University Of Iowa

NIH Research Projects · FY 2026 · 2024-01

Summary Thrombotic events such as myocardial infarction, stroke and deep vein thrombosis are the leading causes of morbidity and mortality in the elderly. The objective of this proposal is to test the usefulness of a calorie-restriction (CR) intervention to diminish prothrombotic effects in aged mice or aged mice with type 2 diabetes and identify a druggable target. Our pioneering studies have established that increased thrombotic susceptibility in aging is associated with platelet hyperactivity due to excessive accumulation of oxidants or the reactive oxygen species. We also observed that the loss in mitochondrial superoxide-dismutase (SOD2) activity within platelets increases mitochondrial-oxidant burden in aged mice and enhances platelet activation and susceptibility to arterial thrombosis. Our novel pilot data shows that sirtuin 3 (SIRT3), a mitochondrial-deacetylase that regulates the activity of several mitochondrial-proteins including SOD2 is markedly decreased in platelets from aged mice and humans. Consistent with these observations, in an ongoing clinical study, we observed that increased platelet activation in healthy older humans and obese-diabetics was overcome by pre-incubation with SIRT3 activator or a SOD2 mimetic, suggesting a Mito-oxidant dependent mechanisms. Recently, we also reported that the increased potential for venous thrombosis during aging is mediated via increased total cell-free DNA and this increase is likely due to elevation in mitochondrial DNA. The damaged circulating mitochondria with cardiolipin surface may also enhance thrombin generation potential and venous thrombosis. So, a major goal of this proposal is to determine a role of damaged-mitochondria that mediates age-associated arterial and venous thrombosis and identify preventive measures. In search for a mitochondria-based protective mechanisms, we conducted a pilot study and employed a 30% CR in aged mice or aged mice with type 2 diabetes for a month. This 30% CR increased platelet-SIRT3, decreased damage in mitochondria, decreased platelet activation and lowered thrombotic susceptibility. We hypothesize that CR protects from age-associated arterial- and venous thrombosis through distinct mitochondrial mechanisms. We will also test that the presence of insulin resistance or type two diabetes exacerbates mitochondrial damage and enhances thrombosis which is also preventable with CR. Aim 1 will focus on platelet SIRT3 activation as a protective mechanism of CR for arterial thrombosis. Aim 2 will test whether CR prevents from circulating damaged- mitochondria and mitochondrial-DNA and lowers the susceptibility to venous thrombosis. Thus, the significance of this proposal is in establishing usefulness of a healthy life-style-intervention to diminish prothrombotic effects in aging and age-associated comorbidity and identify a druggable target downstream to CR for translational application.

NIH Research Projects · FY 2025 · 2024-01

ABSTRACT Glaucoma is the leading cause of irreversible blindness characterize by loss of retinal ganglion cells (RGCs). High intraocular pressure (IOP) increases risk for glaucoma, but glaucoma often occurs at low IOP (i.e., normal tension glaucoma (NTG)). As all current glaucoma therapies target high IOP, there is a critical need for new therapies for NTG that occurs at already low IOPs. We have researched the causes and cures for NTG by studying single-gene forms of disease. We discovered that duplication of the TBK1 gene causes NTG (1 of 3 known single-gene causes). TBK1 encodes a kinase that phosphorylates an autophagy receptor OPTN (which is encoded by another NTG gene) as a key step in stimulating autophagy, a ubiquitous catabolic cellular process that degrades defective organelles (mitochondria) and protein aggregates. We studied fibroblast cells from our NTG patients with TBK1 gene duplications and engineered transgenic mice with an extra dose of the TBK1 gene and showed that these mutations cause glaucoma by increasing TBK1 activity in RGCs (increased transcription, protein abundance, and phosphorylation of OPTN). Moreover, we and others have shown that TBK1 and OPTN mutations that cause NTG are also associated with dysregulation of autophagy. Mitochondria are strongly implicated as a chief target of dysregulated autophagy (mitophagy) in glaucoma. Consequently, we hypothesize that TBK1 gene duplications cause RGC death and glaucoma by dysregulating autophagy and maintenance of mitochondrial function. We plan to test this hypothesis, by creating and analyzing induced pluripotent stem cell (iPSC)-derived retinal ganglion cells with TBK1 gene duplications and their isogenic controls. We propose the following 3-part aim. AIM 1: Determine the influence of TBK1 gene duplication on RGC health and function using NTG patient cell culture, stem cell biology, and genome editing. A. Use CRISPR to correct TBK1 gene duplications in 3 iPSC lines from 3 NTG patients (iPSC lines are in hand). B. Differentiate iPSC lines into RGCs using our 3D retinal organoid protocol (3 mutant and 3 isogenic controls) C. Characterize differences in transcription, autophagy, and mitochondrial function between iPSC-derived RGC lines and isogenic controls using Seahorse, scRNAseq, and autophagy assays. We have already created iPSC lines from 3 NTG patients with TBK1 duplications and have extensive experience with genome editing, iPSC biology, RGC differentiation, and autophagy/mitochondrial assays to facilitate completion of this proposal. We have also published pilot studies of a single iPSC line with TBK1 mutations. TBK1 is at the center of the pathophysiology of a proportion of NTG cases. We have proposed experiments that will gain insight into the mechanisms by which TBK1 mutations cause disease and will facilitate future research to design and test new treatments for NTG patients who already have low IOP.

NIH Research Projects · FY 2026 · 2024-01

PROJECT SUMMARY/ABSTRACT Incidence of heart failure (HF) remains on the rise in the United States despite advances in treatment and prevention. On a cellular level during HF, excitation-contraction coupling and calcium signaling in the heart becomes severely impaired. These mechanisms occur in junctional membrane complexes (JMCs) or cardiac dyads in cardiomyocytes. Cardiac JMCs are the ultrastructure juxtaposed between the transverse tubules (T- tubules) and the sarcoplasmic reticulum (SR) and are integral for facilitating excitation-contraction coupling and calcium signaling. Junctophilin family proteins (JPs) are critical in establishing and modulating JMCs. There are four isoforms of JPs found in excitable tissues (JP1-4). While JP2 has been established as the primary JP isoform expressed in cardiac muscle, our lab has recently found that JP1, normally in skeletal muscle, is also expressed in cardiac muscle. Through examining mouse cardiac tissue, I found that JP1 protein is expressed primarily in the ventricles of the heart and increases during postnatal development until maturity. I have also found that JP1 localizes with JP2 and RyR2, the primary calcium release channel, in the Z-disc. Using a cardiomyocyte specific JP1 knockout mouse model, I found that JP1 knockout mice have a shortened lifespan, reduced cardiac function, and increased incidence of fibrosis and HF compared to control mice. Despite genetic ablation of JP1 in these mice, JP2 protein expression does not change indicating that JP2 is not sufficient to overcome the loss of JP1. To examine whether JP1 expression changes during cardiac stress I used a model of pressure overload-induced cardiac hypertrophy in mice and found that JP1 protein expression decreases with increased cardiac stress. To investigate whether this downregulation could be due to proteolysis by calpain, a stress induced, calcium-dependent protease, I subjected purified recombinant JP1 protein to an in vitro calpain cleavage assay and found JP1 cleaved between amino acid 505 and 510. My primary hypothesis is that JP1 is functionally distinct from JP2 in cardiomyocytes within the cardiac dyad at baseline and in the nucleus after stress-induced cleavage. To test the hypothesis, I will use two novel mouse models developed in our lab: a cardiomyocyte specific knockout of JP1 (JP1cKO), and a global knock-in of 3xHA-tagged JP1. In Aim 1, I plan to examine the functional role of JP1 in cardiac muscle by (1) determining the consequences of depleting JP1 from JMCs by analyzing cardiomyocytes from JP1cKO mice and (2) examining JP1-interacting molecular targets and the potential signaling pathways using proteomics-based approaches and RNA-sequencing. Aim 2 will explore the mechanism of downregulation of JP1 during cardiac stress by further investigating the molecular consequence of calpain proteolysis of JP1. This work will have broad implications for the mechanism of cardiac muscle contraction and calcium signaling in the heart by characterizing a previously unrecognized protein, JP1, in cardiac muscle.

NIH Research Projects · FY 2026 · 2024-01

Knowledge of CFTR function and cell type expression has advanced greatly since its discovery in 1989. Indeed, drug therapies such as Ivacafter and Trikafta restore function for most CFTR mutation classes; however, ~10% of people with CF cannot benefit from these drugs because their CFTR proteins are only partially produced and there is little the drugs can do to help. Here we use an adenosine deaminase fused to a CRISPR-Cas9 nickase (termed an Adenine Base Editor (ABE)) that converts an Adenine to a Guanine which is critical for full length protein production for certain mutations. ABEs do not create double stranded DNA breaks, do not require homologous recombination templates, and are effective in quiescent cells of the airways. We recently reported correction of CFTR nonsense mutations using electroporated ABEs in human airway epithelial cells in vitro as determined by next generation sequencing and correction of chloride current. However, the lung is a challenging organ to correct using gene editing due to an enormous surface area and multiple mechanisms to resist vector uptake. Delivery of the ABE to enough of the appropriate airway cells to be therapeutic is the problem we address in this proposal. Our research group has demonstrated success using many categories of reagents for modifying the genomes of airway cells. In this proposal, we compare two ABE delivery tools: adeno-associated viruses (AAVs) and viral-like particles (VLPs). They each have their pros and cons. We will compare AAV-ABEs and VLP-ABEs in multiple in vitro and in vivo models. We will identify reagents with improved airway progenitor cell targeting. In vivo editing efficiency will be examined in a new reporter mouse model termed Gene Editing Reporter 14 (GER14) and an established GFP transgenic pig model. In addition, in vivo CFTR function will be examined in genetically modified mice with the endogenous mouse Cftr exon 12 replaced with the human CFTR exon 12 with the R553X mutation (termed hEx12R553X). Lastly, we ask if secreted mucins inhibit AAV or eVLP delivery or editing efficiency in models of advanced lung disease. Our goal is to provide a life-long gene repair strategy that could be adapted for a great number of CF causing mutations. The reagents, methods, and data generated by these experiments could be applied to base editing for other monogenic disorders, thereby significantly advancing the gene therapy field.

NIH Research Projects · FY 2026 · 2024-01

Project Summary Orientia tsutsugamushi (O.t.) is the etiological agent of scrub typhus, a devastating disease with a high mortality rate that is transmitted by the bite of certain trombiculid mites or “chiggers”. Over one million individuals are infected annually, however, these statistics may be a gross underestimate of disease incidence as the “tsutsugamushi triangle”, where most cases are reported, encompasses large regions of jungle in rural subtropical environments with limited access to hospitals and diagnostic facilities. Beyond the tsutsugamushi triangle, recent reports have confirmed cases of the disease in South America, Africa, and the Middle East. There is no vaccine and antibiotic resistant strains have been reported. Without treatment, disease associated morbidities including hepatitis, renal failure, myocarditis, encephalitis, multiple organ failure, and death can occur. Despite its significant impact on global health, little is known of the molecular mechanisms the bacterium uses to infect and cause disease in humans. Specifically, the bacterial factors that promote host cell subversion and bacterial pathogenesis remain largely unknown. The key roadblock to a more detailed understanding of how O.t. causes disease lies in the inability to genetically manipulate the pathogen. Overcoming the genetic intractability of O.t. will remove the single largest barricade to extending our operational knowledge of the molecular mechanisms utilized by the bacterium to hijack host cells. Here we propose to develop the first functional system for genetic transformation of O.t. which will present a unique opportunity to overexpress epitope-tagged proteins in O.t. and generate the first site specific O.t. mutants. During the R61 phase, we will develop a shuttle vector and transformation method for use in O.t. (Aim 1) and adapt the TargeTron system to insertionally inactivate select TPR proteins (Aim 2). In the R33 phase, we will use these newly developed tools and mutants to determine whether select TPR are secreted proteins that promote host cell invasion or bacterial proliferation by perturbing the host cell cycle (Aim 3). Our proposed studies will generate the first tools and reagents to genetically alter O.t., a resource that will be invaluable to other Orientia researchers.

NIH Research Projects · FY 2026 · 2023-12

PROJECT SUMMARY Hepatic fuel fluxes underly numerous processes critical to whole-body physiology that become dysregulated in type 2 diabetes (T2D). Among these is the basic biosynthetic process of gluconeogenesis (GNG), which increases during and contributes to the pathophysiology of T2D. Thus, understanding the regulation of the fuel fluxes supplying hepatic GNG is important for understanding both normal and T2D whole-body physiology and metabolism. The TCA cycle anaplerotic (refilling) mechanisms sustaining carbon flux into GNG have been investigated intensively. However, the greatest hypothesized net TCA cycle cataplerotic (withdrawing) pathway, malate export through the mitochondrial dicarboxylate carrier (DiC), has received sparse direct experimental attention in vivo. Elegant, early, ex vivo biochemical studies suggest that mitochondrial malate export through the DiC supports hepatic GNG. However, this idea has been almost totally unaddressed in the molecular era. The overall goal of this application is to understand how the DiC contributes to hepatic GNG in normal and T2D states and the mechanisms by which hepatic DiC function is altered during T2D. Based on our preliminary data, the central hypothesis of this application is that the DiC feeds hepatic GNG by supplying carbon and reducing equivalents, becomes misregulated during T2D and contributes to excessive GNG, and may be disrupted to attenuate T2D. Using mouse genetics, primary cell culture systems, stable-isotope metabolomic tracing, luciferase reporter assays, and proteomic analysis, we will test this hypothesis by pursuing two specific aims: 1) Determine the role of the DiC in hepatic gluconeogenesis (GNG) in normal and T2D states; and 2) Define the mechanistic regulation of DiC abundance and gluconeogenic activity. This research is significant because successful completion will delineate the role of the hepatic DiC as a fundamental regulator of liver and whole-body metabolism and in the elevated GNG of T2D. This research is innovative because it will utilize novel genetic DiC loss- and gain-of-function models to finally test the role of the DiC in linking mitochondrial metabolism and GNG in T2D. The consequence of not doing this research is that the role of the DiC in hepatic GNG and its contribution to T2D will remain undefined.

NIH Research Projects · FY 2026 · 2023-12

ABSTRACT Adverse childhood experiences (ACEs), including abuse, neglect, household and community dysfunction, represent potentially traumatic psychosocial stressors that occur during critical development periods and represent a latent risk factor for future cardiovascular disease (CVD). ACE exposure promotes cardiovascular disease risk in adulthood in a dose-dependent manner, such that adults exposed to 4 or more ACEs have a more than two-fold increased risk of ischemic heart disease and stroke compared to individuals with no ACEs. ACEs are also widespread, with 1 in 6 US adults reporting exposure to four or more ACEs. Identification of modifiable, bio-behavioral mechanisms by which ACEs promote early onset CVD risk is critical to establish effective interventions to improve cardiovascular health in the millions of individuals affected by ACEs. Our recent evidence indicates that young adults exposed to ACEs demonstrate impaired vascular endothelial function that is manifest prior to overt elevations in blood pressure or other CVD risk factors, and that is related to systemic oxidative stress. These vascular impairments were also observed alongside poor sleep, and our strong preliminary data suggests that poor sleep quality, or low sleep efficiency, may mediate the association of ACE exposure with vascular dysfunction. Therefore, we propose that disturbed sleep is a critical, modifiable bio- behavioral pathway by which ACEs promote vascular dysfunction in early life. Here, we propose the critical studies necessary to understand the causal, mechanistic role of disturbed sleep in ACEs-related endothelial dysfunction in young adults. To do so, we propose to 1) objectively quantify the degree to which lower sleep quality contribute to ACEs-related endothelial dysfunction, inflammation, and oxidative stress in young adults, and 2) manipulate sleep quality using an established behavioral sleep intervention, Cognitive Behavioral Therapy for Insomnia (CBT-I), in a cohort of young adults with moderate-to-severe ACE exposure in order to establish the causal, mechanistic role of disturbed sleep in ACEs-related vascular endothelial dysfunction. To accomplish these aims, we will use an innovative translational human approach that includes rigorous at home sleep monitoring using actigraphy and polysomnography, in vivo assessment of endothelial function via flow-mediated dilation testing, and in vitro determination of endothelial cell inflammation and oxidative stress from biopsied endothelial cells. Notably, while vascular endothelial dysfunction contributes to and is prognostic of future CVD, it is also reversable. Thus, outcomes of this study are expected to be of high impact by identifying and understanding how disturbed sleep, a key, targetable bio-behavioral mechanism, contributes to vascular endothelial dysfunction in young adults with exposure to ACEs.

NIH Research Projects · FY 2026 · 2023-09

ABSTRACT Glaucoma is a common cause of severe vision loss, characterized by the progressive loss of retinal ganglion cells. Several large genome-wide association studies (GWAS) for primary open angle glaucoma (POAG) have been performed to date and have discovered over 127 risk factor genes. The mechanism by which these genes lead to POAG is almost completely unknown. Our proposal seeks to 1) translate these risk factor gene discoveries into useful data for patients and their doctors and 2) determine the mechanisms by which risk factor genes confer risk. This proposal will leverage the results of prior GWAS studies along with the unmatched, 20-years of prospective clinical data from the Ocular Hypertension Treatment Study (OHTS) to develop a POAG risk calculator that includes both clinical and genetic risk factors that will be useful to patients and doctors (AIM1A). We will also conduct association studies of the OHTS cohort to identify genetic risk factors for rapid progression of POAG as measured by visual field parameters, with validation in a local Iowa cohort (AIM1B). Identifying genes associated with progression of glaucoma has great potential to personalize and guide glaucoma management. We will also investigate the functional consequences of specific glaucoma risk alleles with a range of complementary technologies. AIM2A will test the effects of risk alleles on gene expression and pathways with studies of genotyped human donor eyes using single-cell RNA sequencing (scRNAseq). Immunohistochemistry and ELISA of genotyped human donor eyes will also be used to determine the effects of risk alleles on the abundance and localization of the proteins they encode. Finally, in AIM2B will use BiT- STARR-seq to locate the specific variants (SNPs) in glaucoma loci that confer risk for glaucoma. These studies will begin to define the precise molecular steps that connect the presence of specific genetic risk factors in one’s genome to the development of glaucoma. Our proposal will lead to improved tools for ascertainment of a patient’s risk for POAG or risk for rapid worsening of POAG that can be readily transferred to clinicians in the form of better diagnostic and prognostic tools. Our proposal will also define the specific mechanisms by which risk factor alleles alter gene expression in key tissues (retinal ganglion cells), which will identify disease mechanisms and new therapeutic targets to facilitate development of targeted treatments. Our proposal has great potential to improve glaucoma care and reduce vision loss.

NIH Research Projects · FY 2024 · 2023-09

Project Summary: Nursing homes are considered one of the most dangerous workplaces in the United States, and the Occupational Safety and Health Administration (OSHA) has emphasized on both inspections and collaborative efforts to improve worker safety in nursing homes. Dangerous work environment can lead to high turnover of workers. In addition, high turnover of staff has potentially adverse consequences for more than a million residents who depend on nursing home workers for their daily needs including bathing, eating, and toileting. Despite the recognition by OSHA on high injury risk and the adverse implication of high turnover on resident quality of care, nursing home worker welfare and safety has not been given enough attention by policymakers and researchers. Although a few recent studies have highlighted the issue of high turnover of nurse staff in nursing homes, the turnover of other essential workers such as dietary and housekeeping staff that earn very low wages has not been examined. We do not yet have a national database of turnover rates for different staff types in nursing homes but some states such as Iowa require nursing homes to report turnover rates for different staff types. In 2015, turnover rates were 61% for dietary staff and 42% for housekeeping staff in Iowa nursing homes. There is limited evidence on the trends and organizational factors associated with high turnover of non-nurse staff in nursing homes. Moreover, the value of lowering staff turnover to improve resident welfare is not understood well. Despite the substantial role played by non-nurse staff on patient care in nursing homes, to our knowledge, no studies have examined non-nurse staff turnover or its impact on nursing home quality. To this end, we have three specific aims: 1) To examine trends in non-nurse staff turnover, overall, and stratified by staff types, 2) To identify risk factors for non-nurse staff turnover, and 3) To estimate the impact of turnover of different staff types on outcomes such as hospitalization rates, falls, pressure ulcers, citations for nutritional issues, and infections. Our findings have the potential to affect national and state policies on the collection of staff turnover data and efforts to lower staff turnover by providing rigorous estimates on the potential causes and consequences of staff turnover in nursing homes. This proposal fits within the Healthcare and Social Assistance sector area under NORA (NAICS code: 62) and focuses on the subsector of nursing and residential care facilities (NAICS code: 623). It addresses several priority goals for strategic goal 7 (7.12A- 7.12D; 7.2A-7.2C) within the NORA priorities. Our study examines how work organization affects both worker and patient safety. Our study also focuses on low-wage occupations such as dietary staff, therapists, housekeeping, and maintenance staff. The manuscripts and presentations from this innovative project will inform state and national policymakers about factors driving staff turnover in nursing homes and are critical to improve Research to Practice (r2p) efforts in the Healthy Work Design and Well-Being (HWD).

NIH Research Projects · FY 2025 · 2023-09

Abstract Problematic alcohol involvement (PAI), characterized by alcohol consumption that leads to or increases risk for acute and/or chronic negative consequences in one or more life domains, poses an enormous public health burden to drinkers, their families, and society. Heterogeneity in PAI’s etiology has been identified as the single most important barrier to progress in remediating this burden. More effective characterization of neurobehavioral traits that increase PAI risk is critical to the development of more effective intervention and prevention efforts. Cognitive science approaches to addiction etiology have suggested PAI risk reflects an interaction between two internal cognitive systems: appetitive motivation for alcohol (AMA) impelling alcohol seeking and use, and executive function (EF) regulating the influence of appetitive drives. This Dual Process Model has been highly influential, but prior studies have failed to support its main interaction hypothesis. We argue this failure is attributable to weaknesses in the rigor of prior research, and propose several innovative refinements to the DPM framework to improve its predictive utility. First, individual differences in AMA and EF have not been adequately characterized in prior studies, most of which represent these constructs with single indicators. Innovations in neuroclinical assessment indicate that neurobehavioral trait constructs are better characterized with measures representing self-report, behavioral, and neurophysiological units of analysis. Second, DPMs assume risk for PAI reflects only the influence of internal processes, implying no role for context. This is a major limitation, as strong evidence indicates that both environmental contexts (e.g., legal constraints; alcohol access) and alcohol exposure (e.g., acute effects of alcohol on EF and AMA) strongly affect the extent to which person-level factors relate to PAI. Finally, given that PAI behaviors are the result of decisions people make while drinking, accounting for heterogeneity in drinking-related decision strategies can improve prediction of PAI, including the role of DPM constructs. We propose a multi-session, within-subject alcohol challenge experiment, combined with online follow-up assessments, structured to take advantage of the abrupt change in contextual factors that occurs with transition to the minimum legal drinking age (MLDA; i.e., 21st birthday). This transition broadly increases risk for PAI within persons, but little is known about the role of specific etiologic factors in this increased risk. Participants (N=220) will be recruited from rural census tracts in mid-Missouri, identified through state administrative and driver’s license databases. Such individuals are broadly underrepresented in PAI research, particularly alcohol challenge research, and tend to experience more chronic (less transient) and severe PAI than their relatively advantaged, college-attending age peers. This sampling approach also will permit over-sampling of individuals at higher risk for DUI arrest (e.g., based on parental history of DUI and area characteristics). Thus, this study will fill important gaps in knowledge concerning PAI etiologic factors in an understudied population.

NIH Research Projects · FY 2025 · 2023-09

Abstract This proposal seeks to determine the regulation of microglial type I interferon signaling following traumatic brain injury (TBI) and to identify how this affects neuroinflammation, neurodegeneration, and the neurocognitive sequelae following TBI. TBI is a major public health problem, representing a leading cause of death and disability from childhood through middle adulthood. Unfortunately, pharmacologic therapies for TBI are non-existant. Treatment strategies that target specific secondary injury cascades like dysregulated neuroinflammation are critically needed. Microglia, the primary immune cells of the CNS, persist in an activated state for months and years following a single TBI and are associated with neurodegeneration. Mechanisms that result in sustained, damaging neuroinflammation vs resolution of inflammation are unclear. Our recently published work, however, demonstrated that the microglial transcriptome at a subacute time point following TBI was highly enriched for type I interferon stimulated genes raising the question of if type I interferons are key signaling molecules resulting in sustained, dysregulated microglial activation. The types and cell sources of type I interferons activating microglia following TBI are unknown. Similarly, the specific effects of type I interferons on sustained microglial reactivity and the subsequent mechanisms of microglial mediated neurodegeneration and neurologic dysfunction following TBI remain unknown. Using a lateral fluid percussion injury model of TBI in mice, combined with sophisticated mouse genetics, our data will uncover cell-specific effects of type I interferon signaling on secondary injury. The hypothesis of this proposal is that multiple type I IFNs drive development of dysregulated microglia subsets, immune cell recruitment, and the subsequent neurodegeneration and neurobehavioral impairments that occur after TBI. In the first aim, we will identify the types and cell-sources of type I IFNs driving persistent microglial and CNS interferon stimulated gene expression following TBI. In the second aim, we will determine the impact of type I IFN signaling on microglial subsets and their transcriptional activation following TBI. Finally, in the third aim, we will determine the mechanisms by which type I interferon activated microglial impact neuropathology and neurocognitive dysfunction following TBI, including through their recruitment of CD8+ T cells. As TBI is a leading, untreatable cause of death and disability in the US, there is a critical need for further study of the specific mechanisms of secondary injury so that ultimately, new therapies may be discovered.

NIH Research Projects · FY 2024 · 2023-09

Project Summary Despite significant overall progress in improving access to dental care among children in the United States in recent decades, substantial disparities in oral health have persisted among publicly insured children. Although Medicaid provides comprehensive dental coverage for all enrolled children, dental benefits for adults are optional and vary across states and over time. There is evidence that reducing Medicaid adult dental benefits (MADB) lowers access to dental care for low-income adults. However, the impact of changes in MADB on access to dental care for Medicaid-enrolled children is not yet understood. Our long-term goal is to improve access to dental care and oral health outcomes among children in low-income families. To achieve this goal, we will leverage natural experiments in states that changed MADB generosity to examine the spillover effect on dental utilization by Medicaid-enrolled children. We hypothesize that expanding MADB generosity results in increases in both preventive and overall dental utilization and a decline in emergency department visits for dental problems by Medicaid-enrolled children. Using a causal inference approach, we will estimate the longitudinal impact of changes in MADB on children’s dental care utilization through these research aims: Aim 1: Determine the impact of changes in MADB generosity on overall dental utilization by children enrolled in Medicaid using administrative data with a quasi-experimental design. Aim 2: Determine the impact of changes in MADB generosity on preventive dental utilization by children enrolled in Medicaid using administrative data with a quasi-experimental design. Aim 3: Determine the impact of changes in MADB generosity on emergency department (ED) dental utilization by children enrolled in Medicaid using administrative data with a quasi-experimental design. Using administrative data, we will generate monthly estimates of these three measures of utilization and use a causal inference framework to assess the changes in child dental utilization trends before and after the MADB policy changes. We will use interrupted time series approach to account for any secular trends and make robust within state comparisons, and supplement it with a differences in difference approach to provide comparison with control states that did not experience a policy change. Our findings will provide evidence on the spillover effect of MADB generosity on children’s access to dental care. This study will be the first to use administrative data to examine the spillover effect of changing MADB generosity on children’s dental care use. The study aligns with the strategic priorities of NIDCR to understand and work to reduce child oral health disparities.

NIH Research Projects · FY 2025 · 2023-09

Individuals who have not had substantial biomedical research experience are less likely to obtain advanced degrees in the field of cancer research. In an attempt to address this problem, the Principal Investigators (Co-Pis) and their colleagues at the University of Iowa (UI) have previously enrolled 182 undergraduate (UR) students during previous 19 summer programs. In order to continue our successful summer programs we are submitting this application for an NCI-funded training program in Youth Enjoy Science Research Education Program (R25 YES). The proposed program will include a research methods training, emersion in an outstanding research project, courses in cancer biology, career counseling, follow-up after the students complete the summer research training program, and community outreach programming. Our plan is to train both undergraduate and high school students to encourage their future participation in cancer research. We will also host faculty from these institutions to expose them to research projects and to learn about development and enhancement of the science curricula at their home institution. We will utilize of the Ul's Department of Educational Policy and Leadership Studies in the College of Education in the development of the course. The goals or objectives of the proposed program include: (a) to recruit students who would not routinely have access to a program that will be designed to provide opportunities to learn about and perform cancer research; (b) to design a training program for these students and their educators that will expose them to cancer research; (c) to provide career counseling about the variety of employment options such as academia, industry, and government or private research centers; (d) to provide one-on-one conversations with heads of departments, directors of interdisciplinary graduate programs, directors of postbaccalaureate programs at University of Iowa, medical school admissions, and directors of the MD-PhD program; (e) to continue training and career counseling between the two years of the training program as well as after completion of the University of Iowa's R25 YES program; and (f) community outreach and extended mentoring. Exposing students and teachers to science and research will support our goal and the goal of the NCI to support cancer research and ultimately long-term survival for cancer patients or their cure by increasing the number and diversity of new scientists engaged in research studies, including clinical trials. The proposed program has a strong likelihood of success based both on our design of the program and our past successes with previous summer research training programs.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY / ABSTRACT Transdermal drug delivery has many benefits including ease of product application for patients and more consistent plasma drug concentrations compared to oral or IV dosing. However, many active pharmaceutical ingredients (APIs) do not meet the strict physicochemical properties needed to permeate through the hydrophobic outer skin layer. For APIs that cannot adequately absorb through skin, microneedles (MNs) temporarily increase skin permeability through formation of epidermal micropores. There are two major routes of API transport to consider with MN treatment: micropores (a hydrophilic environment), and intact skin around the micropores (a hydrophobic environment). Effects of API properties and formulation are well understood for delivery through intact skin, but they need to be systematically considered for MN delivery because of the introduction of the second parallel transport pathway (micropores). In vivo micropore closure time and human skin characteristics also impact API delivery with MNs, and these variables cannot be well simulated in vitro. The long-term goal of this work is to improve API delivery and treatment options for a wide range of health conditions through development of innovative MN dosage forms. The goal for the next 5 years is to determine significant in vitro and in vivo factors impacting absorption of small molecule APIs through MN-treated skin. We will use in vitro permeation tests and in vivo pharmacokinetics studies (in animals and humans) to answer key questions about how API properties, formulation, and in vivo skin characteristics impact permeability and flux through micropore vs. intact skin pathways. The main physicochemical characteristics we will initially investigate include ionization/charge, logP, and pKa. Permeation will be examined in vitro under heated vs non-heated conditions (heat can synergistically enhance skin API permeation), and MN properties (length, number) will be examined for effects on permeation. This will give a mechanistic understanding of the API and MN properties having the greatest impact on micropore permeation. Pharmacokinetics studies in guinea pigs will be completed so that in vivo factors impacting permeation can be assessed and compared to in vitro predictions. A second major area of study will involve pharmacokinetics studies in healthy human subjects. We will enroll a cohort of subjects >50 years of age, along with younger (<50 yrs) control subjects so we can 1) correlate age-related micropore closure estimates with API absorption, and 2) study age-related skin changes on MN API delivery. The combined in vitro and in vivo studies will maintain the high clinical relevance of the work. We expect to identify key physicochemical properties and increase our understanding of structure-permeability relationships and biological aspects impacting API permeability with MNs. This will generate fundamental knowledge that can result in direct clinical applications and supports the NIGMS mission in the areas of drug delivery, absorption, transport, and kinetics.

NIH Research Projects · FY 2025 · 2023-09

ABSTRACT The overarching goal of this research is to promote children’s positive socioemotional pathways and to prevent maladaptive pathways. We elucidate why some children embark on positive paths toward prosocial, internalized, rule-abiding conduct, and robust social competence, whereas others enter maladaptive paths toward callousness, disregard for conduct rules and others’ feelings, antisocial behavior, and impoverished competence. We focus on the parent–child early relationship, formed in the first years of life, as an influential source of the divergent paths, and we longitudinally chart its complex, indirect yet powerful, long-term legacy. Drawing from our extensive research, correlational and experimental, in low- and high-risk families, we propose that although early relationship may not have long-term unqualified, direct effects, it nevertheless serves as a powerful moderator of future parent–child unfolding dynamics. Specifically, early relationship can set the stage for an adversarial, negative cascade. In suboptimal, insecure parent–child dyads, the child’s difficult temperament easily triggers the parent’s negative, harsh, power-assertive control, which, in turn, leads to detrimental child outcomes. In contrast, an early optimal relationship sets the stage for positive, cooperative, effective socialization, and defuses risks of negative cascades. We proposed that parents’ and children’s differing internal representations, expectations, and perceptions of each other (Internal Working Models, IWMs) that characterize suboptimal and optimal relationships and come to guide parents’ and children’s behavior and interactions are the key mechanisms that account for the divergent cascades. We are testing this framework in an ongoing study of 200 community mothers, fathers, and children, richly assessed at 8, 16, 38, and 50-54 months. This application proposes to leverage those massive data to follow up the families at ages 5-6, 7-8, and 9-10. Using state-of-science measures of parents’ and children’s social representations, in Aim 1 we examine how their unfolding IWMs of each other are linked to their relationship quality, how children’s Theory of Mind contributes to their IWMs, how children’s IWMs of the parents generalize to their representations of the social world, particularly hostile attributional biases, and how the child’s IWMs of two parents become integrated in development. In Aim 2, we examine the parent’s IWM of the child as moderating paths from child difficulty to parental control, and the child’s IWM of the parent as moderating paths from parental control to child outcomes. In Aim 3, we embed our model in the dynamics of the family system. Our multi-method, multi-level approach encompasses observational, genetic, and reported measures of the parent’s and the child’s relational information processing, representations, temperament, relationships, parental control, and child developmental outcomes. Variable- and person-centered analyses rely on structural equation modeling to elucidate divergent developmental cascades. We aim to realize a long-advocated – but yet to be accomplished – vision of research integrating relationships, temperament, representation, and behavior in pathways to children’s adjustment.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY/ABSTRACT The long-term objective of the Network of Excellence in Neuroscience Clinical Trials (NeuroNEXT) initiative is to rapidly and efficiently translate advances in neuroscience into treatments for neurological disorders. There are over 1000 different neurological disorders, many of which are associated with significant cognitive and physical disabilities. In the United States, neurological diseases affect over 100 million people of all ages, and they are a significant cause of mortality and disability. While therapeutic options are currently limited or non-existent for many of these disorders, recent exciting advances in the neurosciences and in gene therapies for neurological disorders make it imperative to increase the ability to efficiently perform clinical trials. A central goal of NeuroNEXT is to facilitate, from initial conception through final analysis, high-quality early phase clinical trials and biomarker studies with clear go/no-go decisions in adult and pediatric neurological disorders. This application requests funds for the University of Iowa to continue serving as the DCC for NeuroNEXT. The DCC will continue to support study design, data collection, data management, data sharing, project management, clinical site monitoring, quality management, safety monitoring, and all statistical aspects of trials conducted within the network. The DCC will continue to build on our robust, standardized, and accessible infrastructure in a manner that is explicitly designed to accommodate dynamically changing requirements that naturally occur in clinical trials, and will continue to help NeuroNEXT succeed in providing more rapid evaluations of promising treatments in the neurosciences. Specific Aim 1 is to work closely with the CCC to continue to support a collaborative network community that efficiently solicits and conducts high-quality clinical trials for neurological disorders. Specific Aim 2 is to continue to provide high-quality IT/development, data management, statistical, and protocol coordination support for all network clinical trials. Specific Aim 3 is to expand the pool of experienced clinical Investigators, research staff, and biostatisticians who are prepared to be leaders of multicenter clinical research trials.

NIH Research Projects · FY 2025 · 2023-09

Dynamic Oxygen-Enhanced MRI of Lung Structure and Function, PI: Sean B. Fain. Abstract This project will develop and optimize oxygen-enhanced (OE) imaging of dynamic ventilation of the lungs simultaneously with high resolution acquisition of lung parenchymal anatomy in a single, free-breathing 7-minute acquisition using 3D ultra-short time to echo (UTE) MRI. Advances in 3D UTE MRI now support regional imaging of lung anatomy with CT-like contrast, full chest coverage, and isotropic 1 mm spatial resolution. We will employ advanced motion compensation reconstruction with manifold-based deep learning and UTE center-out k-space trajectories to isolate respiratory motion from T1 changes due to oxygen wash-in and wash-out during free- breathing. The resulting motion compensated reconstruction provides both quantitative ventilation and high- resolution structure in a single 7-minute series. Multiple chronic lung diseases will be studied with this approach to establish utility and repeatability of the method in comparison to quantitative chest CT and hyperpolarized 129Xe MRI. Our preliminary data demonstrates the utility of MRI-only exam of lung structure and ventilation in a manner similar to that provided by nuclear SPECT with technegas and X-ray CT but without ionizing radiation. We hypothesize that 3D UTE MR imaging of ventilation dynamics will capture co-localized structure-function for monitoring ventilation heterogeneity relative to structural features of lung disease, including fibrosis, granulomas, mucus plugging, bronchiectasis, ground glass, and fibrosis. Radiology expert reader studies will be performed with direct comparison of UTE MRI with quantitative chest CT for depiction of structural and functional (derived from static multi-volumetric images) abnormalities, and OE MRI regional patterns of ventilation to hyperpolarized 129Xe MRI. We seek to create an MRI method and protocol for structure-function assessment of chronic lung disease with broad access, no exposure to ionizing radiation (allowing for longitudinal assessment at increased granularity), and using a safe, inexpensive and widely available paramagnetic gas. The specific aims of the project are to: 1) Improve data acquisition efficiency for 3D UTE MRI at clinical field strengths; 2) Develop dynamic OE MRI of oxygen ventilation wash-in and wash-out during free breathing, and 3) Develop OE MRI visualization and analysis tools for regional structure-function associations. The 3D OE MRI approach is inexpensive, uses proton-based contrast and can characterize both structural and functional aspects of chronic lung disease in a manner similar to SPECT/CT without concern for ionizing radiation exposure.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY Major birth defects (BDs) affect morbidity and mortality of tens of thousands of newborns annually in the US. Environmental (i.e. non-inherited) exposures or gene variants are estimated to explain about one-third of BDs that occur. Since 1996, the Iowa Center for Birth Defects Research and Prevention (CBDRP) has played a leadership role in surveillance and research of major BDs. Partnerships with the State of Iowa Department of Health and Human Services and Iowa Registry for Congenital and Inherited Disorders have allowed the Iowa CBDRP timely access to comprehensive surveillance data on deliveries to Iowa resident mothers for enrollment in the National Birth Defects Prevention Study and Birth Defects Study To Evaluate Pregnancy exposureS (BD-STEPS). Using exposure and genomic data collected, the Iowa CBDRP has made substantial contributions to understanding the etiopathogenesis of major BDs, ranging from prevalence estimates to gene- environmental interaction effects, with this work being disseminated in high-impact journals. Findings published support multifactorial inheritance patterns that vary among BDs across system groups and among BD subtypes within a system group. These findings of a heterogeneous etiopathogenesis reinforces the need to continue to investigate major BD phenotypes individually. With this application, the Iowa CBDRP proposes to continue its successful participation in BD-STEPS by timely ascertaining and classifying pregnancies with and without specific major BDs (Aim 1) and developing and evaluating novel modes of exposure data collection (Aim 2). Using data collected, the Iowa CBDRP proposes to expand the traditional epidemiology approach by integrating a multi-omics systems epidemiology approach to more fully investigate risk factors for specific major BDs (Aim 3). The traditional epidemiology approach will be used to examine associations between BD- STEPS exposure data and defects, with international datasets used to conduct confirmation analyses. The systems epidemiology approach will incorporate state-of-the-art genomic studies and machine learning analytics to improve characterization of child phenomes and exposomes and examine gene-environmental interaction effects. Concurrent with the proposed study activities will be training bachelor’s, predoctoral and postdoctoral research scholars to advance the next generation of birth defect researchers (Aim 4). The proposed research and training are not only consistent with BD-STEPS required activities (RFA-DD-23-001) but expands and innovates such activities by improved delineation of exposure pathways and genetic susceptibilities influencing development of specific major BDs. The interdisciplinary expertise of our key personnel and collaborators, along with access to rich exposure and biological data from independent populations, provides the Iowa CBDRP unique resources among all CBDRPs in disseminating novel findings in peer-reviewed publications. Findings generated will provide critical insights into modifiable risk factors for major BDs and be used for family education, treatment improvements, and application of prevention strategies.

NIH Research Projects · FY 2026 · 2023-09

PROJECT SUMMARY/ABSTRACT The neovascular ‘wet’ form of age-related macular degeneration (nvAMD), whereby new blood vessels grow under the retina, requires treatment with anti-vascular endothelial growth factor eye injections typically every 1- 3 months for the remainder of patients’ lives due to persistent fluid, which can be visualized and quantified by optical coherence tomography (OCT) imaging. Recent research has implicated matrix metalloproteinase 9 (MMP9), expressed in immune cells of the retinal blood supply (termed choroid), as a mediator in the formation and persistence of nvAMD. This proposal will elucidate our understanding of this novel immune-based mechanism. Characteristics of patients with high-risk genotype of MMP9 will also be assessed, as they are associated with incomplete response to standard therapies for nvAMD. We will identify genetic regulatory regions that control expression of MMP9 and determine the immunologic activation profiles in patients with nvAMD. We will use artificial intelligence (AI)-based approaches to associate phenotypic features observable on OCT and OCT-angiography with high- and low-risk MMP9 genotypes in patients with nvAMD. Our outstanding team of interdisciplinary researchers will collaborate to employ state-of-the-art genomic and immunologic techniques to study the mechanisms of how MMP9 is implicated in nvAMD and fibrosis. Genetics will aid AI- and machine learning-based analysis of OCT images to learn new features associated with the most challenging patients afflicted with nvAMD. Unraveling the disease mechanisms associated with MMP9 will reveal new targets for therapeutic intervention. This proposal encompasses 5 of the 7 cross-cutting areas of emphasis the NEI laid out in the 5-year Strategic Plan on November 1, 2021. This is the next, necessary milestone in nvAMD that will help to address this major socioeconomic health burden.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY/ABSTRACT Fibrosis and scarring from injury and disease costs $20 billion annually to treat in the US, and even when treated remains a persistent problem that significantly diminishes quality of life. The use of autologous adipose micrografts used in reconstructive surgeries appears beneficial for reducing scar. Furthermore, intradermal adipocytes have also recently gained attention for their role in the early stages of normal wound healing. The objective of this proposal is to determine how human adipocyte lineage cells impact fibroblast-myofibroblast conversion and extracellular matrix (ECM) remodeling in a healing wound. Our long-term goal is to engineer more effective and improved clinical therapies for wound healing. To achieve this objective, we propose three aims that are designed to unravel the communication between adipose lineage cells and fibroblasts. In Aim 1, we will determine how secreted factors from adipocyte linage cells regulate fibroblast-myofibroblast conversion, phenotype, and ECM remodeling. We hypothesize that adipocyte lineage cells differentially regulate inhibition, induction, and reversal of fibroblast to myofibroblast conversion, which in turn impacts the composition of the ECM produced. We also hypothesize that adipokine-induced myofibroblasts (compared to TGF-β1-induced myofibroblasts) produce distinct ECM that results in less scaring. We will test these hypotheses and identify how one-way communication from adipocyte lineage cells regulates fibroblast-myofibroblast activity using a myofibroblast conversion assay coupled with fractionation, proteomics, computational genomics, and inhibition/add-back assays. In Aim 2 we will determine how adipocyte lineage cells coordinate with fibroblasts to remodel ECM. We hypothesize that reciprocal communication between adipocyte lineage cells and fibroblasts- myofibroblasts impacts wound healing by altering cell activity and the composition of the ECM proteins produced by both fibroblasts and adipocyte lineage cells. We will use our 3D adipose spheroids and our 3D in vitro models to interrogate the interactions between the relevant cell types and the composition and mechanics of the ECM produced. In Aim 3, we will complement the in vitro work of Aims 1 and 2 by determining how transplantation of adipocyte lineage cells alter fibroblast behavior and ECM remodeling in a porcine wound model. We hypothesize that wound bed fibroblasts will differentially respond to adipose-based treatments depending on the cellular composition delivered. We will test this hypothesis by assessing the effect of transplanted adipose stem cells, preadipocytes, or adipocytes on fibroblast/myofibroblast behavior and wound healing. Successful completion of these aims will substantially improve our understanding of adipose-fibroblast interactions that will be critical for the development of more effective and improved clinical therapies for wound healing.

NIH Research Projects · FY 2025 · 2023-09

PROJECT SUMMARY / ABSTRACT Alcohol use disorder (AUD) is highly heterogeneous in that its symptoms are highly varied and, often, non- overlapping across individuals. In theory, this phenotypic heterogeneity reflects the influence of multiple underlying causes, or etiologic mechanisms. This etiologic complexity makes AUD treatment very challenging; researchers have identified AUD’s etiologic complexity as the most critical barrier to progress in developing more effective, personalized treatments. Addiction theorists have identified a set of alcohol addiction research domain criteria (AARDoC) functional domains, believed to be important etiologic mechanisms for AUD. Yet, existing models meant to link these mechanisms to AUD-related behaviors and symptoms fail to consider etiology. Hence, many basic questions concerning the etiology of problematic drinking and AUD-related symptoms remain unresolved. The proposed work aims to identify the prospective contributions of functional domain neuro- behavioral indicators to the etiology of heavy drinking (HD) and AUD-related symptoms during adolescence and emerging adulthood, the decade of development when HD and AUD-related symptoms are most prevalent. We will enroll a target sample of 480 adolescents and emerging adults (160 in each of three partially overlapping age cohorts [50% female], pre-screened for elevated HD risk) to participate in a prospective study using an accelerated longitudinal design, which will allow us to characterize trajectories of alcohol involvement and AUD- related symptoms over a 10-year period of development within a five-year study. We will use a neuroclinical assessment approach to comprehensively characterize four functional domains—cognitive control/disinhibition (DIS), reward sensitivity (RS), anxiety (ANX), and incentive salience (IS)—using validated and reliable self- report, behavioral, and neurophysiological measures during each of three waves of data collection (15 months apart). Alcohol involvement, AUD-related symptoms, and social-environment factors will be assessed at 5-month intervals. Using this multi-wave, multimodal approach, we will address three specific aims: (1) characterize the influence of the functional domains on the etiology of alcohol involvement; (2) accounting for the influence of alcohol involvement, characterize the influence of the functional domains on the etiology of AUD-related symptoms; and (3) characterize the influence of HD on functional domain neurobehavioral indicators. Analyses will characterize how various combinations of domain indicators affect latent states of alcohol involvement (volume of consumption; patterns of use) and AUD-related symptoms (numbers of symptoms; clusters of symptoms) and transitions across latent states over time. This work will produce a unique and rich dataset, and its findings will directly inform the development of personalized intervention and treatment strategies that can be deployed to target the functioning of specific domains during periods when their influence is greatest.

NIH Research Projects · FY 2025 · 2023-08

PROJECT SUMMARY I am an environmental epidemiologist and microbiome researcher with a primary research interest in the role of the human microbiome between environmental exposures and human health, including neurological outcomes. The goal of this proposal is to obtain training and acquire the skills needed to continue to build my academic research career by linking early life metal exposure, the gut microbiome and metabolome in childhood, and neurodevelopmental measures of attention. For this proposal, I plan to train with an expert team of mentors, collaborators, and advisors with transdisciplinary expertise in exposomics, exposure biology, neurodevelopment, biostatistics, metabolomics, epidemiology, and microbiome research. The proposed training plan includes clinical and laboratory rotations, coursework, seminars and meetings, conferences, and 1-on-1 tutorials with mentors and advisors, covering research and career development topics. This training will provide skills and knowledge in the areas of neurodevelopment, and exposomics, including exposure biology and multi-omics analysis. Upon completion of this training plan, I will be able to achieve my long-term career goal of becoming an independent researcher focused on the inter-relationships among environmental exposures, the microbiome, and neurodevelopment. To that end, I propose leveraging the use of the established Programming Research in Obesity, GRowth, Environment and Social Stressors (PROGRESS) birth cohort, based in Mexico City, and adding microbiome analysis. Specifically, I will: 1) train with Dr. Arora in exposure biology and analysis of baby teeth as a biomarker of early life metal exposure; 2) train with Dr. Wright, in collaboration with Dr. Shamesh, in neurodevelopmental assessment, principals of neurotoxicology, and their use in epidemiologic studies; 3) train with Dr. Faith in metagenomic analysis, 4) integrate biostatistical assessment of mixtures and high dimensional data, guided by Dr. Gennings; 4) learn about stool metabolomic analysis methodology with Dr. Walker; 5) work with Dr. Tellez-Rojo to facilitate stool sample collection and data analysis in PROGRESS. I will use these training and research activities to integrate the microbiome into an exposomic analysis of neurodevelopment. I will use the research and career development training I receive to establish myself as an independent investigator with a tenure track faculty position, able to secure R01 funding. This research proposal combines innovative measures of exposure, novel data analysis approaches, and high resolution omics analysis to address gaps in the understanding of timing and potential mechanisms by which metal exposures, and the gut microbiome and metabolome, are associated with neurodevelopmental measures of attention. The proposed research and training plan builds the foundation for an independent research career that aims to clarify the links between environmental exposures and neurodevelopment, via the human microbiome.

NIH Research Projects · FY 2025 · 2023-08

PROJECT ABSTRACT MS is an immune-mediated demyelinating disease of the CNS, and despite first line drugs that limit symptoms, disease progresses over time and is incurable. Given its early onset and rise in prevalence for nearly 1 million Americans, MS presents immense health and economic burdens on the United States. Given the success of B cell depletion therapy (BCDT), understanding the pathogenic roles of B cells during disease is of high interest. Because of the CD20-targeted nature of BCDT, it is understood that B cells contribute to MS pathology through mechanisms beyond antibody production, likely through support of autoreactive CD4 T cells. Most animal models of MS (experimental autoimmune encephalomyelitis or EAE) are B cell-independent, which limits impactful insights into pathogenic B:T interactions in vivo. To circumvent this, we have recently developed a B cell-dependent, antibody-independent animal model of MS featuring CD4 T cell immunoreactivity to the extracellular domain sequences of the highly abundant and 100% conserved myelin proteolipid protein (PLPECD). Through rigorous preliminary studies, we have identified B cell-mediated antigen presentation to CD4s through MHC class II as the required pathogenic B cell mechanism in PLPECD-induced EAE, where B cells engage PLPECD through the B cell receptor and are superior vs. non-B cell antigen presenting cells at processing and presenting immunodominant residues from within PLPECD to PLP178-191-reactive CD4 T cells. Further mimicking the sustained pathogenic B cell involvement seen in MS and unlike B cell- independent EAE driven by PLP178-191, BCDT robustly ameliorates established PLPECD disease. B cells’ role in supporting CD4s during neuroinflammation and shaping the diverse T helper cell (Th) profiles observed in MS remains unclear. Understanding these dynamics in MS may prove significant in identifying which patients will respond to BCDT and thus modeling pathogenic B cell involvement in vivo is becoming increasingly important. Our objective is to use this powerful B cell-dependent EAE model to understand how B cells shape the CD4 response and to investigate B cell-mediated support of CD4s in the CNS. These are key aspects of B cell-mediated pathology that are not currently understood and difficult to model appropriately in vivo. Our model put us in a unique position to answer these questions. We hypothesize that B cell-provided cytokines shape a biased Th profile and that CNS B cell:CD4 interactions are critical in promoting demyelination in PLPECD EAE. Specific Aim 1 will determine how B cells shape pathogenic CD4 T cells by testing B cell-provided cytokines’ impact on Th1/17 bias. Specific Aim 2 will determine CNS B cells’ role in PLPECD-induced EAE and visualize B cell-supported neuroinflammation.

NIH Research Projects · FY 2025 · 2023-08

PROJECT SUMMARY/ABSTRACT Breast cancer (BC) is the most common cancer and the second leading cause of cancer death in American women. About 10-20% of breast cancers are triple-negative breast cancer (TNBC), which has a propensity to metastasize, recur, and develop resistance to chemotherapy. TNBC is the only subtype of BC for which there is no targeted therapy. Chemotherapies remain the mainstay of treatment for TNBC, but their clinical efficacy is often limited by resistance. Immunotherapy is emerging as an exciting new treatment option for TNBC patients. While TNBC is more likely to respond to immunotherapy, overall response rate is still low. Developing novel and more effective TNBC therapies is an unmet biomedical need as most of advanced TNBCs do not respond well to current therapies. Epigenetic alterations such as DNA hypermethylation and histone dysregulation have been associated with all stages of TNBC formation and progression. Lysine-specific demethylase 1 (LSD1) is the first identified histone demethylase which specifically demethylates H3K4me1/2. LSD1 is a key component of multiple transcription repressor complexes. Tumors in TNBC patients frequently express higher level of LSD1 compared to other BC groups. Clinically, LSD1 protein overexpression is significantly associated with worse prognosis in TNBC patients, making it an attractive therapeutic target. Our recent study has revealed a new mechanism driving LSD1 protein overexpression in TNBC through HDAC5-mediated posttranslational modification. Treatment with LSD1 inhibitors effectively suppresses tumor progression and sensitizes TNBC cells to chemotherapeutic agents. Furthermore, LSD1 ablation stimulates antitumor immunity and potentiates the efficacy of anti-PD-1 antibody in poorly immunogenic TNBC. LSD1 inhibition leads to reexpression of a key epigenetically silenced tumor suppressor gene, Tissue Factor Pathway Inhibitor 2 (TFPI2), which is required for tumor suppression and responsiveness to immunotherapy. Based on these findings, we hypothesize that LSD1 overexpression facilitates TNBC development and inhibition of LSD1 improves TNBC therapies by inducing TFPI2-mediated cell killing and antitumor immunity. Aim1. Determine the functional roles of LSD1 overexpression in TNBC development; Aim2. Evaluate the in vitro and in vivo therapeutic efficacy of LSD1 inhibition against TNBC; Aim3. Elucidate the immunogenic effects of LSD1 inhibition in TNBC. The results from the proposed studies are expected to provide new mechanistic insights and key preclinical evidence for using LSD1 inhibitors in TNBC. In the long run, these studies may lead to new and improved therapies for patients with relapsed and refractory TNBC.

NIH Research Projects · FY 2024 · 2023-08

PROJECT SUMMARY Zaire Ebola virus (EBOV) infections remain an emerging threat in Central and West Africa with case fatality rates reaching as high as 90%. An FDA-approved, live-attenuated, recombinant vaccine called ERVEBO has shown promise during recent outbreaks in Guinea in 2016 and Democratic Republic of the Congo (DRC) in 2019. ERVEBO stimulates antibody responses directed against the EBOV glycoprotein (GP). “Ring vaccination” is the current emergency immunization strategy that focuses on immunizing direct contacts and geographically proximal populations surrounding the epicenter of an EBOV outbreak. However, emerging data in the DRC show that ring vaccination can be highly porous; nearly 30% of EBOV-infected participants in a recent antiviral trial in the DRC were prior recipients of the ERVEBO vaccine. The mechanistic bases for these vaccine failures are not known. This multi-PI, interdisciplinary project explores the hypothesis that acute malaria impairs EBOV immunization-induced B and T cell responses. In support of this hypothesis, EBOV outbreaks largely occur where Plasmodium falciparum infections are endemic; malaria is common throughout Central and West Africa. Moreover, our collaborative team has developed experimental Plasmodium infection and EBOV vaccination systems to generate preliminary data showing that malaria dramatically impairs EBOV vaccine-induced, virus- specific antibody responses. We have also identified potential strategies to overcome the malaria-associated impairments in vaccine efficacy. In this project we synergistically apply tractable, high-resolution, antigen-specific systems to determine the mechanisms by which Plasmodium infections impact EBOV vaccine-induced humoral and cellular immunity. These new approaches facilitate our long-term goal to define the impact of Plasmodium infection on the efficacy of EBOV vaccine-induced immune responses. Our goal is addressed by three specific aims that test: 1) how Plasmodium infections impact vaccine-induced, virus specific B cell responses; 2) how Plasmodium infections influence the function of helper T cell subsets required for promoting antibody responses; and 3) how malaria affects EBOV vaccine-induced protection against virulent mouse adapted, BSL-4 EBOV challenge. Successful completion of these studies will reveal the mechanisms by which Plasmodium infections impair EBOV vaccine responses and provide clinically applicable approaches to overcome this impairment. Such knowledge gained will inform vaccine strategies that must be rapidly and effectively implemented during EBOV outbreaks.