University Of Iowa

NIH Research Projects · FY 2026 · 2022-12

Project Summary/Abstract Chronic pain afflicts millions of patients, yet treatments are largely ineffective, and development of new, targeted therapies is limited by knowledge gaps in the neurobiology of pain. Despite recent progress, key questions remain about the incredibly complex cellular and functional organization of ascending and descending neural circuits that control pain processing. Answering these questions is essential for developing new, more efficacious, and better targeted therapeutics for acute and chronic pain. The main objective of this proposal is to identify the circuit connections and synaptic mechanisms of a novel group of neurons that we recently discovered in the lateral pons. Situated juxta the A5 noradrenergic cell group, these neurons (which we termed LJA5) express prodynorphin and glutamic acid decarboxylase 1 (GAD1). They are distinct from the A5 noradrenergic neurons as they do not express tyrosine hydroxylase. LJA5 neurons project to all spinal levels of lamina I of the dorsal horn (DH), as well as to the parabrachial nucleus (PB) and the periaqueductal gray (PAG). Notably, we showed that these neurons play an important role in pain regulation. Specifically, chemogenetic activation of LJA5 neurons suppressed capsaicin- and inflammation-induced mechanical pain, but not thermal sensitivity, whereas chemogenetic inhibition of LJA5 enhanced mechanical hypersensitivity during inflammation. Our preliminary data also showed that chemogenetic activation of LJA5 neurons strongly attenuated neuropathic pain both via systemic and intrathecal administration of a designer drug. Collectively, these findings suggest that LJA5 neurons and their projections represent a novel component of descending pain modulation, and also pose many important questions about this novel circuit: 1) Which types of pain are regulated by LJA5 neurons? 2) What are the key projections/outputs of LJA5 neurons? 3) What synaptic mechanisms are utilized by LJA5 neurons? Our central hypothesis is that LJA5 neurons modulate pain processing by controlling synaptic transmission in lamina I of the dorsal horn of the spinal cord. We will test this hypothesis in 3 specific aims by using a multidisciplinary approach that includes chemogenetic manipulation combined with behavioral testing, patch-clamp recording in innovative intact spinal cord preparation and 2-photon Ca2+ imaging in axonal boutons of primary afferent central terminals. Aim 1 will establish the role of LJA5 neurons in mouse models of inflammatory and neuropathic pain. Aim 2 will examine functional significance of the main LJA5 projections. Aim 3 will determine synaptic mechanisms that mediate the antinociceptive effects of LJA5 neurons in the spinal cord. This proposal will define fundamental characteristics of a novel descending pathway involved in pain modulation, including its functional connectivity, synaptic mechanisms and its role in acute and chronic pain states. This work is transformative because it identifies a novel bulbospinal pain modulatory pathway, a significant advance in our basic understanding of pain that may offer alternative approaches to pain control.

NIH Research Projects · FY 2026 · 2022-12

ABSTRACT Sudden unexpected death in epilepsy (SUDEP) is the leading cause of death in patients with refractory epilepsy. SUDEP is second only to stroke in years of potential life lost to neurological disease and is a major public health problem. Several etiologies have been proposed for SUDEP including cardiac and respiratory dysregulation. Another that is postulated is impaired arousal. Seizures impair arousal. Among arousal stimuli, one that may be particularly relevant to SUDEP is CO2. CO2 rises following seizures and is part of the seizure cessation mechanism. Seizures are frequently associated with ictal and post-ictal central and obstructive apneas. Apnea further exacerbates the accumulation of CO2. Impairment of CO2- arousal is proposed as an etiological factor in another sudden death entity, sudden infant death syndrome, which has many parallels with SUDEP. We discovered that seizures impaired CO2-arousal in seizure naïve mice and those that do not have a particularly profound death rate. Whether this is true in mouse models with strong SUDEP phenotypes is unknown. Thus, our goal in this proposal is to determine how seizures in different sleep states impair CO2-arousal in mouse models of temporal lobe epilepsy (TLE) and the genetic epileptic encephalopathy, Dravet Syndrome (DS). In Aim 1 we will determine the extent to which CO2-arousal is impaired in epilepsy models. We will focus on the pilocarpine-TLE model as many patients that die of SUDEP have TLE, and the DS model, as patients with DS have a disproportionately high SUDEP risk. We will also determine whether simply having epilepsy in these models impairs CO2-arousal as a potential easily measurable biomarker for SUDEP risk. In Aim 2 we will determine whether neuronal function, assessed via fiber photometry, of arousal system components in the dorsal raphe nucleus and parabrachial nucleus, two important contributors to sleep-wake regulation and key nodes in CO2-arousal, is impaired by seizures and epilepsy. In Aim 3 we will determine whether optogenetically stimulating a DRN-PBN circuit in DS mice, using a novel mouse model, prior to seizures prevents seizure-induced death lending direct insights into possible therapeutic measures. Since the models employed have known death rates, we will be able to compare findings between mice that die and those that survive making these studies more relevant to SUDEP. Combining these findings with our previous work, we will have a powerful, rigorous, translatable approach to identify convergent and divergent mechanisms across models for how impaired CO2-arousal in epilepsy contributes to SUDEP risk. We expect to be able to leverage these mechanisms to identify at-risk individuals and reduce death from this devastating disease.

NIH Research Projects · FY 2025 · 2022-11

Project Summary/Abstract During pregnancy, viral infections in the mother may have catastrophic effects on her health, or on the viability of the developing fetus. Recently, emerging pathogen outbreaks such as those involving Zika Virus (ZIKV), SARS-CoV-2 or Ebola Virus (EBOV) have highlighted how understudied the role of the placenta is in transmission of viral infections. This project specifically focuses on the cell tropism of EBOV during pregnancy. Ebola Virus Disease (EVD) is caused by infection with EBOV or other members within the Ebolavirus genus. During pregnancy, EVD results in loss of ~100% of fetuses or neonates with or without the additional loss of the mother. Anecdotal data from EBOV outbreaks in Africa suggest that EBOV directly infects placental tissues, thus transmitting virus to the fetal compartment, but rigorous experimental evaluation of placental infection has not been performed; the tropism of EBOV for placental cells, mechanisms of cellular entry, and route of infection from mother to fetus are currently unknown. Aim 1 will examine tropism of EBOV in placental tissues. Further, as EBOV has been shown to bind to and internalize into many cell types via interactions with phosphatidylserine (PS) receptors, Aim 2 studies will evaluate the role of three PS receptors on EBOV infection of the placenta and fetus. These studies will be performed using two low containment EBOV model viruses, rVSV-EBOV-GP-GFP and EBOV ΔVP30, that have been used extensively to understand filovirus tropism and receptor usage. The knowledge gained from these rigorously designed studies will elucidate cell populations within the placenta infected and important for fetal transmission as a first step toward understanding the catastrophic pathogenesis of EVD in pregnancy. Additionally, this work will provide insights for the development of therapeutic treatment options to improve the maternal and fetal outcomes of EVD. The experiences, techniques, mentoring, and concepts in this proposal were specifically tailored to Ms. Hanora Van Ert and her training goals. As a developing researcher passionate about improving the health and wellbeing of pregnant women, Ms. Van Ert is currently completing her studies in the MSTP at University of Iowa under the scientific mentorship of Dr. Wendy Maury and Dr. Mark Santillan receiving individualized training at the intersection of virology, immunology, and reproductive health sciences to supply her passion with the necessary research skills. Additionally, the MSTP, Department of OB/Gyn, and Department of Microbiology and Immunology at the University of Iowa provide ample training opportunities in the forms of seminar series, funding for attending academic conferences, opportunities to meet prominent people in the fields of virology, immunology, and OB/Gyn, as well as a supportive and collaborative research environment. Ms. Van Ert will complete her MSTP training and pursue a research residency in OB/Gyn, clinical Maternal- fetal medicine fellowship, and ultimately tenure track position at an academic medical center to continue investigating the host-pathogen immune response at the maternal-fetal interface within the placenta.

NIH Research Projects · FY 2026 · 2022-11

Influenza A virus (IAV) is a major cause of serious respiratory illness and has been responsible for significant morbidity and mortality in humans worldwide. Seasonal IAV infections lead to approximately 200,000 hospitalizations and 36,000 deaths annually in the United States during non-pandemic years. Furthermore, the IAV pandemics of 1918 (~50 million deaths worldwide), 1957-58 (~1 million deaths worldwide) and 1968-69 (~700,000 deaths worldwide) further demonstrate the impact of IAV on human health. The recent appearance of highly pathogenic H5N2 and the Eurasian highly pathogenic avian H5 viruses in the US as well as the high mortality rate observed in humans infected with the pre-pandemic avian H5N1 (~55-60% mortality rate) IAVs has heightened concerns. Thus, there has been a renewed interest in developing novel and efficacious influenza vaccination strategies that confer broad based protection to combat this significant global public health and pandemic threat. Recent studies have importantly shown that strategies that induce local (i.e. nasal mucosa and lung) tissue-resident memory T and B memory cells in addition to systemic immunity offer the greatest protection against future heterologous IAV encounters. The currently licensed IAV-vaccines by their design do not induce lung resident memory T and B cell responses. Thus, our long-term goal is to develop a protective universal vaccine against pre-pandemic avian IAV that induces lung and nasal resident T and B cells in addition to systemic immunity. We have developed a polyanhydride nanoparticle based IAV vaccine (IAV- nanovax) against seasonal IAV that breaks the cold chain, is needle free, and is biocompatible. This IAV- nanovax has shown efficacy in protection against homologous and heterologous seasonal IAV infections and the ability to induce T cell and B cell responses in the lungs and nasal passages. The HAs from H5 IAV are thought to be poorly immunogenic and require higher doses to be effective when compared to HAs from seasonal IAV thereby limiting vaccine design. Critically, our prior work with polyanhydride nanoparticles has also shown that they can induce robust immunity even at normally suboptimal levels of antigen. Therefore, this proposal will use the combined expertise of the PI and Co-Investigators and robust pre-clinical models to determine if a nanoparticle-based approach will allow for the induction of durable, IAV-specific, lung-resident T and B cell responses and broad-based protection against homologous and heterologous pre-pandemic avian IAV strains using the following Specific Aims: 1) Determine the efficacy of avian pre-pandemic IAV-nanovax in inducing robust local and systemic immunity and conferring protection against subsequent H5 IAV exposures, 2) Determine if apIAV-nanovax confers broad-based protection.

NIH Research Projects · FY 2024 · 2022-11

Project Summary Learning and memory are modulated by dopaminergic circuits, which convey valence and/or arousal signals. This proposal will examine how discrete dopaminergic circuits modulate learning and memory and neuronal plasticity in memory-encoding brain regions in Drosophila. Specifically, it will disentangle the roles of dopaminergic circuits that convey positive valence signals, negative valence signals, and valence-independent arousal signals. In vivo imaging experiments will examine how these dopaminergic neurons drive discrete patterns of plasticity in the mushroom body and downstream valence-coding output neurons that mediate approach and avoidance behavior. Complementary behavioral and optogenetic manipulation experiments will decipher how each of these neuronal subsets modulates arousal, valence, and memory strength. These studies will apply the large genetic toolkit and experimental throughput of the fly toward developing a more comprehensive understanding of how learning and memory alter the flow of information through the brain, to ultimately engage novel behaviors (e.g., conditioned approach/avoidance) following learning. Understanding how memories are encoded in the brain and disrupted in brain disorders is a prerequisite to the rational design of treatments for memory impairment. Results of the present studies will provide guideposts for future research into the molecular biology of memory formation across multiple model organisms, as dopaminergic circuits regulate arousal and memory across taxa. The project will support our long-term goal of understanding of memory down to the single-cell and subcellular levels, contributing to the knowledge base necessary for the rational development of novel treatments for memory impairment.

NIH Research Projects · FY 2026 · 2022-10

Project Summary Neurofibromatosis type 1 is a relatively common monogenetic, multisystemic disorder that affects approximately one in 3,500 individuals worldwide. The causative gene encodes a protein called neurofibromin (Nf1), which essentially acts as a brake on Ras signaling via Ras-GAP activity. Nf1 affects multiple downstream signaling cascades, including central regulators of metabolism. Prior studies have suggested that loss of Nf1 may affect metabolism, but the mechanisms, particularly at the systemic level are unclear. Nf1 effects on metabolic processes may underlie or modulate some of the symptoms of the disease, such as behavioral alterations and cancer predisposition. This project will test the mechanisms underlying how loss of Nf1 affects metabolism in vivo, using the powerful Drosophila model for neurofibromatosis type 1. Upon completion, we will have a clear picture of: (1) the genes and cellular signaling cascades that regulate metabolism in an Nf1- dependent manner, (2) how Nf1 functions in neuronal circuits to regulate metabolism through central control, (3) the neurotransmitters and/or peptides that are involved in the central control of metabolic regulation, (4) how loss of Nf1 mechanistically regulates peripheral energy stores through the effects of novel genes. The highly conserved nature of Nf1 and its signaling functions, as well as fundamental neuronal circuit functional principles, underscores the broad applicability of the results. Overall, this project will contribute to understanding conserved Nf1 functions in metabolism and neuronal function, laying the foundation for research into metabolic effects of Nf1 across organisms and future development of novel therapeutic interventions.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT Photoreceptor cells in the retina are highly polarized and compartmentalized neurons. Most proteins localize to a specific compartment in photoreceptors, and such compartment-specific protein localization is essential for the proper function and survival of photoreceptors. Despite considerable research efforts, however, our understanding of the mechanisms by which photoreceptors achieve compartment-specific protein localization is limited. Related to this, the pathophysiology of retinal degenerations caused by the disruption of these mechanisms is also not sufficiently understood. This is partly because of the lack of easy-to-use means to monitor protein trafficking and confinement in diseased photoreceptors. To address this need, we have developed two transgenic mouse lines, iROSRePT (inducible Reporter for the Outer Segment Renewal and Protein Trafficking) and iRATProx (inducible Reporter for ABCA4 Trafficking and Proximity labeling). In the proposed studies, we will utilize these reporter lines and four disease models representing the disruption of the ciliary gate, intraflagellar transport (IFT), and exocytotic membrane fusion machinery and investigate the precise requirements of these mechanisms for the compartmentalized protein localization in photoreceptors. We anticipate that the outcome of this study will significantly advance our understanding of the mechanisms by which photoreceptor compartment homeostasis is attained and the pathophysiology of retinal degenerations linked to defective protein trafficking and confinement. This knowledge will build a foundation to develop treatments for cilia-related retinal degenerations and assess the efficacy of newly developed therapies.

NIH Research Projects · FY 2026 · 2022-09

Project Summary Although evidence in favor of physical activity (PA) for reducing age-related cognitive decline continues to grow, critical issues for the viability of PA to promote cognitive health are persistently low PA adoption and adherence in the population. Because PA starts declining at middle-age and neurodegenerative pathologies increasing dementia risk start decades before cognitive impairment, there is a pressing need to understand how and why middle-age adults are successful at adopting and sustaining PA. Theoretical and empirical support have identified self-regulatory capacities as critical for acting on our intentions and plans to be more physically active, however causal evidence testing whether and how strongly selfregulatory capacities affect PA behavior change is absent from the literature. Therefore, our objective is to test the causal role of the self-regulation construct, cognitive control, in PA behavior change among inactive middle-age adults. Based on the Temporal Self-Regulation Theory (TST) for PA behavior change, we hypothesize cognitive training designed to improve cognitive control will increase the success of a PA behavior change program, and that training cognitive control with emotionally valenced stimuli will further increase PA adherence at moderate-to-vigorous intensities. Our predictions are based on research showing the importance of cognitive control in PA adherence and maintenance, evidence that negative affective experiences of PA are common and detrimental to future PA initiation and maintenance, and research supporting the modifiability of cognitive control with adaptive cognitive training. We test our overall hypothesis with three aims, including a developmental R61 phase (Aim 1) to develop and refine a computerized training program targeting aspects of motivation, planning, and cognitive control theorized to promote PA behavior change, followed by an implementation R33 phase (Aim 2) with a lab-based, randomized controlled trial (RCT) to determine the extent that cognitive control for emotionally valenced information is a target mechanism for PA behavior change in inactive middleage adults. The RCT implemented in Aim 2 will be tested across two sites to increase breadth of barriers to exercise and generalizability of our sample, which forms the basis for (Aim 3) determining moderators of intervention efficacy for improving PA behavior change. We predict those with poor cognitive control and high negative affective experiences to moderate intensity PA will benefit most from cognitive control training targeted to an affective domain for PA behavior change. Our results will be significant by determining the causal role of cognitive control and affect in sustainable PA behavior change in the critical period of midlife. Success will culminate in a scalable training program to boost PA adherence, setting the stage for personalized and accessible strategies for midlife adults to change their course towards a more physically active lifestyle and a virtuous cycle of sustained cognitive and physical health.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY/ABSTRACT – GREAT PLAINS CENTER FOR AGRICULTURAL HEALTH (OVERALL) The Great Plains Center for Agricultural Health conducts research and provides outreach to reduce the burden of injury and illness facing Midwest agricultural producers, particularly those hazards associated with both large and small row crop and livestock/animal production. This Center will serve a nine-state region: who share these production similarities: Illinois, Indiana, Iowa, Kansas, Minnesota, Missouri, Nebraska, Ohio, and Wisconsin. Our vision of safe and healthy agricultural communities is accomplished through basic and applied research, use of participatory approaches, and theory-driven education and translation activities. Our research projects are designed to address multiple hazards important to both the NIOSH national strategic priorities and priorities for our region’s farming workforce. The research questions being proposed will: reduce the burden of back pain from a lifetime of tractor use, provide resources to help families of aging farmers transition to safer work at multiple stages of dementia, provide tools to help farmers identify and reduce high- risk hazards in their physical environment, reduce injuries from farm vehicles on roadways, and improve the respiratory health of livestock producers and the animals they raise. Research proposes to develop innovative technologies needed to close gaps in knowledge (whole-body vibration), awareness of hazards (safety checklist app), and equipment (air quality systems), and innovative partnerships (e.g., Alzheimer’s Association, state extension health educators) bring new collaborators with unique skills to contribute to agricultural injury reduction. Partnerships throughout the Center projects and cores, including advisory boards and focus group participants, will provide essential information to enhance the adoption of tools being developed and/or tested across the Center portfolio. The lessons learned from our research initiatives will be translated into outreach materials by being incorporated into national educational curricula, disseminated through partners, and incorporated into multimedia discussions (social media, print media-Safety Watch, and FarmSafe podcasts) to aid in the dissemination and uptake of these best practices. The Center will establish and support systems to foster communication, identify and build strategic partnerships, and assess the needs of the agricultural community. Our evaluators will work within all projects and Cores to maximize our ability assess the Center’s contributions to improved health and safety outcomes for our region’s farmers. The Great Plains Center for Agricultural Health has demonstrated its contributions to improving knowledge and practices to protect the health and safety of agricultural workers throughout the Midwest, and we are ideally positioned, with strong regional collaborators, to generate and disseminate evidence-based practices, guidelines, and controls to protect this essential workforce.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT Polygenic scores - summaries of the genomic contribution to risk and resilience for biomedical traits - are an emerging and promising approach for clinical risk assessment and personalized medicine. Minoritized groups, such as gender minorities, do not currently benefit from insights gained via studies of polygenic scores because these groups have not been sufficiently included or characterized in this research. This disparity must be addressed both by inclusion during recruitment as well as consideration of the effects of heterogeneity during analysis. For example, our preliminary data show striking dissociations in suicide risk as influenced by polygenic scores. Notably, these opposing associations are apparent only when gender diversity status (i.e., cisgender; gender-diverse) is modeled, underscoring the imperative to parsing the heterogeneity of such associations by both sex and gender. These results suggest value in genomic research for gender minority groups to understand both innate risk and resilience. The promise of genomic research informed by designated sex, gender, and their interaction (i.e., gender diversity) depends on the ability to rigorously and equitably include and characterize individuals across the spectra of designated sex and gender. Currently, biomedical research relies on checklist or write-in gender identity descriptors, which do not capture the continuous and simultaneous nature of dimensional binary and nonbinary gender experiences and result in statistically underpowered analytics with far too few individuals within each gender self-descriptor category. This perpetuates the exclusion of gender and its intersection with designated sex in genetic research. We propose to close this gap by calibrating and genetically characterizing the Gender Self-Report (GSR), a novel and broadly disseminable method for obtaining multidimensional gender for genomic research and broader research applications. First, we will facilitate partnership between scientists and gender diverse community stakeholders, and reduce paternalistic tendencies in this ethically complex field of research, by building on our established community partnerships with a purposively recruited stakeholder panel (N=50) to provide a final version of the GSR itemset. Next, to advance the dimensional characterization of gender identity and gender diversity in a genomic research context, we will validate the stakeholder-refined GSR in a large sample (N=10,000) of genotyped neurotypical and neurodivergent adults, enriched across broad experiences of gender diversity. Finally, to demonstrate proof-of-principle for how designated sex, gender, and gender diversity contextualize patterns of association with polygenic scores, we will measure key health outcomes (both mental and physical), in a large, genetically informed sample enriched for neurodiversity (e.g., autism) and broad gender diversity. The proposed research will provide value to gender minority groups by seeking a better understanding of how polygenic scores apply specifically to them, and not just the cisgender proportional majority.

NIH Research Projects · FY 2025 · 2022-09

Bartter syndrome (BS) is a congenital renal tubulopathy caused by mutations of transporters impairing NaCl reabsorption in the thick ascending limb of Henle's loop (TAL). Antenatal BS is caused by mutations of NKCC2 or ROMK in the apical membrane of TAL. Classic Bartter’s (cBS) is due to mutations of the basolateral chloride channel ClC-Kb, presenting highly variable phenotypes and renal outcomes. As opposed to the prevailing view that salt wasting in BS is due to loss of function of transporters in mature TAL, we recently reported that the phenotype of cBS in Clc-k2-/- (mouse ortholog of ClC-Kb) mice is mainly due to developmental defects in the inner medulla and TAL hypoplasia. How Clc-k2 deficiency leads to renal tubule hypoplasia is unknown. The growth of renal tubules arises from a positive balance between cell proliferation and cell death. Preliminary data reveal Clc-k2-/- tubular cells are less proliferative and more apoptotic than WT cells. Cell cycle analysis using primary cultured TAL cells reveals more Clc-k2-/- cells reside in the G1 phase than WT cells, suggesting that Clc-k2 deficiency impairs the proliferation and cell cycle of TAL cells. What causes cellular hypoplasia and cell cycle arrest in Clc-k2-/- renal tubular cells is unknown. Mitochondria provide energetics for transport, and mitochondria dysfunction is linked to cell cycle arrest. We hypothesize that decreased transport activity and mitochondrial dysfunction underlies tubular hypoplasia in cBS. To support the hypothesis, Specific Aim-1 will examine that Clc-k2 deficiency causes cell cycle arrest and mitochondrial dysfunction in renal tubular cells via decreasing transport activity. Assays for cell proliferation, cell cycle analysis, and mitochondria bioenergetics will be performed in primary TAL and DCT cells or tubules. Mitochondrial morphology will be examined in tubules of the kidney section of Clc-k2-/- mice. Direct enhancement of mitochondrial functions by expressing PGC1α (peroxisome proliferator-activated receptor coactivator-1α, an activator of mitochondrial biogenesis) transgene or Nrf2 (Nuclear factor-erythroid factor 2-related factor 2, a transcription factor downstream of PGC1α) agonists will be used to rescue Clc-k2-/- mice. Specific Aim-2 will further test the hypothesis using two mouse models with gain-of-function (GOF) transport activity. The effect of GOF mice to rescue cell proliferation and mitochondrial dysfunction caused by Clc-k2 deficiency will be studied. The traditional view of the pathogenesis of BS as salt-wasting in mature renal tubules has led to treatment focused on salt repletion. However, many patients progress to chronic kidney disease despite volume repletion. Our studies will provide new insights into the pathogenesis of BS and provide potential therapeutic considerations targeting mitochondrial function restoration.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY Background and Objectives: Synapses and circuits possess a robust capacity for keeping their outputs stable. Using the Drosophila melanogaster neuromuscular junction (NMJ) as a model synapse, many labs have recently identified dozens of signaling molecules and processes that stabilize synapse function through a non-Hebbian form of homeostatic neuroplasticity called presynaptic homeostatic potentiation (PHP). These findings offer a rich reservoir for discovery science, but at this point we have little understanding of how dozens of discrete homeostatic signaling molecules integrate into coherent system that stabilizes synapse function over time. The objective of this proposal is to solve that problem combining genetics, pharmacology, imaging, biochemistry, and electrophysiology. Ultimately, improved knowledge about homeostatic forms of synaptic plasticity could lead to a better understanding of neurological disorders that occur when synapse stability is lost. Specific Aims and Research Design: This project has two specific aims. We know that PHP at the Drosophila NMJ can be acutely induced in minutes and then chronically maintained for days. The first aim is to define a sequence of events that occurs during the opening minutes of PHP induction. For this aim, we take advantage of a serendipitous finding from a genetic screen: impaired chaperone function in the muscle slows PHP signaling. Using this genetic tool we will delineate an order of processes that occurs as the muscle signals to the nerve and potentiates release. For the second aim, we developed a new pharmacological approach to monitor the transition periods between induction, acute expression, and long-term maintenance of PHP. We will apply this new method to characterize about 25 known genetic conditions that impact the sustained maintenance of PHP. We expect to define distinct PHP signaling modalities. Between our aims, the expected outcome is a model of how a synapse can sustain homeostatic function by integrating multiple signals across phases of time. Health Relatedness: Neurological disorders like epilepsy, ataxia, and migraine are associated with unstable neuronal function. Understanding how synapses work to maintain stability on a molecular level could have pro- found implications for disorders with underlying neuronal instabilities. Yet the signaling events that tightly control levels of synaptic output are poorly understood. The tractable Drosophila NMJ employs homoestatic strategies to stabilize synapse function – such as altering levels of presynaptic calcium influx – that are shared by mammalian central synapses. Taking advantage of the molecular and genetic tools offered by the NMJ promises to shed light on universally conserved mechanisms of how synapses maintain stable function throughout life.

NIH Research Projects · FY 2025 · 2022-09

Project Summary/Abstract Humans possess extraordinary flexibility in our behavior: Given the same environmental input, we can act differently depending on our goals and the context. For example, when facing the same data, we can process them differently depending on our goals (e.g., visualize the data to obtain a figure, perform statistical analysis to test a prediction, or even delete the data if the goal is to free storage space). To date, research on this topic has focused on how such flexible behavior is implemented via cognitive control, which is a set of cognitive mechanisms supporting goal-directed and top-down modulation on information processing in the brain. In other words, much research has been conducted to study how a task is executed. However, less is known about where such task knowledge is from, that is, the mnemonic mechanisms that encode, reinforce, and generalize the neural representations of task knowledge. Understanding these mechanisms is crucial to fully understand human intelligence, as the remarkable abilities of learning and retaining task knowledge promptly and efficiently distinguish humans from other animals and artificial intelligent agents and make us adaptive to this ever-changing world. Furthermore, filling the knowledge gap of how we learn and remember task knowledge is also key to understand, detect and treat task learning deficits that are common in mental disorders such as schizophrenia and attention-deficit / hyperactivity disorder (ADHD). In this project, we will focus on the hippocampus, a central brain structure for learning and memory. To achieve the objective of uncovering the hippocampal contributions to task learning, six experiments are proposed using a combination of behavioral methods and human functional magnetic resonance imaging. Specifically, Aim 1 will identify hippocampal contribution to constructing a task representation by assembling task information and experiences to build a task model. Aim 2 will identify how the hippocampus encodes a new task representation into a memory network of existing representations. Aim 3 will identify how the hippocampus reshapes existing task representations when they become associated with other tasks via compositional relations. This work is expected to identify how the human hippocampus constructs key content of task representations (Aim 1) and organizes multiple task representations in relation to each other (Aim 2 and 3). These findings will further our understanding of how the hippocampus contributes to task learning, cognitive control and adaptive behavior. This work will also have clinical impact in bridging the gap between hippocampal abnormality and task learning deficits, which were separately observed in mental disorders such as schizophrenia and ADHD. Ultimately, this work will have a broad impact in helping detect and treat task learning deficits.

NIH Research Projects · FY 2026 · 2022-09

PROJECT SUMMARY Sleep permeates our early existence: A typical human newborn sleeps 16 hours each day, evenly divided between active (REM) sleep and quiet sleep. The relatively high proportion of active sleep in newborns is the foundation for the decades-old hypothesis that active sleep is important for infant brain development. In considering this hypothesis, it is important to remember that active sleep is a complex state composed of a variety of behavioral and physiological components that emerge and coalesce over development. Thus, it may be that one or more of these components play outsized roles in promoting brain development. One such candidate component is the phasic activity that comprises twitches of the limbs, face, and eyes. Over the past 15 years, research in infant rats has revealed that sensory feedback from twitching limbs triggers discrete and abundant activity throughout the sensorimotor system, cascading from the spinal cord to the brainstem, cerebellum, thalamus, sensorimotor cortex, and hippocampus. In recent years, with funding from the Gates Foundation and an R21 from NICHD, investigations were extended to full-term human infants over the first seven postnatal months. Using behavioral measures and high-density electroencephalographic (EEG) recordings, this research revealed heretofore unknown features about the spatiotemporal organization of infant sleep and twitching and introduced new developmental milestones and hypotheses. Here, we propose to extend these efforts to preterm infants for two reasons: Preterm infants exhibit an even higher proportion of active sleep than newborn full-term infants, and they are at increased risk for deficits in motor skill, cerebral palsy, autism, and other neurodevelopmental disorders. Thus, we will determine whether twitching can enhance our understanding of the origins and time course of atypical developmental trajectories and guide future interventions. Very preterm human infants (<32 weeks postmenstrual age) and mildly preterm infants (32-36 weeks postmenstrual age) in the neonatal intensive care unit (NICU) in the Stead Family Children’s Hospital at the University of Iowa will be recruited to participate in a longitudinal study spanning the period before (Phase I) and after (Phase II) discharge from the NICU. Every two weeks during Phase I, sleep behavior will be recorded along with EEG activity, and respiratory rate. During Phase II, in the same preterm infants as well as full-term age-matched controls, sleep behavior, EEG activity, and respiratory rate will be recorded three times between one week and six months corrected age. Neurobehavioral and motor skill assessments will be performed at one and six months of age and clinical follow-up assessments will be performed at two years of age. Because this study is longitudinal, behavioral and physiological sleep data will be compared in the same infants across Phases I and II and related to developmental outcomes. Ultimately, this project will provide an unprecedented opportunity to understand how sleep and sleep-related behavior contribute to typical and atypical development in early life.

NIH Research Projects · FY 2026 · 2022-09

Telomeres protect chromosome ends in eukaryotes. In the absence of telomerase, telomeres shorten, which eventually leads to senescence. A minority of cells, however, can escape senescence and stabilize telomeres by a recombination process called Alternative Lengthening of Telomeres (ALT). ALT is responsible for telomere maintenance in ~15% of cancers, but it also contributes to stabilization of telomeres in aging or stem cells. Thus, understanding the ALT mechanism is important to identify factors that can influence whether and how it occurs. It is also well known that exposure to various environmental stressors and air toxins influence all aspects of telomere biology and promote telomere-related diseases, including various types of cancer. However, the effects of environmental factors on ALT and telomere dynamics are difficult to study, owing to the absence of experi- mental systems to identify and follow the critical steps responsible for ALT establishment in human cells at the molecular level. A key gap in the study of ALT has been, until recently, the lack of quantitative assays. The goal of this research is to unravel the mechanisms of ALT by identifying genetic, structural, and environmental factors affecting ALT. This research takes advantage of a powerful system in yeast, Saccharomyces cerevisiae, where ALT was originally discovered and where ALT can be followed from the beginning of telomere erosion through formation of survivor cells. This research will employ a unique combination of methods that were recently devel- oped by the applicants that enabled a quantitative study of ALT. This research, so far, has yielded three firsts in the field: (i) a population genetics-based assay that determined the frequency of ALT, (ii) ultra-long sequencing described the detailed structure of individual chromosome ends in ALT survivors, and (iii) a combination of com- putational modeling, Southern blot analysis and PacBio sequencing uncovered “molecular milestones” repre- senting different steps of ALT in large populations of yeast cells. Using these new approaches, this research will determine the effects of genetic, chromosome-structural and environmental factors during the steps of ALT, including: (i) the formation of ALT precursor cell populations initiated by eroded chromosome ends; (ii) the de- velopment of ALT survivors taking place in such populations; (iii) the frequency of ALT outcomes, and (iv) the molecular structure of chromosome ends in ALT survivors. This research will test the effect of environmental stressors on ALT, including oxidative damage from paraquat and the effect of cadmium, a ubiquitous environ- mental pollutant and a type I carcinogen. This will provide insight into molecular effects of environmental factors as well as illuminate new molecular mechanisms of ALT formation, potentially uncovering opportunities for med- ical interventions. Furthermore, this research will establish a robust system to evaluate the influence of other environmental contaminants on ALT, and to utilize telomere erosion and ALT formation as a new type of biosen- sor to assess the effects of various genetic changes and environmental assaults on genetic stability. Overall, this study of ALT in yeast is expected to serve as a roadmap to perform quantitative studies of ALT in humans.

NIH Research Projects · FY 2025 · 2022-09

Project Summary/Abstract The long-term goal of this research is to improve patient safety by establishing simulator training and evaluation of surgical skills as essential components of orthopedic residency programs. Many orthopaedic surgeries involve the challenging integration of fluoroscopic image and video interpretation with skillful tool manipulation to achieve well-defined objectives. Simulation has proved beneficial in this context for surgical trainees, but programs have been slow to embrace this advance, and methods for evaluating operating room (OR) performance of these skills to document improvement have been lacking. Objectively measuring skill in the OR is a critical step toward this goal because it allows skills training to be linked to performance in surgery. This is an important missed opportunity. The proposed research will advance objective measurement techniques that are critically needed to speed improvement in resident performance on technical skills, ultimately reducing costs while enhancing patient safety. The long-term goal will be achieved by partnering with the American Board of Orthopaedic Surgery (ABOS) to more tightly integrate surgical skills training and simulation into pre-certification policies. For this reason, researchers at the University of Iowa are leveraging the skills and experience of existing ABOS grant-funded research groups at the University of Rochester and the University of Texas Health in Houston to pursue this goal. The proposed research approach is based on our multi-institution simulation studies with novel surgical simulators and on our previous, AHRQ-funded, ground-breaking analysis techniques for assessing task- specific, detailed, OR performance. Our central hypothesis is that orthopedic surgical skill competence can be objectively, quantitatively, and reliably measured from behaviors observable in fluoroscopy and video routinely collected in the OR. Our research team is well poised for this work; our core multi-disciplinary team of engineers, surgeons, psychometricians have collaborated for nearly a decade to improve orthopedic residency training. Our team is now partnered with the ABOS to advance simulation as a tool for training orthopaedic residents and assessing performance prior to qualifying for certification. Aim 1 of the proposed research is to measure differences in resident OR performance from objective analysis of surgical imagery, and speed up these measurements. Aim 2 is to determine how differences in simulator training correlate with skills demonstrated in the OR, and use this information to improve training programs. Aim 3 is to identify individual differences in skills among residents, both in the skills lab and in the OR, and use this knowledge to improve individual training. This research is innovative because it demonstrates and disseminates new skill assessment techniques critically needed to hasten improvement in orthopedic resident performance, ultimately reducing costs while enhancing patient safety.

NIH Research Projects · FY 2025 · 2022-09

Project Summary/Abstract The IKS channel is a voltage-gated potassium channel found in the heart. After a cardiac muscle cell is depolarized, the IKS channel opens and allows a potassium current to leave the cell, returning it to resting state. Dysfunction in this channel is associated with numerous acquired and inherited arrhythmias, including long QT syndome, a leading cause of sudden cardiac death. The channel itself is made of two protein components: KCNQ1 (Q1), the pore-forming subunit; and KCNE1 (E1), a single transmembrane accessory protein. When Q1 is expressed alone, the channel opens at a negative voltage and conducts a small potassium current, but with E1 bound, the channel opens at a high positive voltage and conducts a large potassium current. Thus E1 acts as an intrinsic gating regulator of the channel. The IKS channel is also regulated transiently by phosphorylation in response to signaling by the sympathetic nervous system. When phosphorylated, the channel opens more quickly and more often, which leads to an increaed potassium current. This leads to faster cell repolarization and facilitates increased heart rate. However, the molecular mechanisms of both intrinsic regulation by E1 and transient regulation by phosphorylation are not currently understood. The goal of this research is to gain a high- resolution understanding of these mechanisms of regulation. Intrinsically, phenylalanine (Phe) has been shown to play a role in many ion channel mechanisms because despite being overall nonpolar and largely hydrophobic, it has a quadropole moment that creates a negative charge in the center of its aromatic ring. This means that it can behave like an anion and form strong charge-charge interactions with other charged residues, despite being in the hydrophobic core of the channel. Preliminary data has been found that shows a particular Phe residue in the IKS channel participates in a previously uncharacterized charge interaction that slows IKS channel response to voltage, and is thus a key player in intrinsic regulation. This work will characterize this interaction, determine how E1 binding alters it, and determine the specifc role this interaction plays in regulating IKS channel function. As well as intrinsic regulation by E1, Q1 is transiently regulated by phosphorylation at two sites on the N-terminus. Cumulatively, the channel response to phosphorylation is an increase in current, but the mechanism of this increase is unknown. Additionally, the individual roles of the two phosphorylation sites are not understood. This research will use caged serine, a modified serine residue that can only be phosphorylated after photolysis of a bound caging moiety. Encoding caged serine into each of the sites individually and observing the effects of phosphorylation at each site on the channel will provide this key information about phosphoregulation of the IKS channel. This work, which will provide mechanistic explanations of intrinsic and transient regulation of the IKS channel, will pave the way for development of novel therapies for long QT syndrome and other arrythmias attributed to this channel.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT Developmental Language Disorder (DLD) is characterized by difficulties in the ability to learn and use language and is one of the most common neurodevelopmental disorders (prevalence 7-12%1,2). Though problems emerge in childhood, DLD continues into adulthood3-6 and has profoundly negative effects. Adults with DLD are less likely to seek post-secondary education7-9, may have extended bouts of unemployment9, and have higher rates of depression10. Yet, DLD in adulthood is severely under-researched. An understanding of the language profile is crucial as language abilities in adulthood impact well-being, income, and job performance11. Additionally, there is a clear need to better understand the mechanisms that mediate language abilities in adults with and without DLD. Doing so will help explain theories of DLD12-15 (speed of processing and working memory accounts) and expose a wider range of individual differences in language ability. Examining competition – the activation of competing linguistic representations as speech unfolds – is an ideal approach to exploring these mechanisms. Competition is a fundamental component of language, is well-documented in typical adults16-19, and critically, distinct aspects of competition can be linked to each theoretical account20,21. Our overall objective is to characterize the long-term outcomes of DLD in adulthood (Aim 1) and to identify specific cognitive mechanisms mediating these outcomes (Aim 2). To address our objectives, we utilize a large, pre-existing dataset and participant pool from one of the most comprehensive examinations of DLD to date: the Iowa Longitudinal Study22. We will re-recruit subjects with DLD (n=150) and with typical language (TL; n=250) from this historic cohort, who are now adults (30–34 years old). In Aim 1, we leverage retrospective language measures from kindergarten through 10th grade and collect new outcome measures in adulthood to characterize the long-term outcomes of DLD. We predict that adults with DLD will diverge from adults with TL in language skills that are more complex and higher-level language skills that are important for communication in the workplace11. Further, we predict a fanning effect: some children with DLD will “catch up” to their TL peers in adulthood, some will show evidence of a decline, and others will show stable trajectories. In Aim 2, we measure real-time competition across language modality and level using eye-tracking in the Visual World Paradigm23. According to speed of processing accounts15,20, adults with DLD may be slower than their TL peers to activate competitors and targets. According to working memory accounts21,24, adults with DLD will show sustained competitor activation. Further, we predict that measures related to the dynamics of competition (speed of activation and timing of competitor suppression) will account for variation in language outcomes in adults across the ability spectrum. The proposed work would represent the largest and most comprehensive characterization of language abilities in adults with (and without) DLD to date, inform theories of DLD and general theories of language processing, and provide foundational knowledge toward clinical models of prevention and long-term intervention for adults with DLD.

NIH Research Projects · FY 2025 · 2022-09

The rate of mortality by suicide is approximately twenty times higher in psychiatric disorders as compared to the general population. Among psychiatric disorders, bipolar disorder (BD) has the highest rate of attempts (~40%), which is 2-3 times higher than in major depressive disorder (MDD). While neural circuits underlying suicidal behavior have been proposed, these have emerged largely based on studies of MDD and exclude the cerebellum. Work from our group and others have implicated the cerebellum in suicidal behavior, impulsivity, and bipolar disorder suggesting that it may play a key role in suicidal behavior. In this study, we propose to study a putative suicide risk circuit (SRC) that includes the cerebellum to prospectively evaluate the connectome of this neural circuit and metabolism in the nodes of the proposed SRC. To assess the SRC, brain imaging coupled with measurements of suicidal behavior, psychiatric symptoms, and personality traits will be acquired in a sample of 300 subjects with a psychiatric disorder (BD I and MDD) with 75 having a prior suicide attempt and 75 without a prior attempt for each psychiatric diagnosis. Seventy-five matched controls will also be acquired. Brain imaging will include multi-modal MR imaging to study anatomy (T1, T2), functional (task based fMRI), connectome (resting state fMRI and diffusion imaging) and metabolism (MRS and T1ρ). This data will be used to answer the following aims: Aim 1) Does the SRC differentiate suicide attempter from non- attempter in BD? and Aim 2) Does the SRC differentiate suicide attempter / non-attempter in MDD in the same way as BD? This work will increase our understanding of the brain circuits implicated in suicidal behavior, how the cerebellum may be involved in these circuits, and what metabolic differences are associated with suicidal behavior. In addition, the study will reveal if the same neural circuit (i.e. the SRC) plays a significant role in suicidal behavior across disorders or if there are different neural circuits involved across disorders. The goal of this project is to better undertand the neurobiology of suicidality to help identify those at risk for a future suicide attempt. We anticipate that this study will reveal new targets for treating subjects at risk for suicidal behavior. .

NIH Research Projects · FY 2024 · 2022-09

The rate of mortality by suicide is approximately twenty times higher in psychiatric disorders as compared to the general population. Among psychiatric disorders, bipolar disorder (BD) has the highest rate of attempts (~40%), which is 2-3 times higher than in major depressive disorder (MDD). While neural circuits underlying suicidal behavior have been proposed, these have emerged largely based on studies of MDD and exclude the cerebellum. Work from our group and others have implicated the cerebellum in suicidal behavior, impulsivity, and bipolar disorder suggesting that it may play a key role in suicidal behavior. In this study, we propose to study a putative suicide risk circuit (SRC) that includes the cerebellum to prospectively evaluate the connectome of this neural circuit and metabolism in the nodes of the proposed SRC. To assess the SRC, brain imaging coupled with measurements of suicidal behavior, psychiatric symptoms, and personality traits will be acquired in a sample of 300 subjects with a psychiatric disorder (BD I and MDD) with 75 having a prior suicide attempt and 75 without a prior attempt for each psychiatric diagnosis. Seventy-five matched controls will also be acquired. Brain imaging will include multi-modal MR imaging to study anatomy (T1, T2), functional (task based fMRI), connectome (resting state fMRI and diffusion imaging) and metabolism (MRS and T1ρ). This data will be used to answer the following aims: Aim 1) Does the SRC differentiate suicide attempter from non- attempter in BD? and Aim 2) Does the SRC differentiate suicide attempter / non-attempter in MDD in the same way as BD? This work will increase our understanding of the brain circuits implicated in suicidal behavior, how the cerebellum may be involved in these circuits, and what metabolic differences are associated with suicidal behavior. In addition, the study will reveal if the same neural circuit (i.e. the SRC) plays a significant role in suicidal behavior across disorders or if there are different neural circuits involved across disorders. The goal of this project is to better undertand the neurobiology of suicidality to help identify those at risk for a future suicide attempt. We anticipate that this study will reveal new targets for treating subjects at risk for suicidal behavior. .

NIH Research Projects · FY 2025 · 2022-08

PROJECT SUMMARY Dysregulated metabolic function and chronic inflammation are prominent features of obesity in both humans and animal models. Brown adipose tissue (BAT) plays a critical role in metabolic adaptation in response to stresses including overnutrition, wherein the metabolic adaptation is disrupted and inflammatory stress is elevated. However, there remains a key knowledge gap in the interplay between inflammatory and metabolic cues in BAT during overnutrition. Obesity-associated chronic inflammation is characterized by excessive nitric oxide (NO) production and aberrant protein cysteine nitrosylation (S-nitrosylation). Our preliminary data showed that diet- induced obesity (DIO) elevates BAT protein S-nitrosylation, including uncoupling protein 1 (UCP1). This aberrant BAT NO bioactivity is in part due to downregulation of alcohol dehydrogenase 5 (ADH5), the major denitrosylase modulating cellular nitro-thio redox balance. Moreover, we showed that BAT Adh5 deletion suppressed UCP1- dependent mitochondrial respiration, worsened glucose intolerance and increased BAT inflammation in mice with DIO. All of these defects were improved by restoration of Adh5 expression in the BAT. These data provide the first evidence that ADH5 plays a protective role in the BAT against metabolic stress. Thus, we hypothesize obesity compromises ADH5-regulated cellular nitrosative homeostasis in the thermogenic adipose tissue, contributing to obesity-associated metabolic dysfunction. We will test this hypothesis by completing two specific aims. In Aim 1, we will define the mechanism by which obesity suppresses ADH5 expression and its pathophysiological significance in obesity. In Aim 2, we will determine the molecular mechanisms underlying ADH5-mediated BAT metabolic homeostasis. The regulation of BAT metabolic function by nitro-redox signaling and the contribution of this regulation to metabolic dysfunction in obesity are new and unexplored concepts. Accomplishment of this project will provide first insights into the mechanisms by which aberrant NO signaling links BAT inflammatory cues to metabolic dysfunction and new avenues for developing of therapeutic targets to ameliorate BAT dysfunction in the context of obesity.

NIH Research Projects · FY 2025 · 2022-08

Project Summary: Percutaneous liver tumor ablation procedures are performed with ultrasound (US), computed tomography (CT) and CT-fluoroscopy guidance. Each image guidance modality has strengths and weakness. US provides superior tissue contrast and real-time guidance; however, suboptimal or absent acoustic windows, motion (respiratory, bulk), and poor applicator conspicuity compromise precise tumor targeting and a widening skill gap limit widespread adoption. The larger field-of-view (FOV) provided by CT is critical for evaluating proximity of non-target anatomy; however, poor tissue contrast, limited access along the X-Y-Z axis, motion, and non- real-time guidance limit accurate tumor targeting. Magnetic resonance imaging (MRI) provides superior soft tissue contrast but presents challenges with the comparatively long acquisition times for real-time guidance and limited access within the scanner bore. Image fusion technologies attempt to harness the strengths of each modality. Unfortunately, current image fusion technologies are unable to sufficiently account for liver deformation, bulk respiratory and patient motion which compromise targeting. Further, they do not address challenges associated with US. An Academic Industrial Partnership is proposed to develop and validate a solution comprised of two primary elements. The first is a combined platform with a novel MRI compatible hands-free US and fast 3D deformable image registration for optimized simultaneous US and virtual MR image guidance in the ablation setting. The second element will be development of advanced MRI methods for real- time treatment monitoring through MR Thermometry in combination with near-real-time assessment of ablation zone margins using advanced diffusion and perfusion MRI. This solution will improve primary efficacy and reduce local recurrence after ablation of liver tumors. This proposal will focus on microwave ablation however the solution is applicable for a wide variety of thermal therapies.

NIH Research Projects · FY 2025 · 2022-08

The objective of this proposal is to determine how high fat diet (HFD)-induced lipid dysregulation links obesity with increased breast cancer (BC) risk. Epidemiologic studies have confirmed that obesity increases the risk and mortality of BC, but the molecular mechanisms of obesity/breast cancer associations remain largely unknown. Our recent studies demonstrate that lipid chaperone A-FABP (adipose fatty acid binding protein, also known as FABP4) promotes obesity-associated BC by intracellular regulating pro-tumor activity in tumor associated macrophages (TAMs) and extracellular enhancing aggressive phenotype of BC cells. Thus, A- FABP might represent a new factor linking dysregulated lipid metabolism with obesity/BC risk. Moreover, we observed that obesity can be induced by consumption of different types of HFDs, including saturated fats (e.g. cocoa butter) or unsaturated fats (e.g. olive oil, fish oil). However, only cocoa butter HFD-induced obesity was associated with increased A-FABP expression and mammary tumor growth. These observations suggest a novel concept that not all HFD-induced obesity is tumorigenic. Given the undefined links underlying obesity- induced BC risk, we hypothesized that HFDs rich in saturated fats promote BC risk through A-FABP-mediated immune and metabolic regulations. As such, A-FABP offers a novel therapeutic target and biomarker for obesity-associated BC risk. Three complementary but independent specific aims are designed to address our central hypothesis. Aim 1 is to determine the molecular mechanisms by which different HFDs upregulate A- FABP for BC risk. We will identify which HFDs promote mammary tumor risk and further dissect how the “bad” fat drives intracellular A-FABP expression in TAMs and promotes extracellular A-FABP secretion from adipocytes. Aim 2 is to delineate the downstream metabolic mechanisms of HFD-upregulated A-FABP in BC risk and immunotherapy. We will delineate how intracellular A-FABP in TAMs regulates the FA oxidation (FAO)/HIF2α/PD-L1 pathway for immune suppressive function, followed by delineation of how extracellular A- FABP reprograms lipid metabolic profile in BC cells to enhance their aggressive phenotype. We will further evaluate if blocking A-FABP activity with our unique humanized antibodies improves A-FABP-induced metabolic dysregulation and reduces obesity/BC risk. Aim 3 is to evaluate A-FABP as a biomarker for obesity- associated BC in humans. We will determine the function of A-FABP in peripheral monocytes and measure the levels of soluble A-FABP in serum and A-FABP expression in tumor stroma using specimens collected from lean and obese women with or without BC. Successful completion of this proposal will determine the “bad HFDs” in promoting BC risk and identify the molecular and metabolic mechanisms by which A-FABP mediates the pro-tumorigenic activities in HFD-induced obesity/BC risk.

NIH Research Projects · FY 2024 · 2022-08

Project Summary/Abstract I am currently a mentored assistant research scientist in the Department of Obstetrics and Gynecology at the University of Iowa. My long-term goal is to establish an independent research program to improve outcomes for women with ovarian and endometrial cancer through the safe delivery of effective and personalized treatment regimens. I intend to pursue an independent tenure-track faculty position at a competitive research institution, building upon my postdoctoral training in novel RNA-based therapeutics and current work in personalized medicine for gynecologic cancers. The objective for my independent K22 research program is to mechanistically distinguish between common missense p53 mutants and develop synthetic RNA-based therapeutics to overcome the deleterious functions. Approximately 40-50% of all observed p53 mutations are single nucleotide variants that result in missense mutations that change a single amino acid. Missense mutations not only abrogate canonical DNA binding and interaction with co-factors, but also confer new activities, including transcription of non-canonical targets and new in protein:protein interactions. Studies will focus on endometrial and ovarian cancer given the widespread occurrence of p53 mutations in these cancer types and the extremely poor 5-year survival of patients with p53 mutant cancers. I hypothesize that missense mutations in p53 that result in protein hyperstabilization activate different transcriptomic signatures that can be perturbed using novel RNA aptamers evolved to be specific for each mutant p53. This work leverages a highly innovative strategy to isolate native p53 from patient-derived organoid (PDO) cultures, which allows for study of p53 mutations in their endogenous environment. First, I will determine the mechanism(s) of survival and chemoresistance for recurrent p53 mutants by calculating the DNA binding specificity native p53 mutants and link this to genomic localization, gene regulation and chemosensitivity. This work includes a novel approach to isolate p53 mutant proteins from PDO models to capture the native protein conformation, posttranslational modifications and splice isoforms. Next, I will restore chemosensitivity in models with p53 mutants by perturbing the deleterious activity of individual p53 mutants with mutant-specific RNA aptamers. These studies will provide the first comprehensive assessment of the functional consequences of a large panel of p53 mutants using native protein and establish the feasibility of developing aptamer-based tools that inactivate p53 mutants. This project merges my graduate work in cell signaling, postdoctoral training in RNA aptamer-based therapeutics and more recent work in translational studies of gynecologic cancers. With funding from this K22 award and my multi-disciplinary collaborators, I will be well-equipped to establish an independent research program to improve outcomes for women with gynecologic cancer.

NIH Research Projects · FY 2025 · 2022-08

Abstract Interferon (IFN) and IFN stimulated genes (ISGs) are a critical component of innate immune responses that help in the early control of viral infections. Transcriptional upregulation of IFN and ISGs is tightly controlled through negative regulators under basal conditions to maintain homeostasis, as aberrant expression of IFN and ISGs can promote diseases like chronic viral infection, cancers, neurodegeneration, and diabetes. Through a genome- wide CRISPR/Cas9 screen to identify proviral host factors and regulators of innate immunity, we identified Capicua (CIC) as a novel regulator of innate immune responses, as CIC knockout (CIC KO) cells demonstrated transcriptional upregulation of IFN and ISGs under basal conditions and showed restricted replication of different RNA viruses. Based on studies in drosophila and cancer models, CIC, a highly conserved DNA-binding transcriptional repressor, binds to a specific octameric sequence (CIC binding site or CBS) near target genes and constitutively represses gene transcription. Through promoter motif analysis of ISGs upregulated in CIC KO cells identified by RNA-Seq, we observed the presence of CBS motifs in several, but not all, upregulated ISGs, including RIG-I, MDA-5, IRF3/7/9, IFNβ, IFNγ, IFIT1, TRIM22, MX1, and ISG15. Preliminary Assay for Transposase-Accessible Chromatin (ATAC)-Seq analysis confirmed increased chromatin accessibility near the putative CBS containing IFIT1 and TRIM22 promoters in CIC KO as compared to control A549 cells. In addition, we validated direct CIC binding to IFNβ and ISG promoters (STAT1, IFIT1,TRIM22) by chromatin immunoprecipitation-qPCR (ChIP-qPCR) analysis. Moreover, the CIC-ATXN1/L co-repressor complex is rapidly degraded during influenza virus infection, suggesting that degradation of the CIC-ATXN1/L complex may be an integral part of the innate immune response pathway. Importantly, the innate immune regulatory function of the Cic-Atxn1/l co-repressor complex is conserved in the murine model. Based on these findings, we propose a novel model for the transcriptional regulation of IFN and ISGs: under basal conditions, the CIC-ATXN1/L co- repressor complex binds to CBS and represses ISG transcription; however, virus-induced proteasomal degradation of CIC-ATXN1/L relieves these target genes from active repression, thereby allowing for robust transcription through IRFs and STATs. The goal of this proposal is to gain mechanistic insights into the transcriptional regulation of IFN and ISG promoters by the CIC-ATXN1/L co-repressor complex, ultimately providing a new paradigm for the regulation of cell autonomous antiviral responses. PHS 398/2590 (Rev. 06/09) Page Continuation Format Page