University Of Iowa

NIH Research Projects · FY 2026 · 2025-05

While there have been great advances in HIV therapies over the past decades, the only true cure so far has been through hematopoietic stem cell transplant (HSCT) of cells naturally lacking the HIV co-receptor CCR5. Due to the toxicity of conditioning regimens prior to HSCT and significant morbidity due to graft-verses-host disease (GVHD), allogeneic HSCT is not a viable option for most patients. With the advent of CRISPR, modification of a patient’s own cells to prevent GVHD is now possible, but thus far the efficiency at which cells have been modified was not enough to prevent viral rebound in the absence of anti-retroviral therapy. In line with the rationale for combination anti-retroviral drug therapy to target multiple steps in replication, we have developed a multi-factor knock-out/knock-in strategy for editing CD34+ hematopoietic stem and progenitor cells which allows greater than 90% deletion of CCR5, as well as up to 50% allelic knock-in of two inhibitory peptides targeting either fusion (C46V2o) or uncoating (a human-rhesus chimeric TRIM5a). When using this strategy to edit primary human CD4+ T cells, the efficiency of knock-out/knock-in is sufficient for complete inhibition of CCR5-tropic virus (BaL), and an average of more than 700-fold inhibition of CXCR4-tropic virus (NL4-3) when tested across 5 different primary human T cell donors. While these data are extremely promising, there was a large disparity in the efficiency of inhibition of CXCR4-tropic replication by the human- rhesus TRIM5a, with some T cell donors showing greater than 1,000-fold inhibition in the hRhTRIM5a knock- in condition, and some showing little or no significant inhibition of replication. This observation was in spite of sustained levels of allelic knock-in and hRhTRIM5a RNA expression throughout the course of the infection, and no resistance mutations observed in the infectious virus at endpoint. We therefor propose here to study the underlying T-cell/TRIM/virus interactions which may be uniquely donor specific and influence viral replication including T cell cytokine expression, viral reactivation, and expression of endogenous cellular factors that may act as a dominant negative to our inhibitory hRhTRIM5a in addition to investigation of other TRIM-related factors for inhibition such as TRIMCyp. Although the final goal is to develop a therapy through transplantation of edited hematopoietic stem and progenitor cells, this proposal is entirely based on editing and investigating the mechanisms of restriction in differentiated primary human target cells such as CD4+ T cells, monocytes, and macrophages in order to better understand the biology within our editing platform and allow eventual improvement of the platform. Successful completion of this proposal will allow a better understanding of mechanisms of restriction in primary cells where often these mechanisms have been predominantly studied in immortalized cell lines. In addition, completion of this proposal to improve our therapeutic platform will widen the pool of patients potentially able to be treated using genetically modified autologous HSCT such as patients later in infection that may have predominantly CXCR4-tropic virus.

NSF Awards · FY 2025 · 2025-05

The 2025 conference on Stem Cells, Cell Therapies, and Bioengineering in Lung Biology and Diseases at the University of Vermont in Burlington, 7-10 July 2025, will mark the 20th anniversary of this important biennial meeting. Over the past two decades, this conference has provided a platform for advancing research and collaboration in lung biology. This meeting stimulates discussion and debate by international leaders, new and emerging investigators, and trainees in the field. Meeting participants will also develop and publish guidelines for investigators and funding agencies on basic and translational research in stem cells, lung biology, and bioengineering. Junior investigators and trainees will involved in conference organization, oral presentations, chairing activities, discussions, and poster sessions. In 2025 the High School Science, Technology, Engineering, and Math (STEM) Experience will be expanded. This program will cultivate the next generation of scientists by providing an opportunity for high school students to engage with scientists from around the world and learn about state-of-the-art scientific techniques. The repair and regeneration of diseased lungs using stem cells or bioengineered tissues represent transformative therapeutic strategies for a range of lung diseases and critical illnesses. Advances in preclinical models have demonstrated the potential for stem cell-based approaches to modulate immune responses and facilitate tissue repair following lung injury. Additionally, bioengineering research using artificial and acellular matrices as scaffolds for three-dimensional lung and airway regeneration has made strides, including promising transplantation attempts in large animal models. These efforts are complemented by a growing understanding of how endogenous lung stem and progenitor cells differentiate and contribute to repair and remodeling during lung development and after injury. In addition to attracting leading figures in lung biology and disease, the conference regularly features renowned stem cell experts from other disciplines, facilitating cross-pollination of ideas and innovation. For the 2025 conference, scientific symposia have been chosen to reflect the most cutting-edge advances in the field and will include sessions focusing on the integration of omics with function, matrix-cell dynamics, and immune cell contributions to repair and regeneration. Sessions will explore how inflammatory and mechanical cues influence stem cell regulation and highlight technological breakthroughs in bioengineering, such as precision-cut lung slices, hydrogels, and biomechanical modulation of lung function. These cutting edge basic and translational science studies are rapidly enhancing fundamental understanding of lung repair and regeneration, and the planned conference will continue to help shape further progress. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2025 · 2025-05

Patients admitted to healthcare facilities are routinely exposed to dangerous pathogens and antimicrobial-resistant organisms, leading to secondary infections unrelated to the primary reason for hospital admission. These infections, broadly termed healthcare-associated infections (HAIs), impose a significant health and economic burden, with an estimated 4.5 HAIs per 100 hospital admissions and annual costs ranging from $28 to $45 billion dollars. The health and economic burden of HAIs can be mitigated by deploying timely and effective interventions. This project aims to develop novel computational approaches to identify effective intervention strategies by leveraging electronic medical records data. The proposed intervention strategies will also incorporate predictions about potential future infections and the previously unobserved missing/asymptomatic infections inferred by machine learning algorithms. The algorithms developed will lead to practical resource recommendations before and during an HAI outbreak and help mitigate a wide range of secondary effects (e.g., the development of antimicrobial resistance organisms). A significant outcome of the project will be the formulation of a novel class of online optimization problems and the resulting suite of online resource algorithms with worst-case approximation and hardness guarantees. The project paves the way for the principled incorporation of machine learning-based epidemic forecasting models with intervention algorithms; it does so by developing learning-augmented online algorithms whose performance demonstrably improve with forecasting accuracy. It also tackles the problem of inferring unobserved infection events (e.g., asymptomatic cases and surface contamination) and incorporates them into the online intervention algorithms. Advances will be made in the evaluation of data-driven algorithms in HAI intervention, inference, and forecasting tasks by setting standardized benchmarks. The real-time recommendations made by the online algorithms developed as part of this project will lead to near-optimal resource strategies to reduce the risk of HAI and secondary outbreaks. High-school, undergraduate, and graduate student training at the intersection of computer science and public health will produce workforce with exceptional computational skills and the ability to solve critical public health problems. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2026 · 2025-05

PROJECT SUMMARY/ABSTRACT Duchenne Muscular Dystrophy (DMD) is a severe muscle-wasting disease caused by mutations in the dystrophin gene (DMD). Beyond the characteristic muscle wasting, patients with DMD are often plagued by significant skeletal comorbidities, including severe osteoporosis and consequent fracture. Although an estimated 60% of patients are affected by these comorbidities, there are no recommended prophylactic treatments. This is partially due to poor understanding of the pathogenesis of osteoporosis in DMD. Osteoporosis results when bone resorption by osteoclasts outweighs bone deposition by osteoblasts. Our lab has identified a novel myokine, fibroblast growth factor 21 (FGF21), which is increased in the serum of DMD patients and mouse models. Neutralization of FGF21 prevented osteoporosis in a mouse model by reducing the number of osteoclasts. Furthermore, in vitro studies demonstrated that FGF21 directly promotes osteoclastogenesis. However, the mechanism of FGF21 signaling to osteoclasts remains unknown. Aim 1 will investigate the role of FGF21 signaling to osteoclasts in bone homeostasis in a mouse model of DMD by using osteoclast-specific FGF21 receptor knockout mice. Aim 2 will explore how FGF21 potentiates osteoclastogenesis by evaluating signaling mechanisms and the effect on precursor fusion, an essential step in differentiation. Since preliminary data also suggest that FGF21 promotes osteoclast activity, Aim 3 will examine how FGF21 affects secretion of bone-resorptive proteins via the lysosome. These carefully and rigorously designed studies will provide valuable insight into how FGF21 promotes osteoporosis in DMD and advance our understanding of osteoclast biology. Successful completion of this project will establish FGF21 as a critical regulator of bone homeostasis and a promising target for treatment of osteoporosis in this patient population. The project and training plan described herein were developed specifically for Ms. Hurley-Novatny. The experiences, skills, mentoring, and topics of this proposal were designed to ensure that Ms. Hurley-Novatny achieves her training goals and becomes a successful, independent physician-scientist in the field of orthopedics and bone research. Under the guidance of Dr. Hongshuai Li and Dr. Matthew Potthoff, Ms. Hurley- Novatny will receive the mentorship she needs to develop as a scientist and the technical training to become a researcher in musculoskeletal biology. The MSTP, Department of Orthopedics, and Department of Anatomy and Cell Biology provide ample training opportunities in the form of seminars, collaborations, opportunities to present research, and research support both financially and otherwise. The University of Iowa has an unparalleled history of research in the field of orthopedics, providing Ms. Hurley-Novatny a unique training environment to achieve her goal of becoming a surgeon-scientist in the field of orthopedics at a major academic medical center.

NIH Research Projects · FY 2026 · 2025-05

PROJECT SUMMARY/ABSTRACT Oral health disparities by income emerge early in life in the United States, with disproportionately higher rates of untreated dental decay dental decay and unmet dental care needs more among low-income children and those in Medicaid. Poor oral health is associated with reduced school performance and lower psychosocial wellbeing. However, the extant evidence is mostly based on contemporaneous and self-reported measures of oral health, dental services use, and academic outcomes. Despite the conceptual relevance of preventive dental services and the clinical evidence on their effects on oral health, there is no causal evidence on effects of preventive dental care on academic achievement including the magnitude of effects and their timing (at what ages effects develop and oral health interventions are most effective over the child’s life). Policymakers recognize the links between oral health and school health and achievement. Some states have enacted dental screening requirements before kindergarten. However, there is no causal evidence on the effects of such policies on dental services use and academic achievement. Adding this evidence is critically important to guide policymaking by accounting for the full benefits of preventive oral health services and screening programs including those accruing to school achievement. This study leverages unique population-based dataset linking birth certificates, Medicaid enrollment/claims files, and school test scores for children enrolled in Medicaid in Iowa. The study will identify the causal effects of preventive dental services including comprehensive oral exams, prophylaxis, fluoride application, and dental sealants on children’s academic achievement including the ages of the children when these effects are most pronounced (based on age when dental services are received and on school age when academic achievement is measured). Examining the timing of effects is critical to identifying developmentally sensitive ages when these services become more effective. The study will also identify the causal effects of requiring a dental screening certificate before kindergarten on subsequent dental services utilization and academic achievement. The study will address key gaps in the literature and provide evidence of direct implications for policymaking aiming at addressing the adverse impacts of poor oral health on the children’s academic performance and overall wellbeing.

NIH Research Projects · FY 2025 · 2025-04

PROJECT SUMMARY/ABSTRACT This S10 proposal is for the purchase of a modern transmission cryogenic electron microscope (cryo- EM) to advance biomedical research at the University of Iowa. The instrument will be housed in a central location easily accessible by all university research groups. The cryo-EM instrument will be integrated into the Carver College of Medicine Protein and Crystallography Facility (PCF) that has provided researchers access and training in cryo-EM, X-ray crystallography, and additional techniques for a combined 15+ years. Through institutional investments in PCF staff training, computational resources, new faculty hires, and ancillary cryo-EM equipment, the PCF has established a workflow and provided expertise for cryo-EM structure determination. However, there is no dedicated cryo-EM instrument at Iowa to screen sample quality or determine high-resolution macromolecular structures. Thus, rapid optimization of samples and collection of datasets for cryo-EM structure determination is not possible and has a substantial negative impact on timely research project progression for numerous investigators. The acquisition of a Thermo Scientific Glacios Cryo-TEM instrument would overcome these limitations and transform the ability of University of Iowa researchers by completing our local workflow and eliminating the sample optimization bottleneck. This will increase research output by collecting high- quality data on the Glacios and only using the National Cryo-EM Centers for select samples that require Krios data collection. The proposed used Glacios microscope offers significant cost savings over the same configuration on a new instrument. It has a user-friendly software interface, high sample throughput, automated tasks, AI-optimized data collection, and high-resolution data collection which makes it the ideal single particle cryo-EM instrument for our user community. Structure determination by our investigators would enhance a diverse collection of NIH-funded projects that address biomedical research questions in areas such as ion channel function, eukaryotic and prokaryotic signal transduction, DNA repair, vision, membrane transport, cancer therapeutics, viruses, and drug delivery systems. Structural studies in these areas will also provide substantial training opportunities for students and postdoctoral scientists that will add to the core group of independent instrument users. The proposed Glacios cryo-EM will significantly impact biomedical research at the University of Iowa by increasing research productivity, enhancing scientific collaboration, boosting research grant competitiveness, and advancing efforts to attract new investigators and students.

NIH Research Projects · FY 2025 · 2025-04

Project Summary/Abstract Funds are requested to purchase a VisualSonics Vevo F2® high resolution research ultrasound imaging system for the Cardiovascular Phenotype Core at the University of Iowa, which has been in continuous operation since 1997. In the time since, the Core has provided over 40,000 imaging studies in mice and rats, and has introduced a number of novel methods for assessment of organ structure and function in vivo. In 2010, a Vevo 2100 system was purchased with NIH funds. Usage of that instrument has exceeded projections by > 50%; reaching > 4000 studies per year, in support of researchers from the University of Iowa College of Medicine and College of Dentistry. The vendor no longer supports or services the Vevo 2100 line of instruments. Vevo 2100 equipment failures have been rare but consequential, including meltdown of its central processing unit in 2021. Accommodation of new users, along with existing users, would not have been, and will not be, possible without prompt expert vendor service. The requested instrument will reside in Cardiovascular Phenotype Core space, which is already configured for that purpose, administered by the PI (Weiss). The University of Iowa Carver College of Medicine (UICCOM) will fund the instrument's service contract for 5 years after the initial one-year warranty expires, and will maintain necessary space and infrastructure without charges to the Core. The Cardiovascular Research Center will provide funds for faculty, a trainee, and one research assistant. Critically, all fixed costs of operation of the instrument will be underwritten by the institution. Users of the instrument will thus only be charged for variable costs of operation, which will be limited to sonographic technologist hourly wage plus consumable supplies necessary for imaging studies. In summary, funds are requested to support the urgent need to replace an obsolete, heretofore highly productive, high-resolution ultrasonograph, to support 15 NIH-funded research projects representing 6 departments in the University of Iowa Carver College of Medicine, one department each in the UI Colleges of Dentistry and Engineering, and the School of Engineering at University of Colorado-Boulder.

NIH Research Projects · FY 2026 · 2025-04

PROJECT SUMMARY / ABSTRACT Heart disease is a leading cause of morbidity and mortality, and the incidence of heart failure (HF) is growing, while costing our public healthcare system over $30B per year. HF coincides with significant perturbations in cardiac gene regulation, mitochondrial function, and ion channel expression and activity, and my research program embodies three independent projects related to these pathological processes. For Project #1, our goal is to define microRNA (miR) targeting events and their biological-relevance in heart and to understand the clinical significance of SNPs that alter cardiovascular miR functions. MiRs play key roles in cardiac biology and disease; however, identifying their targets is critical to understanding how. There remains a lack of empirical miR targeting data from primary tissues, and this has slowed the translational impacts of miR research. We address this by employing high-throughput methods to generate global miR-target interactomes in heart tissues. Our hypothesis is that heart disease is influenced by rewired miR-target interactions and SNPs perturbing these interactions. We are filling knowledge gaps regarding the mechanistic targets of cardiac miRs by defining and comparing miR binding sites in nonfailing and diseased human hearts. Our data point to clinically-relevant SNPs that may modulate cardiomyopathy- and arrhythmia-related miR- target interactions, and we functionally test these interactions and determine if SNPs are linked to clinical outcomes in patients. For Project #2, our goals are to better define the function of mitoregulin and establish its role in cardiac ischemia-reperfusion (IR) injury. Upon IR, mitochondrial Ca2+ overload and ROS trigger mitochondrial permeability transition (mPT) pore opening to induce cell death. We previously discovered that a heart/muscle-enriched long “non- coding” RNA encodes a mitochondrial microprotein that increases mitochondrial respiration and Ca2+ retention capacity, while reducing ROS; we named this protein mitoregulin (Mtln). The precise molecular function of Mtln remains unclear, and Mtln’s influence on cardiac stress has not been adequately studied. We are working to address our central hypothesis that 1) Mtln protects against cardiac IR injury by delaying Ca2+- and ROS-triggered mPT and/or by maintaining mitochondrial membrane integrity, and 2) Mtln overexpression (OE) will blunt cardiac IR injury and HF, providing a new therapeutic avenue. For Project #3, our goals are to define regulatory paths controlling SCN5A (encodes the primary cardiac Na+ channel, NaV1.5) and to characterize non-canonical actions of NaV1.5 beyond its known role in conduction. We discovered a SNP that bolsters miR-mediated silencing of SCN5A and associates with lower cardiac NaV1.5 levels, conduction slowing, and increased non-arrhythmic death in HF patients. Our central hypothesis is that lower SCN5A/NaV1.5 expression elevates one’s susceptibility to subclinical cardiac pathologies that transform into significant risk in the setting of HF. We are defining two novel modes of SCN5A/NaV1.5 regulation and examining underappreciated roles for NaV1.5 in cardiomyocyte metabolism and oxidative stress to better understand how lower NaV1.5 could worsen HF. Overall, these projects will advance our mechanistic understanding of heart disease at both the molecular and genetic level, with significant potential to someday improve clinical care.

NIH Research Projects · FY 2026 · 2025-04

Project Summary/Abstract The Developmental Studies Hybridoma Bank (DSHB) has existed since 1986 as a resource to facilitate the sharing of open-source monoclonal antibodies for research. The central mission of DSHB is four- fold: 1) to maintain and distribute monoclonal antibodies (mAbs) and hybridomas shared by the inventing scientists, 2) to provide this service at cost and without regard for commercial popularity, ensuring the availability of rarely used antibodies in smaller fields of research, 3) to provide accurate characterization and validation information on the antibodies and outstanding customer support, and 4) to pursue innovative methods for developing, using, and sharing mAbs. DSHB is an academic unit and has been self-supported by user fees since 1997. Whereas the savings afforded researchers and funding agencies have been substantial, the DSHB's at-cost pricing model has limited funds available for developing new ideas and practices to improve the open-source distribution of mAbs. The recent availability of mAbs from non-rodent hosts (e.g., rabbits), a dramatic increase in the overall number of mAbs (including recombinant antibodies) coupled with the awareness that many are poorly validated, and a flood of information on the use and limitations of these innumerable reagents has opened several opportunities for DSHB to expand and extend our services to these high-priority areas. The aims of this proposal are (1) to better develop new open-source non-rodent hybridoma methodology, (2) to perform antibody validation for large, less well-characterized collections of mAbs to improve model organism research, and (3) to further develop DSHB web-based informatics and expand purified antibody product offerings. We expect these proposed studies will enhance the rigor, reproducibility, and translatability of model animal research by developing technologies, tools, and resources that will enable more effective use of mAbs in animal models across a broad range of research areas.

NIH Research Projects · FY 2026 · 2025-04

Project Summary / Abstract Virtually all neuropsychiatric disorders display sex bias, with the prevalence, age of onset, and clinical symptomology differing significantly between males and females. Historically, female subjects have been excluded from preclinical research, with male-only studies in neuroscience research outnumbering those of females by 5.5:1, a discrepancy attributed to concern over the impact of estrus cycling on population homogeneity or the assumption that sexual dimorphism is absent, such that results obtained with males extrapolate to females. However, accumulating evidence demonstrates that genetic/gonadal sex and sex hormones influence several critical brain functions including the release, reuptake, and signaling response to the neurotransmitter dopamine (DA). DA is heavily implicated in substance use disorders and, notably, when compared to men, women typically report experiencing greater subjective effects from psychostimulants, are nearly twice as likely to abuse these drugs, and display enhanced dopamine release in the nucleus accumbens (NAc), a critical component of the mesolimbic dopaminergic reward circuit, following psychostimulant exposure. Similarly, in rodents, there are robust sex differences in the behavioral impacts of the psychostimulant amphetamine with females exhibiting greater amphetamine place preference scores but blunted hyperlocomotion relative to males. Though existing evidence points to a critical role for gonadal hormones in determining differential psychostimulant responses in females, the neural substrate(s) linking steroid hormone signaling to synaptic DA availability remain unelucidated. We recently described a sex-biased and region-specific functional coupling between the dopamine transporter (DAT), a direct target of psychostimulants and primary mediator of synaptic DA clearance. More specifically, the ability of D2 autoreceptors (D2ARs) to promote phosphorylation and surface trafficking of DAT occurs in the dorsal striatum, the critical efferent target of nigrostriatal dopamine neurons heavily implicated in DA-driven locomotor behavior but is absent in the reward- linked NAc of male mice. In contrast, a functional coupling between D2AR and DAT is observed only in the ventral striatum (NAc) in females. The central hypothesis of this proposal is that gonadal hormone signaling differentially enhances D2AR-dependent DAT regulation in the nigrostriatal vs mesolimbic dopaminergic circuits translating into sex-selective impacts of AMPH on synaptic DA and behavior. First, we will establish if/when the actions of gonadal hormones in the brain define sex-biased patterns of D2AR effector coupling and the specific hormone/receptor signaling involved. Second, we will investigate how circuit-specific DAT regulation via D2AR shapes amphetamine-dependent DA outflow and behavior in a sex-biased manner. Ultimately, delineating the mechanisms contributing to the sex biased architecture of the dopaminergic circuitry is imperative and has the potential to identify novel therapeutic targets for the treatment of mental illnesses including psychostimulant dependence for which there are currently no available pharmacotherapies.

NIH Research Projects · FY 2026 · 2025-04

ABSTRACT Age-related macular degeneration (AMD) is a major cause of vision loss, affecting millions of individuals. Two major loci (ARMS2/HTRA1 and CFH) drive most of the risk for AMD. The genes responsible for the pathophysiology of these loci are expressed in ocular cells where they exert their effects. New developments in genome editing have greatly increased the efficiency of editing and reduced the likelihood of off-target events; and the application of these technologies (base editing and prime editing) offers promise for the permanent reduction of risk, especially important as a means to slow or arrest disease progression. Whereas existing treatments for exudative and atrophic AMD can be employed only following neovascularization or geographic atrophy, genome editing to diminish AMD risk in eyes with high-risk genotypes could be employed in a preventative manner. This grant application seeks to address the technological and tissue-specific considerations for editing the pathogenic alleles in two AMD loci (10q26 harboring ARMS2/HTRA1, and 1q harboring CFH). In this multicenter program with experts in retinal stem cell biology, AMD pathology, base editing and prime editing, virus like particle packaging, and gene expression, we will conduct base editing and prime editing of risk loci in human cells including iPSC-derived RPE, retina, and choroid, and donor retinal and choroidal explant cultures. Upon completion, this research program will address fundamental science questions regarding the impact of ARMS2 A69S gene on HTRA1 expression and the role of CFH variants in complement deposition. Additionally, it will shed light on translational science questions, exploring the efficacy of advanced gene editing for modifying critical risk genes in AMD.

NIH Research Projects · FY 2026 · 2025-03

PROJECT SUMMARY/ABSTRACT Maintaining the appropriate volume of airway surface liquid is crucial for respiratory health. Airway epithelia control airway surface liquid volume by balancing the absorption and secretion of liquid. Recent studies re- vealed that ionocytes perform most of the salt and liquid absorption, and secretory cells perform most of the secretion. Both cell types use cystic fibrosis transmembrane conductance regulator (CFTR) channels to control Cl– movement across the apical membrane. This raises the question—how do airway epithelia balance the contributions of two neighboring cell types that perform opposing functions through the same apical channel? The long-term goal is to establish new regulatory nodes between ion transport, its regulation, and physiological function and then target these mechanisms in pulmonary diseases. The overall objective of this application is to define the mechanism by which PGE2 balances absorption and secretion to target novel mechanisms that control airway surface liquid volume. The central hypothesis is that PGE2 production balances ASL absorption and secretion by regulating ionocytes and secretory cells simultaneously. The central hypothesis will be tested by pursuing the following specific aims posed as questions: 1) How does PGE2 balance airway epithelial Cl– absorption and secretion? 2) How does PGE2 modify the ASL? The proposed research is innovative because it tests the hypothesis that PGE2 turns off CFTR in ionocytes while turning on CFTR in secretory cells. The mechanism is the first example of how one molecule can control CFTR in two neighboring cells performing op- posing functions. The proposed research is significant because results identify new targets for therapeutic in- terventions in pulmonary diseases.

NIH Research Projects · FY 2026 · 2025-03

Project Summary Antioxidant proteins which regulate reactive oxygen species (ROS) are often considered to be protective in the setting of intestinal inflammation and disease, such as inflammatory bowel disease (IBD). Glutathione peroxidase 1 (GPx1) is a ubiquitously expressed selenoenzyme and potent antioxidant which decreases cellular hydrogen peroxide (H2O2). However, our previous studies using Gpx1-/- mice have indicated that unlike many antioxidants whose loss exacerbates murine colitis, loss of GPx1 confers striking protection from dextran-sodium sulfate (DSS)-induced colitis. Further investigation has suggested GPx1’s protective effect may be due, at least in part, to increased intestinal proliferation and stem cell activity which may help to prevent or repair colitis- induced injury. However, specifically testing epithelial-dependent roles for GPx1 has been hindered by the lack of necessary mouse models, as a mouse model capable of cell type-specific deletion had not yet been developed for Gpx1. Although we have improved our understanding of GPx1’s epithelial roles by use of ex vivo intestinal organoids, which indicate that the observed changes in intestinal cell growth are epithelial-cell intrinsic, our reductionist models cannot truly investigate the contribution of these findings to a complex, multifaceted disease setting such as that observed during IBD. To address this need and better leverage our previous research results, we have now generated a novel Gpx1 floxed mouse model. These mice have subsequently been crossed with the Vil-CreERT2 line, allowing us to restrict Gpx1 loss to intestinal epithelial cells. For the first time, these mice will allow us to specifically investigate GPx1’s epithelial-dependent contributions to intestinal homeostasis, stem cell biology, and experimental colitis as implicated in our K01-funded studies. These contributions will be examined in two specific aims. The first aim will clearly delineate epithelial-dependent versus -independent effects for GPx1 in intestinal homeostasis, mucosal immunology, and colitis. The second aim will complement broader epithelial-based studies by more specifically determining how GPx1 loss alters intestinal stem cell biology. This aim will also begin to explore potential mechanisms by which GPx1 may affect intestinal homeostasis and disease, with initial studies focused on roles in the WNT signaling pathway. Together, these studies will address a major technical limitation, characterize novel research reagents, and provide clarity and preliminary feasibility for future studies as we work to build upon our previous findings on GPx1 in the intestine. Thus, this award will provide greater means to expand my independent research program focused on understanding mechanisms by which GPx1 and antioxidant proteins contribute to intestinal health and disease. As previous research has indicated that optimal ROS levels are key to maintaining intestinal health, ultimately these studies will advance our knowledge of IBD pathobiology and therapeutic options to reduce development, diminish severity, and promote intestinal healing in the setting of intestinal inflammation and IBD.

NIH Research Projects · FY 2026 · 2025-03

PROJECT SUMMARY / ABSTRACT Heart failure (HF) is a leading cause of morbidity and mortality worldwide. HF is associated with high risk of fatal ventricular arrhythmias and sudden cardiac death. The pathophysiological basis of systolic HF includes the impaired function of existing cardiomyocytes and their loss. While cardiac regeneration is a hot area of study to regenerate new cardiomyocytes, it is equally important to continue improving our understanding of the mechanisms underlying cardiomyocyte damage and dysfunction in HF. At a cellular level, defects in excitation- contraction (E-C) coupling or Ca2+ signaling is a hallmark of HF. Cardiomyocyte E-C coupling occurs in specialized ultrastructural domains, i.e., cardiac dyads or junctional membrane complexes (JMCs), which are established by physical couplings between the transverse tubules (T-tubules) and the terminal cisternae of sarcoplasmic reticulum (SR). The Junctophilin family proteins (JPs) are critical in establishing and organizing JMCs. There are four isoforms of JPs found in excitable tissues (JP1-4). It is generally accepted that JP2 is the only JP isoform found in cardiac muscle, responsible for the formation of JMCs. Our lab recently found that JP1, enriched in skeletal muscle, is also expressed in cardiac muscle, although at a much lower level in both human and mouse heart tissues. To dissect the function of JP1 in the heart, we generated cardiac specific JP1 knockout (JP1cKO) and 3xHA-tagged JP1 knockin (3xHA-JP1KI) mouse models. Our preliminary data demonstrate that JP1 co-localizes with JP2 and RyR2, the primary Ca2+ release channel, in the Z-disc. JP1cKO mice progressively develop systolic HF and die prematurely. 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 in the heart. This led us to hypothesize that JP1 is required for normal cardiac function and is functionally distinct from JP2 in cardiomyocytes within the cardiac dyad. The goal of this proposal is to unravel the physiological function of JP1 in the heart and its potential significance in heart disease. With the necessary unique mouse models already available in our laboratory, we will test the hypothesis using a multidisciplinary approach in three specific aims: Aim 1, to investigate the functional role of JP1 in cardiac muscle by determining the structural and functional consequences of depleting JP1 from cardiomyocytes; Aim 2, to define the mechanism by which JP1 regulates cardiomyocyte structure and function by illustrating JP1-interacting molecular targets and signaling using candidate and unbiased proteomics-based approaches, co-immunoprecipitation, and high resolution imaging. Aim 3, to investigate the mechanism and significance of JP1 dysregulation during cardiac stress. Accomplishment of this project will have fundamental implications for furthering our understanding of the mechanisms underlying cardiac function and its regulation by characterizing a previously unappreciated protein, JP1, in the heart.

NIH Research Projects · FY 2026 · 2025-03

PROJECT SUMMARY/ABSTRACT Fuchs endothelial corneal dystrophy (FECD) affects 6.1 million Americans over 40 years of age, is the leading indication for corneal transplant surgery, and although it can be diagnosed early it requires corneal transplantation because no medical therapy can prevent its progression. Genetic factors, abnormal cell-matrix interactions, and oxidative stress induced by ultraviolet light-A (UVA) all contribute to FECD, but the precise mechanisms inciting cell death remain poorly understood. We present novel and compelling evidence that ferroptosis, a nonapoptotic oxidative cell death resulting from iron-mediated lipid peroxidation, is a key driver of corneal endothelial cell (CEC) death in FECD. Specifically, increased reactive iron (Fe2+) concentrations, lipid peroxidation, and expression of transferrin receptor (TFR1, a ferroptosis-specific marker) occur in corneal endothelial tissue from FECD patients. These data are foundational and guide us to further investigate drivers of ferroptosis in FECD. Pathological extracellular matrix features in FECD (e.g., guttae) are recognized risk factors associated with premature CEC death in FECD. Exciting preliminary data indicate that FECD patients have increased TFR1 expression surrounding guttae compared with adjacent cells, which compels us to study the role of guttae in driving ferroptosis. Additionally, UVA exposure is a risk factor that drives FECD progression and triggers ferroptosis in other exposed tissues (e.g., skin). Exciting preliminary data demonstrate that UVA increases Fe2+ concentrations, which leads directly to Fenton reactions and oxidative damage in lipid membranes with subsequent ferroptosis-induced cell death in FECD. Our overall goal is to develop a preventive medical therapy (e.g., eye drops) that preserves corneal function, protects against vision loss, and increases quality of life without surgery for FECD patients. To achieve this, we will target iron-mediated lipid peroxidation that is involved in FECD progression. Our team is well positioned to conduct this research due to our expertise in ferroptosis and animal models of FECD and phamacoengineering. The overall objectives of the proposed research are to investigate how guttae and UVA drive ferroptosis in FECD and develop anti-ferroptosis drug therapy using in vitro and in vivo FECD models. Our central hypothesis is that FECD genetics predispose CECs to ferroptosis, and that guttae eruption through the CEC monolayer and UVA exposure both drive aberrant reactive iron to accumulate in cells and cause ferroptosis. Guided by strong preliminary data, the hypothesis will be tested through the following specific aims: 1) Define the interplay between guttae and ferroptosis in FECD; 2) Determine how UVA mediates ferroptosis in FECD; and 3) Modulate ferroptosis to mitigate cell loss in FECD. The approach is innovative in that we will utilize well-characterized human tissues and primary cells from FECD patients, multiple animal models of FECD, and state-of-the-art technical assays. The proposed research is significant because it will clarify the mechanism of how genetic and environmental risks lead to premature cell death through ferroptosis in FECD, and advance targeted drug therapy to prevent or delay its progression.

NIH Research Projects · FY 2026 · 2025-03

ABSTRACT The ability to generate patient induced pluripotent stem cells (iPSCs) has revolutionized the field of regenerative medicine. Like many researchers, we foresee a future where autologous stem cell-derived tissues will be used to treat a wide variety of neurodegenerative disorders, including inherited retinal degenerative blindness. One of the greatest hurdles for autologous cell replacement is that validated patient-specific therapeutics are difficult to produce using traditional manufacturing strategies, which are designed for large scale production of a single product to treat a large patient population. Despite this, decades of human transplant experience (spanning many organs and tissue types) have consistently shown that immunologic matching has a significant impact on graft function and longevity. To enable autologous cell production, we recently reported development of a robotic cell culture platform with imaging capabilities, that can automatically identify, pick, weed, and feed patient-derived cells over the life of a manufacturing campaign. While a step in the right direction, several hurdles to efficient production of autologous cell therapeutics remain. First, despite the use of well characterized iPSC generation protocols, significant variability between patient iPSC lines with respect to retinal differentiation capacity exists. For iPSC lines that lack the ability to efficiently differentiate into the target cell type, production costs and time are significantly increased. Second, traditional methods used to evaluate the clinical suitability of a product are inherently destructive and require repeat sampling throughout the manufacturing campaign (e.g., repeat immunohistochemical analysis during differentiation to track development). Furthermore, it is impossible to evaluate the clinical product intended for human use when destructive release tests are utilized (e.g., impossible to perform RNA-seq analysis on the actual graft that will be placed in a human). In this application, we propose two specific aims which are designed to address each of these major manufacturing hurdles. In Aim 1, we use imaging data collected during iPSC generation and retinal derivation to train an AI algorithm to select lines with a propensity for retinal differentiation. This would eliminate the wasted time and resources that are currently associated with using lines that are difficult to differentiate into retinal tissue. In Aim 2, we will correlate developmental transcript expression with unbiased imaging data collected throughout the retinal differentiation process, to identify morphological cues that can be used to train our robotic platform to assess organoid maturity and enable non- destructive clinical release testing. Upon completion of this program, we believe that we will have generated image analysis algorithms that can be used with automated cell culture platforms to enable production of patient iPSC lines with excellent retinal differentiation capacity, permit tracking of photoreceptor cell development, and allow for the non-destructive qualification of the actual product intended for clinical use.

NIH Research Projects · FY 2026 · 2025-02

Aspergillus fumigatus (Afu) is the primary human filamentous fungal pathogen and has been designated by the World Health Organization as one of the four most critical fungal pathogens impacting humans. The patient population at risk to Afu infections continues to rise owing to increased utilization of immune suppression for organ transplants, viral infections and cancer chemotherapy. While our need to understand the pathogenesis of aspergillosis is high, the experimental tools for genetics in Afu are limited. Most genetic approaches rely on changing the level of the gene of interest using disruption or overproduction strategies. Doxycycline-regulated promoter systems have also been used but these also only indirectly effect the level of the target protein by altering its mRNA production. Here we describe a new approach to analysis of the transcriptional control of virulence in Afu by regulated proteolysis of transcription factors already known to be important in pathogenesis. We have expressed an anti-GFP nanobody (GFPNb: single chain antibody from camelid species) fused to a mammalian E3 ligase enzyme (Rnf4) and found this fusion protein to trigger proteolysis of GFP fusion proteins in Afu. Expression of this fusion protein is under the control of a doxycycline-inducible promoter system allowing us to trigger protein degradation with the addition of doxycycline. We propose to accomplish two goals in this application. First, we will prepare GFP fusion proteins to 8 different transcription factors already known to be required for virulence when deleted from the cell. We will test all of these fusion proteins for localization, expression, phenotype and the ability to be degraded by the GFPNb-Rnf4 fusion protein. We will select up to 4 of these fusion proteins to test in a mouse infection model based on their retention of a normal in vitro phenotype and sensitivity to the GFPNb-Rnf4-mediated degradation. Immunosuppressed mice will be infected with these strains and then treated with doxycycline at different times to determine if the acute proteolytic removal of a factor of interest has an impact on fungal burden/survival. We will also confirm loss of a given factor after doxycycline treatment by western analysis of proteins from the mouse lung. This approach of directly degrading a transcription factor of interest, coupled with the ability to do this in a time-dependent manner, represents a fundamentally different means of interrogating the progression of mammalian infection by Afu.

NSF Awards · FY 2025 · 2025-02

The powder compaction process is commonly used to turn loose powder into a solid form. During this process, fine powder particles are placed into a mold and then pressed together using a machine. Pressure is applied to compress the powder into a solid shape. Powder compaction is widely used in the pharmaceutical industry to produce tablets. When compacting smaller tablets, multiple tablets can be made simultaneously in a single compaction cycle using multi-tip tooling, which contains more than one mold. However, the compaction process becomes increasingly challenging as the size of the tablets decreases, especially with the use of multi-tip tooling. This research investigates how laboratory-generated data can be combined with computational modeling to produce high-quality tablets. Specifically, the study will use computational modeling to simulate how powders behave within the mold and assess the impact of multi-tip tooling design on tablet quality. The goal is to facilitate the use of smaller tablets in personalized medicine to advance national health. Additionally, this project supports education and diversity by training graduate and undergraduate students, including those from underrepresented minorities and women. The use of multi-tip tooling enables the compaction of multiple products simultaneously in a single cycle, offering significant efficiency gains. Multi-tip tooling is especially promising for producing small-sized, high-volume components with simple geometries, such as pharmaceutical mini-tablets. However, this advancement introduces new challenges in optimizing process parameters, as the configuration of multi-tip tooling differs significantly from traditional tooling setups. To fully capitalize on the potential gains from using multi-tip tooling, a thorough understanding of both the formulation and tooling configurations is essential. This project aims to use experimentally validated numerical analysis with the Discrete Element Method to gain fundamental insights into the micromechanical behavior of powders when used with multi-tip tooling, focusing on both formulation and tooling design aspects. This EPSCoR Research Infrastructure Improvement (RII): EPSCoR Research Fellows project would provide a fellowship to an assistant professor and training for a graduate student at the University of Iowa. Our goal is to obtain accurate, realistic, and computationally efficient estimations to guide the rational selection of compaction parameters, considering both the powder micromechanical behavior and tooling design. The proposed research will yield substantive new knowledge with respect to the use of multi-tip tooling for powder compaction and provide a powerful tool for designing multi-tip tooling with applications in many technological areas. Thus, the knowledge generated by this study has far-reaching implications that transcends the pharmaceutical industry, addressing a wider range of scientific challenges across various industries where powder compaction holds relevance. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2026 · 2025-02

Abstract Overview. This study will characterize clinically relevant novel phenotypes related to osteoporosis and sarcopenia in chronic obstructive pulmonary disease (COPD) by leveraging established datasets from nationwide lung studies, artificial intelligence, and image and data analytic methods. This project will build normative models of distinct phenotypic aging and deliver chest CT-based automated measures of breathing-related diaphragm deformation between two lung volumes (ΔLV) and other static features of thoracic bone and muscle. Data of healthy never-smoking participants of the Multi- Ethnic Study of Atherosclerosis Lung (MESA Lung) and Genetic Epidemiology of COPD study (COPDGene) will be used to develop normative models of phenotypic aging, while COPDGene data will be analyzed to uncover distinct pathways of thoracic musculoskeletal aging and characterize relevant phenotypes in COPD. Methods. Osteoporosis phenotypes will be characterized using spinal bone density, strength, and fractures, while sarcopenia in mass and function phenotypes will be defined using pectoral muscle mass and ΔLV metrics of lung diaphragmatic surface deformation, respectively. Static bone and muscle metrics will be computed from an inspiratory chest CT scan, while the ΔLV metrics of lung diaphragmatic surface deformation will be derived over inspiratory and expiratory scans. Aim 1 involves development and validation of CT-based automated methods to drive static metrics of spine, individual vertebrae, and pectoral muscles and ΔLV metrics of lung diaphragmatic surface deformation between inspiration and expiration. Aim 2 to characterize different phenotypic groups in COPD by (i) developing normative aging models for different phenotypic metric groups, (ii) establishing a method to compute the phenotype-specific age of each participant, and (iii) determining participant’s phenotype(s) based on their chronological and phenotypic ages. Aim 3 to characterize the association of body-size, spirometric and radiographic markers, and clinical outcome metrics with different phenotypic groups and to explain the prevalence and overlaps of these groups with COPD severity. Novelty. (i) Establishment of clinically relevant osteoporosis and sarcopenia phenotypes in COPD. (ii) Development of normative models for different phenotypic aging. (iii) Development of methods to determine phenotype-specific age of individual participants. (iv) CT-based automation of ΔLV metrics of breathing-related lung diaphragmatic surface deformation. Strengths. Established longitudinal data repositories, multi-disciplinary expertise of the research team, the PI’s experience with artificial intelligence and quantitative analysis in lung and musculoskeletal imaging, and strong preliminary data. Deliverables and Significance. Characterization of new phenotypes will facilitate the understanding of mechanistic associations of osteoporosis and sarcopenia with different risk factors and comorbidities in COPD and their impacts on disease progression and clinical outcomes. Normative models of phenotypic aging will offer references to distinguish different pathways of musculoskeletal aging and study their impacts on various pulmonary and cardiac diseases. Automation of CT-based measures of bone, muscle, and lung diaphragmatic surface deformation will enable translation of the study outcomes to large research and clinical collections of chest CT scans.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY Three decades of research on social neuroscience have identified a set of brain regions supporting human social cognition; however, critical gaps in our understanding of the causal mechanisms between these regions prevent the explanation and treatment for disorders affecting social cognition, such as autism. Our long-term goal is to describe the relations among brain regions and the networks that determine social behavior in health and disease The overall objective for this application is to sharpen our understanding of the causal role for the medial prefrontal cortex (MPFC) in the brain network for social cognition. The central hypothesis is that a unique combination of the lesion method, well-established behavioral assessments, and high-quality neuroimaging data will provide evidence that MPFC is a causal node in the brain network for social cognition. The rationale for this project is that identifying MPFC’s role in orchestrating brain regions and producing social behavior is likely to provide a strong basis for the development of neurobiological models of social cognition in conditions where the function is impaired (e.g., autism, schizophrenia). Guided by supportive preliminary data, the following three specific aims will be pursued: In Aim 1, the predictive value of MPFC lesion location on social cognitive profiles will be determined by using multivariate lesion symptom mapping in Iowa Neurological Patient Registry participants who will complete a new social task battery extending currently limited social cognitive characterization. In Aim 2, we will evaluate MPFC function during social behavior with a well-established social inference task during 7T functional magnetic resonance imaging (fMRI) in individuals with lesions to regions i) in MPFC, ii) in the social brain network outside of MPFC, iii) outside of the social brain network and iv) healthy adults (matched for e.g., age, sex, IQ). In Aim 3, MPFC’s role in the social brain network will be validated by comparing how lesions to MPFC impact a graphical network model derived from densely sampled resting state 7T fMRI data from large neuroimaging datasets and well-matched new collected healthy samples. The research proposed in this application is innovative, because it combines big data (> 2000 subjects with and without focal brain lesions) with novel applications of lesion approaches towards understanding the neurobiology of human social cognition. The proposed research is significant, because it is expected to provide a strong scientific foundation for the development of neurobiological models of social cognition in conditions where the function is impaired.

NIH Research Projects · FY 2026 · 2025-01

Abstract Dementia is associated with staggering economic, social, and personnel costs. Strikingly, most dementia cases have co-pathologies beyond the dominant type. Research suggests these non-dominant protein aggregates can impact cognition, symptoms, and progression. These findings motivate our central hypothesis that individual pathologies uniquely contribute to the degenerative trajectory of clinical dementia. Our long-term goal is to build a model of degenerative dementia that is inclusive of co-pathology. Accomplishing this goal requires identifying pathologies and determining how each pathology contributes to neurodegeneration, including synapse loss, cell loss, and clinical symptoms. To accomplish this goal, we will first determine the relationship between pathology in cutaneous nerves and pathology in the brain of individuals that died with dementia, including Alzheimer’s disease or related disorders. There is strong evidence tying alpha-synuclein aggregates in peripheral nerves with Parkinson’s disease. However, it is unknown how this clinically available biomarker relates to brain pathology in patients with dementias. In Aim 2, we will use both unbiased stereology as well as high throughput screening methods to evaluate the relationship between brainstem pathology and cell loss. Pathology in brainstem projection neurons such as norepinephrine producing locus coeruleus, serotonin producing dorsal raphe, and acetylcholine producing pedunculopontine nucleus have the potential to substantially impact the brain and its response to pharmacological treatment. Although substantial evidence supports the premise that these regions show pathology, few studies have compared how different pathologies impact cell morphology or death. Finally, in Aim 3 we will directly quantify how specific pathologies impact cortical synapse loss. Synapse loss is considered one of the strongest predictors of cognitive deficits. However, studies are not definitive on which pathologies best predict this loss. We will use converging techniques, including Golgi-staining and high- resolution immunofluorescence to increase generalizability and reproducibility when addressing this critical question. Underdiagnosis of co-pathologies impacts care and research and undermines our understanding of neurodegenerative dementias. These aims address the role and pattern of co-pathology and begin to bridge the gap from the gold standard brain autopsy to clinically available peripheral biomarkers.

NSF Awards · FY 2025 · 2025-01

With support from the Chemical Structure and Dynamics (CSD) program in the Division of Chemistry, and the Established Program to Stimulate Competitive Research (EPSCoR), Professor Aditi Bhattacherjee of the University of Iowa is investigating the early steps in the activation of single-site transition-metal catalysts when irradiated with visible light. Catalysts are molecules that facilitate chemical reactions but they themselves are not consumed during the reaction. Understanding catalyst function is a challenge, as their chemical behavior oftentimes involves the motion of charge between the catalyst's subcomponents on very short time scales. Professor Bhattacherjee and her students will utilize femtosecond X-ray transient absorption spectroscopy to watch photoexcited charges as they move through a catalyst, determining both the time scales and pathways they take. Their discoveries could lead to a better understanding of single-site catalysts used extensively in polymerization reactions. The education plans will weave professional skill development into chemistry curricula by developing a content-context balanced pedagogy that is mindful of diverse student experiences. The central objective of this proposal is to identify the intersystem crossing rates and photochemical branching ratios in functionalized metallocene catalysts with atomic site specificity to inform a fundamental photophysical understanding of organometallic spin crossover concerning light and heavy atoms in multi-coordinating ligand fields. The charge transfer steps and spin-orbit coupling mechanisms in the photoactivation of these molecular catalysts will be resolved at the level of the frontier orbitals and the constituent metal and ligand spheres. Atom-specific spectroscopies such as X-ray spectroscopy have the potential to resolve the charge transfer steps precisely. The energy gaps and orbital mixing coefficients will be measured directly in different charge transfer excited states, which can be compared with electronic structure calculations. The project will contribute to the advancement of tabletop ultrafast X-ray spectroscopy. The results will also enable a mechanistic understanding of photocatalysis with earth-abundant metals and be of wide interest in fields encompassing non-adiabatic chemical dynamics, excited-state charge transfer, and photoredox chemistry. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2026 · 2025-01

Summary Nonsense mutations are point mutations that convert an amino acid encoding codon into a premature termination codon (PTC). The consequences of PTCs are nonsense-mediated mRNA decay, and for transcripts that persist, premature translation termination. PTCs are a major cause of human disease, accounting for >10% of all described disease-causing genetic alterations, including nearly all forms of hereditary eye disease (retinal disease, corneal disease, glaucoma, congenital cataract, retinoblastoma, syndromes impacting vision, and others). There has been a longstanding appreciation that therapies promoting readthrough of PTCs might be broadly useful for treating many different diseases which are all fundamentally initiated by PTCs. In recent years there has been a surge in PTC therapy research using modified suppressor tRNA molecules. In this approach, tRNA with altered anticodons recognizing stop codons are utilized to base- pair to the PTC and allow translation. Remarkably, suppressor tRNAs appear to recognize PTCs and natural termination codons distinctly. Recently, we have conducted screens for anticodon engineered tRNA (ACE- tRNA) with a high suppression activity. Here, we utilize a newly generated strain of transgenic mice that ubiquitously express one of these, Arg-TGA ACE-tRNA, to assess its in vivo efficacy, safety, and activity in multiple ocular cell types. To achieve this, Specific Aim 1 will assess the ability of transgenic encoded ACE- tRNA-ArgUGA to rescue the pigment dispersing iris disease caused by the GpnmbR150X PTC mutation and the retinal degeneration caused by the Rd3R107X PTC mutation. Control cohorts will also be characterized with a variety of assays to test whether there is phenotypic evidence of damage induced in ocular tissues by presence of the ACE-tRNA-ArgUGA. To complement these experiments, Specific Aim 2 uses a reporter strain of mice to characterize whether there are any ocular cell types with unique features changing their sensitivity to ACE-tRNA-ArgUGA. Broadly, the results from SA1 and SA2 will form a rigorous initial test of in vivo efficacy, safety, and cellular activity of ACE-tRNA in the eye. The combined outcomes allow many opportunities to observe the degree to which this ACE-tRNA-ArgUGA can provided sustained correction of PTC mutations in the eye, whether it induces any additional detectable abnormalities, and whether there are any yet unknown biological processes regulating protein translation in some cell-types that impact how ACE-tRNA function.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY Bacterial gene expression can be regulated at nearly every conceivable point across the central dogma of molecular biology where DNA is transcribed into RNA, which is in turn translated into proteins. Following transcription, mRNA transcripts are subject to regulation by so-called post-transcriptional regulators, which includes both RNA-binding proteins (RBPs) as well as short (50-500 nt), non-coding RNA transcripts, referred to as small RNAs, or sRNAs. The post-transcriptional regulation networks comprised by the sRNAs and associated RBPs are thought to allow bacteria to rapidly respond to changing environmental conditions by enhancing plasticity within gene expression networks. Research in my lab combines large-scale, genome-wide analyses facilitated by next generation sequencing approaches with classic bacterial genetics techniques. We utilize these approaches in a complimentary fashion; we leverage the sequencing results to inform the genetics experiments, and vice versa. Ultimately, we use the data to generate and test hypotheses about novel regulatory networks and regulatory mechanisms, with a particular focus on the RNA binding proteins, sRNAs and post- transcriptional regulatory networks in the gram-negative bacterium, Acinetobacter baumannii, a notorious opportunistic human pathogen. A. baumannii is a frequent cause of ventilator associated pneumonia, skin and soft tissue infections, and other hospital acquired infections. These organisms are amongst the most problematic species in the context of antibiotic resistance, where extensively drug resistant isolates present an urgent crisis to our healthcare system, especially considering that contemporary isolates also demonstrate increased virulence properties relative to ancestral strains. In this proposal, I present two major research projects in my laboratory, which address key concerns for understanding bacterial pathogens through the lens of understanding post-transcriptional regulation. Post-transcriptional regulatory networks remain largely unstudied in A. baumannii. We previously performed experiments to identify and characterize key RNA-RNA interactions and identified hundreds of RNA-RNA interactions, suggesting the existence of a robust post-transcriptional regulatory network in A. baumannii. In this proposal, we propose to systematically dissect relevant RNA-RNA interactions that are mediated by a key RNA binding protein, called Hfq, and further characterize how this essential protein integrates RNA-mediated regulation with cellular RNA processing enzymes. Collectively, the results of these studies will provide a comprehensive picture of the sRNA regulatory landscape in a significant bacterial pathogen and will enhance our understanding about the post-transcriptional regulatory network coordinates gene expression in the context of human disease.

NIH Research Projects · FY 2026 · 2024-12

Project Summary / Abstract Hypertension is an important public health challenge in the United States due to its high prevalence and strong association with stroke, myocardial infarction, congestive heart failure, and end-stage renal diseases. Although effective therapies for hypertension exist, many adults with hypertension remain resistant to treatment. This makes hypertension an important medical and public health issue and highlight the need to identify new mechanisms of hypertension. The cilium is an evolutionary conserved organelle that serve as a sensor of extracellular cues and the transduction of those cues into cellular signaling ultimately affecting a wide range of cellular and physiological processes. The goal of this project is to study the role of neuronal primary cilia in hypertension, autonomic dysfunction and body fluid imbalance. We recently discovered that neuronal primary cilia regulate blood pressure and that hypertensive animals display abnormally elongated cilia in the supraoptic nucleus, a key brain nucleus for blood pressure and fluid homeostasis. We also uncovered a novel role for the renin-angiotensin system in the control of primary cilia. Based on these exciting findings, we propose to test the novel hypothesis that neuronal primary cilia contribute to hypertension, fluid imbalance and sympathetic nerve activation through a mechanism that involve Angiotensin II type 1a receptor (AT1aR) signaling. We will evaluate how conditional ablation of cilia in the supraoptic nucleus affects blood pressure, drinking, urine volume, sodium concentration, plasma osmolality regional sympathetic nerve traffic and neuronal activity in hypertensive mice. We will also examine how these parameters are affected by AT1aR deficiency in the supraoptic nucleus. Finally, we will probe the mechanisms underlying cilia regulation by the AT1aR. The current proposal should fundamentally advance our understanding of the neuronal mechanisms that mediate regulation of blood pressure, sympathetic traffic and body fluid balance in health and disease. Insights into these mechanisms may make it possible to selectively interfere with the hypertension and other cardiovascular risks.