University Of Iowa

NIH Research Projects · FY 2025 · 2020-07

ABSTRACT Parkinson’s disease (PD) is the most common hypokinetic movement disorder. It is characterized by an over- inhibited motor system, which results in bradykinesia, tremor, and stiffness of gait. Deep-brain stimulation (DBS) is the primary neurosurgical treatment of PD. It commonly targets the subthalamic nucleus (STN) of the basal ganglia, the primary inhibitory node in the motor system. One purported mechanism underlying STN DBS is that it reduces STN’s inhibitory influence, thereby moving the pathologically inhibited motor system back to a more typical excitation-inhibition balance. However, this is only one of the many competing explanations for the mechanism underlying STN DBS’ therapeutic effect. To adjudicate between these explanations, there is a critical need to precisely assess the inhibitory motor control function of the STN. The premier method to assess inhibitory motor control is the stop-signal task (SST), in which inhibition is needed to periodically stop an already-initiated action. Converging evidence shows that STN activity correlates with the ability to successfully stop an action in the SST. As such, the SST is the ideal method to assess the mechanism(s) underlying STN DBS, as it links STN activity with its purported function. The prediction is straightforward: if DBS indeed reduces STN’s inhibitory influence, it should impair SST performance. However, existing work has produced a highly contradictory, paradoxical picture of STN DBS’ effect on motor inhibition: while some studies do show the predicted pattern, other studies show the exact opposite: DBS seemingly improves action-stopping. The current proposal asks a very simple question: How can this be? Previous work has not been able to answer that question, partially because small sample sizes prevented the investigation of individual differences, and partially because methods to assess some key variables did not exist. The core hypothesis of the current work is that DBS’s effects on motor inhibition are mediated by DBS lead placement (Aim 1), cortico-STN connectivity (Aims 1 & 2), and/or task-related cortical dynamics (Aim 3). We propose to investigate these aspects in a large sample of N=100 PD patients, who will perform two sessions of the SST – On and Off DBS. In Aim 1, we will reconstruct the exact location of the implanted STN leads, as well as the associated structural connectivity to the rest of the brain, and test whether changes in motor inhibition are related to systematic variation in either. In Aim 2, we will investigate the same for cortico-STN functional / effective connectivity, as assessed using EEG and a new method for concurrent task-related LFP recordings. In Aim 3, we will focus on local cortical activity measured by EEG to test if secondary network-effects of DBS contribute to different effects on motor inhibition. If successful, this work would provide unprecedented insights into STN’s role in inhibitory motor control, resolve an unaddressed paradox in existing work, lay the methodological groundwork for the comprehensive, causal study of cortico-subcortical circuits and human behavior, and provide novel insights into the mechanism(s) underlying STN DBS.

NIH Research Projects · FY 2026 · 2020-06

PROJECT SUMMARY All cells and organisms are subjected to mechanical forces. These forces are sensed by cell surface receptors, such as the epithelial (E)-cadherin, which links cells to their neighbors. E- cadherin responds to force by activating signaling pathways inside the cell. These pathways trigger the formation of new cell-cell adhesions and stimulate the rearrangement and reinforcement of the actin cytoskeleton. These actin cytoskeletal rearrangements are energetically costly. We discovered that the energy required to fuel the cytoskeletal rearrangements is provided by AMP-activated protein kinase (AMPK). AMPK is a master regulator of metabolism. It is activated when force is applied to E-cadherin and signals for ATP. The ATP fuels the cytoskeletal changes necessary for cells to resist external forces. In this renewal, our goal is to advance the paradigm for how mechanotransduction and metabolism are coordinated. We will address how: (1) force stimulates glucose metabolism, (2) glycolysis localized to regions of the cell where the actin cytoskeleton is reinforced, (3) actin is polymerized to allow for cytoskeletal reinforcement, and (4) the uncoupling of metabolism and mechanotransduction affects physiology. When this work is complete, significant advances in understanding of the linkages between mechanotransduction and metabolism will emerge. Our proposed animal studies examining uncoupling mechanotransduction and metabolism in vivo have the potential to provide new insight into physiology. The use of sophisticated assays will provide a novel understanding of the relationship between metabolic enzymes and the actin cytoskeleton. These studies, combined with our conceptually innovative observation that PFK-M is the only force sensitive PFK1 isoform, have the potential to call for a re-evaluation of published work and are key to informing the nature of the defects that might be present in disease.

NIH Research Projects · FY 2024 · 2020-04

Most healthy individuals are persistently infected with the human polyomavirus JC (JCPyV) without significant consequences, yet in immunocompromised hosts, JCPyV can cause an often fatal disease -- progressive multifocal leukoencephalopathy (PML). There is currently no effective treatment to prevent or treat PML and novel immune-based therapies are urgently needed to decrease the morbidity and mortality associated with PML. Classically, natural killer (NK) cells are viewed as nonspecific effector cells of the innate immune system that play critical roles in defense against viral infections. Unexpectedly, it was recently demonstrated that besides their ability to rapidly eliminate virus-infected cells without the need for prior antigen sensitization, NK cells also exhibit adaptive immune functions. Different forms of adaptive capabilities have been identified among human NK cell subpopulations, including reports of true antigen-specific memory NK cells as well as adaptive NK cells with enhanced antibody-dependent functions. Adaptive NK cell subsets are endowed with potent anti-viral properties and protection mediated by virus-specific memory NK cells has been demonstrated in mouse models. Importantly, increasing evidence suggest that NK cell can mediate anti-viral responses to JCPyV. In particular, our preliminary data now show potent responses to JCPyV peptides by NK cells, isolation and cloning of single JCPyV-specific NK cells, and reduced JCPyV-specific antibody-dependent cellular cytotoxicity in PML patients. Based on these data, we hypothesize that NK cells and antibodies eliciting NK cell responses play a role in JCPyV pathogenesis. Specifically, we propose to build on our preliminary data to investigate the overarching hypothesis that specific subsets of NK cells can mediate potent anti-viral responses against JCPyV and protect immunocompromised patients against PML, through two focused independent Aims: (i) Define mechanisms crucial to control of JCPyV by NK cells; and (ii) Evaluate the role played by JCPyV-specific antibodies in modulating NK cell activity against JCPyV. If successful, the results of these innovative studies will contribute new knowledge of human immune response against JCPyV and provide the rationale to develop novel immunotherapeutic approaches to harness NK cell function against JCPyV.

NIH Research Projects · FY 2026 · 2020-04

PROJECT SUMMARY/ABSTRACT Project Title: “Birth to Three – Cavity Free: Effectiveness of a Psychoeducational Intervention for ECC Prevention” Early childhood caries (ECC) is a potentially painful and debilitating disease, which represents a significant public health problem among young children. There are profound disparities in ECC experiences such that children from minority and low-income families suffer a disproportionate share of the disease burden. The likelihood of parents of high-ECC risk young children seeking prevention in dental facilities is low; therefore, there is a need to increase preventive dental opportunities where these children already seek health care services. In particular, there is an urgent need to develop and evaluate ECC behavioral interventions for use in public health settings attended by high-risk children. Many authors recommend early implementation of oral health education as one means of preventing ECC. However, major issues discussed in the oral health promotion literature involve a lack of effectiveness among programs based on education alone, as well as a lack of high quality preventive interventions using evidence-based psychological and behavioral strategies. Our research team has been the first to introduce to the ECC prevention arena the self-determination theory (SDT) of motivation, internalization, and healthy functioning, proven effective in promoting positive behavioral changes in several other fields, including oral health care. We have demonstrated that SDT has great promise as a motivational approach by providing evidence, based on results from our R21 (R21-DE016483) study, of the effectiveness of SDT in changing several desirable oral health behaviors for ECC prevention. Building upon the rigor of our previous experience and formative research work in the past several years, we propose a Stage II NIH Model research project that will compare the efficacy of autonomy-supportive videotaped oral health messages framed by SDT to more traditional neutral videotaped messages. We intent to recruit 634 pregnant mothers enrolled in Iowa Women, Infants and Children (WIC) Supplemental Nutrition Programs and follow them until their future child is 36 months old. The primary outcome of interest will be children's caries status. Secondary outcomes will be changes in children's oral health behaviors conducive to better oral hygiene and dietary habits, as well as lower levels of dental plaque and mutans streptococci.

NIH Research Projects · FY 2025 · 2020-04

Project Summary Chlamydia trachomatis (C.t.) is the most common bacterial cause of sexually transmitted infections worldwide. Infections in women often go untreated due to subclinical inflammation masking symptoms, allowing the infection to persist and potentially leading to severe complications such as pelvic inflammatory disease, ectopic pregnancy, and infertility. Identifying and characterizing C.t. factors that drive pathogenesis and subvert host defenses is critical for developing effective therapeutics or vaccines. C.t. replicates within a specialized, host- derived vacuole called the inclusion, which is extensively modified by type III secreted effector proteins known as inclusion membrane (Incs) proteins. These Incs are essential for shielding the bacterium from immune detection and maintaining the inclusion as a replicative niche. Despite their importance, fundamental gaps remain in our understanding of how Incs coordinate interactions with host and bacterial factors to maintain inclusion integrity, prevent fusion with degradative compartments, and promote fusion with nutrient-rich vesicles. Our recent work has identified two key Inc proteins, CpoS and IncC, as critical for inclusion stability and bacterial replication. During the prior funding period, we demonstrated that CpoS forms a functional tetramer, engages multiple Inc proteins via its CC2 domain, and interacts with Rab GTPases through its CC1 domain. We hypothesize that CpoS and IncC coordinate a network of Inc-Inc and host interactions to construct and maintain a replicative niche that evades host immune defenses while supporting bacterial replication and nutrient acquisition. In Aim 1, we will determine how Rab-CpoS-Inc interactions mediate homotypic and heterotypic inclusion fusion, leveraging structural insights from Cryo-EM to evaluate how CpoS mimics SNARE proteins to drive membrane fusion. In Aim 2, we will elucidate how IncC disrupts host trafficking pathways to protect the inclusion from degradation and enable bacterial survival. This work will address critical questions in the field, advancing our understanding of C.t. pathogenesis and informing the development of innovative therapeutic strategies to combat persistent infections, ultimately reducing the global health burden associated with this pathogen.

NIH Research Projects · FY 2025 · 2020-02

PROJECT SUMMARY Cryptococcus neoformans is an important cause of human fungal infections and is one of the most common causes of infection-related deaths in people living with HIV/AIDS. As an environmental fungus, C. neoformans must be able to adapt to mammalian physiology in order to cause disease and only a minority of C. neoformans strains can do so. Three traits have been identified that distinguish many pathogenic C. neoformans strains from non-pathogenic strains: 1) capsule formation; 2) melanization; and 3) the ability to grow at mammalian body temperature. Recent studies of large collections of C. neoformans strains indicate that these three traits do not explain the observed variation in virulence because many closely related strains that express the three major traits still show significant differences in virulence. In the previous funding period, we have discovered that the ability to tolerate carbon dioxide (CO2) concentrations of mammalian host is required for C. neoformans virulence. We also identified protein kinase and transcription factor networks involved in CO2 tolerance. Finally, quantitative trait loci (QTL) mapping and in vitro evolution of low CO2 fitness isolates to identified modulators of CO2 fitness and revealed that multiple pathways and genes contribute to variation in CO2 fitness. In the next funding period, we propose to determine how these virulence-associated regulatory pathways interact to balance positive and negative responses to host CO2 stress (Aim 1). We will also identify and characterize the downstream physiologic and cell biologic processes by which the regulatory networks modulate CO2 tolerance (Aim 2). Our preliminary data indicate that CO2 tolerance may have independently emerged multiple times across the Cn genomic complexes. In Aim 3, we will identify and characterize the mechanisms through which this trait is expressed in clinical isolates with an initial focus on loss of Rim pathway function. Our mechanistic characterization of this emerging Cn virulence trait is expected to provide new insights into Cn pathogenesis and to identify novel pathways and targets for antifungal drug development.

NIH Research Projects · FY 2026 · 2019-09

Sudden unexpected death in epilepsy (SUDEP) is the leading cause of death in patients with refractory epilepsy and is two to four times as common as sudden infant death syndrome (SIDS). Because the mechanisms responsible for SUDEP have not been clearly defined, there are no specific treatments to prevent it. Observations from human and animal studies indicate that seizure-induced respiratory arrest typically precedes asystole, and that many patients experience varying degrees of respiratory depression, autonomic dysfunction, and impaired arousal following seizures. There is a fundamental gap in understanding how seizures do this, and why only a small fraction of seizures lead to death. Serotonin (5-HT) neurons are central CO2 chemoreceptors (CCR) that regulate breathing, autonomic function, and arousal. Patients with low interictal CCR are more hypercapnic following a generalized tonic-clonic seizure (GTCS), and seizures may depress CCR, an effect that can be measured with the hypercapnic ventilatory response (HCVR) test. Impairment of CCR by seizures may contribute to autonomic dysfunction and impaired arousal after GTCS, increasing the risk of SUDEP. The long-term goal is to develop new treatments to prevent SUDEP by elucidating the mechanisms responsible for seizure-induced respiratory depression, autonomic dysfunction, and impaired arousal. This knowledge will also lead to novel biomarkers to identify patients at highest risk. The objective here is to characterize the relationship between seizure-induced blunting of CCR and postictal respiratory depression, autonomic dysfunction, and impaired arousal. The central hypothesis is that seizures inhibit 5-HT neurons in a subset of patients, and this leads to impairment of the ability to detect hypercapnia and to initiate respiratory, autonomic and arousal responses to restore blood gas homeostasis. This hypothesis has been formulated based on human and animal data from the applicants’ own laboratories and will be tested with the following Specific Aims. (1) Characterize the role of CCR in postictal respiratory control and the acute effect of seizures on CCR. (2) Determine how CO2 affects interictal autonomic function and how this is altered by seizures. (3) Determine whether seizures impair the ability of CO2 to hasten recovery of consciousness in the postictal state. Patients admitted to the epilepsy monitoring unit for video EEG study will undergo HCVR testing during the interictal period and several times after seizures to determine the time course of impaired CCR and its effect on cardiorespiratory and autonomic function. We will also measure the effect of CO2 on arousal from sleep and on recovery of consciousness after GTCS, and relate these measures to CCR. This approach is innovative because it is the first to directly examine how inhibition of serotonin neuron function by seizures affects the respiratory, autonomic, and arousal response to CO2. The proposed research is significant because identifying these mechanisms may lead to novel treatments to prevent SUDEP.

NIH Research Projects · FY 2025 · 2019-08

Invasive Aspergillosis caused by azole resistant Aspergillus fumigatus (Afu) has a mortality rate exceeding 60%, making this a clinical problem of acute significance. Extensive analyses of resistant isolates of Afu have shown that the majority of resistance-causing mutations are linked to the cyp51A gene encoding the enzyme that is the target of azole drugs. The most common forms of resistant Afu contain two types of mutations: one that causes a substitution mutation within the coding sequence for the enzyme while a second alteration is a duplication of a short segment of the cyp51A promoter region. The most common allele is referred to as TR34 L98H cyp51A in which the TR34 designation indicates a tandem repeat of 34 bp of the promoter region. In the previous grant period, we identified a transcription factor called AtrR that binds to an element within the 34 bp region of cyp51A. AtrR binding is required for the elevated cyp51A expression driven by the TR34 promoter element as well as the enhanced voriconazole resistance this mutant gene provides. We provide genetic evidence that the function of the two 34 bp regions in the TR34 cyp51A promoter do not have similar functions and the appearance of the second 34 bp repeat triggers enhanced chromatin accessibility as measured by Assay for Transposase- Accessible Chromatin using sequencing (ATAC-seq). Our data indicate that the differential behavior of the 34 bp elements in cyp51A are essential for the unique elevated transcription supported by this mutant promoter. Previous models that the simple duplication of the effect of this 34 bp region was sufficient to explain its behavior are incorrect and the upstream 34 bp has now acquired a genetically distinct behavior that explains the elevation in cyp51A transcription. We believe that this behavior is likely reflective of the normal function of the 34 bp region and will test this idea using ATAC-seq on the wild- type promoter in the presence and absence of azole treatment. We will also examine the role of both the AtrR binding sites and the presence of AtrR in the observed increased chromatin accessibility seen in the TR34 cyp51A promoter. We have used biochemical purification of AtrR to identify proteins that associate with this factor and found that several of these are involved in chromatin modification. We will determine the role of these proteins in control of cyp51A promoter accessibility and expression of the TR34 cyp51A gene. We will purify the transcription factor SrbA that is also known to bind to the 34 bp element region and examine its role in chromatin accessibility and recruitment of coactivators. Finally, we will determine the role for cyp51A mutants in the azole response in a murine model using isogenic strains of Afu. Additionally, we will use an enrichment strategy to measure Afu transcription in infected lungs +/- azole drug challenge. This work will illuminate the function of a key region of the cyp51A promoter that is associated with 80% of azole resistant Afu isolates.

NIH Research Projects · FY 2025 · 2019-06

Project Summary/Abstract While outcomes have substantially improved for many types of cancer, endometrial cancer (EC) incidences and deaths are on the rise, with the five-year survival rate worse today than three decades ago; owing largely to the ineffectiveness of current treatments. As a tumor is exquisitely sensitive to the growth promoting effects of estrogen and the growth limiting effects of progesterone, hormonal therapy for EC using progestins has been a traditional choice for treatment. It is highly effective in the short term; however, responsiveness wanes over time due to loss of progesterone receptor (PR) expression, and recurrences are common. There is a critical need to identify strategies to improve or restore responsiveness to progestin therapy, and we propose that molecularly enhanced progestin therapy will make a major positive impact on survival of patients with EC. The objective of this application is to identify the molecular mechanisms driving the downregulation of the PR in EC patients and identify novel strategies to further enhance the effectiveness of progestin therapy. We will develop molecular agent combinations with progestins that will significantly enhance tumor cell differentiation in vitro and improve survival in mouse xenograft models of human EC. Our central hypothesis is that targeting PR repressors will enhance the expression of PR, the most important tumor suppressor in the endometrium, thereby improving response to progestin therapy. This hypothesis stems from our strong recently published data in EC cells that PR expression is downregulated through distinct molecular mechanisms, and epigenetic modulators potently increase PR expression and tumor suppressor activity. We have now identified additional PR suppressors that have the potential to more clearly define the multiple mechanisms of PR inhibition in EC, setting the stage for new therapeutic opportunities. In Aim 1, we will determine the impact of epigenetic modulators on PR expression and activity in EC patients from the clinical trial NRG-GY011 results. In the extended period we will evaluate the potential new endpoint markers. In Aim 2, we will enhance PR expression using small molecular drugs and test drug efficacy using in vitro and in vivo EC models. In the extended period, we will identify and validate novel drug and target sites for PR regulation such as Topoisomerase II inhibitor. In Aim 3, we will identify novel PR downregulation mechanisms using genome-wide gene silencing. In the extended period, we will focus on novel PR repressor PHB2 and SETDB1. At the completion of these studies, we will have better understanding of mechanisms of hormonal resistance and integrate innovative molecular therapies into enhanced progestin therapeutic regimens in preclinical and clinical studies, and as well as design future EC clinical trials. Therefore, these studies will have a strong impact on the treatment of EC.

NIH Research Projects · FY 2026 · 2019-04

ABSTRACT The accurate and timely DNA replication program is a prerequisite of a stable genome. This research project is built around our discovery that the human DNA repair protein RAD52 performs an important and previously unknown function in supporting DNA replication. RAD52 promotes genome integrity through several biochemical and cellular functions. As a gatekeeper of replication forks, RAD52 binds to and stabilizes stalled or damaged replication forks protecting them from reversal by SMARCAL1, and subsequent MRE11-dependent degradation. RAD52 absence promotes RAD51-dependent recruitment and activation of DNA polymerase alpha/primase and an alternative replication restart pathway that leaves behind large ssDNA gaps. Finally, upon prolonged replication stress, RAD52 cooperates with MUS81 nuclease to cleave the fork producing DNA breaks that can be repaired by homologous recombination. Depletion or pharmacological inhibition of RAD52 results in fork restoration with large ssDNA gaps, genome instability, and selective toxicity in cancer cells displaying BRCAness phenotype or defects in ATM (ataxia–telangiectasia mutated) serine/threonine kinase. We discovered that the replication fork structure enables a unique head-to-head arrangement of two undecameric RAD52 rings. This organization juxtaposes the RAD52 DNA binding sites creating an extended positively charged surface. In addition to DNA binding, this surface is involved in the interaction with MUS81 nuclease. We propose that the two-ring RAD52 structure plays two distinct functions: (1) it enables a dynamic DNA strand exchange reaction which locally remodels the stalled fork, and (2) it positions MUS81 for cleavage. The transition between a spool-like two-ring arrangement to a single ring is a switch between fork protection and mutagenic single-strand annealing activities of RAD52. Our goal for the next funding period is to develop a comprehensive mechanistic understanding of the RAD52 function at the replication fork. In Aim 1 we will combine cryo-electron microscopy (CryoEM), single-molecule biophysics, super-resolution microscopy, biochemical and cell-based analyses with computational modeling to build the structure-activity relationship (SAR) of the nucleoprotein complexes containing fork DNA, RAD52 and ssDNA binding protein RPA. In Aim 2 we will dissect functionally of the MUS81-RAD52 axes. We will map the MUS81 cleavage sites on stalled replication forks protected by RAD52 and RPA, will describe the architecture of the RAD52/fork/MUS81 complexes, and will investigate the MUS81-RAD52 interactions in cells using super- resolution microscopy and proximity labelling. In Aim 3 we will test the hypothesis that RAD52 phosphorylation by c-ABL tyrosine kinase at Y104 prevents formation of the two-ring RAD52-fork structure and switches the RAD52 function from fork protection to single-strand annealing. By completing the proposed studies, we will learn how RAD52 functions at replication forks, how it contributes to genome stability, and will obtain SAR crucial for development of novel, function-specific inhibitors.

NIH Research Projects · FY 2026 · 2019-04

ABSTRACT Efficient genome maintenance is a double-edged sword. Accurate repair of DNA lesions and damaged replication forks promotes genome stability, which is a key to avoiding cancer, aging and neurodegenerative diseases associated with DNA repeat expansion. The same mechanisms that maintain genome integrity in healthy cells, allow cancer cells to acquire a more aggressive character and develop resistance to radiation and chemotherapy. Untimely deployment and/or dysregulation of the DNA repair machines may further destabilize the genome or may result in the accumulation of geno- and cytotoxic repair intermediates. Significant gaps remain in our understanding of the molecular events that funnel the intermediates of otherwise accurate repair into “rogue”, genome-destabilizing mechanisms. This research program emphasizes the molecular and structural mechanisms by which the intermediates of DNA metabolism bound by Replication Protein A (RPA) are channeled into DNA repair, protection of DNA replication forks, homologous recombination, DNA damage tolerance and signaling. Our central hypothesis is that the activities of the RAD51 recombinase, the ssDNA-binding protein RPA, recombination mediators BRCA2 (in human) and Rad52 (in yeast), and DNA repair helicases are finely tuned by a variety of factors, which include posttranslational modifications, interacting partner proteins, specific DNA structures and DNA lesions. These factors affect the protein configurational dynamics and critical protein-protein interfaces. Understanding how the protein plasticity and kinetics of assembly of the macromolecular machines of DNA repair will show us new ways to selectively manipulate the activities of RAD51, RPA and multifunctional DNA helicases and polymerases in DNA replication and repair. We are leveraging and building the tools of single-molecule biochemistry, biophysics, structural and chemical biology. Our unique perspective on the formation, activities and regulation of the nucleoprotein complexes orchestrating genome maintenance is rooted in our ability to visualize microscopic configurational dynamics of and sort individual human DNA repair proteins with their native posttranslational modifications, and to probe and separate activities associated with different surface-tethered proteins and nucleoprotein complexes at the single- molecule level. Our goal is to provide an entirely new outlook on how the cell balances the assembly and activities of the molecular machines that can repair, but also destabilize, the genome, and to be able to alter this balance with new chemotherapeutics targeting cancer and neurodegenerative diseases.

NIH Research Projects · FY 2026 · 2019-04

Project Summary Lung transplantation has become the standard of care for end-stage lung diseases with no available medical therapy. However, long-term survival after lung transplantation is affected by development of chronic lung allograft dysfunction (CLAD), which is progressively fatal and for which there is no effective therapy. Almost 50% of recipients die within five years due to the development of obliterative bronchiolitis (OB) in the allograft, an obstructive type of CLAD. We developed a novel orthotopic lung transplant model in the ferret that closely models human CLAD. Utilizing this model, we showed for the first time that airway submucosal glands (SMGs), a facultative airway stem cell niche, and basal stem cells (BSCs) are depleted in human and ferret OB allografts. We also showed that in human and ferret OB allografts, there is a predominance of Keratin 14 (Krt14) positive BSCs and a loss of Keratin 15 (Krt15) BSCs. This loss of Krt15 in BSCs decreases their proliferative capacity. Additionally, with our collaborator (Dr. Wa Xian), we demonstrated a dominance of pro-fibrotic and pro- inflammatory BSC clones in end-stage lung diseases like chronic obstructive pulmonary disorder, idiopathic pulmonary fibrosis, and cystic fibrosis. We and other investigators have shown that glandular myoepithelial cells (MECs) are a facultative stem cell niche that repairs airway injury and contributes to surface airway BSC and differentiated epithelial cells. The role of these glandular MECs in the transplanted lung remains unclear. Since the bronchial SMG SC niche is not conserved in widely used rodent models, there has been a critical gap in our ability to study and understand how alterations in the bronchial SMG SC niche contribute to impaired epithelial regeneration in the allograft and lead to CLAD. To address, we generated novel ACTA2-CreERT2:ROSA-TG ferrets to lineage label MECs. The overall objective of this proposal is to understand mechanisms leading to CLAD pathogenesis, identify BSC signatures for early detection of CLAD, and identify potential therapeutic targets promoting CLAD. We hypothesize that tight regulation of region-specific airway surface epithelium and SMGs is disrupted in transplanted lungs, which promotes suboptimal regeneration resulting in the expansion of abnormal surface BSCs, the depletion of stem cells in the SMGs, and thus the exacerbation of inflammatory responses that drive CLAD. We will achieve these objectives by addressing the following specific aims: 1) Determine the multipotency of glandular MECs to regenerate surface BSC with abnormal stem cell properties in CLAD. 2) Identify how Keratin 14/15 interact with Stratifin (Sfn) to modulate p63 levels and direct BSC fates. 3) Determine the effect of persistent Krt14 expression coupled with the loss of Krt15 and the emergence of inflammatory basal cell clones on the development of OB in human allografts. The expected outcomes will provide an understanding of the mechanisms leading to the development of CLAD, and identify novel ways to detect, prevent/treat CLAD.

NIH Research Projects · FY 2024 · 2018-09

Project Summary Up to 95% of premature infants undergo red blood cell (RBC) transfusion while in the intensive care unit, yet it is unknown whether more restrictive or more liberal transfusions will lead to optimal brain development. The Transfusion of Prematures (TOP) Trial is a multi-center study funded by the NHLBI and supported by the NICHD Neonatal Research Network (NRN). The primary objective of the TOP Trial is to assess survival and rates of neurodevelopmental impairment at 22-26 months corrected age in extremely low birth weight (ELBW) infants that are randomized to either liberal or restrictive RBC transfusion thresholds. The trial began enrollment in December 2012 and reached the target sample size of 1,824 infants on time in April 2017. Although major deficits in motor and cognitive function may be detected at 22-26 months of age, these infants are too young to assess cognitive, behavioral, and coordination skills that, if impaired, can lead to problems with academic skills, motor performance or adaptive functioning in home or school environments, conditions that are far more prevalent in this population and create substantial morbidity for the children and their families. Because optimal transfusion management is a critical knowledge gap in neonatology, the currently proposed TOP 5 Study will assess functional neurodevelopmental outcomes of infants randomized to two different transfusion thresholds in the TOP Trial at 5 years corrected age. The TOP 5 Study Clinical Coordinating Center (CCC) is led by Co-PIs Dr. Peg Nopoulos, who has a ten-year history of studying the outcomes of premature infants exposed to liberal or restrictive transfusion, and Dr. Sara DeMauro, who has extensive experience conducting multicenter outcomes studies in collaboration with the NICHD NRN and the Data Coordinating Center (DCC) at RTI International. The DCC PI Dr. Abhik Das also leads the DCC for the NICHD NRN. The NRN has a superb track record in school-age outcomes research and history of productive collaboration with NHLBI. Thus, the TOP 5 Study has been thoughtfully designed to leverage existing successful research infrastructure, relationships, and resources in order to reduce redundancy and ensure success. The results of the TOP 5 study will provide evidence about which approach to neonatal transfusion (liberal or restrictive) minimizes damage to vulnerable neuronal circuits and, in turn, which transfusion strategy will improve both short and long-term outcomes for these vulnerable premature infants.

NIH Research Projects · FY 2024 · 2018-09

Abstract/Summary Proton arc therapy has lagged behind photon arc therapy, which is now commonplace in the clinic, due mostly to slow proton energy switching times which make treatment durations impractical. Fast energy modulation systems are now clinically available, and, by applying delivery optimization tools that intelligently change beam energy as the gantry rotates, proton arc therapy is on the verge of becoming a clinical reality that can improve plan delivery speed and robustness to range uncertainties relative to conventional fixed field proton therapy. Without dynamic lateral beam collimation, however, proton arc therapy tumor dose conformity will be inferior to fixed field collimated proton therapy plans. This is a major problem especially for brain and head and neck cancer patients whose normal tissues could be spared significant radiation dose using beam collimation. The long-term goal is to develop the next generation of pencil beam scanning (PBS) proton therapy delivery systems that maximize the achievable tumor dose conformity, robustness, and delivery speed. The overall objective of this proposal is to develop dynamically collimated arc-based PBS by expanding the dynamic collimation system (DCS) technology developed in the first part of this R37 proposal, providing the capability for rapid, tumor-conformal delivery of dose distributions that are more robust to uncertainties in Bragg Peak position placement than those delivered with fixed field proton therapy. The rationale for this project is that superior treatment plans will result from the combination of energy-specific collimation and rotational arc delivery than either of the individual technologies, thus improving the quality of care of proton therapy. Guided by strong preliminary data from our in-silico treatment planning studies and constructed DCS prototype, development of collimated proton arc therapy will be carried out by pursuing three specific aims: 1) develop arc-based treatment planning and delivery methods for dynamically collimated proton therapy, 2) enhance the clinical DCS prototype to perform proton arc treatments, and 3) adapt existing treatment verification methodologies for quality assurance. Under specific aim 1, established multi-field treatment planning techniques, both dose calculation and optimization, will be extended to include the optimization of trimmer and energy sequencing for the case of a rotating gantry. Under specific aim 2, real-time feedback mechanisms will be incorporated to monitor and synchronize gantry angle to the sequencing of the high-speed trimmer blades. Under specific aim 3, experimental and computational techniques will be developed and demonstrated to enable successful commissioning of dynamic collimated proton arc therapy. The research proposed in this application is innovative because it represents a new combination of two promising and synergistic technologies: dynamic collimation and proton arc therapy. This contribution is expected to be significant since the two technologies together can preserve conformity improvements gained by collimated PBS while adding the robustness and delivery speed benefits achievable with arc therapy, improving treatment outcomes.

NIH Research Projects · FY 2026 · 2018-08

The University of Iowa Regional Coordinating Stroke Center (UIRCC) is ideally positioned to develop, promote, and efficiently conduct high-quality, multi-site clinical trials on stroke prevention, treatment, and recovery, in line with the overarching goal of the NIH StrokeNet. The University of Iowa (UI) is the only academic medical institution and comprehensive stroke center in the state. The UI has led and participated in landmark clinical trials in cerebrovascular disease for over 50 years such as the Org 10172 in Acute Stroke Treatment (TOAST) study, the NIH Stroke Scale and the TOAST classification of stroke subtype. Over the past 10 years, the UI has served as the UIRCC, bringing together our tradition of multidisciplinary, cutting-edge, cerebrovascular research. We demonstrated our ability to recruit patients by becoming the top enroller in the first StrokeNet trial, DEFUSE-3, and enrolling patients from the Upper Midwest—a region that experiences unique geographic barriers for timely trial participation. The UIRCC is committed to holistically expanding stroke care by increasing the number of stroke interventions that effectively improve patient outcomes, and by expanding the number of patients who can benefit from such interventions. We have a strong commitment to remain a productive member of StrokeNet. Our first aim is to further advance the UIRCC as an efficient and adaptive infrastructure to recruit patients from the Upper Midwest. The restructured UIRCC comprises eight trial-ready geographically linked hospitals that will efficiently conduct critical trials and provide access to patients living in 119 counties from seven states, 56% of whom reside in rural zip codes and traditionally have been excluded from trials due to geographic barriers. Our second aim is to continue to develop innovative interventions to expand stroke care. We plan to reproduce our uric acid success story, the first cerebroprotectant to succeed through the NIH Stroke Preclinical Assessment Network (SPAN). Our group will now be leading the testing of uric acid through the new StrokeNet Thrombectomy Endovascular Platform (STEP). We will continue to build on our multidisciplinary research infrastructure, including our group of investigators and trainees. This includes the UI SPAN translational team, as well as clinical researchers and industry partners. This infrastructure will continue to leverage dedicated UI pilot funds to ignite the development of interventions, with an emphasis on those that are simple and therefore widely applicable to stroke patients.

NIH Research Projects · FY 2026 · 2018-05

PROJECT SUMMARY/ABSTRACT Age-related hearing loss is a substantial national problem due to its high prevalence and significant quality- of-life consequences. Although hearing aids (HAs) are the primary choice for the management of age-related hearing loss, the adoption rate of HAs is quite low. A commonly reported barrier to HA uptake is that HAs ob- tained in the traditional healthcare pathway, which requires multiple visits to a licensed professional (e.g., audi- ologists) for hearing loss diagnosis and the month(s) long process of fitting and fine-tuning HAs, are not afford- able or accessible. We refer to this healthcare pathway as the AUD pathway. As an alternative pathway to im- prove affordability and accessibility of hearing healthcare, the over-the-counter (OTC) pathway uses a direct- to-consumer service-delivery model and enables users to self-determine hearing loss, self-fit the HA, and self- manage device without professional support. Although the OTC pathway has become popular, little is known about how potential users determine the right pathway for themselves (OTC vs. AUD) and how the OTC pathway impacts users’ long-term well-being. To guide potential users in their selection of one pathway over the other and to improve well-being of people with hearing loss, our project aims to characterize the entire OTC patient journey and compare that with the AUD journey, from pathway selection, through patient outcomes and consequences of unsuccessful HA expe- riences. Aim 1 seeks to understand the decision-making processes used by individuals to choose between the OTC and AUD pathways (i.e., what factors they consider and what personal characteristics drive the decision). Aim 2 seeks to determine the long-term (12 months) course of OTC outcomes (e.g., HA satisfaction) com- pared with that of the AUD pathway and to establish the individual differences (e.g., finger dexterity) that ex- plain the variance in long-term outcomes of each pathway. Aim 3 seeks to understand what people think and do following unsuccessful OTC experiences and whether this is the same as for the AUD pathway (Aim 3). To achieve our aims, we will conduct a two-site (Iowa and Vanderbilt), mixed-methods, longitudinal study in which participants choose their preferred healthcare pathways (OTC vs. AUD). Participants will purchase HAs up-front, but will be able to return the devices for a refund within the manufacturers’ trial periods. After selecting a pathway (Aim 1), participants will be contacted at 1-, 6- and 12-months post-fitting. At each contact, depend- ing on the status of HA usage we will assess HA outcomes (Aim 2) or the engagement with hearing healthcare behaviors (e.g., adopting new HAs, Aim 3). Our project will generate crucial data that will help us develop tools (e.g., decision aids) that adults with hearing loss can use to make informed decisions regarding hearing healthcare, and help ensure that the OTC pathway has a positive impact on their well-being.

NIH Research Projects · FY 2026 · 2017-09

Abstract Cognitive symptoms of Parkinson’s disease (PD) include deficits in attention, working memory, and reasoning. These deficits affect up to 80% of PD patients and lead to mild cognitive impairment (PD-MCI) and dementia in PD (PDD). There is a critical need to better understand cognitive impairment in PD to develop new targeted treatments. Our long-term goal is to define the mechanisms of PD-related cognitive impairment. PD involves diverse processes such as dopamine and acetylcholine dysfunction, synuclein aggregation, and genetic factors. During the past funding period, we linked PD-related cognitive impairment to dysfunction in frontal midline delta (1–4 Hz) and theta (5–7 Hz) rhythms, which our work has established as a marker of cognitive control. However, it is unknown why PD patients have deficits in these low-frequency brain rhythms. Our preliminary magnetic resonance imaging (MEG) and magnetoencephalography (MRI) implicate the anterior midcingulate cortex (aMCC) as a potential source of frontal midline delta/theta rhythms. In the next funding period, our objective is to determine the mechanisms and predictive power of delta/theta rhythms in PD, which will help to better understand the pathophysiology of PD-related cognitive impairment. Collaboration between the University of New Mexico (UNM) and University of Iowa (UI) that will bring together MEG, MRI, longitudinal EEG, and adaptive subthalamic (STN) deep-brain stimulation (DBS). We will test the overall hypothesis that frontal midline delta/theta dysfunction contributes to cognitive impairments in PD. In Aim 1, we will determine the structural basis for delta/theta rhythm deficits in PD. In Aim 2, we will determine the predictive power of delta/theta rhythm deficits in PD. In Aim 3, we will determine how tuned low-frequency STN DBS impacts cortical activity and cognition. Our results will have relevance for basic-science knowledge of the fundamental pathophysiology of cognitive impairment in PD and related dementias. Because this proposal will study patients with PDD, our findings are directly relevant to Alzheimer’s-related dementias (ADRD).

NIH Research Projects · FY 2025 · 2017-09

Bipolar disorder (BD) is a frequently devastating psychiatric illness that is challenging to diagnose and treat. In our prior work, we have shown that metabolism, function, and morphology of the cerebellum is different in participants with BD as compared to controls. Furthermore, we have observed a relationship between mood and cerebellar metabolism as well as function in a cross-sectional design. These findings support the growing body of literature that the cerebellum is involved with BD. However, there has been a sparsity of studies undertaken that have attempted to follow participants with BD and observe changes in brain metabolism and function associated with fluctuations in mood. We are proposing to conduct a two-year longitudinal study of 170 participants with a diagnosis of BD type I as well as 90 matched controls to study changes in cerebellar metabolism and function associated with mood fluctuations. The participants with BD will receive a brief weekly mood assessment to identify changes in mood where subjects will be assessed using neuroimaging. Brain imaging will include metabolic, functional, and anatomical imaging with the data used address the following aims. Aim 1) Does cerebellar metabolism change with mood in BD? We hypothesize that the cerebellum plays a significant role in maintaining the euthymic mood state (i.e. plays a compensatory role) and when the cerebellum is no longer able to serve this compensatory role depressive or manic mood states develop. To test this hypothesis, we will assess changes in cerebellar metabolism (31P, 1H MRS, and T1ρ) associated with mood changes. Aim 2) Does cerebellar function and connectivity vary with mood in BD? To test the function of the cerebellar vermis, we plan to perform two task-based fMRI studies (backward masking emotional faces and go/no-go). In addition, resting state functional connectivity will allow us to explore connectivity of the cerebellum with the emotional control network. We hypothesize that connectivity between the vermis and nodes of the emotional control network will vary with mood state and expect increased connectivity in the euthymic state and decreased connectivity during exaggerated mood states (depression/mania). Furthermore, we expect that as mood ratings for depression and mania increase subjects with BD will exhibit a greater number of incorrect responses and failure to activate the cerebellum. These results would provide further evidence that the cerebellum is playing a compensatory role to maintain mood. In addition, this study may provide information leading to novel therapy trials in the future.

NIH Research Projects · FY 2026 · 2017-09

Project Summary Maintaining internal environment constancy is essential for life. The circumventricular organs (CVO’s) of the brain including organum vasculosum of the lamina terminalis (OVLT) and the subfornical organ (SFO) lack a blood-brain-barrier, function as central sensors to provide feedback regulation for maintaining osmotic equilibrium. Neurons in CVO’s detect changes in osmolality and transduce the signals into action potentials (AP’s) travelling down the axon projecting to magnocellular neurons in the paraventricular (PVN) and supraoptic nuclei (SON). Upon activation by AP’s, magnocellular neurons synthesize antidiuretic hormones (ADH), which is transported via axon process to the posterior pituitary gland and released into blood circulation herein. ADH acts on kidney to effect free water reabsorption. While hyperosmolality also stimulates thirst and drinking, separate neuronal networks are involved. Together, renal free water reclamation and drinking restore serum osmolality in response to water deprivation. The molecular identity of the osmolality sensor(s) in the OVLT/SFO neurons remains elusive. With-no-lysine [K] kinases WNK1-4 are protein kinases in which gain-of-function mutations of WNK1 and 4 in humans cause familial hypertensive and hyperkalemic disease called pseudohypoaldosteronism type II (PHA2). Our preliminary results strongly support the central hypothesis that WNK1 functions as a central osmosensor for osmolality regulation of ADH release. Mice with neuronal conditional knockout (cKO) of Wnk1 exhibit phenotypes of partial central diabetes insipidus (DI). WNK1 activate downstream oxidative-stress responsive-1 kinase (OSR1) and related SPAK (Ste20-related proline/alanine-rich kinase). WNK1-OSR1/SPAK kinase cascade regulates many ion channels and transporters. To support the hypothesis, Specific Aim-1 will test the hypothesis that OSR1 and/or SPAK acts downstream of WNK1 to regulate osmolality-induced ADH release. Control and mice with genetically altered WNK1 kinase cascade will be studied in metabolic cage under free water access and water restriction. Urine volume, urine and serum electrolyte, osmolality, ADH and copeptin levels will be measured. Specific Aim-2 will test the hypothesis that activation of Kv3.1b voltage-gated K+ channels by WNK1 increases AP firing in OVLT/SFO neurons leading to stimulation of ADH release by hyperosmolality. Electrophysiological recording of freshly isolated individual OVLT/SFO neurons, native neurons in situ in acute brain slice, and HEK cells expressing recombinant channels and WNK1 cascades will be performed. The proposed studies will reveal novel findings that an intracellular protein functions as a sensor for extracellular osmolality and provide fresh insights into how body maintains osmotic equilibrium in health and into disease processes that affect total body water homeostasis.

NIH Research Projects · FY 2026 · 2017-08

Title: Life-Long Phenotypic Correction of CF Airways Project Summary/Abstract: Cystic fibrosis (CF) is caused by mutations in the cystic fibrosis conductance regulator (CFTR) gene. Knowledge of CFTR function and cell type expression has advanced greatly since its discovery in 1989, with notable discoveries in the last 5 years. While significant advances have been made with small molecule modulator therapies to restore function for most CFTR mutation classes, ~10% of people with CF have not benefited from these strategies. We have a demonstrated track record of using many categories of viral and non-viral based reagents for gene delivery to the airways. Our goal is to achieve life-long correction from a single dose of aerosolized viral vector. As such, efficient delivery of a therapeutic gene or gene editing machinery to airway progenitor cells is critical. In this proposal, we advance two gene therapy technologies and compare their pros and cons. Both strategies take advantage of the impressive transduction efficiency and large packaging capacity of Adenoviral (Ad)-based vectors. Two potential drawbacks of Ad vectors are transient expression and immune response, both of which will be addressed. In Aim 1 we engineer Ad-based viral vectors with improved progenitor cell targeting and correction. We compare chimeric vectors based on Ad5 with fibers from species B adenoviruses. In addition, we show that Ad has the capacity to penetrate airway mucus barriers but investigate mucolytics that may further improve vector delivery. In Aim 2 we contrast efficiency of CFTR functional correction using gene delivery and gene editing in vitro. To achieve gene correction, will use an adenine base editor (ABE) delivered with Ad (Ad-Cas9-ABE) to correct CFTR in cells. As a proof of principle, we focus on the CFTR nonsense mutation R553X. This mutation results in premature termination codon and does not respond to any small molecule modulator. Following vector delivery, we will confirm gene editing using a combination of next generation sequencing and Cl- current measurements. The achieved levels of phenotypic correction will be compared to the PB/Ad-CFTR gene addition strategy. We hypothesize that regardless of gene therapy strategy, a maximum threshold level of Cl- current is achievable. This current is similar in heterozygous (CFTR+/-) or wild-type (CFTR+/+) cells. In Aim 3 we contrast gene delivery and gene editing efficiency in pig airways. We will generate a CFTRG551D/R553X compound heterozygous pig model to screen leading vectors designed to correct a CF mutation for which no current small molecule treatments are available. Our goal is to provide a life-long gene repair strategy that could be adapted for a great number of CF causing mutations. This proposed research is highly innovative. The reagents, methods, and data generated by these experiments could be applied to gene addition or base editing for other monogenic disorders, thereby significantly advancing the gene therapy field.

NIH Research Projects · FY 2025 · 2017-07

PROJECT SUMMARY / ABSTRACT There is an ongoing and urgent need to devise better approaches to prevent, treat, and ultimately reverse diabetes, which requires constant training of highly qualified cohorts of investigators. The Diabetes Research Training Program at the University of Iowa is mentoring and launching the next generation of investigators who will address this critical need by focusing their scientific efforts on diabetes. We seek, in this competing renewal, ongoing support for 6 postdoctoral positions. Both physician and PhD scientists will be trained and most will be appointed to the program for 2 years. There are no other diabetes research training opportunities in the state. The program is built on the exceptional strength and depth of the University of Iowa Fraternal Order of Eagles Diabetes Research Center (FOEDRC), which is driving innovative strategies aimed at further understanding the pathophysiology of diabetes and its complications and at devising novel preventative and curative strategies. Training in basic and clinical scientific investigation will be augmented by a core curriculum in diabetes, metabolism, grant writing, and research ethics. Trainees involved in clinical and/or translational diabetes research will enroll and complete a Master of Science in Translational Biomedicine. All trainees will receive tailored career mentoring to ensure a successful transition to their next career stage. Training will be underpinned by synergistic scientific collaborations across multiple disciplines of relevance to diabetes research, as embodied by the Center. Mentored research opportunities available in the Program span basic and mechanistic investigation across a broad range of model organisms to translational and epidemiologic studies in humans. Program mentors comprise 44 diverse and interactive faculty with vigorous diabetes research programs, stable extramural funding, and robust training records. They span 5 colleges, 12 departments and 8 clinical divisions. Training will leverage existing synergies between collaborating investigative teams. Recruitment of outstanding trainees will be secured by the broad reach of the FOEDRC. Qualified physician scientist trainees will be recruited from the Physician Scientist Training Pathway and from clinical fellowship training programs in Internal Medicine, Obstetrics and Gynecology, and Pediatrics. PhD scientist trainees will be selected from national pools recruited into Program mentors’ labs. The training program will be administered through the FOEDRC, and will be overseen by an executive committee comprising 2 (multi) principal investigators and 5 co-directors, all of whom are seasoned investigators with uncompromising commitment to mentoring trainees.

NIH Research Projects · FY 2026 · 2017-04

Project Summary/Abstract C3 glomerulopathy (C3G) is an aggressive ultra-rare kidney disease that occurs at any age and carries the highest risk for irreversible renal failure of the known glomerular diseases. It is defined by underlying complement dysregulation and characterized by predominant complement C3 deposition on kidney biopsy. Two major disease subgroups are recognized–dense deposit disease (DDD) and C3 glomerulonephritis (C3GN)–although overlapping clinical and pathological features suggest that C3G is more appropriately considered a disease continuum. Dysregulation of the alternative pathway (AP) of complement is fundamental to disease manifestation although terminal pathway dysregulation is also common. The estimated renal half-life in C3G patients is 10 years and in patients who progress to end-stage renal disease (ESRD), transplant decisions, including timing and post-transplant medical therapy, are overshadowed by the fact that disease recurrence remains a major medical issue. The challenges faced by clinicians in caring for C3G patients reflect our incomplete understanding of both the underlying pathophysiology and natural history of this disease. These knowledge gaps impact treatment decisions and the development of disease-specific therapies. In this grant, we propose to: • Specific Aim 1. To study the role of genetic variation in C3G • Specific Aim 2. To define the characteristics of autoantibodies in C3G by epitope mapping • Specific Aim 3. To develop and clinically validate predictive models of disease outcome in C3G Completing these specific aims will significantly refine our insight into the pathogenesis of C3G, improve clinical care of these patients, and lay the foundation for effective and personalized treatments for this disease.

NIH Research Projects · FY 2026 · 2016-09

PROJECT SUMMARY Plasmodium infections and the disease malaria remain global health emergencies. Plasmodium parasites replicate within and cause the destruction of host red blood cells, which triggers inflammation and causes the symptoms of malarial disease. Parasite-specific antibody responses that develop following infection are critical for controlling parasite burden and limiting disease severity. CD4+ helper T cells are essential for coordinating these protective antibody responses. However, sterilizing anti-Plasmodium immunity rarely develops, even following repeated infection. We hypothesize immune failures are mechanistically linked to aberrant or inefficient Plasmodium-specific effector CD4+ T cell development and function. One of the most critical challenges to developing new immune-based therapies or vaccines against Plasmodium is understanding the mechanisms by which Plasmodium-specific CD4+ T cells develop, function and regulate humoral immunity following infection. In the continuation of this project, we apply powerful new cellular and molecular genetic approaches that enable direct, high-resolution analyses of Plasmodium-specific CD4+ T cells. These new approaches facilitate our long-term goal to understand the mechanisms governing the development and function of Plasmodium- specific CD4+ T cell responses. Our goal is addressed by two specific aims that have evolved to test: 1) how parasite-derived molecules influence cell antigen presenting functions and priming of Plasmodium-specific CD4+ T populations; and 2) how host physiological perturbations and constraints on cellular metabolism regulate CD4+ T cell development and function. Our innovative conceptual and technical advances and mechanistic approaches enable us to establish additional new paradigms for understanding and enhancing CD4+ T cell-dependent anti- Plasmodium immunity. Understanding cellular and molecular events governing CD4 T+ T cell responses during malaria will enable us to identify and develop new immune-based strategies to limit Plasmodium pathogenesis and disease burden.

NIH Research Projects · FY 2026 · 2016-09

Abstract SARS-CoV-2, the cause of COVID-19, continues to cause widespread infection and morbidity, but death rates have decreased as most of the world has either been infected or vaccinated or both. However, at the same time, it has become clear that many patients have developed long term sequelae. These sequelae, called PASC (Post Acute Sequelae of COVID-19), affect many organ systems even though virus is found at autopsy in nearly all studies only in the respiratory and gastrointestinal tracts. One common symptom of acute COVID-19 is anosmia. Recovery of olfactory function is often incomplete in patients. However, only supporting (sustentacular) cells and not olfactory sensory neurons are infected in patients, raising the question of how infection of a supporting cell could have such profound effects on olfaction. In addition, neurological disease is also present in many patients with PASC. Many of these manifestations reflect ongoing inflammation (usually observed on autopsy), but the basis of these inflammatory changes is unclear since virus cannot be detected. We have isolated a mouse- adapted virus that causes severe acute respiratory disease in infected mice, as well as persistent signs of disease is the lungs and brains months after the acute infection has resolved. Infected mice develop anosmia, as well as long term behavioral abnormalities and defects in neurotransmitter expression, but virus is not present in the brain. Our central hypothesis is that ongoing inflammation is a major contributory factor in the observed dysfunction in the olfactory and neurological systems. This hypothesis will be addressed in the following specific aims: Specific Aim 1: To understand the relationship between sustentacular cell infection and olfactory dysfunction and to understand the basis of chronic changes in the brains of SARS-CoV-2-infected mice. Acute and chronic changes in olfactory pathways will be probed using electrophysiological and olfactory measurements. These changes will be related to sustentacular function and gene expression. Parts of the brain, such as the substantia nigra, which are affected in neurodegenerative diseases such as Parkinson’s disease will be studied. Our preliminary results show changes in neurotransmitter expression in the substantia nigra several months after infection, supporting this hypothesis. Specific Aim 2: To examine the role of viral macromolecular products and infiltrating inflammatory cells in the brains of mice infected with SARS-CoV-2 in the development of PASC. Using molecular and immunological approaches, we will assess whether inflammation results from extrapulmonary SARS-CoV-2 infection and rapid clearance within the first days of infection (‘hit and run’). Using a mouse in which infiltrating myeloid cells can be readily identified, we will analyze localization, function and gene expression of these infiltrating myeloid cells, as well as of T cells that infect the brain chronically. The ultimate goal is to relate these changes in inflammation to the neurological/olfactory dysfunction that we characterize in Aim 1.

NIH Research Projects · FY 2025 · 2016-09

PROJECT SUMMARY The goal of this National Eye Institute (NEI) P30 competitive renewal is to continue to enhance the research activities of NEI-funded investigators at The University of Iowa. This will be achieved by providing a suite of Core Services, performed by skilled technical staff and supervised by experienced NEI-funded faculty members, employing standardized, automated procedures when possible. We have modified the Core Services in this renewal application to continue to provide the most up-to-date state-of-the-art services to meet the evolving needs of NEI-funded faculty at The University of Iowa. The combination of centralization (allowing specialization and concomitant increases in efficiency) and the breadth of services available from the various Cores are expected to continue to provide utility and increased productivity to all vision researchers at The University of Iowa. The University of Iowa NEI P30 program is aimed at supporting three primary service cores: 1) Molecular Imaging Core; 2) Rodent Phenotyping Core; and 3) Bioinformatics and Biostatistics Core. The capabilities of and research activities provided by these cores all require specialized equipment and highly trained staff that would be inefficient and costly to duplicate in every laboratory seeking to employ these techniques.