadillo tissues. Aim 2 will generate and validate the necessary reagents to perform SWGA on M. leprae DNA extracted from invasive (skin biopsy) and non-invasive (nasal swab) human samples.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY/ABSTRACT The purpose of the proposed Career Development Award (K01 SERCA) is to provide Dr. Anna Fagre with protected time and collaborative mentorship so that she can build a strong foundation for her career as an independent investigator. Dr. Fagre’s proposed K01 career development training plan builds on >7 years of experience studying bats (order Chiroptera), arthropod vectors, and arboviruses in both the field and the lab. Importantly, it integrates additional training in three focus areas: (1) development of atypical animal models for complex studies involving host-arbovirus-vector interactions, (2) advanced training in multi- omics analysis, and (3) application of comparative approaches in immunology and stress physiology to Chiropteran models of infection and disease. Dr. Fagre’s proposed K01 research outlines the development of a bat model to characterize the impacts of thermal stress on host-arbovirus-vector interactions, incorporating novel datalogging technologies to minimize animal handling while leveraging methodologies used in the fields of stress physiology and environmental health. Dr. Fagre will explore these questions with 3 specific aims: (1) development of a Chiropteran model for assessing innate immunity and inflammasome activation in bats exposed to thermal stress, (2) characterization of ambient temperature’s impact on host-vector interactions in bats following subdermal inoculation with mosquito salivary gland extract, and (3) quantification of ZIKV infection dynamics and host responses at different temperatures in cell lines derived from taxonomically diverse bat species. The anticipated results will contribute to ecoimmunology and ecophysiology studies by expanding our knowledge of how heat stress affects bat immunology and host-virus interactions. In leading these studies, Dr. Fagre will be uniquely poised to build out an independent research program interrogating the transmission dynamics of emerging arboviruses, harnessing a combination of in vivo and in vitro methods supplemented with field-based studies leveraging her past epidemiologic training. Not only will the proposed aims build a framework within which to characterize the impact of environmental stress on host-virus-vector interactions and viral infection outcomes, but results will aid in the identification of valuable diagnostic biomarkers for heat stress and viral infection. The K01 SERCA award would provide Dr. Fagre with continued funding and dedicated time to strengthen her research portfolio and build her own research program before applying for tenure-track faculty positions.

NIH Research Projects · FY 2026 · 2024-07

PROJECT SUMMARY Bronchopulmonary dysplasia (BPD) is the most common morbidity of prematurity with over 10,000 infants diagnosed each year in the United States. Underdeveloped lungs at birth, the need for ventilatory support, which may result in ventilator-induced lung injury, and other factors result in significant structural and functional pulmonary abnormalities associated with short- and long-term lung disease. Effective management includes an individualized approach to choosing the right ventilation strategy and medications. Two important techniques include the use of a long i-time ventilatory strategy and administration of bronchodilators. However, each of these treatments has some associated risk, and there is currently no real-time method to assess their effectiveness on improving ventilation and perfusion. Electrical impedance tomography (EIT) is a noninvasive, non-ionizing real-time functional imaging technique with no harmful side effects, suitable for patients of any age. We hypothesize that EIT images and derived measures can provide real-time, actionable information to guide the clinician's treatment strategy in infants with BPD, and satisfy this unmet need. In this proposal we will carry out the following specific aims: (1) Build a new Adaptive Current Tomography system designed for 3D imaging of very young intubated infants including novel 3-D reconstruction algorithms to provide real-time images of regional ventilation, pulsatile perfusion and quantitative EIT-derived measures. (2) Assess the ability of EIT images and derived measures to determine the effectiveness of long i-time ventilation strategies while they are being applied. An EIT Visualization Platform will be developed to enable the clinician to visualize the images and output measures to assess treatment effectiveness. (3) Determine what combination of EIT- derived measures provides the best assessment of bronchodilator responsiveness (BDR) in infants with or at risk of BPD after the first administration of bronchodilator therapy. We will evaluate the efficacy in Aims 2 and 3 in a study with infants enrolled from the Level IV NICU at Stanford University Medical Center and the University of Colorado Hospital Neonatal Intensive Care Unit.

NIH Research Projects · FY 2026 · 2024-07

Hypoxia is a common feature of gestational complications. Even when the origins of hypoxia vary (i.e., environmental hypoxia versus hypoxia limited to the placenta), there are shared, intermediate features in placental physiology that are also common across mammals. These commonalities suggest that there are some fundamental effects of hypoxia on placental development that negatively impact gestational outcomes. We recently demonstrated that evolutionary adaptations to chronic hypoxia in high elevation environments protects placental and fetal growth in deer mice act on mechanisms shared with humans. These results point to conserved processes that are involved in both mediating the negative impacts of hypoxia on fetal outcomes and preventing these effects. In this proposal, we address three key questions, the answers to which will significantly advance our understanding of how placental responses to hypoxia contribute to adverse gestational outcomes (particularly fetal growth restriction) and the genetic and cell type-specific mechanisms that underlie variation in susceptibility to these complications. First, we ask how individual placental cell type responses hypoxia contribute to tissue-level outcomes, including identifying the cell types that are responsible for tissue-wide transcriptional signatures associated with fetal growth under hypoxia. To answer this first question, we generate cell type-specific transcriptomes from placental tissues using single-nuclei RNAseq from highland-adapted and non-adapted mice experimentally acclimated during gestation to hypoxia or normoxia. Second, we interrogate the cis-regulatory variation that explains cell type-specific transcriptional variation by linking allele-specific gene expression to chromatin conformation in placentas from F1 hybrid crosses of the aforementioned populations, again in an experimental framework. Finally, our third aim asks how these cell type-specific hypoxia responses contribute to organizational remodeling of the placental exchange structures by combining in vivo histological approaches and in vitro experimental approaches focused on cell-autonomous function of a single, important placental cell type. The proposed aims thus combine experimental approaches, cutting-edge sequencing analyses, and molecular and cellular biology with the broader goal of resolving the genetic and developmental processes by which natural genetic variation alters placental function and influences fetal outcomes. The results of this research will advance our understanding of conserved physiology that shapes placental development and pregnancy in mammals, thereby supporting broader research focused on gestational health and disease in humans.

NSF Awards · FY 2024 · 2024-07

This project aims to develop new mathematical theory and statistical tools that will enable monitoring for changes in complex systems, for example global trade networks. Comprehensive databases containing details of trade between almost all countries are available. Detecting in real time a change in the typical pattern of trade and identifying countries where this change takes place is an important problem. This project will provide statistical methods that will allow making decisions about an emergence of an atypical pattern in a complex system in real time with certain theoretical guarantees. The project will also offer multiple interdisciplinary training opportunities for the next generation of statisticians and data scientists. The methodology that will be developed is related to sequential change point detection, but is different because the in-control state is estimated rather than assumed. This requires new theoretical developments because it deals with complex infinite dimensional systems, whereas existing mathematical tools apply only to finite-dimensional systems. Panels of structured functions will be considered and methods for on-line identification of components undergoing change will be devised. All methods will be inferential with controlled probabilities of type I errors. Some of the key aspects of the project can be summarized in the following points. First, statistical theory leading to change point monitoring schemes in infinite dimensional function spaces will be developed. Second, strong approximations valid in Banach spaces will lead to assumptions not encountered in scalar settings and potentially to different threshold functions. Third, for monitoring of random density functions, the above challenges will be addressed in custom metric spaces. Fourth, since random densities are not observable, the effect of estimation will be incorporated. The new methodology will be applied to viral load measurements, investment portfolios, and global trade data. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-06

PROJECT SUMMARY This F32 application is intended to support Dr. Rachel Doser’s postdoctoral research and training, and to effectively prepare her for a career as an independent researcher. Dr. Doser is a new (<6 months) postdoctoral research associate in Dr. Tom LaRocca’s Healthspan Biology Lab at Colorado State University. Under this fellowship, Dr. Doser will investigate a novel contribution of mitochondria-derived double-stranded RNA (mt-dsRNA) to neuroinflammation in aging and Alzheimer’s disease (AD). The rationale for the proposed studies is that dsRNA is a potent activator of innate immune/inflammatory signaling, and mitochondria are a major source of dsRNA, as bidirectional transcription of the circular mitochondrial genome results in the formation of mt-dsRNAs. Although mt-dsRNA-induced immune activation has been shown to contribute to certain cancers and autoimmunity, it has not been studied in the central nervous system, or in the context of aging or age-related neurodegenerative disease. However, mitochondrial dysfunction has been shown to correlate with increased mt-dsRNA formation, and dysfunctional mitochondria are a key hallmark of both aging and AD that may precede neuroinflammation, especially in metabolically demanding neurons. Thus, investigating the downstream effects of mitochondrial dysfunction in neurons due to aging and/or AD will advance our understanding of how aging increases the risk of AD and neurodegeneration in general. The preliminary data presented in this proposal suggest that mt-dsRNAs are released with older age and in AD, and that they correlate with increased expression of dsRNA immune sensors. Therefore, the proposed research is designed to test the hypothesis that age/AD-related mitochondrial dysfunction leads to release of mt-dsRNAs that instigate an innate immune response. First (Aim 1), Dr. Doser will use both in vitro and in vivo organismal models to determine if age/AD-related mitochondrial dysfunction contributes to mt- dsRNA release. Then (Aim 2), she will use similar approaches to determine if mt-dsRNAs cause neuroinflammation. Finally (Aim 3), Dr. Doser will investigate if /how mt-dsRNA levels correlate with cognitive function or AD pathology in humans. Together, these aims will detail the hypothesized mechanism linking age/AD-related mitochondrial dysfunction to neuroinflammation and assess its relevance to human health/function, and they may identify novel targets for preventing or limiting inflammation. The proposed studies will provide Dr. Doser with extensive new training in translational research using multiple models. She will leverage and expand her current skillset, including by learning new cutting-edge techniques such as bioinformatics, human neuronal cell culture and reprogramming, measurements of mitochondrial function and clinical/data science. The relevant expertise of Dr. Doser’s sponsor, Dr. LaRocca, and consulting mentors Drs. Karyn Hamilton and Chris Link, will support successful completion of the proposed aims and provide Dr. Doser with an exceptional, interdisciplinary and translational research training experience.

NIH Research Projects · FY 2026 · 2024-05

PROJECT SUMMARY This proposal is to develop a generalizable method to make genetically-encoded biosensors to monitor the dynamics, abundance, and positions of specific post-translational modifications (PTMs) of proteins. The strategy is based on our previous success in generating avidity-driven biosensors for distinct types of ubiquitin (Ub)- modified nucleosomes; each sensor is comprised of a nucleosome-recognizing Anchor, a Ub-binding UBD domain, and a Linker developed to maximize affinity and specificity. For Ub-modified proteins in particular, development of antibodies or other site-specific detection reagents has been notoriously difficult. Our goal is to extend the avidity-based strategy to engineer sensors able to detect heretofore intractable molecular targets such as the multiple and functionally distinct Ub signals found on nucleosomes. To accomplish this, our team will develop rational protein design technology that embraces multi-valent binding with tunable molecular flexibility. In Aim 1, we will design and test Linkers that deliver tunable domain geometry and motion. We will adapt new machine-learning algorithms to design fusion proteins that fix constituent Anchor and UBD domains in conformational space to maximize avidity and specificity. Furthermore, we will explore two new approaches to expand the Anchor repertoire and broaden applicability of the avidity strategy. In Aim 2, we will adopt the splitFAST fluorogen system to install a transferrable Anchor-binding tag on the substrate, and In Aim 3 we will develop customized Anchors from a yeast-display DARPin library. The efficacy and utility of the new sensors will be evaluated in vitro and in cells where they will be used to probe signaling associated with DNA damage repair pathways. In Aim 4, we will test and optimize conditions to use the sensors in genomic applications such as CUT&RUN assays. The reagents we develop in this project will allow researchers to probe otherwise invisible live-cell processes that are difficult or impossible to image with existing technologies. Our innovative approach directly addresses the challenge of binding to a highly flexible multi-domain protein target. As such, the resulting technology and design workflow will find application for diverse ubiquitinated targets, and more generally for binding targets that would otherwise be inaccessible due to high flexibility.

NIH Research Projects · FY 2026 · 2024-04

Project Summary – Burkina Faso ICEMR Overall Project This project will establish the Burkina Faso ICEMR, intended to bring together local and international experts in the host, parasite, and vector, under one umbrella to define the spatiotemporal heterogeneity of malaria across the Sudan, Sudan-Sahel, and Sahelian zones of Burkina Faso and within differing landscape environments (urban, rural, and migrant/gold-mining camps). In this process, parasite (species and genetics) and host data (including age and clinical status) will be tightly linked to the complexity of vector species (primary and secondary), their locations near transient or permanent water sources, and translational studies of vector competence, transmission dynamics, and drug and insecticide resistance. The malariology experts in this ICEMR are long-term collaborators and comprise field and lab scientists, clinicians, biostatisticians, and data specialists at the four key institutions, Institut de Recherche en Sciences de la Santé, Institut des Sciences et Techniques, Colorado State University and Yale School of Public Health, as well as at partner institutions inside Burkina Faso and the U.S. Together they will form an Administrative Core to provide organizational capacity for the entire project, to manage budgets, facilitate the science, enhance communication within and outside of the project, and foster capacity building among all partners. We will also form a Data Management Core, which will provide infrastructure and oversight for the coordination of data collection and analysis occurring across ICEMR institutions in Burkina Faso and in the U.S. And we will initiate two interconnected research projects that will seek to reassess malaria epidemiology in the country. The first project will characterize the epidemiology of all human species across study sites via testing of mosquito blood meals and conducting longitudinal household-based cohorts over three years. The goal of these studies will be to carefully characterize the epidemiology and clinical impact of mono- and mixed species infections and to understand the performance of current diagnostics for detecting the symptomatic and asymptomatic reservoir of infection. We will also utilize samples to characterize their ex vivo and genetic resistance profiles to current and promising antimalarials. The second project will assess the natural vector bionomics and parasite transmission across spatiotemporal gradients, perform laboratory experiments of parasite transmission to validate our field findings using wild type mosquitoes and parasites, and also expose these infected mosquitoes to sub-lethal insecticide concentrations or drugs that the vectors are likely to encounter during the extrinsic incubation period in the field to determine if this influences parasite transmission. With the data from these projects and support from the Cores, we ultimately hope to inform country- wide policies, and build the local skill set and resources to sustainably advance malaria control across the country and identify specific strategies to help better control malaria in the greater West African region.

NIH Research Projects · FY 2026 · 2024-03

Summary Cardiovascular disease is the leading cause of death worldwide and psychosocial stress is a significant predictor of disease incidence and severity. However, the specific neurobiological mechanisms that mediate the cardiovascular consequences of stress are largely unknown. Therefore, the current proposal will determine how the cortical circuits responsible for cognitive appraisal of stress regulate physiological stress responses. Recent studies in rats identified a population of cells in the infralimbic (IL) region of the medial prefrontal cortex that integrate endocrine and autonomic responses to stress in a sexually-divergent manner. Further, stimulation of IL glutamate neurons and chronic stress interact sex-specifically to affect cardiac structure and function. While the activity of excitatory IL projection neurons is critical for regulating the deleterious effects of chronic stress, the pathways used by these cells to modulate reactivity of autonomic and endocrine systems remain to be determined. Preliminary data indicate that IL projections to the posterior hypothalamus (PH) reduce male stress responses but enhance female stress responses. These excitatory IL projections target both inhibitory GABAergic and excitatory glutamatergic neurons in the male and female PH. Further, chronic variable stress upregulates PH gene expression related to glutamate and GABA signaling in males but not females. Altogether, these findings led to the hypothesis that sex-specific IL glutamatergic signaling to the PH differentially engages glutamatergic and GABAergic cells to regulate cardiovascular and endocrine stress reactivity, as well as the consequences of chronic stress on vascular stiffness and cardiac hypertrophy. This hypothesis will be tested in 3 aims. First, circuit signaling will be examined with patch-clamp slice electrophysiology in male and female rats. Specific experiments will determine how IL glutamate signaling targets genetically-defined postsynaptic PH GABAergic and glutamatergic cells as well as chronic stress- induced circuit plasticity. Second, in vivo optogenetic activation of IL synapses in the PH following chronic variable stress will interface with measures of heart rate, blood pressure, and neuroendocrine stress responses. This approach will isolate circuit effects to normalize or exacerbate stress reactivity. Third, a combinatorial viral approach will be used to retrogradely inhibit IL projections to the PH with Cre-dependent tetanus toxin. In addition to cardiovascular telemetry and hypothalamic-pituitary-adrenal hormones, cardiac hypertrophy and microvascular function will be assessed in chronically-stressed male and female rats to determine circuit regulation of stress-induced cardiac and vascular dysfunction. Collectively, these experiments will determine specific circuit and cellular pathways linking cognitive appraisal with physiological stress responses. Further, this investigation is poised to not only increase understanding of brain-body interactions and sex-based health disparities, but also identify potential targets to mitigate cardiovascular risk.

NIH Research Projects · FY 2026 · 2024-03

Nontuberculous mycobacteria (NTM) are environmental bacteria that cause serious pulmonary infections in people with underlying lung diseases. One of the most common NTMs associated with pulmonary disease is Mycobacterium abscessus (Mabs). Mabs is highly drug resistant, and it is the most difficult NTM to treat. The therapeutic use of bacteriophage (phage), which are viruses that infect and kill bacteria, is a potential alternative to antibiotics for treating drug resistant bacteria including Mabs. Recent compassionate use cases where NTM disease was treated with phage, highlight the potential for phage to be a new therapeutic option for Mabs infection. At the same time, the recent cases emphasize the need to better understand and improve the approach of using phage to treat Mabs infection. Phage dosing in the recent Mabs treatment cases was performed without knowledge of in vivo phage exposure over time (pharmacokinetics, PK) or the impact of phage on the Mabs burden (pharmacodynamics, PD). For any bacterial infection, knowledge of phage PK/PD, which differs from traditional small molecule PK/PD, is minimal. This R01 proposal addresses this significant gap in knowledge. Using a Mabs isolate and phage from recent clinical cases and a mouse lung infection model, we will perform the first in vivo PK/PD studies of phage therapy to treat Mabs infection. Time course PK/PD data, along with pathology, spatial analysis of phage and Mabs localization in the lung, and respiratory function data, will be used to develop an in silico quantitative systems pharmacology (QSP) model that describes in vivo interactions between phage, Mabs, and the host. With the aid of phage PK/PD knowledge we will also systematically investigate different phage dosing strategies to optimize phage treatment and improve therapeutic outcome. In Aim 1, we will evaluate PK/PD of phage delivered via the intravenous (IV) route or intratracheal (IT) route. In Aim 2, we will evaluate the PK/PD of liposome-encapsulated phage. Liposome encapsulation may help target phage to macrophages, which serve as an intracellular niche for Mabs, and it may shield phage from host clearance or inactivation. In Aim 3, we will evaluate the effect on phage PK/PD of combining phage with antibiotic drug treatment. The results of these Aims will provide mechanistic insights into interactions between phage, Mabs, host, and antibiotic and they will support our long-term objective of using QSP modeling to promote translation of phage therapy for Mabs infection to successful use in the clinic.

NIH Research Projects · FY 2026 · 2024-02

PROJECT SUMMARY/ABSTRACT There is growing recognition that timing of behaviors, such as eating, sleeping, and physical activity, have a significant impact on human health and disease risk. For example, when people are awake at the “wrong” time of the day (i.e. during the biological night), a mismatch occurs between behavior and biology, termed circadian misalignment. Shift workers experience repeated bouts of circadian misalignment and are at higher risk of cardiometabolic disease compared to people who work days. However, the mechanism(s) by which shift work and associated circadian misalignment increase disease risk are not clear. As a result, there have been few attempts to develop strategies to improve health in shift workers. Data from our group and others demonstrate that sleep and circadian disruption impair vascular endothelial function and insulin sensitivity, two important risk factors for future development of cardiovascular disease (CVD) and Type 2 diabetes (T2D), together termed cardiometabolic disease. Furthermore, sleep and circadian disruption increase lipids in plasma, including free fatty acids and ceramides, a class of lipids associated with cardiometabolic disease risk. We also find increases in specific lipid species in muscle, such as 1,2 diacylglycerols during circadian disruption, which can directly impair tissue-specific insulin signaling. Lastly, plasma lipids are elevated in shift workers; however, strategies that specifically target lipids in shift workers have not yet been conducted. Using a circadian-based eating model (time-restricted eating; TRE), we and others can consistently reduce lipids in circulation. TRE is also associated with reduced blood pressure and improved glucose homeostasis in healthy, lean adults during inpatient conditions of simulated night shift work. Thus, the clear next step is to translate this strategy to improve cardiometabolic disease risk in free-living shift workers who are neither young nor lean. The overall objective for this project is to improve the cardiometabolic health of free-living shift workers. Our central hypothesis is that eliminating food intake from the biological night (via TRE) will improve blood pressure, vascular function, insulin sensitivity, and glucose homeostasis in shift workers. To test our hypothesis, we will conduct a randomized crossover study (4-week TRE vs. 4- week controlled eating) in 50 non-rotating night shift workers (25F; >18y) with overweight or obesity. At the end of each outpatient condition, we will conduct a rigorous 2-day inpatient assessment to determine the impact of TRE and associated reductions in plasma and muscle lipids on blood pressure, vascular function, circulating markers of endothelial health, whole body and muscle-specific insulin sensitivity, whole body glucose homeostasis, and muscle insulin signaling. Successful completion of the proposed study will identify a targetable mechanism to counter impairments in vascular function and insulin sensitivity during shift work. The knowledge to be gained supports cost-effective programs to minimize CVD and T2D in populations at elevated risk, including anyone working nonstandard hours. These populations include workers in healthcare (nurses, doctors, paramedics), emergency services (military personnel, police, firefighters), security, transportation (pilots and truck drivers), manufacturing (our commercial partner), and hospitality, as well as the many individuals with sleep and circadian disorders.

NIH Research Projects · FY 2025 · 2024-02

PROJECT SUMMARY This proposal outlines a training program for independent scientist with a focus on mRNA localization using genetics and molecular biology. The research outlined in this proposal will develop a platform to evaluate local translation, a process of whose disruption results in many neurological diseases and cancer. The phenomena directing local translation to axon termini and the Endoplasmic Reticulum (ER) have been described. Scientists have recently made the novel discovery that local translation also occurs at plasma membranes. For example, in Caenorhabditis elegans, ezrin/radixin/moesin (erm-1) mRNA localizes to plasma membranes with its encoded protein, a plasma membrane-actin cytoskeleton linker that will coordinate cell shape changes. erm- 1/ERM-1 undergoes translation-dependent localization and local translation directed by its N-terminal encoded FERM domain. However, neither the mechanisms directing their localization nor the reasons for their local translation are understood. The proposed research aims to use erm-1 as a model to understand how mRNA localization to the plasma membrane arises mechanistically, functionally links to protein production, and impacts gene expression. Based on preliminary evidence, the hypothesis is that cytoskeletal components interact with the translating complex to direct it to the plasma membrane, similar to co-translational transport of secretory proteins to the ER. This model will be explored from a genetics and molecular biology perspective with 3 specific aims. The first aim will test whether erm-1/ERM-1 localization occurs through diffusion or directed transport mechanisms and explore which cytoskeletal components are required. The second aim will develop a live imaging tool to better resolve the kinetics of erm-1 translation. The third aim will identify the effector proteins involved in this process. Since impaired mRNA localization in neurons and other cell types causes disease but studying mRNA localization in disease-specific models is challenging, this project will achieve our long-term goal of characterizing novel mRNA transport pathways that have the potential to be generalizable in human health.

NIH Research Projects · FY 2026 · 2024-02

Project Summary Lauren Shomaker, Ph.D., is a licensed clinical psychologist and clinical researcher with a strong track record of NIH grantsmanship, scholarship, and commitment to mentoring in patient-oriented research (POR) at the intersection of mental health and cardiometabolic health. Dr. Shomaker's POR program is embedded in developmental and prevention science frameworks, with a particular focus on intervening at sensitive windows such as adolescence that are ripe with opportunity for altering lifecourse trajectories of mental/behavioral health and cardiovascular disease (CVD) risk. In particular, her work centers on developing more targeted approaches than “one-size-fits-all” lifestyle approaches for type 2 diabetes (T2D) and CVD prevention, through intervening with underlying, stress-related risk factors, especially in adolescents facing heightened social adversity. Dr. Shomaker's current, key projects as PI (R01DK132557, U/R01AT011008, R01DK111604, USDA2022- 4152037651): (1) Span the behavioral trials spectrum, (2) Are single and multisite, (3) Occur in diverse settings, (4) Address social/psychological/behavioral underpinnings of T2D/CVD risk, and (5) Have a common thread of applying an interdisciplinary, team science approach to mental health and T2D/CVD prevention science. Dr. Shomaker's long-term career goals are to lead a cutting-edge POR program in mental/behavioral health and T2D/CVD prevention that has sustained funding and effective partnerships, and to support a legacy of scientists who will optimize feasible, acceptable, effective, and sustainable interventions for mental/behavioral health and T2D/CVD prevention, to reduce population-level health disparities. An NHLBI K24 Midcareer Investigator Award in POR provides the ideal mechanism to alleviate her clinical, teaching, and service responsibilities, and allow Dr. Shomaker to augment her POR capabilities by filling two gaps, dissemination and implementation (D&I) science and computer/information technology (CIT) tools for interventions, both of which are pivotal for advancing the impact and reach of her work. The K24 also would provide an opportunity to obtain new, formalized training in mentoring, with a particular emphasis on diversity, equity, and inclusion, to bolster inclusive excellence in mentoring diverse, beginning clinical investigators. The K24 training aims, to gain formalized knowledge/skills in (1) D&I science, (2) application of CIT tools to T2D/DVD preventative interventions, and (3) mentoring skills, are supported by an exceptional midcareer mentorship and advisory team (D&I science: Mentor Bethany Kwan, PhD, Co-mentors Russell Glasgow, PhD, Jesse Owen, PhD; CIT tools: Mentor Joshua Smyth, PhD; Mentorship: Mentor Randi Streisand, PhD, Co-mentor Stanley Szefler, MD; Advisors: Matthew Hickey, PhD, Paul MacLean, PhD). This K24 proposal leverages the institutional resources of Colorado State University, University of Colorado Anschutz, and the CTSA-supported Colorado Clinical & Translational Sciences Institute. Specific aims are to (1) Augment expertise in D&I science and CIT intervention supplements, and (2) Provide skillsful mentoring to beginning clinical investigators in the conduct of POR in mental/behavioral health and T2D/CVD prevention.

NIH Research Projects · FY 2026 · 2023-12

Project Summary Cardiovascular diseases are the leading cause of death globally and pathologies are exacerbated by chronic stress. Stress-cardiovascular comorbidities are more prevalent in females, however, the neurobiological mechanisms linking stress to cardiovascular outcomes are not well understood and could inform underlying cardiovascular susceptibility and resilience. The prefrontal cortex and hypothalamus are key regulators of stress and cardiovascular output; therefore, this proposal will test the hypothesis that cortical- hypothalamic neural circuitry mediates the sexually divergent cardiovascular consequences of chronic stress. Chronic psychosocial stress increases the incidence of cardiovascular diseases. The prefrontal cortex (PFC) is critical for stress appraisal, and both mood disorders and chronic stress are associated with altered PFC function. Recently, the sponsor’s lab reported that optogenetically activating the predominantly glutamatergic projection neurons from the rat infralimbic PFC (IL) produces sexually divergent effects on the stress response. However, the IL does not directly innervate neuroendocrine or preganglionic sympathetic neurons to mediate these effects; therefore, intermediate neurocircuitry must be involved in translating sex- specific cortical processing into sex-specific cardiovascular outcomes. The posterior hypothalamus (PH) is a major target of IL projection neurons and IL-PH projection neurons are stress-activated, however, neuroanatomical studies examining IL-PH circuitry have been performed exclusively in males. Pharmacological inactivation of the PH in vivo restrains acute stress responses while pharmacologically activating the PH exacerbates acute stress responses. PH activity also regulates cardiovascular function: pharmacologically activating the PH increases blood pressure and heart rate and pharmacological inactivation of the PH robustly blocks stress-induced increases in heart rate. The proposed experiments hypothesize that the PH is an intermediate synapse for sexually divergent outcomes and that stress-induced plasticity in this region mediates cardiovascular consequences of chronic stress. To test this hypothesis, I will learn viral-mediated circuit- and cell-type-specific slice electrophysiology to investigate synaptic plasticity within the IL-PH circuit after chronic variable stress in males and females. Additionally, I will learn pulse wave velocity and pressure myography to investigate vascular stiffness and reactivity in vivo and ex vivo, respectively, to determine the necessity of the IL-PH circuit for the detrimental consequences of chronic stress on vascular function. I have assembled a mentorship team with accomplished neurophysiologists and cardiovascular biologists to ensure the technical and career development training necessary for this project and, ultimately, starting my independent laboratory. These mentors will be critical for navigating my postdoctoral fellowship and preparing to run an independent research program studying unique dimensions of cardiovascular resilience.

NIH Research Projects · FY 2025 · 2023-11

Project Summary / Abstract COVID-19 is caused by a bat-borne coronavirus, SARS-CoV-2, that has resulted in more than 1 million deaths in the USA. Experimental and bioinformatic analyses suggest cricetid rodents, but not murid rodents, may be susceptible to the virus. We determined that two such cricetid rodents, the North American deer mouse (Peromyscus maniculatus) and California deer mouse (P. californicus) are susceptible, with some of the latter species developing severe disease that required euthanasia. This raises the concern that spillback from humans or susceptible domesticated animals, such as mins (Neogale vison), to North American cricetid rodents could occur and lead to establishment of SARS-CoV-2 in secondary reservoir hosts in the New World. Several rodent species in Europe and Asia have been found to harbor coronaviruses but, surprisingly, there are no reports wild rodents in the New World have been examined for coronaviruses, even though they are found in several bat species. Moreover, because of disease that occurred in California deer mice, it is possible that it or other North American rodents could serve as new pathogenesis models for COVID-19. To examine these possibilities, we will survey cricetid rodents in Colorado for the presence of coronaviruses, which we have detected at a site in Utah, and generate genome sequences of these viruses for phylogenetic analysis and important domains (e.g., protease cleavage sites). We will also determine the T cell response of deer mice vaccinated against SARS-CoV-2 using single-cell RNA seq analysis and compared to unvaccinated deer mice. This will lay the foundation of using deer mice as a long-lived small animal model (8 years), which cannot be replicated with Syrian hamsters or human ACE2 laboratory mice (2 years). Together this work will determine coronavirus diversity in North American rodents, and whether the deer mouse can serve as a durable immunity model for SARS-CoV-2.

NIH Research Projects · FY 2026 · 2023-11

Many species depend on social activity to thrive, and social impairment is a fundamental symptom of several mental diseases. According to research, synaptic signals and activity can control social behavior. Yet, it is still unclear how synapse control and social behavior are related. δ-catenin functions as an anchor for the glutamatergic AMPA receptor (AMPARs) to regulate synaptic activity in excitatory synapses. Several families with autism have been identified to have mutations in the δ-catenin gene, which results in a loss of δ-catenin functions at excitatory synapses and is thought to be the etiology of autism in people. Recent studies, including our own, suggest that the loss of δ-catenin functions significantly decreases cortical neurons' inhibition to increase excitation. Our new data further reveal that δ-catenin deficiency disrupts social behavior in mice. These new findings strongly support the scientific premise that δ-catenin is critical for glutamatergic synaptic activity and social behavior. Nonetheless, the δ-catenin-mediated link of synaptic, cellular, and neural substrates to social behavior is understudied. The significance of the current proposal is thus predicated on filling this gap. Postsynaptic glutamatergic activity in excitatory and inhibitory cells in the medial prefrontal cortex (mPFC) regulates cellular excitation and inhibition to control prefrontal rhythmic activity patterns, which is essential in the control of social behavior in humans and animals. However prior research on the neural basis of social behavior has mostly concentrated on top-down mPFC projections to various subcortical regions. Hence, in these earlier studies, glutamatergic inputs to prefrontal neurons were mainly disregarded. Therefore, it is unclear how the postsynaptic glutamatergic activity of prefrontal cells relates to social behavior. Research suggests that parvalbumin-positive (PV+) inhibitory interneurons in the mPFC receive direct glutamatergic inputs from multiple subcortical regions to provide feedforward inhibition onto prefrontal pyramidal neurons and control network activity. This is important for the regulation of social behavior. Our preliminary data suggest that δ-catenin deficiency impairs prefrontal neural activity at cellular and network levels to induce social deficits. Therefore, existing data and our new findings lead us to hypothesize that δ-catenin is important for glutamatergic activity in PV+ inhibitory interneurons to provide feedforward inhibition onto pyramidal neurons in the mPFC, which regulates prefrontal activity at the cellular and network levels to ensure normal social behavior. We will employ two new mouse models of δ-catenin deficiency - δ-catenin KO mice and human ASD-associated δ-catenin glycine 34 to serine (G34S) mutant mice – to examine if δ-catenin controls pathway-specific glutamatergic activity in PV+ interneurons and feedforward inhibition onto pyramidal neurons in the mPFC (Aim 1), to determine whether δ-catenin deficiency disrupts prefrontal network activity during social interaction (Aim 2), and to address if altering prefrontal δ-catenin expression is sufficient to affect social behavior (Aim 3).

NIH Research Projects · FY 2024 · 2023-09

Modified Project Summary/Abstract Section Project Summary / Abstract Bats are reservoirs, or suspected reservoirs, of many zoonotic viruses, including SARS, SARS2 and MERS coronaviruses, Nipah and Hendra viruses, and Ebola and Marburg viruses. Little is known about how these viruses circulate in their bat reservoirs, principally because of a lack of bat colonies that can be used for the development of experimental infection models. To address this deficiency, we will capture horseshoe bats and Indian flying foxes, respective reservoir hosts of Nipah virus and SARS-related coronaviruses, in Bangladesh where they will be quarantined and provided veterinary care as they adapt to captivity. Bats will be shipped to CSU to establish the breeding colonies as a resource for investigators who study these viruses. We will generate primary cell cultures and immortalized cell lines from various tissues and freeze live bone marrow that will be useful for studying how these viruses infect bat cells, and how the viruses may modulate the innate immune responses. Recombinant cytokines will also be produced for the research community, including those for generating macrophages and dendritic cells (GM-CSF, Flt3L), T cells (IL-2) and for in vivo modulation of the adaptive immune response (IFNγ, IL-4). Moreover, we will generate monoclonal antibodies for use in cytokine detection assays and flow cytometry of immune cell subsets and in vivo neutralization. Finally, we will perform experimental infection studies of Nipah virus, SARS-CoV-2 and the SARS-related coronaviruses, BANAL-52 and BANAL-236, to study the infection kinetics, virus distribution and transcriptomic, proteomic and metabolomic profiles of bats during infection, and escalation and resolution of the immune response. Tissues, cells and sera from naïve and infected bats will be archived in a biobank that will be made available to the research community. The establishment of this resource will lead to a better understanding of how bats host highly pathogenic viruses without disease and may shed light on events that increase spillover risks to humans. In turn, this information could lead to development of mitigation strategies to prevent future virus spillover and uncover new strategies for therapeutic treatment of coronavirus and Nipah virus diseases.

NIH Research Projects · FY 2025 · 2023-09

Bats are reservoirs, or suspected reservoirs, of many zoonotic viruses, including SARS, SARS2 and MERS coronaviruses, and Ebola, Sudan and Marburg viruses. Little is known about how these viruses circulate in their bat reservoirs, principally because of a lack of bat colonies that can be used for the development of experimental infection models. To address this deficiency, we have established a breeding colony of Jamaican fruit bats (Artibeus jamaicensis) and will establish a breeding colony of Egyptian fruit bats (Rousettus aegyptiacus) as a resource for investigators who study these viruses. Egyptian fruit bats are the principal natural reservoir of Marburg virus and Sosuga virus, both of which are human pathogens. Jamaican fruit bats are the best studied bat model for infectious diseases and we have demonstrated that they are a model organism for Ebola and Marburg virus infections. We will generate primary cell cultures and immortalized cell lines from various tissues from these species and freeze live bone marrow that will be useful for studying how these viruses infect bat cells, and how the viruses may modulate the innate immune responses. Recombinant cytokines will also be produced for the research community, including those for generating macrophages and dendritic cells (GM-CSF, Flt3L), T cells (IL-2, IL-7, IL-15) and for in vivo modulation of the adaptive immune response (IFNγ, IL-4). Moreover, we will generate monoclonal antibodies for use in cytokine detection assays and flow cytometry of immune cell subsets and in vivo neutralization. Finally, we will perform experimental infection studies of with ebolaviruses, Sosuga virus, and the SARS- related coronaviruses, BANAL-52 and BANAL-236, to study the infection kinetics, virus distribution and transcriptomic, proteomic and metabolomic profiles of bats during infection, and escalation and resolution of the immune response. Tissues, cells and sera from naïve and infected bats will be archived in a biobank that will be made available to the research community upon virus inactivation. The establishment of this resource will lead to a better understanding of how bats host highly pathogenic viruses without disease and may shed light on events that increase spillover risks to humans. In turn, this information could lead to development of mitigation strategies to prevent future virus spillover and uncover new strategies for therapeutic treatment of coronavirus and Nipah virus diseases.