Univ Of North Carolina Chapel Hill

NIH Research Projects · FY 2025 · 2021-09

Project Summary/Abstract Increased access to cesarean sections (c-sections) has contributed to the decline of maternal mortality in sub- Saharan Africa (SSA); however, as the rate of c-sections has increased, so has the rate of c-section related complications. While women who deliver vaginally in rural SSA often receive postpartum follow-up care in their homes from community health workers (CHWs), most programs require that women who deliver via c-section return to health centers or hospitals for follow-up because of the increased complexity of their care. Facility- based follow-up can be financially catastrophic and physically burdensome for mothers, leading to delays in care and increased risk for morbidity. The overall goal of this proposal is to develop a safe mobile health (mHealth) tool to support CHW-led home- based follow-up for women delivering via c-section in rural Rwanda at postoperative days (PODs) 5 and 10. In the first R21 phase, we will develop a software library for an existing photo-based surgical site infection (SSI) diagnostic algorithm to run locally on a smartphone (without internet or cell network connection) (Aim 1a). We will work with Insightiv Technologies to develop the comprehensive mHealth-CHW tool, including incorporating the SSI diagnostic algorithm software library and integrating three CHW usability assessments into the design phase (Aim 1b). Finally, we will test the usability and acceptability of the new tool in a group of 30 CHWs (Aim 2). When we achieve 80% usability and acceptability, we will continue to the second phase. The R33 phase of the grant will study the validity of the mHealth-CHW tool for c-section follow-up (Aim 3) by prospectively following 450 women delivering via c-section and implementing the mHealth-CHW tool and follow- up in their homes at PODs 5 and 10. The women will then return to the hospital at POD 16 for a physical examination and we will compare diagnoses and complications identified via the mHealth-CHW tool to those from the physical exam. We will then evaluate the time-to-diagnosis for c-section complications by randomizing 1350 women to follow-up by the mHealth-CHW tool versus standard of care (Aim 4). Finally, we will assess the acceptability of mHealth-CHW follow-up in 40 women who have delivered via c-section through focus group discussions (Aim 5). We will also conduct two research training series, one in quantitative methods and one in qualitative methods, to strengthen our team's ability to lead mHealth research in the future.

NIH Research Projects · FY 2026 · 2021-09

Migraine affects one in three women of childbearing age in the United States (U.S.). Women with migraine have up to a five-fold increased risk for comorbid depression. The postpartum period is an especially challenging time for women with migraine, as rapid hormonal shifts, fatigue, exhaustion, and increased external stressors are known triggers for both migraine and postpartum depressive symptoms. If women’s migraine self-management style (MSMS) primarily involves disengagement (i.e. social withdrawal, avoidance, wishful thinking), the implications for developing depressive symptoms and synergistically worsening symptoms across conditions may be more significant. These risks may be exponentially more harmful to women who have lower levels of personal resources (PR) that have been shown to facilitate effective chronic disease self-management in other non-migraine disease states, as they experience the greatest burden from migraine and postpartum depressive symptoms. A syndemic is characterized by the (1) higher prevalence of these co-occurring disorders, and (2) synergism that exists across symptoms and self-management strategies of one condition (migraine) that facilitates developing a comorbid condition (depression) are influenced by (3) social contextual factors (social and/or economic disadvantage in the face of postpartum demands). Despite the high prevalence of these disorders individually, the increased risk for comorbid migraine-depressive symptoms in the postpartum period, and the potential impact on women and their families, there is a paucity of research in this area. The proposed study will use a syndemics framework to understand the relationships, mechanisms, and interactions among migraine and depressive symptoms, disengagement, and level of PR over the first 3 months postpartum. This will be accomplished by the following: Aim 1: Examine the relationships among (a) migraine symptom severity, (b) depressive symptom severity, (c) level of disengagement as a migraine self-management style, and (d) level of personal resources, over time. Aim 1a: Test the extent to which disengagement mediates the relationship between migraine symptom severity and depressive symptom severity. Aim 1b: Determine whether the level of personal resources moderates the extent to which disengagement mediates the migraine-depressive symptom relationship. Aim 2: Compare maternal quality of life among women who develop the syndemic factor (comorbid migraine and moderate-to-severe postpartum depressive symptoms) and women who do not. The applicant has outlined a comprehensive training plan to be completed under this award. Outcomes of the study and training plan will provide the applicant with skills and data to pursue a career development award on prevention-oriented interventions targeting syndemic conditions that may be useful in disrupting the development of postpartum depressive symptomatology among women with migraine.

NIH Research Projects · FY 2024 · 2021-09

Abstract Allogeneic hematopoietic stem cell transplantation (allo-HSCT) is the preferred approach for the treatment of patients with malignant diseases of the bone marrow (BM) and congenital BM failure syndromes. The efficacy of allo-HSCT is dependent on the anti-tumor activity of the conditioning therapy and the donor graft. The efficacy of allo-HSCT is dependent on the ability of donor bone marrow/stem cells to replace the host immune system. However, it is clear that not all immune cells are generated from adult bone marrow cells. B1b lymphocytes and Langerhans cells are generated from fetal cells and not reconstituted by donor bone marrow. Additionally, our group and others have shown that innate lymphoid cells (ILC), which generate cytokines similar to T lymphocytes but don’t express germ-line encoded receptors, are not completely reconstituted after stem cell transplantation. The mechanism for the absence of ILC2 cells in the GI tract post allo-HSCT is not clear as these cells routinely are found in the bloodstream of patients and are reconstituted after autologous stem cell transplantation. Our group has found in mice that fetal liver derived ILC2 precursor cells can reconstitute the GI tract indicating that the niche can support ILC2 cells. This has led us to hypothesize that the inflammatory response found after allo-HSCT from the GvH response limits the development of ILC2 cells. ILC2 cells have significant plasticity, mediated by epigenetic changes in critical lineage-specific loci that leads to the development of ex-ILC2 cells, which are functionally similar to ILC1 or ILC3 cells. Our group has found that ILC1 cells, which generate IFN-γ, exacerbate GI tract GvHD. Furthermore, the pro-inflammatory cytokines, IL-12 and IL-1β, which are increased post allo-HSCT especially in the presence of the GvH response, convert ILC2 to ILC1 cells. Our group has found in ILC2 cells that H3K9me1/2 marks, which maintain gene transcription of gata3, cmaf and rora, critical for ILC2 maintenance and function, are enhanced by inhibiting lysine specific demethylases. The first specific aim focuses on enhancing the function of ILC2 cells in vivo to treat GvHD by inhibiting Jumonji containing and lysine-specific demethylases. Bronchiolitis obliterans syndrome (BOS) is a significant complication of allo-HSCT. T cells that generate IL- 17A, Th17 cells, are critical to the pathogenesis of BOS. One risk factor for BOS is an antecedent viral infection of the lung, although the mechanism for this is not clear. Quite recently our group has found that viral infection in the lung converts lung ILC2 cells to ILC3-like cells that generate IL-23 critical for the expansion of Th17 cells. The second specific aim of this proposal focuses on the use of demethylases given with ILC2 cells to prevent and/or treat post-viral BOS. Completion of the goals of this project will greatly enhance our understanding of the mechanisms important for the loss of ILC2 cells post-transplant, and provide new approaches to utilize ILCs to treat GvHD.

NIH Research Projects · FY 2025 · 2021-09

Health-related social needs, particularly food insecurity, housing instability, and transportation barriers, are associated with poor outcomes for people with type 2 diabetes mellitus (T2DM). In particular, these factors are associated with worse glycemic, blood pressure, and LDL cholesterol control, which significantly increases the risk of macrovascular and microvascular complications. Thus, there is a growing call for interventions to address these needs and improve T2DM outcomes. However little research has assessed whether changes in health-related social needs are associated with changes in T2DM outcomes. This knowledge gap hampers efforts to develop effective interventions. Addressing it will provide much- needed evidence on which patients to screen for which social needs, and on which interventions targeting social needs are most likely to improve T2DM outcomes. The pathways linking social needs and T2DM outcomes are likely characterized by interactions between individual- (e.g., needs, age, sex), clinic- (e.g., clinic characteristics; specifics of the intervention), and area-level factors (e.g., local economic conditions; community resources). Given this complexity, a multi-pronged approach is needed. We will combine 3 methods of investigation: 1) longitudinal, multi-level, regression analysis; 2) innovative machine learning methods to detect novel combinations of factors associated with heterogeneous response to health- related social needs and health-related social needs interventions while avoiding spurious findings; and 3) thoughtful qualitative investigation. Such an analysis has never before been possible, because the needed data elements have not been united. This proposal seeks to answer whether improvements in specific health-related social needs are associated with improvements in specific clinical outcomes, in what circumstances, and which approaches to addressing health-related social needs, if any, best improve outcomes. We will leverage what we believe to be the nation’s largest dataset of patient-reported health-related social needs, clinical outcomes, and community- and clinic-level data. We will examine whether changes in health-related social needs are associated with changes in hemoglobin A1c, systolic and diastolic blood pressure, and LDL cholesterol. Further, we will evaluate whether clinic-based interventions seem to improve these outcomes, and if there are important variations in these interventions that are associated with different responses to the intervention. The proposed work will yield important, previously unavailable evidence on how to refine and improve health-related social need interventions. Specifically, it will help us understand better how better to care for a population at high risk for T2DM complications. Overall, this project will substantially advance NIDDK’s mission to improve care for individuals with T2DM.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY In this study we seek to understand how genetic factors influence the risk of developing obsessive-compulsive disorder (OCD) in Latin American individuals. OCD and related disorders are of major public health importance owing to their profound personal and societal costs. Little is known for certain about their etiology, and treatment, detection and prevention strategies are not optimal or directed by knowledge of pathophysiology. In other psychiatric disorders (e.g., schizophrenia, bipolar disorder, and autism), genomics has begun to deliver fundamental knowledge about genetic architecture, identify specific loci for biological follow-up and localize pathways altered in disease. We intend to realize these same advances for OCD by markedly increasing and diversifying the worldwide sample size for genomic analysis, in a first step toward elucidating the fundamental biology of this condition. Three overlapping areas will be investigated in this project. First, we will collect the world’s largest ancestrally- diverse sample of OCD cases (N = 5,000 individuals from Latin America). To do this in an efficient and cost- effective manner, we will take advantage of a network of OCD clinics we have established across Latin America, in addition to clinics in the USA and web-based recruitment. The phenotypic data collected will include a detailed clinical characterization including comorbidities and OCD symptom dimensions. Second, we will genotype all 5,000 samples on the Illumina Global Screening Array (genotypes for >10,000 matched controls will be available). This will allow us to, in collaboration with the Psychiatric Genomics Consortium, discover genomic loci harboring common variation associated with OCD. Third, we will fine-map genome-wide significant loci and calculate individual polygenic risk scores (PRS) as a measure of genetic liability to OCD. We expect the new inclusion of ancestrally diverse samples to improve our fine-mapping ability, to yield more accurate PRS in non-European samples and ultimately to reduce health disparities when OCD genomic findings are used clinically. Overall, this study will improve our understanding of the causal mechanisms implicated in OCD, with a view towards improving clinical outcomes and reducing chronicity and societal costs.

NIH Research Projects · FY 2025 · 2021-09

Project Summary The ability of therapeutic drugs to access specific organs strongly depends on the nature of the blood-tissue barrier at said organs. The prostate and brain, for example, have tight barriers with no intercellular gaps, and most drugs cannot permeate them at sufficiently high levels to be therapeutically effective. Thus, understanding the relationships between drug chemical structure, dosing regimen, and organ penetration is crucial to the development of new and effective drug therapies. Motivated by this, the long-term goal of this program is to establish a measurement standard of molecular transport parameters affecting the passage of therapeutic agents across biological barriers in vivo. Specifically, the objective of this proposal is to demonstrate that electrochemical, aptamer-based (E-AB) sensors – an emerging sensing platform with the ability to continuously measure the levels of specific molecules in the body – can support continuous monitoring of molecular transport from blood to liver, prostate and brain. The proposed measurements will determine the transport parameters of seven therapeutic agents (three aminoglycoside, three β-lactam and one glycopeptide antibiotics) across four biological barriers (i.e., blood-liver, blood-prostate, blood-brain and blood-cerebrospinal fluid). These drugs were chosen because, although effective at treating infections across organ barriers, they cause dangerous side effects driven by their narrow therapeutic window, making their precise dosing an important medical challenge. The central hypothesis of this work is that achieving spatially and temporally resolved drug measurements in blood and target organs will produce unprecedented permeability data that will guide new therapeutic drug development toward the creation of permeability-enhanced therapeutics and more effective dosing regimens. This hypothesis will be tested by pursuing three specific aims: 1) Determine the kinetics of drug uptake in the liver via continuous, seconds-resolved E-AB measurements; 2) Determine the transport kinetics of antibiotics through the prostatic barrier; and 3) Determine the transport kinetics of prophylaxis antibiotics delivered from blood to the brain. The proposed research is significant because it will define the structural and transport characteristics necessary for therapeutic agents to penetrate targeted organs and propel the study of other therapeutics beyond the families of antibiotics considered here. Thus, this work will develop foundational knowledge and generate the necessary resources for other researchers and industries – working on drug development, in-vivo testing and clinical dose scaling – to advance the field of therapeutics. The proposed research will have an immediate positive impact as it will establish a better understanding of therapeutic drug transport within compartments in the body. Longer term, this work will have established the groundwork necessary for the in-vivo evaluation of molecular transport across tight biological barriers.

NIH Research Projects · FY 2025 · 2021-09

Abstract Integrative analysis methods are in great needs as multimodal multi-cohort neuroimaging data rapidly emerge in neuro science. In Alzheimer's Disease (AD) studies, many research relies on multimodal neuroimaging data to identify key image biomarkers for the early diagnosis of AD. Despite great endeavors in data collection, there still lacks rigorous statistical methods and efficient computational tools to properly integrate big neuroimaging data in a statistical model and carry out inference to address practical problems. Important problems such as missing data and adjustment for between-subject heterogeneity still remain unsolved. In this proposal, we propose to build two integrative models, one handles multimodal data and the other handles longitudinal multi-cohort data. They will be built under a generic M-estimation framework that covers many widely used statistical models as its special cases. We will provide various inference tools for these models and develop efficient algorithms to solve the M-estimation problem in presence of block missing values. In Aim 1, we propose a factor-adjusted integrative model for multimodal data and provide a complete set of inference tools. These tools can test the significance of one whole data modality as well as the significance of multiple linear combinations of predictors from one or more modalities. In Aim 2, we provide a powerful computational tool to handle block missing values of multimodal data. Such a tool does not need to perform ad-hoc imputation on missing values, but rather relies on an innovative mini- batch gradient descent algorithm to yield a good estimator. In Aim 3, we will develop an interactive factor model to jointly model longitudinal data coming from multiple cohorts. We show that such a model includes the standard random effects model as a special case and is more flexible modeling the longitudinal data and accounting for the between-subject heterogeneity. The proposed research will likely transform how we analyze neuroimaging data and enhance our understanding of Alzheimer's Disease and its relation to public health.

NIH Research Projects · FY 2025 · 2021-09

Abstract Black women have the highest rates of obesity in the United States and are at heightened risk for type 2 diabetes mellitus. While achieving clinically significant weight loss of 8-10% may reduce risk, Black women have disparate behavioral weight loss treatment outcomes. Untreated binge eating may be a contributing factor. Indeed, nearly 30% of Black women with obesity report binge eating behaviors; those who binge eat are likely to regain weight at a faster rate, drop out of behavioral weight loss interventions (SBWL), and have poorer health outcomes. Black women, however, are far less likely to engage in mental health treatment for binge eating. Furthermore, treatment for binge eating is often not available in primary care and community- based settingsplaces where Black women are more likely to receive treatment for their eating and weight- related concerns. Given the barriers Black women face in accessing in-person treatment for binge eating, digital treatment platforms may provide the opportunity to construct a culturally-relevant and accessible treatment option. Current intervention research to treat binge eating among Black women is scarce. To fill this gap, I desire to become a leading health disparities independent investigator with a research program focused on the development, implementation, and evaluation of interventions to prevent and treat binge eating and obesity, but require additional training. The NIH Mentored Patient-Oriented Research Career Development Award will provide protected time to seek this training and further my skills and ability to implement the proposed research. My short-term training goals are to develop advanced expertise in: (1) the design and implementation of clinical trials for obesity; (2) using digital health tools in clinical trials for binge eating and obesity; (3) implementation science theory and methods; and (4) development and submission of independent, investigator-initiated research grants. I have developed a strong training plan and mentoring team, led by Carmen Samuel-Hodge, PhD (co-primary mentor), Cynthia M. Bulik, PhD (co-primary mentor), Dori Steinberg, PhD (co-mentor), and a team of expert consultants. RESEARCH STRATEGY: The objective of this study is to modify a validated intervention to develop a digital health tool to treat binge eating and prevent weight gain in Black women, and examine the feasibility and preliminary efficacy of this tool in a pragmatic clinical trial. I will engage Black women who binge eat (BMI > 30 kg/m2) to identify barriers and facilitators to detecting and treating binge eating, adapt a validated eating behavior and nutrition intervention guided by stakeholder input, and use Semblie, a UNC-hosted, no-cost online platform that provides tools to build, deliver, and track digital health interventions to construct the tool. This training and research plan will form the basis for a future multicenter clinical trial testing the efficacy of the digital health tool for improving binge eating and weight outcomes among adults who binge eat, to be proposed in an R01 grant application before the end of the K23 award.

NIH Research Projects · FY 2024 · 2021-09

SUMMARY Severe Acute Respiratory Syndrome Coronavirus-2 (SARS-CoV-2; CoV2) is the first highly pathogenic and highly transmissible human coronavirus that is the causative agent for the worldwide COVID-19 pandemic. As of November 2020, 50 million cases of CoV2 infection worldwide and 1.25 million deaths have been reported. The U.S. accounts for the majority of cases, 9.7 million (20%) and deaths 235,000 (19%), and COVID-19 is expected to add an $8 trillion burden to the U.S. health care system. A particularly challenging aspect of clinical management is the variable patient response to CoV2 infection. Some infected individuals report few symptoms whereas others display severe disease characterized by hypoxia, acute respiratory distress syndrome, and multi-organ involvement that can lead to death. A pro-inflammatory ‘cytokine storm’ in COVID- 19 patients promotes derangements in vascular function and blood composition. Elevated fibrinogen and D- dimer (the breakdown product of fibrin clots) track with significant elevations inflammatory markers (e.g., IL-6, C-reactive protein), which significantly and positively correlate with poor patient outcomes. Autopsy studies of COVID-19 patients have revealed intravascular and extravascular fibrin deposits in lung tissue and other organ systems. A current critical knowledge gap is the molecular basis of how persistent fibrin deposits develop and whether they are functionally linked to the pathophysiology of severe COVID-19 disease. Our central hypothesis that an insufficiency in the plasminogen activation (PA) system is a trigger point for transition of COVID-19 from mild to severe disease due accumulating, proinflammatory, and tissue-damaging fibrin deposits within the lung and other organ systems. To test this hypothesis, our research team developed a mouse-adapted CoV2 virus that replicates key immunological and hematological aspects of COVID-19 in humans. This unique tool will be used in conjunction with mice carrying single or combined deficiencies or functional mutations in fibrinogen or PA system components to define the natural course of hemostatic changes following infection and elucidate functional contributions of coagulation and fibrinolytic factors to the host response. Specifically, we will determine (i) the differences in local and systemic activity of host factors that control fibrin(ogen) deposition, stabilization, and dissolution following mild vs. severe CoV2 infection; (ii) how PA deficiency promotes severe disease following CoV2 infection characterized by exacerbation of local and systemic inflammatory, organ damage, and host mortality; and (iii) the mechanisms linking fibrin(ogen) to exacerbation of host inflammatory responses and induction of severe disease following CoV2 infection. The proposed studies will provide novel insights into the contribution of the plasminogen/fibrinogen axis to the CoV2 pathobiology, illuminate key mechanisms coupling deficiencies in PA system components to CoV2- mediated thrombophilia, tissue damage, and loss of organ function, and provide essential proof-of-principle data to facilitate translation of findings into new treatments for COVID-19.

NIH Research Projects · FY 2025 · 2021-09

Project Summary/Abstract Lung cancer 5-year survival rates drop from 61% for early stage diagnosis to just 6% for late stage diagnosis. Currently, fewer than 1 in 5 cases are diagnosed at an early stage. The increasing frequency of chest CT scans and changes in lung cancer screening guidelines are expected to increase the number of incidentally discovered lung lesions, representing an opportunity for earlier lung cancer diagnosis. Bronchoscopy is currently the safest, least invasive, and least expensive diagnostic option, but its poor diagnostic yield greatly limits its procedural benefit. Even when advanced techniques like radial endobronchial ultrasound and electromagnetic navigation are used, the diagnostic yield is just 50-60%. This is primarily due to challenges with intraoperative localization of the bronchoscope prior to needle deployment. Additionally, access to these techniques is limited because they require expensive equipment and unique expertise. Efforts relying on the bronchoscope's built-in camera require no additional equipment or specialization, but have struggled with generalizability across individuals in part due to limited data availability and assumptions about airway features. The objective of this proposal is to improve the success rate of traditional bronchoscopes by addressing limita- tions in intraoperative localization using a data-driven model that is robust to differences in human anatomy. This work has potential for significant public health benefit by (1) increasing early lung cancer detection, (2) reducing morbidity and mortality by reducing the number of invasive procedures, and (3) making minimally invasive bron- choscopy more accessible in areas without expert bronchoscopists. The proposed work will be accomplished via two Specific Aims. In Aim 1, a dataset will be generated of virtual and real bronchoscopy videos with video-frame matched six degrees-of-freedom poses (position and orientation in three-dimensions) of the bronchoscope's dis- tal tip. This data will be made publicly available as the first large dataset of its kind to promote future research and reproducibility. In Aim 2, a real-time bronchoscope localization model will be developed using advances in ma- chine learning, including deep neural networks, that have shown success in camera localization for non-medical applications. These models will regress the pose of the bronchoscope's distal tip using current and past video frames of the bronchoscope's built-in camera. The clinical utility of the system will be evaluated in simulation, 3D printed lung phantoms, and ex-vivo porcine lung experiments. The research, tightly coupled clinical experience, and associated training plan will provide a unique interdisciplinary skill-set in computer science, medical robotics, and procedural medicine. The outstanding research and clinical environment for this training at the University of North Carolina at Chapel Hill ensures exceptional preparation for a career conducting cutting-edge research as a physician-scientist in medical robotics.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY Dr. Heather McCauley is mentored by Dr. James Wells at Cincinnati Children’s Hospital Medical Center (CCHMC), the largest pediatric research institution in the nation. CCHMC ranked 2nd in NIH support in 2017 and is home to an NIDDK-sponsored Digestive Diseases Research Center with a vibrant community of physicians and scientists. CCHMC provides a very supportive training environment for young investigators. Dr. McCauley proposes an innovative cross-disciplinary research plan which bridges developmental biology, stem cell and organoid medicine, gastroenterology and nutrition, endocrinology, and metabolism. Enteroendocrine cells (EECs) are specialized intestinal epithelial cells which secrete more than 20 bioactive peptides to regulate satiety, gut motility, glucose homeostasis, nutrient absorption, and whole-body metabolism in response to ingested nutrients. While the systemic targets of EEC hormones are well known, such as the brain and the pancreas, the role of EEC peptides in regulating the function of the intestine itself is surprisingly understudied. Our lab has developed a unique, high-throughput human model system to test the roles of individual EEC peptides on intestinal function by generating EEC-deficient human intestinal organoids from pluripotent stem cells. We recently used this model system to show that some EECs regulate ion-coupled nutrient absorption in neighboring cells. Because EECs are nutrient-sensing cells, we considered that they might coordinate other intestinal responses to nutrient availability. Most EEC peptides signal via G-protein coupled receptors which activate second messengers to modulate the function of their target cell. These second messengers are intimately linked to ancient regulators of cellular metabolism, such as mammalian target of rapamycin (mTOR), which are known to play essential roles in many intestinal functions. This proposal will test the hypothesis that EECs regulate cellular metabolism in the stem cell niche (Aim 1) and in the differentiated enterocyte (Aim 2), thus impacting both intestinal homeostasis and efficiency of nutrient absorption. As EECs are now common targets for the treatment of type 2 diabetes, understanding how EECs affect intestinal metabolism and function will inform how dietary or pharmacological manipulation of EECs will impact gut health, nutrient absorption, and whole-body metabolism. To ensure success of the proposed research strategy, Dr. McCauley requires additional mentored time to become fluent in metabolic and metabolomic assays. Dr. McCauley has obtained a co-mentor, Dr. Kenneth Setchell, an international expert in using Mass Spectrometry to understand nutrition- and digestive disease- related changes in metabolism. Dr. McCauley proposes a rigorous yet feasible schedule of formal and informal training opportunities in metabolomics. At the conclusion of the mentored portion of her training, Dr. McCauley will be established as a fully independent, leading young investigator in the field of gastrointestinal metabolism, a critically understudied niche in the GI community.

NIH Research Projects · FY 2025 · 2021-09

The BRAIN Viral Vector Service and Distribution Core will serve as a resource for the maintenance, propagation, and distribution of viral vectors that are used by neuroscientists for neural circuit identification and manipulation, as well as for preclinical translational studies. High-quality viral vectors are essential tools to mark and manipulate neuronal circuits in live animals and identify long- range synaptic connections. Moreover, adeno-associated virus (AAV) vectors are the de facto standard for gene therapies. Existing commercial viral vector sources do not offer end-to-end services, from assisting with viral vector design, rigorous manufacturing and quality controls, troubleshooting with clients to enhance vector performance in the brain, and a mission focused on neuroscience. The Core Director (Dr. Kim Ritola) will translate her ten years of providing viral tools to the Howard Hughes Medical Institute neuroscience community towards creating a BRAIN Viral Vector Service and Distribution Core capable of producing and distributing AAV, rabies, and lenti virus tools to neuroscientists. The Core will have two main purposes: 1) Produce and disseminate affordable, high-quality AAV, rabies, and lenti viral vectors to the neuroscience community including newly developed vectors, custom preps and curated stocks and 2) Collaborate with BRAIN researchers to optimize vector design and prep conditions for optimal performance and serve as an educational resource for viral vector production. This Core will provide investigators with an extensive choice of vector capsid or envelope, paired with custom viral construct, including those with traditionally difficult to package genomes, titers and volumes appropriate for experiment type, rigorous quality controls, and affordability.

NIH Research Projects · FY 2025 · 2021-09

ABSTRACT Thousands of genetic loci are associated with human traits or disease risk, and these loci each typically contain tens to hundreds of variants, most of which are non-coding and lack direct evidence of effects on genes. Experimental tests of genomic variants are needed to identify functional effects, which can be specific to one sex, tissue, and/or perturbed environmental context. Testing effects of risk variants on gene regulation requires an ability to quantify the potentially modest consequences of thousands of alleles in a carefully controlled study. Our overarching goal is to systematically characterize the impact of human genetic variation on gene regulation via massively parallel reporter assays (MPRA). We will select variants based primarily on genome-wide association studies (GWAS) for common diseases and complex traits relevant to the brain, liver, lung, muscle, and/or heart. We will examine all plausible functional candidates at prioritized GWAS loci to provide data for tests of regulatory variant prediction algorithms, positive control variants, and variants prioritized based on regulatory element annotations. The gene regulatory effect of ~500,000 variant alleles will be interrogated in five organs (brain, liver, lung, muscle and heart) using systemic circulation of adeno-associated viral (AAV) MPRA libraries. We will repeat this experiment in a perturbed inflammatory state to evaluate gene-environment interactions. As a result, we will comprehensively characterize variant effects on regulatory function by analyses of variants in the physiological conditions of multiple tissues, in both sexes, with and without perturbation typical of disease environments. Selected variants will be edited into human pluripotent cells for validation. As members of the Impact of Genomic Variation on Function (IGVF) Consortium, we will generate a regulatory variant catalog for the community, and enable future studies through data collection and predictive models. Successful completion of these aims will provide ~10 million allelic effect data points that encompass tissue-, sex-, and perturbation- specific regulatory effects. We will work with the IGVF consortium to finalize selection of variants, organs, and perturbations to generate a comprehensive catalog. The expertise of the study investigators in GWAS, statistical and computational genetics, human genomics, AAV delivery, and mouse physiology make achievement of these aims feasible and likely highly informative to understand how genomic variation impacts human health and disease.

NIH Research Projects · FY 2024 · 2021-09

Project Summary Enhancers play a vital role in initiating and maintaining expression of cell specific transcriptional programs. A key feature in enhancer activation and gene regulation is changing the local chromatin state to facilitate binding of transcriptional co-regulators to enhancers. Chromatin-modifying enzymes that are associated with enhancer function are commonly mutated in human craniofacial disorders including histone methylases, demethylases, acetylases, chromatin readers, and chromatin remodeling proteins. Many of these chromatin-modifying factors biochemically associate and may co-regulate enhancer and transcriptional activation events within neural crest, stem cells from which anterior facial bone and cartilage are derived. We have modeled several of these human craniofacial disorders through tissue specific knockout of chromatin-modifying factors in mouse neural crest cells. The experiments outlined in this proposal will characterize de novo enhancer activation by chromatin profiling of primary cranial neural crest cells undergoing osteoblast differentiation. Neural crest cells deficient in chromatin-modifying machinery will be analyzed molecularly for disruption of enhancer chromatin states and to identify cooperative functions between these factors. This proposal will utilize a combination of biochemical, genetic, and low cell number genomic approaches to elucidate the role of chromatin-based enhancer activation in craniofacial disorder pathogenesis.

NIH Research Projects · FY 2025 · 2021-08

ABSTRACT Chronic kidney disease of uncertain etiology (CKDu) has reached epidemic proportions in Costa Rica’s Guanacaste province (Chorotega region), where it has become a leading cause of premature death. CKDu has a high burden among young, male agricultural workers, but women and children are also affected, and its causes remain largely unknown. In response to the RFA for Field Epidemiology Sites for the Chronic Kidney Diseases of UnceRtain Etiology in Agricultural Communities Research Consortium (CURE Consortium), we propose to recruit, enroll and follow 400 participants with evidence of CKDu and 400 control participants for comprehensive assessments of individual risk factors and contextual exposures for CKDu. We propose an Epidemiology Field Center in Liberia, Chorotega region, as part of a collaboration among the University of North Carolina at Chapel Hill, the University of Costa Rica and local nephrologists in the Chorotega region. Our study design includes a large case-control study with nested substudies and state-of-the art exposure assessment methods to identify risk factors and cause(s) of CKDu. We will conduct clinical assessments and collect extensive biological and environmental/occupational samples and data. In addition to analysis of a wide range of self-reported lifestyle/demographic, environmental, and occupational factors and environmental measurements in the case-control study, we will conduct 1) a nested family study to assess familial aggregation/genetic susceptibility; 2) a prospective study among controls of serial targeted and non-targeted contemporary environmental/occupational chemical exposures (exposomics) measured in urine and drinking water in relation to measures of acute kidney injury (AKI) and new-onset CKDu; and 3) a prospective field study among a representative sample of controls employed in agriculture and construction to investigate seasonal workplace exposures through repeat measures of targeted and non-targeted chemical exposures (exposomics, including but not limited to pesticide, metals, and combustion products), assessed in urine and via silicone wristband samplers, in relation to measures of heat stress-related AKI. We propose to assemble a resource of sufficient breadth, depth and size to permit informative assessments of a wide range of risk factors, including environmental/occupational exposures and health contexts, with sufficient power to identify one or more causes of CKDu that could be targeted for prevention, intervention or policy.

NIH Research Projects · FY 2025 · 2021-08

PROJECT SUMMARY Contamination of drinking water and foods with inorganic arsenic (iAs) represents a major public health risk in the U.S. and worldwide. Exposure to iAs has been linked to cancer, diabetes, cardiovascular, respiratory and neurological diseases. Humans and most other mammalian species have developed mechanism for detoxification of iAs, which involves a two-step conversion of iAs to methyl-As (MAs) and dimethyl-As (DMAs) and excretion of the methylated metabolites in urine. In mammals, iAs methylation is catalyzed by orthologs of a single enzyme, arsenic methyltransferase (AS3MT). An impaired capacity to methylate iAs, e.g., due to AS3MT polymorphism, has been linked to increased risk of diseases associated with iAs exposure. Mechanisms underlying the adverse effects of iAs exposure have been extensively studied using laboratory models. However, laboratory research has been hindered by substantial differences between laboratory animals and humans in their capacity to metabolize iAs. In particular, laboratory mice have been shown to methylate and detoxify iAs much more efficiently than humans, making it difficult to reproduce in mice some of the adverse phenotypes reported in population studies, specifically cancer and diabetes. The ultimate goal of the proposed research is to develop novel mouse models, in which iAs metabolism resembles that in humans and in which iAs-associated diseases can be studied at environmentally relevant iAs exposure levels. We have recently generated a new mouse strain in which the Borcs7/As3mt locus was humanized by syntenic replacement. AS3MT expression in tissues of the humanized (Hs/Hs) mice resembles that in human tissues and differs from expression of mouse As3mt: AS3MT expression is lower in livers and much higher in adrenals. Notably, the different pattern of AS3MT expression in tissues of Hs/Hs mice is associated with low efficiency of iAs detoxification and with the profiles for iAs and its methylated metabolites in tissues and excreta that are consistent with those reported in humans. The goals of this project are: (1) To characterize susceptibility of Hs/Hs mice to adverse effects of iAs exposure, focusing on the diabetogenic effects, (2) to generate a new mouse strain expressing AS3MT haplotype that has been linked to impaired iAs methylation and risk of iAs-induced diseases in human cohorts, and (3) to explore association between AS3MT expression in adrenals and adrenal function. The proposed research will generate and validate unique mouse models for iAs toxicology. These models will make it possible to study adverse effects of iAs at environmentally relevant exposure levels and in context with human-like metabolism of iAs and AS3MT polymorphism. Using these models will markedly improve translatability and impact of laboratory studies focusing on iAs induced diseases.

NIH Research Projects · FY 2025 · 2021-08

Summary/Abstract The molecular mechanism of pathogenesis of preeclampsia (PE) is largely unknown, and effective prevention and treatment strategies remain elusive. PE is a pregnancy-associated hypertensive condition and complicates approximately one in 20 pregnancies in the US and is a leading cause of pregnancy-related maternal mortality and neonatal morbidity/mortality worldwide. Endothelin-1 (ET-1) is a vasoconstrictive peptide of 21 residues, and single nucleotide polymorphisms (SNPs) in EDN1, coding for a precursor for ET-1, are associated with PE. Edn1H/+ mice in which Edn1 expression is elevated to 3X normal have normal blood pressure, despite elevated circulating ET-1. However, Edn1H/+ dams develop full spectrum of PE-like phenotypes in their late pregnancy. In addition, the embryos from Edn1H/+ dams, regardless of their Edn1 genotypes, lag in development during early implantation stage with disoriented ectoplacental cones. We reported that nicotinamide (amide form of vitamin B3, Nam), inhibitor of ET-1 downstream of ADP ribosylcyclase, ameliorates the PE-like phenotypes in two separate mouse models of PE, and our preliminary data show that Nam decreases urinary albumin excretion and increases the number of survival fetuses when Edn1H/+ dams were treated during the entire pregnancy. These observations have led us to hypothesize that PE in Edn1H/+ dams originates from abnormal placentation caused by a high maternal ET-1 expression at early implantation stage, and that Nam can correct this damage and protect dams from later PE development. Accordingly, Specific Aim 1 will test this hypothesis by dissociating early effects from later effects of Nam on PE of Edn1H/+dams by treatments with this vitamin starting at different gestational stages and for different durations. Pregnancy outcomes including blood pressure, urinary albumin and fetal number and weight will be determined at 18.5 day post coitus (dpc). In addition, the expression of components of ET-1 system and Nam’s effects on them at implantation stage will be examined. Specific Aim 2 will investigate the mechanism of effects of ET-1 and Nam on differentiation from human trophoblast stem cells into designated trophoblast cells by using 2- and 3- dimension culture system. Pharmacological dose of ET-1 alone, or ET-1 plus Nam will be added to the specific conditioned medium. Cells will be examined by their morphology, motility, and expression of markers of different types of trophoblasts. Specific Aim 3 will investigate the mechanism of effects of ET-1 and Nam on impaired uterine decidualization and angiogenesis. The markers of endometrial stomal differentiation, the structure of blood vessels and the expression of vascular endothelial growth factor (VEGF) in uteri at the implantation stage of pregnancy will be examined. Primary cultured endometrial stromal cells will be treated with pharmacological dose of ET-1 alone, or ET-1 plus Nam, and the markers of differentiation and the expression of VEGF will be determined. The proposed research will broaden and deepen our understanding regarding the role of the maternal genetic factor ET-1 on PE and identify a potential intervention strategy for PE.

NIH Research Projects · FY 2025 · 2021-08

Abstract Since its inception in 2011, the Martin Delaney Collaboratory program has made important advances towards a cure for HIV. In response to the Martin Delaney Collaboratories (MDC) for HIV Cure Research RFA, we seek to continue to advance the field by discovery of successful modalities to cure HIV infection. We will expand our expertise and work toward a better understanding of persistent HIV infection, the discovery of novel approaches to disrupt latency, methods to clear the HIV reservoir, and identification of strategies to control viral rebound. By building on the significant advances that we have made to develop, implement, and execute a suite of pre-clinical experiments that represent the most advanced and novel concepts, we will continue to pursue our central unifying hypothesis that reversing HIV latency such that viral proteins are expressed, in parallel with interventions that speed the clearance of cells emerging from latent infection, will ultimately lead to eradication of persistent HIV infection. In parallel to the efforts to clear the infection, we will pursue interventions to prevent rebound of viremia after ART interruption. We will leverage a broad portfolio of tools from both academic and industry partners, and apply new discoveries, demonstrating proof-of-concept for clinical initiatives. We will engage academic scientists and clinicians, industry investigators, and the community to a) define novel targets to destabilize proviral genomes that persist despite antiretroviral therapy (ART) b) define novel approaches to block proviral establishment c) develop and deploy novel effectors to clear viral reservoirs, d) delineate effective strategies to prevent rebound viremia that might emanate from such reservoirs after ART is discontinued and e) create bridges to the community to improve the understanding of and access to HIV cure research and clinical trials. Our initial efforts will focus on biology discovery to illuminate new host targets for latency reversal, and the validation of the novel biological concept of latency prevention. Universal strategies for proviral control or clearance will be developed and tested, including those based on HLA-E targeting, eCD4, and CD4 mimetics. Our major recent advance in latency reversal via NF-kB signaling will be further developed in both non-human primate and humanize mice models, in combination with candidates to clear infected cells. We envision an iterative process with insights gained in ex vivo and pre-clinical studies, carried forward to enhance the next step in clinical development and, importantly, fed back to scientists to validate assays or hypotheses, and explore new directions. As we have done in the past, we will develop human clinical trials to address questions and test concepts developed in our work through funding mechanisms distinct from CARE. We are dedicated to working together in a nimble program, with our research direction following our discoveries. Together we will catalyze advances that will ultimately lead to the eradication of HIV infection.

NIH Research Projects · FY 2025 · 2021-08

ABSTRACT Cardiovascular disease (CVD) exacts a disproportionate toll on rural African American communities in the Southeast. Implementing and scaling a proven-effective, evidence-based program (EBP) is essential to mitigate growing disparities in CVD risk among rural communities. We previously adapted PREMIER, an EBP, into Heart Matters and conducted a randomized controlled feasibility trial at seven host sites in two rural counties in eastern North Carolina (NC), largely populated with African Americans with high CVD burden. As with PREMIER, systolic blood pressure and self-reported physical activity and dietary behaviors significantly improved in the intervention arm compared to controls after 6 months. Heart Matters’ success, however, was dampened by critical implementation barriers at the organizational level, including limited readiness, partial collaboration between stakeholders, and low organizational efficacy to implement an EBP, which reduced fidelity and penetration of Heart Matters. To address implementation barriers, our research team will investigate the implementation and effectiveness of the Heart Matters EBP by scaling to five rural counties in Eastern NC. Guided by the Consolidated Framework for Implementation Research, our overall objective is to partner with organizations to scale and test Heart Matters implementation to other rural African American communities, and support translation of evidence to practice in eastern NC. In year 1, we will collaborate with our longstanding community-academic coalition to identify and recruit eligible organizations from our study setting (Edgecombe, Franklin, Nash, Vance, and Warren Counties). We will engage organizations (n=60) in formative research using concept mapping to identify and map contextual implementation factors affecting EBP implementation in rural African American communities. We will use these findings to refine existing training protocols and develop an organizational collaborative called “Collaborate and Leverage Evidence in an African American Rural Network” or Co-LEARN. In year 2, we will identify Co-LEARN sites (n=18) and employ participatory systems science methods to develop an implementation blueprint through: 1) shared learning aimed at training and capacity building and 2) shared action planning aimed at continuous quality improvement of implementation strategy at the site-level. The objective of Co-LEARN is to increase organizational readiness, strengthen network collaborations, and enhance organizational efficacy to implement a CVD EBP. In years 3-4, we will employ a hybrid type II implementation effectiveness design to conduct a cluster randomized controlled trial (n=486). We will evaluate outcomes of implementation (e.g., acceptability, adoption, penetration), CVD biomarkers (e.g., blood pressure, cholesterol, physical activity) and cost effectiveness of Heart Matters. Our long-term goal is to increase acceptability, adoption, and penetration of CVD EBPs in rural United States by building organizational readiness and capacities to implement sustainable and cost-effective EBPs to mitigate CVD disparities.

NIH Research Projects · FY 2025 · 2021-08

Abstract In the first years of life, when the brain is rapidly developing, children are disproportionately exposed to xenobiotics, including phthalates. However, the immaturity of the blood brain barrier cerebrovasculature and xenobiotic metabolism and excretion pathways render the infant brain more vulnerable to toxic compounds. Despite growing evidence of associations of prenatal phthalate exposures with diverse aspects of neurobehavioral development, few studies have assessed the role of early life exposure to phthalates on neurodevelopment. Furthermore, the findings on prenatal exposure have been paradoxical, suggesting that phthalate exposure accelerates the maturity of functional networks in infancy but is maladaptive in later life. Our objective is to examine the extent to which phthalate exposures change structural and functional brain development at a critical window of vulnerability (from birth to age 5), and to reconcile the paradoxical findings by tracking a variety of social, behavioral and developmental outcomes through longitudinal evaluation. We propose to leverage the University of North Carolina Baby Connectome Project (BCP), the goal of which is to map normative brain development in early life using serial structural (sMRI) and resting-state functional (rsfMRI) magnetic resonance imaging paired with age-appropriate developmental assessments. In a pilot study, we found that higher early life exposure to monobenzyl phthalate (MBzP) is associated with larger cortical gray matter volumes in regions of the frontal cortex that direct language processing and executive function, as well as dysregulated functional connectivity in the primary visual, default mode, and sensorimotor networks. While this pilot established a strong scientific premise for further study, it had a limited sample size and only measured a subset of relevant phthalates. To provide a comprehensive and unbiased understanding of the phthalate and exposomic landscape in early life, we propose to extend our analysis to 19 phthalates and phthalate replacements and to evaluate the unbiased, untargeted exposome. For a more in-depth developmental perspective, we also propose to examine the longitudinal relationship between early life toxicant exposures and sMRI, rsfMRIs and developmental inventories. We plan to increase enrollment by 50 children, resulting in a final sample size of approximately 250 children contributing approximately 540 scans. Our group has pioneered the quantitative characterization of spatiotemporal brain development in early infancy and includes a unique assemblage of expertise in environmental epidemiology, infant brain imaging, early brain development, toxicology, biostatistics, and child psychiatry that will ensure successful completion of this work. This study has important public health significance because phthalate exposures are ubiquitous, largely unregulated in the US, and more extensive and impactful to infants than to adults. Imaging biomarkers will provide crucial information on the mechanism of phthalate neurotoxicity that will guide regulatory action to protect children from maladaptive developmental outcomes.

NIH Research Projects · FY 2025 · 2021-08

ABSTRACT Fetal brain abnormalities (FBA) are one of the most common prenatal sonographic abnormality detected and account for ~20% of birth defects posing a substantial burden on the health care system. FBA can be isolated or syndromic and have vast phenotypic heterogeneity. The paired approach of prenatal diagnosis using ultrasound to characterize aberrant phenotypes with genetic analysis to determine causal lesions has improved the ability to accurately counsel families about diagnosis, prognosis, and recurrence risk. Recently, prenatal exome sequencing (ES) has been applied in cases of lethal or multiple fetal abnormalities to determine a molecular diagnosis that otherwise could not be identified with traditional testing. Our group and others using ES have shown a diagnostic rate of 23.6% in cases of multiple fetal abnormalities, but only 2.6% in isolated FBA abnormalities, indicating a need to improve diagnostic capabilities for FBA. We posit that the overabundance of unresolved fetal cases is due to a gap in our understanding of the repertoire of genotypes underlying prenatal FBA and limitations of population genetics to establish causality of rare variants in novel candidate genes. Our team who is at the forefront of prenatal genetic diagnostics and in vivo zebrafish modeling of human disease will overcome the current challenges of diagnosing prenatal FBA. We will intersect exome- and genome-wide variation with a relevant model system (zebrafish). We hypothesize that we will 1) generate initial discoveries directly relevant to human brain development by modeling novel candidate FBA genes in zebrafish; and 2) improve prenatal diagnosis for FBA using whole genome sequencing (WGS) and deep phenotyping. We will: 1. Perform bioinformatic analysis of 200+ clinically ascertained fetuses with FBA and their parents using a tiered filtering strategy on already available parent-fetus trio exome data 2. Perform WGS on 114 prospectively enrolled fetuses and their parents paired with comprehensive prenatal and postnatal phenotypic data to further characterize genotype/phenotype of FBA; 3. Establish relevance of candidate genes to FBA development and determine variant pathogenicity using genome-editing and phenotyping tools in zebrafish. Our work will expand the understanding of molecular processes governing human brain development, establish a clinical-research hybrid platform readily applicable to FBA and other anatomical defects detectable by fetal imaging, build an animal model of aberrant FBA development with potential for future use in therapeutic target identification. Our immediate results will improve counseling/management of prenatally diagnosed FBA and lead to future work to develop novel therapeutic and preventative strategies for FBA.

NIH Research Projects · FY 2025 · 2021-08

Project Summary/Abstract Child maltreatment (CM) is a pervasive public health problem and there is a critical need for brief, effective, scalable prevention interventions. Problematic parent-child relationships lie at the heart of CM. Parents who maltreat their children are more likely to have punitive parenting styles characterized by high rates of negative interaction and ineffective discipline strategies with over-reliance on punishment. Harsh, reactive parenting can contribute to the development of child behavioral problems which, in turn, can increase parental stress and lead to escalation of negative parenting behaviors, including abusive and neglectful parenting. Thus, parenting interventions that strengthen parent-child relationships, teach positive discipline techniques, decrease harsh parenting, and decrease child behavioral problems hold promise as CM prevention strategies. Challenges in engaging parents, particularly low-income and minority parents, and a lack of knowledge regarding effective implementation strategies, however, have greatly limited the reach and impact of parenting interventions. To address these challenges, our interdisciplinary team from the University of North Carolina and Children’s Hospital of Philadelphia collaborated in implementing and evaluating, Child Adult Relationship Enhancement in Primary Care (PriCARE)/Criando Niños con Cariño. PriCARE/Cariño is distinct from other programs because it was (1) developed and iteratively adapted with input from racially and ethnically diverse families, including low- income families and (2) designed specifically for implementation in primary care with inclusion of strategies to align with usual-care work flow to increase uptake and retention. In randomized controlled trials (RCTs), PriCARE has demonstrated success in engaging parents, promoting positive parenting, reducing parent stress, and improving child behaviors. The objective of this project “Child-Adult Relationship Enhancement in Primary Care: Effectiveness Trial of a Primary Care Based Parenting Intervention to Prevent Child Maltreatment” is to conduct a multi-site RCT of PriCARE/Cariño plus Usual Care vs. Usual Care among 1932 parents and their 2- 6-year-old children with Medicaid/CHIP/no insurance from 6 clinics. We will examine the effectiveness of PriCARE/Cariño on reducing suspected CM, in addition to reducing proximal CM related outcomes (child behavior problems, parent stress, harsh/neglectful parenting). By testing proximal outcomes in addition to the more distal outcome of CM, this study will help to elucidate the mechanisms of change and advance the science of CM prevention. Finally, in this study we will also identify PriCARE/Cariño implementation determinants and examine their relationship to implementation outcomes. If successful, results will be used to seek funding from state agencies and payers to sustain PriCARE/Cariño in primary care, while continuing to refine and study implementation and dissemination strategies, with the ultimate goal of achieving levels of exposure sufficient for breaking the intergenerational transmission of CM. Inclusion of consent for longitudinal data collection will facilitate future longitudinal studies of the impact of PriCARE/Cariño on additional outcomes.

NIH Research Projects · FY 2025 · 2021-08

ABSTRACT Stemness is a functional cellular attribute defined by properties of multipotency (the ability to produce all differentiated cell types in a tissue) and self-renewal (the ability to produce new stem cells). In adult epithelial tissues, stemness can exist in: (1) dedicated stem cells occupying a defined niche, and (2) differentiated cells that exhibit plasticity to drive regeneration following injury. Stem cells must balance self-renewal with differentiation, and differentiated cells must balance lineage specific identity with plasticity to participate in regenerative responses. My lab is interested in understanding how chromatin modifications and their associated enzymes interact with site-specific transcription factors (TFs) to establish cell identities while simultaneously facilitating alternative cell fates. We focus on intestinal stem cells (ISCs), which drive the renewal of the intestinal epithelium approximately once a week throughout adult life, as a model system. Over the next five years, my research program will explore how TFs and chromatin modifying enzymes contribute to an exit from the stem cell state as ISC differentiate. Current models propose that rapid differentiation and plasticity in the intestinal epithelium is regulated by chromatin landscapes that are highly similar across all intestinal epithelial cell types, regardless of differentiation stage. Recent studies in my lab have challenged this model using a Sox9EGFP mouse model that allows us to isolate ISCs, progenitors, and differentiated cells with a single reporter transgene. We quantified changes in chromatin accessibility and 5-hydroxymethylcytosine and identified significant chromatin dynamics at genomic loci consistent with enhancers. Our analyses identified novel candidate TF-chromatin interactions and raise exciting questions about the importance of chromatin regulation in adult epithelial stem cells. Our future research will build on this progress to understand: (1) how stem cell- associated chromatin is decommissioned by TFs in differentiation, (2) how lineage-specific regulatory programs are “primed” in ISCs, and (3) how chromatin modifying enzymes bridge regulation of self-renewal and differentiation. The approach proposed here will establish a long-term research program, with broad potential to expand into studies of plasticity/de-differentiation, chromatin regulation of epithelial responses to environmental challenges, and development. Our goal is to pursue a basic mechanistic understanding of chromatin regulation in stemness that will have foundational relevance to human health and disease, including epithelial responses to inflammation, infection, injury, aging, and tumorigenesis.

NIH Research Projects · FY 2025 · 2021-08

Epilepsy is a devastating disease affecting over 50 million people worldwide (WHO). About 30% of patients do not respond positively to medication and are diagnosed as having drug resistant epilepsy (DRE). DRE causes significant costs, morbidity, and mortality. The most effective treatment is to surgically remove the seizure onset zone (SOZ), the region from which seizure activity is triggered. The localization of the SOZ is essential for surgical success. Unfortunately, surgical success rates range from 30%-70% because there is no reliable biomarker of the SOZ. We propose to develop a combined intracranial EEG-fMRI biomarker of the SOZ while the patient is not seizing or at “rest”. One may ask, “how does one identify where seizures start in the brain without ever observing a seizure, and if this is possible why have previous methods failed?” The fundamental limitation of current computational approaches for both resting state fMRI (rs-fMRI) and intracranial EEG (rsiEEG) SOZ localization lies in the fact that they compute static measures from observations produced by a dynamic epileptic network. We believe that a computational method that can provide a characterization of how the observations are dynamically generated in the first place, and how internal network properties can trigger seizures or prevent seizures will be successful in SOZ localization. Therefore, we will construct dynamical network models (DNMs) in this study. DNMs are generative models that capture how every network node (location of centralized network signal processing and transfer) interacts with every other node dynamically. DNMs uncover internal properties including bandwidth, stability, controllability, system gain, and most important to this application - connectivity. We propose that when a patient is not having a seizure, it is because the SOZ is being inhibited by neighboring nodes (brain regions). We thus will apply DNM algorithms in a novel manner to identify two groups of network nodes from rs-fMRI and rs-iEEG: those that are continuously inhibiting a set of their neighboring nodes (denoted as “sources”) and the inhibited nodes themselves (denoted as “sinks”). Thus, in line with the most recent advancement in precision medicine, for each patient, we will build DNMs customized to identify and quantify, via a score, key sources and sinks, optimized to localize the primary causative SOZ nodes in the epileptogenic network and their connectivity properties. We will leverage functional imaging data while patients are “at rest” in a study population of children with DRE who are undergoing epilepsy surgery evaluation. Specifically, we will construct DNMs from rs-fMRI and rs-iEEG data and test our novel “source-sink” hypothesis that may point to the SOZ when patients are not seizing. If successful, the proposed DNMs could significantly increase surgical candidacy and improve surgical outcomes by increasing the yield of surgically actionable results and precision of SOZ localization. Furthermore, by removing the need to capture seizures, this novel dynamic network model-based SOZ localization biomarker may substantially reduce invasive monitoring times, avoiding further risks to patients and reducing costs to hospitals.

NIH Research Projects · FY 2025 · 2021-08

ABSTRACT Scientific Approach: Patients with severe dentofacial disharmonies (DFD; jaw disharmonies) seek orthognathic surgery and orthodontic care to address issues with mastication, esthetics and speech. Preliminary data from thousands of DFD patients generated by our colleagues at UNC-CH indicate speech concerns surpass impaired chewing function as a motivator for surgery. Due to limited studies, we, as providers, are unable to give evidence- based recommendations as to whether our orthodontic and surgical interventions will address speech disorders associated with DFD. Despite limited data, patients undergo invasive jaw surgery in hopes of speech improvement. To address this gap in knowledge, we have gathered surgical records and pre-operative audio recordings of patients with DFD to quantitatively characterize their speech. In this proposal, we present preliminary data identifying anatomical thresholds for significant articulation errors. Based on these data, we hypothesize that speech distortions correlate with severity of anterior-posterior and vertical jaw disharmonies, and that corrective surgery yields long-term improvement in speech. We propose the following aims to address this hypothesis and explore the relationship between correction of jaw disharmonies and speech. Aim 1: To examine the relationship between underbite (Class III) and open bite jaw disharmonies, and speech distortion. Aim 2: To assess short- and long-term effects of corrective orthognathic surgery on speech distortion in patients with underbite and open bite. Training approach: Dr. Jacox is a dual-trained clinician-scientist, who completed dental school in parallel with graduate school at Harvard School of Dental Medicine (HSDM- DMD) and Graduate School of Arts and Sciences (GSAS- PhD). In addition, she is a certified diplomate of the American Board of Orthodontics. Her research program and focus of this application was inspired by a patient seeking treatment `to help her talk normally', whereby an evidence-based treatment option was not clear. The mentoring plan for this K08 award is designed to ensure Dr. Jacox will be a well-rounded and successful independent clinician-scientist following completion of the activities in this proposal. Dr. Jacox has assembled a strong mentoring team to guide her professional development and the establishment of her independent research program. Two co-mentors and four consultants from diverse yet appropriate scientific and professional backgrounds will assist in the completion of the training and scientific activities. Scientific interactions as well as didactic training in speech science and technologies of speech science will add to Dr. Jacox's strong foundation in clinical orthodontics and the biology of dentofacial disharmonies. These growth opportunities will allow for successful completion of the proposed experiments, while the professional development activities will complement the training activities and provide Dr. Jacox with skills necessary to thrive in academics and launch her independent research program.