University Of Iowa

NIH Research Projects · FY 2025 · 2021-04

Abstract Aberrant redox homeostasis has been proposed to contribute to the pathophysiology of type 2 diabetes (T2D). However, the mechanisms are poorly understood. Redox systems are regulated by pro-oxidants, such as reactive oxygen species (ROS) and antioxidants such as glutathione. Patients with T2D have elevated levels of ROS and lower levels of glutathione. Attempts to reverse this redox imbalance in T2D using redox-modulating drugs or infusion of antioxidants has shown promise in reversing insulin resistance in preliminary studies, but ultimately have failed in clinical trials due to their short half-lives and delivery challenges. New methods and a better understanding of redox mechanisms in T2D are needed to address an underlying pathophysiology of T2D that is not currently effectively addressed by current modalities. ROS possess an unpaired electron, making them paramagnetic and capable of interacting with externally applied electromagnetic fields (EMFs). We recently identified a unique set of EMF parameters that rapidly modulate ROS and redox homeostasis. When applied to mouse models of T2D and human cells, EMFs were found to exert remarkable effects on glycemia and insulin sensitivity, reversing glucose intolerance and insulin resistance in three days, without adverse effects. We also found that application of EMFs altered the metabolic flux of glucose, increasing glucose incorporation into glycogen and reducing fatty acid levels. Scavenging paramagnetic ROS or preventing redox adaptations by infusing oxidizing redox solutions (GSSG) abolished these striking therapeutic effects. These findings lead us to hypothesize that EMFs target ROS to induce an NRF2-mediated redox response that is insulin sensitizing in part by altering the fate of glucose. Therefore, the goal of this project is to elucidate the redox and metabolic mechanisms underlying the insulin sensitizing effects of EMFs. We will test our hypotheses in two specific aims: 1) Determine the redox mechanisms that mediate the insulin-sensitizing effects of EMF-therapy; and 2) Determine the mechanisms by which EMF-therapy or redox modulation redirects the metabolic fate of glucose to improve insulin sensitivity. The use of EMFs as a redox- and glycemia-modulating modality provides an unprecedented opportunity to study the role of redox in T2D pathophysiology and to advance the understanding of a novel, insulin sensitizing phenomenon. We will identify specific metabolic changes that occur in response to EMF exposure and determine mechanisms by which the redox state regulates hepatic metabolic flux and insulin sensitivity. This work will identify a novel mechanistic link between two previously disconnected fields of inquiry, static EMFs and glycemic regulation and will bridge redox biology with glucose metabolism. Successful completion of this work will lay the foundation for the future clinical development of a wearable device that emits EMFs to target redox systems for the noninvasive management of T2D.

NIH Research Projects · FY 2025 · 2021-04

Project Summary / Abstract Deficits in cognitive function and memory are a debilitating aspect of neurodegenerative disease resulting in long-term disability, enormous suffering to individuals and their families, and significant socioeconomic cost. Currently, more than 16 million Americans live with cognitive impairment, and this number is expected to continue rising as the number of individuals affected by Alzheimer's disease and related dementias is predicted to double by 2060. Long-term memory consolidation requires the induction of gene expression in a specific temporal pattern. Newly synthesized transcripts are translated and folded into functional proteins and then trafficked to the correct cellular location. Dysfunction in any of these steps can lead to memory impairment and may be dysregulated in disease conditions, although the precise mechanisms by which this occurs are unclear. My long- term goal is to determine the specific molecular mechanisms through which the Nr4A sub-family regulates gene expression to control long-term memory in order to develop therapeutic tools to treat AD, which I will pursue as an independent investigator at a research-focused institution. The overall objective of this proposal is to determine the mechanisms that link transcriptional regulation and protein folding during memory formation and how the disruption of these processes contributes to cognitive impairment in Alzheimer's disease. My central hypothesis is that Nr4A regulates genes in specific cell types to facilitate protein folding and memory formation. I will test this through the following three aims: Specific Aim 1 (K99): identify cell-type specific transcriptional signatures of memory and identify direct gene targets of Nr4A during memory consolidation using a single-cell RNA sequencing approach; Specific Aim 2 (K99): determine the role of an Nr4A target, the protein folding chaperone Hspa5, in memory consolidation and identify its downstream protein targets; Specific Aim 3 (R00): determine the effect of activating Nr4A transcription factors on memory deficits and gene expression in a mouse model of Alzheimer's disease. This project provides training in cutting-edge research skills, including computational analysis of single cell data and chromatin enrichment in memory research. The University of Iowa is home to experts on memory, computational psychiatry, neurodegeneration, and molecular biology, and the collaborative environment provides an ideal setting in which to obtain the necessary skills that will allow me to transition into a successful independent research career. As such, during the mentored (K99) phase, I will engage in activities designed to prepare me to successfully achieve independence, including training in scientific presentations, laboratory management skills, grant writing tools, scientific peer-review, and interview/application preparation. Collectively, this award will provide me with the cutting-edge skills and expertise in molecular biology, neurodegeneration and behavioral pharmacology necessary to ensure a strong technical and conceptual foundation to start my independent laboratory investigating mechanisms for the treatment of Alzheimer's disease.

NIH Research Projects · FY 2025 · 2021-04

Project Summary/Abstract 1 Cystic fibrosis is one of the most common autosomal regressive genetic disorders in the United States: 2 approximately 1 out of 32 Caucasians and 1 out of 61 African Americans is a CF carrier. A hallmark of this 3 disease is recurrent respiratory infections. However, cystic fibrosis requires two copies of the CFTR 4 mutation. Traditionally, one non-mutated CFTR gene was thought to be sufficient for maintaining health. 5 However, a few small studies and preliminary work by our group have demonstrated that CF carriers may 6 be at increased risk for respiratory infections, including recurrent sinusitis, pneumonia, and atypical 7 mycobacterial infections, than non-CF carriers. Given an estimated population of 15 million CF carriers in 8 the United States, if carriers are more likely to acquire respiratory infections, the attributable burden of 9 respiratory infections and corresponding antimicrobial use attributable to the CF-carrier state may be 10 substantial. Thus, there is a critical need for population-based investigations to precisely determine the 11 attributable risk of the CF-carrier state for respiratory infections. 12 The goal of our research is to determine the role of the CF-carrier state on the risk for acquiring respiratory 13 infections. Due to the frequency of genetic testing for CFTR mutations for genetic counseling, it is possible 14 to identify CF carriers in some existing population-based data sets. Our group has recently demonstrated 15 that CF carriers are at significantly greater risk for respiratory infections compared to age and sex matched 16 controls. However, more work is needed that considers other patient characteristics as well as patients' 17 specific CF mutations. In addition, the biological plausibility of these findings needs to be established. Thus, 18 we will determine the natural history of respiratory infections for CF carriers using administrative claims 19 data; determine the risk for respiratory infections among CF carriers in a genotyped cohort using data from 20 electronic medical records, and assess airway epithelial function in CF carriers compared to non-carriers. 21 This work is significant because respiratory infections are a major cause of morbidity and mortality; CF 22 carriers are common and can be identified by current genetic counseling methods; and there are CFTR- 23 modulating therapies available that may be cost-effective treatments for CF carriers with excessive 24 respiratory infections. This work is innovative because all previous work in this area has been done using 25 small cohorts of patients, and we will have access to a large cohort of CF carriers and controls. Also, we will 26 be able to identify the CF carriers using diagnosis and procedure codes in administrative data and the 27 medical record an in one cohort we will be able to identify specific CFTR mutations. 28 Given the number of CF carriers in the general population, the methods and models we develop will have 29 broad implications for other diseases beyond respiratory infections.

NIH Research Projects · FY 2025 · 2021-04

Project Summary/Abstract The purpose of this Mentored Patient-Oriented Career Development Award (K23) is to support my short-term career objective of determining if dysfunction of the autonomic nervous system (ANS) is an early pathological feature of HD by quantitatively characterizing functional connections between brain regions that regulate the ANS in children with the gene expansion that causes HD using magnetic resonance imaging (MRI). I will also investigate unique physiologic measures of ANS function and early effects on the vascular system in these participants. ANS dysfunction has been described in adult patients with HD, but it has been thought that this is a secondary complication of neurodegeneration. However, I recently discovered that children carrying the HD gene expansion that causes HD (GE children) exhibit symptoms consistent with enhanced sympathetic tone decades prior to their predicted motor onset. These findings indicate that ANS dysfunction may be one of the earliest manifestations of neurodegeneration in HD. As a result, the ANS may be a therapeutic target for disease modification of HD, but more information is required. The ANS is highly regulated by cortical brain regions that comprise the Central Autonomic Network (CAN), and cortical thinning and atrophy have been well-described in HD. However, there are no published reports that have objectively characterized the integrity of the functional connections in the CAN in HD. I will perform resting-state and tasked functional MRI on GE children to characterize the function of the CAN at different stages of the disease. This experiment will test the specific hypothesis that quantitative changes in functional integrity of the CAN are apparent decades prior to the predicted motor onset of HD. Additionally, I will explore physiologic measure of ANS dysfunction including baroreflex sensitivity (BRS) and how this relates to the function of the vascular system early in the disease course of HD. Specifically, I will measure aortic stiffness and carotid artery compliance while also measuring cerebral blood flow using arterial spin labeling to test the hypotheses that relative to healthy control children, GE children will demonstrate increased aortic stiffness, decreased BRS, and decreased cerebral blood flow. These experiments will provide vital information regarding when ANS dysfunction occurs in HD, the underlying mechanisms causing the dysfunction, and if these changes have negative effects on the cardiovascular system early in the disease course. I have a unique background that positions me well to be a successful translational scientist. Further training is required in sophisticated neuroimaging methods, neurodevelopment and neurobiology, as well as biostatistics. The proposed integrated research, world-class mentorship team, and didactic training programs will ensure my short-term and long-term success. Additionally, the proposed research and training plans support my long-term career goal to be an independent translational pharmacist studying the structure and function of the brain in patients with HD to advance therapeutic strategies.

NIH Research Projects · FY 2025 · 2021-04

PROJECT SUMMARY There is growing evidence that limited access to high-quality cancer treatment is one of the main drivers of higher cancer mortality rates among rural cancer patients. Our analyses of Iowa Cancer Registry data indicate that 40% of rural patients with breast and colorectal cancers receive most or all definitive treatment in rural hospitals that do not collect or monitor data on their quality of cancer care, and are not accredited by the American College of Surgeons Commission on Cancer (CoC). Our data also shows these patients are less likely to receive guideline-concordant care. Given patients' needs and preferences to receive cancer care locally, a promising strategy to improve quality of cancer care and outcomes in rural populations is to intervene directly with the community hospitals in these areas. New evidence has demonstrated effectiveness of this approach: the Markey Cancer Center Affiliate Network (MCCAN) was formed by the University of Kentucky (UK) Markey Cancer Center to improve quality of cancer care in their own rural, low-resourced state, one that leads the nation in cancer incidence and mortality. Over the last decade MCCAN has facilitated the sharing and diffusion of resources and best practices throughout their network. As a result, affiliates markedly improved performance on established, cancer care quality measures and expanded their services (e.g., psychosocial and survivorship support). They were also almost 3 times more likely to obtain CoC accreditation than their matched controls. However, the MCCAN model has not been rigorously defined, evaluated or tested in any other setting. We propose to adapt this successful health system-level intervention for Iowa, establishing the Iowa Cancer Affiliate Network (I-CAN). Although there are similarities between Iowa and Kentucky's populations that suggest the MCCAN model may be a good fit, there are also significant differences in healthcare infrastructure and resources that require careful adaptation of the intervention prior to its implementation in order to retain its effectiveness. We will use novel, rigorously developed, theory-based implementation science methods to identify MCCAN's core functions (i.e., what makes it effective), study the implementation process and evaluate how I-CAN performs in a new context. We have identified 4 rural, Iowa hospitals to participate in this intervention trial and developed expert support teams to assist key stakeholder groups within each hospital. Through interviews and qualitative analyses, we will assess determinants and outcomes of the implementation process, and perceived value of the CoC accreditation standards and the intervention itself as a way to improve the quality of cancer care for their patients. We will compare compliance with treatment-related quality measures and the proportion of CoC standards of cancer care implemented in target and control hospitals, pre- and post-intervention using a difference-in-difference estimator. This work could lead to dissemination of similar models across rural settings thereby improving quality of care, reducing rural disparities in cancer outcomes and giving rural hospitals an avenue to demonstrate their quality of care.

NIH Research Projects · FY 2025 · 2021-04

Abstract Cognitive symptoms of Parkinson’s disease (PD) can affect up to 80% of PD patients and lead to enormous societal cost. These symptoms involve impaired executive functions such as working memory, attention, behavioral flexibility, and timing, and can progress to psychosis, hallucinations, and dementia. There are few therapies that improve cognitive function in PD. Thus, there is a critical need to better understand the fundamental mechanisms of cognitive dysfunction that occurs in PD. Our long-term goal is to elucidate the mechanisms by which cognitive dysfunction occurs in PD patients in order to develop new targeted treatments. PD involves death of midbrain dopaminergic neurons in ventral tegmental area (VTA). These neurons send mesocortical projections to key cognitive cortical areas such as the prefrontal cortex. It is unknown how VTA dopamine neurons are involved in cognitive processes that malfunction in human PD patients. Our goal in this proposal is to harness cell-type specific rodent models to characterize VTA dopamine neuronal function in cognitive processes relevant to PD. Our preliminary data demonstrates interval- timing variability correlates with PD-related cognitive dysfunction. Our rodent research demonstrates that VTA dopamine is necessary for interval timing, prefrontal timing-related modulations and prefrontal 4 Hz rhythms. Here, we will combine optogenetics, fiber photometry, and neuronal ensemble recordings in transgenic mice to interrogate VTA dopamine projections with cell-type-specificity and millisecond resolution. We will test the overall hypothesis that VTA dopamine neurons engage cognitive processing in the prefrontal cortex. In Aim 1 we will determine how silencing VTA dopamine neurons impacts cognitive processing. In Aim 2 we will define how VTA dopamine neuron dynamics predict cognitive processing. In Aim 3 we will determine how stimulating VTA dopamine neurons impacts cognitive processing. This proposal is broadly significant in determining when and how VTA dopamine engages prefrontal cognitive processing. Although this is a basic-science proposal focused on VTA dopamine neurons, our results will guide new therapies for human PD. This work could contribute to biomarkers for PD and for other Alzheimer’s disease and related dementias (ADRDs) such as dementia with Lewy bodies (DLB).

NIH Research Projects · FY 2025 · 2021-03

PROJECT SUMMARY Pathogen recognition receptors and their associated adaptors are essential for the prompt detection of pathogens and the subsequent initiation of effective host responses and restoration of homeostasis. Over the last decade, Nod-like receptors (NLRs) have been on the forefront of innate immune research, and several groundbreaking studies have been published, to which my research has contributed. Leishmania spp. are parasites with global health importance, yet the role of NLRs and their adaptors during Leishmania spp. infection has been an understudied area. Receptor interacting protein kinase 2 (RIPK2) is an essential adaptor downstream of cytoplasmic sensors NLRC1 and NLRC2, and play an important role in a wide variety of clinical settings including bacterial, viral and fungal infections, as well as non-infectious inflammatory diseases such as multiple sclerosis, inflammatory bowel disease and metabolic diseases. However, the role of RIPK2 during cutaneous leishmaniasis remain unknown. In this grant, we propose to investigate the precise cellular and molecular mechanisms involving RIPK2 during Leishmania major (L. major)-induced cutaneous disease. Our preliminary work has already elucidated several exciting features of the role of RIPK2 during L. major infection. We found that RIPK2- deficient mice are highly susceptible to L. major infection. Unexpectedly, mice deficient in NLRC2 or both NLRC1/NLRC2 are dispensable in L. major infection suggesting a novel sensor that function upstream of RIPK2. Moreover, RIPK2 interacted with CARD9 to provide protection against L. major. Based on these preliminary data, we propose that a novel Dectin-1/RIPK2/CARD9 signaling axis modulates anti-leishmanial immunity. Successful completion of this project will yield a better understanding of RIPK2 biology in general, unveil a novel pathway involving RIPK2 in signal transduction, and potentially identify therapeutic targets to ameliorate Leishmania spp.-associated pathology. 1

NIH Research Projects · FY 2025 · 2021-03

PROJECT SUMMARY/ABSTRACT Breast cancer has the highest incidence of cancer for women in the U.S. and across the world. Despite advances in technology—from film-screen images to Full Field Digital Mammography (FFDM) and now to Digital Breast Tomosynthesis (DBT)—the yearly miss rate has remained stubbornly stable, ranging between 10-30% at screening. Technology alone is not reducing errors of omission; we need to understand the specific challenges faced by the human readers interpreting the images, and the specific errors that they lead to. Satisfaction of Search (SOS) refers to the fact that, after having detected a first lesion in a case, the miss rate for additional lesions in the same case is substantially elevated. This specific type of error has been shown to account for 30% of misses in the domains of Radiology where it has been studied, including chest radiography and Computed Tomography. And yet, it has never been studied in the domain of breast cancer. Thus, there is a critical need to determine how SOS contributes to errors in breast cancer screening. In the present project we will determine the rates of occurrence and the underlying causes of SOS in FFDM and DBT. We have devised a novel method that overcomes limitations of previous methods and that is optimized for use in FFDM and DBT. Previous approaches to studying SOS involved the photographic addition of artificial lesions to images, which is not feasible for breast imaging. Instead, we will construct a database of naturally occurring cases that is structured for studying SOS. This will involve the collection of multiple-lesion cases and controlled single-lesions cases, where the former are matched with the latter on key diagnostic dimensions, such as lesion type, lesion size, and breast density. In two main experiments (one with FFDM and one with DBT), radiologists will read cases from the experimental set, marking the locations and diagnoses for benign and malignant lesions. Signal-detection analyses over dual- and single-lesion cases will be used to estimate the rate of SOS. Eye position and pupil diameter will be tracked as participants read each case. These data will allow us to assess the prevalence of different known causes of SOS: (a) premature termination, in which search following first lesion detection is less comprehensive compared with single-lesion control cases; (b) perceptual set, in which, after having detected a first lesion, participants are biased to find subsequent lesions with similar perceptual features, leading to reduced sensitivity in the detection of perceptually dissimilar targets; and (c) resource depletion, in which the demands of maintaining information about a first-detected lesion in memory reduce available perceptual/cognitive resources, thereby reducing the efficiency of subsequent search. Understanding the rates and underlying causes of SOS in breast cancer detection will lay the foundation for planned future work to develop training programs and best practices that mitigate the specific causes of SOS errors and thereby reduce miss rates in breast cancer screening.

NIH Research Projects · FY 2025 · 2021-01

Heart failure (HF) is the most common reason for hospitalization and death among those older than 65 years, and statistic is projected to grow as our population ages. The socioeconomic impact of HF on our health care system is enormous. Development of innovative approaches to the treatment of HF is therefore a top research priority. Although inflammation and immune activation have been implicated in the pathophysiology of HF over the past two decades, the progress for development of new pharmaceutical agents targeting this mechanism was stagnant, especially given that several anti-cytokine clinical trials targeting a single effector cytokine at the peripheral manifestations of HF did not produce clinical benefits. Obviously, the inflammatory mechanisms underlying the pathogenesis of HF have not been challenged. The proposed project studying a role of brain interleukin (IL)-17A (previously known as IL-17) in advancing central inflammation, sympathetic activation and cardiac dysfunction will address the need for a better understanding of the inflammatory mechanisms in HF and provide a novel anti-cytokine approach in treating this devastating disease. The research plan was developed based on the intrinsic property of IL-17A and our compelling preliminary data: 1) IL-17A is a kay inflammatory regulator bridging immune responses and tissue inflammation; 2) It boosts the expression of a broad spectrum of inflammatory mediators in the brain and in the peripheral tissue and cells; 3) Systemic and central administration of IL-17A induced dramatic and long-lasting increases in blood pressure, heart rate and renal sympathetic nerve activity to the levels not seen by other pre-inflammatory cytokines; 4) levels of IL-17A in the plasma, cerebrospinal fluid, and paraventricular nucleus of hypothalamus (PVN, a key cardiovascular and autonomic center of the brain) are higher in a rat model of HF vs. in sham-operated (Sham) animals; and 5) Its receptor, IL-17RA, is highly expressed in the PVN and substantially upregulated in HF. Using a multifaceted approach including electrophysiology, molecular biology, immunocytochemistry, pharmacology, biochemistry and neuroscience in Sham and HF rats, this project will: 1) identify the role of IL-17A in advancing central inflammation in HF; 2) determine the inflammatory mechanisms whereby IL-17A triggers sympathetic activation in HF; 3) evaluate the protective effect of central interventions targeting the IL-17A signaling, alone or in combination with other cytokines in HF. The proposed research will target a master regulator of inflammation rather than a single effector cytokine as a novel anti-cytokine strategy in treating HF, and consider the synergistic actions of multiple cytokines as a potentially more effective means of ameliorating HF. The proposed studies will characterize a previously unrecognized role of brain IL-17A in sympathetic activation and test its potential as a target in treating cardiac dysfunction of HF. Completion of this research project will provide important insights into the anti-inflammation therapeutic strategy in HF and may carry the implication for other cardiovascular disorders like hypertension and metabolic diseases like obesity or diabetes.

NIH Research Projects · FY 2026 · 2021-01

PROJECT SUMMARY: Halogenated compounds, including legacy pollutants (e.g., chlorinated ethenes (CEs), polychlorinated biphenyls) and emerging contaminants (e.g., 1,2,3-trichloropropane), are frequently en- countered at Superfund sites. A common bioremediation strategy for halogenated pollutants in groundwater and sediments is anaerobic reductive dehalogenation by organohalide-respiring bacteria (OHRB). Although effective, OHRB-driven bioremediation strategies are often incomplete in field applications. An emerging reme- diation strategy involving amendment of pyrogenic carbonaceous matter (PCM; e.g., activated carbon (AC)) to the subsurface offers a potential solution to problems with OHRB-driven bioremediation. Recent research high- lights the potential for PCM to promote synergistic interactions among OHRB and the auxiliary microbial com- munity and subsequently improve OHRB-driven bioremediation efficacy. However, the underlying mechanisms of how PCM properties best support microbial network interactions, and thereby enhance OHRB performance and contaminant biodegradation remain unknown. These unknowns limit our ability to optimize OHRB perfor- mance in bioremediation strategies where PCM is used. This proposal is aimed at closing the knowledge gap concerning specific surface effects of PCM on the performance of pollutant-degrading microorganisms, espe- cially OHRB. The central hypothesis is that key PCM properties will shape microbial community structure and drive the expression of metabolic functions associated with reductive dehalogenation processes. Elucidat- ing positive impacts between PCM and OHRB will allow for the development of tailored PCM that foster syner- gistic microbial network interactions and facilitate more effective and sustainable bioremediation. The hypothe- sis is based on preliminary data showing that OHRB-driven CE biotransformation performance was improved in the presence of biochar, OHRB were attached to carbon surfaces, and that PCM-like tunable polymer net- works can be successfully synthesized. Guided by these preliminary data, we will test the hypothesis by 1) providing a tunable platform for synthesis of PCM-like polymer membranes where surface charge and redox- active properties are varied individually, 2) quantifying the effects of PCM surface properties on microbial net- work interactions and subsequent performance of an organohalide-respiring mixed culture and, 3) developing tailored PCM for enhanced anaerobic bioremediation and contaminant mixture retention and validating material performance in microcosms. The proposed research is innovative because we will use a tunable platform to change material surface properties and employ advanced molecular microbial ecology tools to assess the im- pacts of these properties on microbial community structure, function, and activity including OHRB. Outcomes of this project will benefit human health and realize economic benefits by reducing human exposure to halo- genated pollutants in the environment and demonstrating the potential for more effective and sustainable re- mediation approaches that combine tailored PCM and OHRB.

NIH Research Projects · FY 2025 · 2020-12

PROJECT SUMMARY Sudden unexpected death in epilepsy (SUDEP) is the most common cause of death in patients with refractory epilepsy. Findings from SUDEP cases and from animal models of seizure-induced death suggest that central apnea is critical for SUDEP. The mechanisms underlying seizure-induced inhibition of breathing during SUDEP are unclear, and therefore, no known preventive strategies exist. Candidate: Dr. Brian Dlouhy, M.D. is a neurosurgeon at the University of Iowa whose clinical practice focuses on the surgical treatment of epilepsy. This proposal will be critical for his continued development as a neurosurgeon-scientist. This proposal will allow his research to build upon his established interest in how the amygdala modulates the neural control of respiration, will produce novel scientific results, and will provide critical training for Dr. Dlouhy. Dr. Dlouhy’s ultimate goal is to become an R01-funded academic neurosurgeon-scientist with an independent research program exploring neural control of respiration. Environment: The University of Iowa provides a rich training environment for Dr. Dlouhy, and he has the full support from the Carver College of Medicine and his Chair in the Department of Neurosurgery, Dr. Matthew Howard. Dr. Dlouhy has combined a unique team of mentors, each leaders of their respective fields and who possess expertise instrumental to Dr. Dlouhy’s research plan: amygdala circuitry, neural control of respiration, and human brain neurophysiology. Dr. George Richerson, is an expert in the field of respiratory neurophysiology; Dr. John Wemmie is a leader in the field of amygdala neurobiology; Dr. Matthew Howard is an expert in human electrophysiology. Research: Using intracranial recordings in epilepsy patients, Dr. Dlouhy previously found that apnea occurs when seizures propagate to the amygdala. Electrical stimulation of the amygdala can lead to apnea that is not associated with air hunger (dyspnea) or urge to breathe. Volitional control of respiration is spared during stimulation-induced apnea; subjects can speak normally and breathe when prompted. The underlying neural mechanisms by which the amygdala influences the brain’s respiratory control network to mediate these effects or regulate normal breathing are unknown. This proposal aims to identify and characterize neural activity within the amygdala and its functional connections with brain respiratory control sites. Dr. Dlouhy will also study how experimental neuromodulation of the amygdala affects volitional versus automatic breathing, and the perception of dyspnea. These aims will be accomplished using a combination of direct electrophysiological recording and stimulation techniques, electrical stimulation concurrent with fMRI, and respiratory physiology experiments. The proposed scientific research plan, the excellent mentorship team of Drs. Richerson, Wemmie, and Howard, and the enthusiastic support of the candidate’s institution and department will enable Dr. Dlouhy to launch a successful career as an independent neurosurgeon-scientist.

NIH Research Projects · FY 2024 · 2020-09

Project Summary/Abstract Increasing antimicrobial resistance (AMR) is one of the most urgent public health threats. In 2019, the Center for Diseases Control and Prevention (CDC) estimated that infections with AMR affect at least 2.8 million people and are associated with at least 35,900 excessive deaths annually in the US. This threat is particularly problematic among Gram-negative rod (GNR) pathogens in which high-rates of resistance to last-line antimicrobials have emerged globally, while our efforts to develop new antimicrobials have stumbled. To decelerate the emergence of AMR among GNR pathogens, it is essential to guide clinicians away from choosing unnecessarily broad-spectrum antimicrobials. An antibiogram, a facility-level summary of antimicrobial susceptibility data, is a common local reference tool which clinicians use when choosing empiric therapy. However, antibiograms have major limitations. First, little is known about how clinicians are currently using them when making empiric therapy decision. Second, antibiogram data is aggregated at the facility-level, and data may be skewed based on the type of practice or geographic area. Lastly, but most importantly, an antibiogram does not consider any patient-level factors. Therefore, there are strong, and pressing needs to understand 1) how an antibiogram is used by the clinician, 2) how much an antibiogram reflects the risk of AMR for individual patients and 3) how we can overcome limitations of antibiogram to optimize empiric therapy and reduce AMR. The overall goal is to create a novel, real-time personalized antibiogram (“Smart Antibiogram”) to overcome current limitations of antibiogram and to optimize clinician choice of empiric therapy for GNRs by providing a “predicted risk of AMR” based on a machine learning model incorporating patient- and facility-level data. This goal will be accomplished through (a) Master of Science in Health Informatics coursework, (b) a Mentorship Advisory Committee, (c) carefully selected conferences and workshops, and (d) a mentored research study. Our specific aims are to (1) Characterize the current use of antibiograms in clinical practice and measure the acceptable risk of resistance when clinicians make empiric therapy decisions for Gram-negative bloodstream infections and urinary tract infections within diverse clinical settings; (2) Assess the accuracy of currently-used antibiograms to predict the risk of resistance for individual patients in a large retrospective microbiology cohort for GNR infections; (3) Develop a machine learning model to predict the individualized risk of AMR for patients infected with GNR pathogens and validate prospectively and externally. This will lead to the future development of a personalized decision support tool (“Smart Antibiogram”). The expected outcomes of this AHRQ K08 Award will be the comprehensive understanding of the effectiveness and limitations of antibiogram, and the informatics toolkits to develop Smart Antibiogram. At the end of this K08 Award, the candidate will be well-prepared to become an independent investigator with expertise in AMR and health informatics, with specific strength in AMR prediction model decision-support system.

NIH Research Projects · FY 2024 · 2020-09

PROJECT SUMMARY/ABSTRACT There are no pharmacological treatments for cytotoxic edema, which is a common consequence of multiple brain insults, including hypoxic-ischemic and traumatic brain injury, stroke, metabolic derangements, and seizures. Hypoxic-ischemic encephalopathy (HIE), with an incidence of 1.5 of every 1,000 live births, is a type of brain damage in newborns caused by oxygen deprivation and limited blood flow. HIE is associated with seizures, and both correlate with long-term morbidity, including cerebral palsy, cognitive delay, epilepsy, vision loss, and deafness. HIE and neonatal seizures result in cytotoxic edema, which is characterized by the accumulation of water, chloride (Cl-), and other ions. The mechanisms of water movement that make neurons swell during the neonatal period are unknown. There is a critical need to determine how water moves into neurons that result and perpetuate neuronal swelling during the neonatal period, as there are no direct treatments for cytotoxic edema at this age. Knowing the pathways of water movement in neurons is the first step to develop innovative ways to treat cytotoxic edema, which will prevent neuronal cell death and improve the treatment of neonatal seizures. Neurons do not have water channels to allow the movement of water. Multiple pathways have been described in different cell types, but it is unknown which ones participate during the neonatal period. Our long- term goal is to identify the mechanisms of neuronal swelling in the developing brain and how this swelling results in neuronal death. Our central hypothesis for this proposal is that specific cation-chloride cotransporters (CCCs) move water, along with Cl-, in and out of neurons during cytotoxic edema in the neonatal period. This hypothesis is based on our data demonstrating the linked movement of water and Cl- in neurons during cytotoxic edema. We will test our hypothesis through two specific aims. Aim 1 will determine the pathway of water movement into neurons during swelling in the neonatal period. Aim 2 will determine the paths of water movement out of cortical neurons that prevent progressive swelling during the neonatal period. We will use multiphoton imaging techniques to measure changes in neuronal size and their Cl- concentration during swelling in different transgenic mouse lines expressing both Cl- sensitive and insensitive fluorophores, in vitro, and in vivo, while altering the CCC function either pharmacologically or through genetic manipulation. Also, we will use a novel deep learning algorithm to analyze the changes in neuronal size during swelling. Our studies will uncover fundamental mechanisms on how neurons swell and what mechanisms prevent progressive swelling during early brain development. Our results will have a broad impact as they will open new research avenues on neuronal volume regulation in the newborn and will guide the development of drugs targeting cytotoxic edema, which are currently lacking. Moreover, our results will apply to other severe brain injuries in children that are associated with cytotoxic edema and elevated neuronal Cl- concentration, including trauma, and stroke.

NIH Research Projects · FY 2024 · 2020-09

Project Summary Chlamydia trachomatis (C.t.) is the leading cause of non-congenital blindness and the most prevalent sexually transmitted bacterial infection in the world, which if left untreated can result in severe consequences. All chlamydiae are obligate intracellular bacteria and thus gaining entry into a host cell is essential for the pathogen to complete its replicative cycle and proliferate. Despite the critical nature of the invasion process, the molecular mechanisms and details regarding how C.t. forces its way into non-phagocytic cells remains a large knowledge gap. A major premise of the current model suggests that invasion is facilitated by delivery of prepackaged conventional type III secretion system (cT3SS) effector proteins into the host cell prior to pathogen entry. We hypothesize that a subset of these cT3SS effectors coordinate active subversion of the host cytoskeleton through direct manipulation of key regulators of actin dynamics. New data from our laboratory indicates that the cT3SS effector protein TmeA binds to the nucleation promoting factor N-WASP, both of which we show are essential for invasion. In Aim 1, we will mechanistically determine how TmeA regulates N-WASP and determine whether this interaction promotes key membrane features that likely aid in bacterial entry such as; membrane ruffling, pedestal formation, and filopodial dynamics. Given the essential nature of invasion to bacterial proliferation and survival, C.t. likely employs multiple measures for invasion. In Aim 2, we will evaluate the complex interplay between TarP, TepP, and TmeA and reveal the ultimate molecular effects of their effector function and describe how the seemingly disparate pathways targeted by these effectors converge to assure chlamydial invasion. Furthermore, we will determine whether other cT3SS effectors, only expressed in the invasive EB form of chlamydia, are necessary for invasion of non-phagocytic cells. Detailed characterization of the bacterial and host proteins required to promote reorganization of the actin cytoskeleton during C.t. invasion will provide a holistic view of how intracellular pathogens, such as C.t., coordinate subversion of cytoskeletal regulators to invade host cells.

NIH Research Projects · FY 2024 · 2020-09

Abstract: Duchenne Muscular Dystrophy (DMD) is a severe, progressive muscular disease that affects both muscle and bone. To date, effective therapies for DMD are limited. Studies have shown that reduced nitric oxide (NO) bioavailability resulted from secondary loss of neuronal nitric oxide synthase (nNOS) in the absence of dystrophin is a key contributor to disease progression. Restoring NO homeostasis via dietary nitrite and nitrate representing a novel therapeutic approach due to its ability to be converted to NO in low oxygen and ischemic states that can bypass nNOS. The purpose of this study is to test the efficacy of inorganic nitrite and explore its mechanism of action on both skeletal muscle and bone. Our preliminary data, based on a severe dystrophic mouse model (dKO-dystrophin/utrophin double knock out), demonstrated disrupted NO homeostasis in dystrophic muscle and more excitingly, oral administration of nitrite significantly improved the life span and a series of pathological changes in dystrophic mice, both in skeletal muscle and bone tissues. The mechanisms underlying these improvements deserve further investigation to provide important preclinical and mechanistic information for identifying novel therapeutic targets. We hypothesize that inorganic nitrite administration improves both muscle and bone pathologies in DMD by enhancing NO signaling pathways in dystrophic muscle and by modulating the expression and secretion of bone- regulating myokines. We will test this hypothesis in three specific aims. Aim 1: To test the hypothesis that inorganic nitrite restores nitrate/nitrite pool in dystrophic mice and improves muscle/bone pathologies and preserves muscle function. Aim 2: To test the hypothesis that nitrite affects skeletal muscle via myoglobin-mediated NO-cGMP signaling pathway. Aim 3: To test the hypothesis that in addition to increased mechanical loading, nitrite affects bone homeostasis via modulating the expression and secretion of bone-regulating myokines from dystrophic muscle. We anticipate that these findings will provide a novel, safe and low-cost therapeutic approach benefiting both muscle and bone for the currently untreatable DMD. Completion of these aims will advance our knowledge of novel mechanisms for the pathogenesis of bone abnormalities in DMD through bone-regulating myokines as well; which may uncover new potential therapeutic targets. Importantly, our findings may have profound translational implications not only to DMD but also to other neuromuscular diseases that lack normal NO signaling pathway function.

NIH Research Projects · FY 2024 · 2020-09

ABSTRACT Adverse pregnancy outcomes, including gestational hypertension and preeclampsia, affect 10-20% of pregnant women and are major risk factors for future maternal cardiovascular disease (CVD). Though moderate-to- vigorous intensity physical activity (MVPA) during pregnancy can reduce these risks, less than 25% of pregnant women meet MVPA guidelines. Identification of additional modifiable behaviors that could reduce maternal health risks during pregnancy is needed to inform clinical interventions and practice. Remarkably, little is known about the role of sedentary behavior or sleep in adverse pregnancy outcomes that increase future CVD risk. This is a critical research gap because sedentary behavior and sleep are increasingly recognized as CVD risk factors in the general population, independent of MVPA. Unfortunately, existing studies in pregnancy largely focus on MVPA rather than the emerging paradigm that considers the full 24-hour behavioral cycle (i.e., sedentary behavior, sleep, light-intensity physical activity, and MVPA), lack repeated measures, use suboptimal assessment methods, and fail to measure clinically relevant outcomes. Clarifying how sedentary behavior and sleep influence pregnancy outcomes could inform intervention targets and guidelines to improve cardiovascular health during and after pregnancy. The overall goal of the proposed study is to examine the associations of sedentary behavior (Aim 1) and sleep (Aim 2) with hypertensive disorders of pregnancy and other adverse pregnancy outcomes that increase future CVD risk. Novel statistical methods will be used to determine optimal 24-hour behavioral patterns (Aim 3) during pregnancy for cardiovascular health. For this, 500 women in early pregnancy will be recruited to take part in a multi-site (Universities of Iowa and Pittsburgh) cohort study. Women will wear three state-of-the-art devices for seven days in each trimester of pregnancy to assess sedentary behavior (activPAL micro 3), sleep (ActiWatch Spectrum Plus), and physical activity (ActiGraph GT3X). Hypertensive disorders of pregnancy and other adverse outcomes will be obtained through medical chart abstraction. We hypothesize that women with a high sedentary behavior pattern, short sleep duration pattern, and poor sleep quality pattern across trimesters will be more likely to have hypertensive disorders of pregnancy (primary), gestational diabetes, preterm birth, small-for-gestational age infants, high blood pressure, and poor glycemic control (secondary). Further, we hypothesize that statistically reallocating time spent in sedentary behavior for physical activity (LPA or MVPA), but not sleep (among adequate or high duration sleepers), will be associated with lower incidence of these adverse outcomes. In direct response to the NHLBI Research Priority 3.CC.04, this project will use advanced methods and models to assess and characterize the 24-hour behavioral cycle during pregnancy to better understand differences in health. This project will inform guidelines on optimal 24-hour behavioral patterns in pregnancy, as well as lay the foundation for future studies testing behavioral interventions in pregnancy to achieve optimal outcomes and improve women’s long-term cardiovascular health.

NIH Research Projects · FY 2024 · 2020-09

OVERALL COMPONENT PROJECT SUMMARY Cystic fibrosis (CF) is a common life-shortening genetic disease that causes progressive lung failure due to recurrent infections and airway obstruction. While our knowledge of CFTR function has advanced greatly in the 30 years since the discovery of the gene, treatments for the disease remain suboptimal and CF remains progressive and fatal. Advances with small molecule CFTR modulator therapies have helped restore protein function for many mutations, but approximately 10% of people with CF have not benefited from these strategies, including people with nonsense and splicing mutations. The central theme of this proposal is developing new molecular therapies to prevent or treat CF lung disease. The goal of our three projects and four cores is to exploit the power of our in vitro and animal models to address questions fundamental to lung disease pathogenesis and to use this knowledge to inform new therapeutic strategies to complement CF defects, including gene repair and the addition of a small molecule that forms anion channels. The three closely interrelated Projects will work together to accomplish the following goals: 1) To restore CFTR function using targeted single nucleotide editing. We hypothesize that cells in the surface airway epithelium, including those with progenitor capacity, can be targeted to repair CFTR mutations using base editing. 2) To understand the mechanisms of amphotericin B (AmB)-induced anion secretion in airway epithelia and to test the hypothesis that AmB can restore CF host defenses in vivo. AmB is a small molecule that forms anion channels. 3) To determine how CFTR expression in pulmonary ionocytes and ciliated cells regulates properties of the airway surface liquid that are crucial for clearance and innate immunity. The development of effective gene therapies for cystic fibrosis lung disease must be guided by a clear understanding of pathophysiologic mechanisms of disease and the relevant cellular targets for CFTR gene replacement or editing. The Project Leaders and their teams have outstanding track records of collaborative CF research, and here they sharpen their focus to a common goal. Their highly creative research is supported by four cores that provide innovative infrastructure and services. Through these studies we hope to accelerate the development of new therapeutics for CF lung disease. !

NIH Research Projects · FY 2024 · 2020-09

Project Summary Fibromyalgia (FM) is a complex condition characterized by widespread pain and fatigue that is associated with sleep dysfunction and reduced function that affects 2-4% of the population (Heidari et al., 2017). Current 2016 diagnostic criteria are by symptomology only, as there are no validated chronic pain biomarkers to assist with diagnosis, or treatment evaluation endpoints (Wolfe et al., 2016). Diagnosing FM often takes years with patients seeing multiple physicians, which delays treatment (Choy, 2010). This delayed diagnosis and treatment initiation would be dramatically reduced with the identification of FM biomarkers. The long-term goal of this line of research is to identify unique biomarkers for FM to improve the diagnosis and/or develop therapeutic targets for individuals with widespread pain. Using a semi-targeted metabolomics approach, our preliminary data from women with FM (n=59), compared to healthy controls (n=38), show 18 potential candidates that differ significantly between cohorts with several metabolites showing good-excellent sensitivity (>90%) and specificity (>90%). The primary goal of this proposed research is to assess and validate candidate metabolic biomarkers in a new, larger cohort of individuals and compared to other chronic pain populations. The proposed study will use a multi-site, cross-sectional design to identify and characterize metabolic biomarkers, biosignatures, and their associations with multiple symptomology domains to address the following two specific aims: Aim 1: We will characterize diagnostic test metrics for candidate biomarkers using receiver operating curves (ROCs), i.e. sensitivity and specificity, and test-retest reliability, to correctly identify individuals with FM from healthy controls and other chronic pain conditions: osteoarthritis, carpal tunnel syndrome, and rheumatoid arthritis. Aim 2: We will determine associations between putative metabolic biomarkers and multiple self-reported symptom domains in those with FM: a) pain; b) fatigue; c) sleep; d) physical function; e) psychological factors, and f) disease impact/disability. We have identified several promising metabolic biomarkers that may serve as diagnostic or within-disease phenotype identifiers. Once completed, we will examine potential mechanistic and therapeutic targets for the candidate biomarkers in subsequent studies. These novel studies have the potential to identify a diagnostic, and potentially a therapeutic, biomarker of FM associated with cell metabolism. To accomplish this study, we have developed a strong multidisciplinary and multi-site team, leveraging blood samples and phenotype data collected as part of an on-going funded study, as well as additional data collection for repeatability analyses. The study team has the necessary expertise in human, basic science and metabolomics investigations to successfully complete these aims.

NIH Research Projects · FY 2024 · 2020-08

Project Summary/Abstract Healthcare-associated infections (HAIs) are a major source of mortality and morbidity and affect about two million patients each year. Within hospitals, pathogens like Clostridioides difficile and methicillin-resistant Staphylococ- cus aureus (MRSA) are routinely transmitted to and among hospitalized patients: also of particular concern are multidrug resistant organisms (MDROs), because HAIs caused by these pathogens are increasingly difficult to treat. These infections can be amplified in hospitals, transmitted to other hospitals, long-term or skilled-care facil- ities, and then, eventually, to the community at large. Developing effective interventions to prevent the spread of HAIs remains an important public health goal, and demands some means by which the effectiveness of proposed interventions (or combinations thereof) can be efficiently and inexpensively compared. In fields where experi- ments are not possible, mathematical models and simulations can yield insight into how a system responds to the intervention under study. The overarching theme of this project is to overcome existing barriers for modeling the spread of HAIs. We hypothesize that high-fidelity models derived from complex, fine-grained data can be used to understand the acquisition and transmission of HAIs within and across healthcare facilities. Simulations based on our models can be used to compare alternative interventions and provide effective and practical guidance for how to reduce the transmission of MDROs and other pathogens capable of causing HAIs.

NIH Research Projects · FY 2024 · 2020-08

Gene regulatory evolution is a major driver for phenotypic divergence. While this has been well-studied in development, its importance in non-developmental processes is far less understood. The overall vision of the lab is to fully characterize the evolutionary rewiring of gene regulatory networks (GRNs) for major stress responses in opportunistic yeast pathogens, and elucidate the contribution of such changes to the survival and virulence in the host. The lab previously discovered a significant difference in how a commensal and opportunistic yeast pathogen, Candida glabrata, regulates its Phosphate Starvation (PHO) response compared with its non-commensal relative, Saccharomyces cerevisiae: the commensal has dramatically expanded its PHO response targets both in number and in function, which was attributed to derived changes in its master transcription factor (TF) by being less dependent on the co-TF. The goal of the lab in the next five years is to determine the genetic and mechanistic bases of this novel mode of TF evolution, i.e. acquiring new targets by reducing co-TF dependence, and the effect of such evolution on stress resistance to phosphate starvation as well as combinatorial stresses in the host. Understanding how this model stress response evolved by itself and in its interaction with other stress responses will begin to elucidate the principles for stress response evolution in commensal yeasts in general. To reach this goal, three Directions will be pursued: 1) Elucidate the mechanisms of a novel mode of TF evolution and its impact on the downstream response, using biophysical and fitness assays; 2) Dissect the crosstalk between stress responses and how it evolved in commensal yeasts, by determining the interaction of PHO response with oxidative stress and general stress responses in C. glabrata and S. cerevisiae; 3) Determine how the PHO network evolved in other commensal yeasts and related non- commensals, using transcriptome profiling coupled with genome-wide Chromatin-IP. Research proposed in this application is innovative because the evolutionary approach will identify key changes in the wiring of the stress response GRNs underlying host adaptation. The proposed research is significant because it will both shed light on the general principles for GRN evolution, and will provide a conceptual framework for developing novel antifungal strategies targeting stress responses.

NIH Research Projects · FY 2024 · 2020-08

PROJECT SUMMARY/ABSTRACT Candidate: Aaron Scherer, PhD is a social psychologist who utilizes insights from the psychological sciences to create innovative preventive healthcare interventions, primarily for health risks that affect adults aged ≥50. Dr. Scherer’s long-term career objective is to become an independent investigator leading multidisciplinary research teams in the design and evaluation of innovative, patient-centered interventions to optimize the delivery and utilization of preventive health services for middle-aged and older adults. Research Context: Age-associated changes in immune function and chronic conditions coupled with suboptimal influenza vaccination rates (50%) result in adults aged ≥50 accounting for 95% of the 50,000 influenza-associated deaths that occur every year. Unfortunately, vaccine messaging strategies that health organizations currently utilize to increase vaccine uptake have been ineffective and, in some cases have worsened vaccine attitudes. Motivations to satisfy psychological needs such as managing threats, reducing uncertainty, and achieve social goals may bias how people process vaccine-related information and vaccine outcomes. “Motivational fit”, an alignment with a motive that undermines adult vaccine uptake, may be a more effective mechanism of behavior change to target with vaccine interventions than current approaches. Specific Aims: 1) Identify which motives have the strongest associations with vaccine outcomes for adults aged ≥50; 2) Collaborate with vaccine-hesitant adults aged ≥50 to create influenza vaccine messages that have motivational fit; 3) Conduct a pilot study to evaluate the feasibility of a vaccine messaging efficacy study. Research Plan: To accomplish these aims, Dr. Scherer will use a national, demographically-diverse online sample of adults aged ≥50 to identify four motives that are most strongly associated with vaccine uptake (i.e., largest effect sizes) for adults aged ≥50 and test whether scales measuring these motives need to be vaccine- specific. He will then collaborate with vaccine-hesitant adults aged ≥50 to develop and test influenza vaccine messages targeting each of the four motives. Finally, he will conduct a pilot study with a clinical population to evaluate the feasibility of conducting planned efficacy studies of the motivational fit vaccine messages. Career Development Plan: Dr. Scherer will develop 1) foundational content knowledge in the aging process to engage in aging research, and expertise in 2) psychometrics; 3) patient-centered design; and 4) health services research with adults aged ≥50. Dr. Scherer’s training goals will be supported by close mentorship from an interdisciplinary team; advanced didactic coursework; and other career development opportunities. Environment: The University of Iowa offers an ideal environment for Dr. Scherer to pursue his training; with mentorship from well-established experts, additional guidance from an advisory committee, and a department dedicated to his long-term success in becoming an independent investigator in healthy aging research.

NIH Research Projects · FY 2024 · 2020-08

Project Summary Tissue specific regeneration requires the function of progenitor cells that can differentiate to repair diseased or injured tissues. Affected tissues can contain stem cells and their growth and patterning are controlled by transcription factors and signaling pathways. Stem cells derived from stem cell niches contribute to the regeneration of mature tissue types in many different organs, including the trachea, lungs and teeth, amongst others. These niches are formed in developing embryos, and must be maintained throughout life by symmetric cellular divisions that produce daughter pluripotent stem cells. Another equally important behavior of the cells in a stem cell niche is the production of differentiated daughter cells by asymmetric cell division, which then take the place of damaged cells in regenerative organs, in order to allow the organ to continue to function. However, the roles of specific microRNAs (miRs) in these processes are unclear. miRs have become appreciated as playing a role in stem cell differentiation. The ability to coopt miR expression in order to reprogram and control the differentiation of naive cells into different cell types will be an important tool required to create artificial organs and repair diseased tissues, saving millions of lives and public dollars. We hypothesize that miR-26b is acting in part to regulate the stem cell niche of the lower and upper incisor by regulating Lef-1 expression, and that this mechanism may be common to many tissues that house stem cell niches. Thus, the absence of miR-26b expression prior to E14.5 allows Lef-1 expression in the oral epithelium (placode) and LaCL (stem cell niche) during development. However, after E14.5, miR-26b expression in the dental epithelium restricts Lef-1 expression and then other factors (Sox2) regulate stem cell maintenance. In order to determine how Lef-1 and miR-26b is affecting gene expression during development and cells we will pursue three aims: 1) to use our murine models to understand in vivo stem cell proliferation and differentiation by Lef-1 and miR-26b OE; and use miR-26b OE to rescue the Lef-1 OE phenotype and define the role of miR-26b OE in dental development; 2) validate molecular interactions between miR-26b, Lef- 1 and known factors involved in DESC proliferation and differentiation; 3) isolate the dental epithelial cells from WT, Lef-1 OE, miR-26b OE and rescue mice to identify new genetic pathways using RNA-Seq, ChIP-Seq. and Single cell Seq.

NIH Research Projects · FY 2024 · 2020-08

Enteric infections remain the second leading cause of diarrheal morbidity and mortality globally in children, despite significant improvements in access to latrines and safe water sources in high disease burden countries. Our prior research has demonstrated that the “enteric pathome” - i.e. the communities of viral, bacterial, and protozoan pathogens transmitted by human and animal feces in the environment - ingested by children living in low-income urban neighborhoods of Kenya is taxonomically complex and varies by exposure pathway. Our preliminary data indicates that the risk of multi-pathogen infection is elevated among those 6- month old Kenyan infants fed cow’s milk and living in urban households with dirt floors, shared latrines, and cohabitating domestic animals. This suggests multiple aspects of societal development beyond latrines and water sources are required to reduce complexity in pathogen transmission in high burden countries. We hypothesize that joint modeling of enteric pathome agents across households and neighborhoods that represent contrasts in urban societal development will show that development leads to pathome evolution from complex to simple community structures, and thus lower detection frequencies for individual pathogen taxa. Understanding the evolution in pathome complexity induced by societal development and subsequently identifying effective intervention strategies is challenging since field experiments are expensive to implement, difficult to generalize to other settings, are only informed by existing, limited observational data, and sufficiently sophisticated statistical and mathematical pathome modeling tools are not currently available. Our proposal aims to (1) develop spatiotemporal and trajectory statistical models to understand the complex exposure risks for infants from the enteric pathome; (2) collect environmental, behavioral, spatial, economic, and microbial data to characterize the enteric pathome along pathways for disease diffusion and the intersection of humans and animals with these pathways; and (3) develop and validate agent-based models (ABMs) for predicting which social and environmental urban developmental interventions (e.g. animal penning, building latrines or drains, concrete floors) best prevent multipathogen transmission to infants in high disease burden countries using established Kenyan study sites as a model. Our interdisciplinary team includes a biostatistician, infectious disease epidemiologists, microbiologist, computational scientist, behavioral researcher, and urban development geographer.

NIH Research Projects · FY 2026 · 2020-07

Abstract Hearing impairment is common and, in severe cases, results in significant communication, cognitive, and social challenges; cochlear implants (CIs) effectively restore hearing and mitigate many of these deleterious consequences. However, an chronic inflammatory/immune response, universally observed following CI that includes a foreign body response (FBR) to the electrode array biomaterials, has been linked with diminished performance. Results from our current funding period led to major advancements in understanding this inflammatory FBR, to effective therapies that mitigate this response, and, most recently, to revolutionary discoveries of a functional lymphatic network and adaptive immune responses in the cochlea; these are the focus of the new funding period. We now have direct evidence for a robust lymphatic network in the cochlea that proliferates in response to CI and interacts with multiple cellular components of an adaptive immune response. These findings lead us to propose a model for the development of functional, ‘ectopic lymphoid structure’ in the scala tympani surrounding an electrode array. The overall goal of the proposed studies is to investigate the response of this newly described lymphatic system, including proliferation and immune cell trafficking, to CIs and to evaluate the systemic immune response to CIs in healthy and noise-damaged cochleae and in the contralateral cochlea. Aim 1 will characterize the spatial and temporal response of cochlear lymphatics to implantation in mouse models and human cadaveric specimens and determine the role of lymphangiogenesis in post-CI inflammatory responses. Aim 2 explores the effects of common cochlear injuries, including noise damage and hair cell ablation, on cochlear lymphatic and inflammatory responses and the extent to which these injuries prior to CI exacerbate inflammation, fibrosis, and hearing loss following CI. Finally, Aim 3 seeks to determine the contribution of cochlear inflammation following CI to the response of the contralateral ear to CI. Results from this highly novel conceptual work will provide evidence of functional lymphatic systems in the cochlea, identify therapeutic targets and windows for CI, and exert profound impact across multiple fields including cochlear immunology and homeostasis, responses to injury and disease, gene therapy, and the FBR to biomaterials.

NIH Research Projects · FY 2024 · 2020-07

PROJECT SUMMARY / ABSTRACT Parkinson's disease (PD) is an increasingly prevalent progressive neurodegenerative disorder that leads to disabling motor, speech, and cognitive symptoms that reduce quality of life for patients and their families. As the prevalence of PD increases, so too does the use of deep brain stimulation (DBS) as a surgical treatment option when medications fail to control motor fluctuations or side effects from medications limit their efficacy. As many as 15% of the more than 1 million American PD patients are felt to be candidates for DBS. The subthalamic nucleus (STN) is the most common target utilized for DBS and limb motor outcomes after STN-DBS show reliable and consistent improvement in properly selected patients. On the other hand speech motor outcomes after STN-DBS are unpredictable and more patients experience deterioration of speech than those that notice improved speech. An estimated 40% of STN-DBS patients notice worsened speech and with declines significant enough to reduce quality of life. Furthermore, few effective treatments exist to provide sustained improvement in speech performance for those with impaired speech. In order to optimize and develop better treatments for those living with PD, it is necessary to better understand the role of STN in speech as contemporary mechanistic models of human speech production omit STN. Here we study 80 PD patients undergoing bilateral STN-DBS along with 40 non-surgical PD control subjects to address existing critical knowledge gaps. We will document speech performance of both groups through functional communication ability (i.e. objective measures of intelligibility and a validated survey tool for quantifying communication participation) and instrumental speech (i.e. acoustic) measures pre-surgery, 6- and 12-months post-surgery with all subjects off PD medications and with the STN-DBS subjects evaluated with stimulation ON & OFF at both post-operative timepoints. Direct neuronal recordings will be obtained during DBS implantation surgery during both speech and limb motor tasks to define speech-specific STN neurophysiology. In addition, we will perform regression analyses using baseline patient factors and intraoperative physiology findings to identify factors leading to the speech outcomes we will document, and test the feasibility of low frequency DBS in reversing DBS- induced speech declines. To our knowledge, this data will be the first of its kind to evaluate patients before, during, and after surgery with the goals of both directly defining the role of STN in speech and predicting speech outcome after STN-DBS. Such knowledge will provide mechanistic insights that cannot be obtained using other techniques and will guide development of new treatments of impaired speech in PD.