Univ Of North Carolina Chapel Hill

NIH Research Projects · FY 2025 · 2024-08

Abstract Aging exhibits remarkable individual heterogeneity as exemplified by the varying health trajectories and outcomes experienced by people of the same chronological age. Much of this variability is due to different live experiences and environments. Understanding the impact of environmental factors on aging is essential because it holds the key to unlocking strategies for promoting healthier and more equitable aging experiences for individuals across diverse backgrounds. The F99 phase of this project explores the interplay between environmental adversity, epigenetic shifts, and aging, using a cohort of 1,000 dogs from the Dog Aging Project. The goal is to understand how environmental factors modulate the epigenome, potentially accelerating aging and influencing health and lifespan. Specifically, I am analyzing associations between age, sex, weight, and site-specific and global DNA methylation changes, alongside validating a DNA methylation-based 'epigenetic clock' associated with chronological age, and assessing whether environmental adversity mirrors molecular aging. At the end of the F99 phase, I will have revealed mechanisms behind environmentally-induced aging in dogs, and gained training and expertise that will serve as a bridge to the postdoctoral, K00 phase. In the K00 phase, my research will expand to explore molecular determinants of aging in women, with an emphasis on understanding how specific environmental adversities impact their aging processes. Utilizing data from extensive cohort studies like the Study of Women's Health Across the Nation study (SWAN), a long-term cohort study that investigates the physical, psychological, and social aspects of women's health during the menopausal transition and beyond, involving a diverse group of women across different racial and ethnic backgrounds. I aim to uncover how epigenetics, and gender-specific environmental adversity intertwine to define aging trajectories in women, with the goal to identify high-risk groups and develop targeted interventions, thereby ameliorating age-related disparities and enhancing health and longevity in women.

NIH Research Projects · FY 2025 · 2024-08

SUMMARY/ABSTRACT Hypospadias, a urethral opening on the underside of the penis, is one of the most common birth defects in boys (1 in 250 newborns). Left untreated, hypospadias may affect quality of life, cosmetic appearance, urination, sexual function, and fertility. Currently, parents must make this important, preference-sensitive treatment decision with neither evidence-based guidance nor a clearly superior option. Consequently, 55-71% of parents experience decisional regret regardless of whether they choose surgery which is strongly associated with decisional conflict. We aim to reduce parental decisional conflict and regret by improving information delivery through use of a shared decision-making (SDM) tool called the Hypospadias Hub. It was developed and pilot-tested at two U.S. sites by our team. The objective of this proposal is to conduct a type 1 hybrid effectiveness-implementation study of the Hypospadias Hub Intervention. Our two-arm randomized controlled trial will assess its effectiveness to: 1) improve quality of SDM during hypospadias consultations as measured by parent report (primary outcome), and observer report (secondary outcome), 2) decrease decisional conflict and regret, and increase decision-relevant hypospadias knowledge (secondary outcomes). We will also assess barriers and facilitators to its future implementation. Our aims are 1. Compare the effect of the Hypospadias Hub Intervention vs. control on the quality of SDM (primary outcome). We will assess how providers influence parents’ choices using qualitative content analysis of transcripts of the consultations. Parents will be randomized to the respective arms of the study. Hypothesis: The quality of SDM during the consultation will be higher in the intervention than the control group. 2. Compare the effect of the Hypospadias Hub Intervention vs. control on decisional conflict, decision-relevant knowledge, and decisional regret (secondary outcomes). Hypothesis: Decisional conflict and regret will be lower, and knowledge higher in the intervention vs. control group. 3. Assess barriers and facilitators to future implementation using constructs from the Consolidated Framework for Implementation Research. We will use qualitative interviews with a diverse sample of parents, providers, nurses, and clinic staff to explore recommendations for implementation, and potential future implementation strategies. Evidence of the effectiveness of the Hypospadias Hub Intervention to improve quality of SDM and decision quality for parents making decisions about hypospadias, combined with insights gained from our implementation assessment, will support wider adoption, and inform development of tools for other preference-sensitive decisions, which will improve child health and quality of life.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY/ABSTRACT Hidradenitis suppurativa (HS) is a chronic inflammatory skin disease that has a devastating effect on quality of life. HS affects up to 1.2% of people and disproportionately affects women and Black people. The age of onset is typically in adolescence/young adulthood and symptoms persist for decades. More than half of patients have affected family members, consistent with a strong genetic contribution to HS. Variants implicated in familial cases of HS have been reported in four genes, three of which encode subunits of the γ-secretase complex, but these likely affect less than 5% of patients with HS, raising the question: what other variants may be at play? In our recent genome-wide association study (GWAS) of 760 patients and meta-analysis with additional cohorts, we identified significant risk loci near SOX9 and KLF5, which play important roles in follicular and epidermal maintenance, wound healing, and inflammation. We hypothesize that larger sample sizes will identify additional significant risk loci that will substantially improve our understanding of disease pathogenesis and identify potential treatment targets. We further hypothesize that dysregulated SOX9 and KLF5 expression will be found in the hair follicles, epidermis, and cutaneous tunnels in skin from HS patients. In this study, we propose to build upon our recent GWAS to examine the genetic basis of HS in an expanded cohort of 2,000 diverse patients with HS at the University of North Carolina at Chapel Hill. We continue to recruit participants from one of the highest volume HS subspecialty clinics in the country. Approximately 50% of our cohort is composed of Black patients, and we will examine potential differences in the genetic architecture based on ancestry, sex, and phenotypic diversity. We also will perform GWAS in the All of Us cohort and meta-analyze GWAS results with the FinnGen, UK Biobank, and BioVU biobanks, and other cohorts as described in the research plan. This meta-analysis will include more than 12,000 total subjects. Identifying genetic variants that show evidence for association with HS disease status will provide candidate genes for further validation and biological study to develop new treatments and possibly prevent disease. At the recently discovered GWAS loci near SOX9 and KLF5, we will analyze gene expression patterns in HS-affected tissue using RNA-seq and immunohistochemistry. To identify candidate genes for new loci, we will analyze transcriptome data to identify variants associated with both HS and gene expression and integrate with existing chromatin data. To identify HS risk variants that affect gene expression, we will evaluate skin cell transcriptome and epigenome data and perform reporter assays in keratinocytes. By identifying dysregulated gene expression patterns in the epidermis, hair follicles, and cutaneous tunnels of HS-affected tissue, we will learn how risk variants contribute to follicular disruption, inflammatory responses, chronic wounds, and tunnel formation, which will impact our understanding of HS pathogenesis and therapeutic strategies.

NIH Research Projects · FY 2025 · 2024-08

Helping adolescents learn about contraception is important for promoting shared decision-making about reproductive health. Yet decision-making can be overwhelming amidst an increasingly vast health information environment—or “infosphere”—driven by the expansion of the internet. This is particularly true for adolescents who have 1) high rates of social media use; 2) low levels of contraceptive knowledge; and 3) limited skillsets and lack of supportive relationships to help navigate sensitive health information online. Clinicians who care for adolescents can fill a critical gap in helping patients engage in shared decision-making about contraception. Yet studies show that clinicians often avoid extensive engagement when discussing patient-identified internet information due to lack of time and training. As young people are increasingly exposed to health information online, research is needed to understand how clinicians can support adolescent patients’ emerging development and contraceptive choices, particularly as they integrate components of their digital health infosphere into the contraceptive decision-making process. Discussion of a patient’s infosphere can be incorporated into adolescent-tailored patient-centered contraceptive counseling (PCCC), an approach that centers shared decision-making. To help pediatricians provide PCCC that incorporates adolescents’ digital infospheres, our objectives are: 1) collect and review content and communication strategies related to the adolescent digital contraception infosphere, including conducting a relevant qualitative content analysis of social media sites; 2) develop a Counseling about Contraception Digital Information (CCDI) framework and training module to help clinicians incorporate adolescents’ online information-seeking into PCCC; and 3) use mixed methods to evaluate pediatrician-reported acceptability, appropriateness, and self-efficacy related to the CCDI module within the context of a larger training pilot called Adolescent-Centered Counseling and Empowerment Skills for Success (ACCESS). This project fills a crucial gap by conducting, to our knowledge, the first content analysis of adolescent-focused contraception information online. Furthermore, the study will develop a modern, adolescent-adapted, and stakeholder-informed PCCC training for clinicians that addresses this population’s digital contraception infosphere. This study has potential future adaptations in many other areas of health that are also impacted by complex digital infospheres.

NIH Research Projects · FY 2026 · 2024-08

PROJECT SUMMARY Each year, nearly 2 million Americans receive a cancer diagnosis, with surgical removal being the primary treatment approach for solid tumors. Nowhere are the challenges and benefits of high precision surgery more apparent than in the brain where maximal resection is the primary therapeutic approach and where healthy tissue must be preserved. For glioblastoma multiforme (GBM), one of the deadliest and treatment-resistant malignancies, the extent of surgical resection provides the best indication of overall survival which is why establishing maximum safe boundaries is a fundamental neuro-surgical objective. Despite two decades of relentless improvements in surgical techniques, the median survival for GBM remains at 16 months using best- practice radiosurgery with temozolomide. Advancements in diagnostic and imaging capabilities must be matched by surgical practices to be effective. Consequently, as imaging modalities such as MRI, CT, or ultrasound have improved in resolution, sensitivity, and specificity, so has the precision of surgical tools, such robotic surgery systems or navigation systems, and minimally or non-invasive methods such as laparoscopies, radio-frequency ablation, radiosurgery, and high intensity focused ultrasound. However, these methods struggle to achieve a resolution beyond 2 mm, which is a significant hurdle when precision is crucial for the resection of complex tumors near critical structures. In this application, UNC-Chapel Hill and Caltech will collaborate to develop non-invasive transcranial volumetric super-resolution imaging, targeted contrast agents, and image-guided focused ultrasound surgery. Our proposal focuses on combining these techniques to resolve the persistent challenges of a) identifying tumor boundaries b) targeting them with ultrasound, and c) establishing interaction between imaging and therapy systems to avoid and/or target critical microvasculature. Advancements in imaging resolution (10 µm) will be matched by the co- registered focused ultrasound beams (750 µm diameter, 50 µm positional accuracy). New ultrasound array designs combined with super-harmonic sequences will enable the targeting of GV contrast agents as well as freely circulating microbubble contrast agents that quantitatively image the intra- and extra-tumoral microvascular environment. A programmable ultrasound scanner platform will control both the proposed therapy array and a 3D imaging array which will allow high-precision volumetric targeting and monitoring as well as enabling sophisticated feedback between thermoablation and its effect on tumor microvasculature. If successful, this project will thus enable effective surgical interventions in the highest mortality tumors.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY/ ABSTRACT Family planning prevents unwanted pregnancy and reduces maternal and child mortality in low-resourced settings; however, women in these settings encounter unnecessary medical barriers to contraceptive care. Inappropriate medical contraindications (IMCs) occur when providers deny eligible women their preferred contraceptive method without an evidence-based medical rationale. This medical barrier to family planning use is difficult to identify using traditional survey methods and has been understudied for the last 20 years. The applicant’s previous research suggests non-preferred method use is one indicator of IMCs, as 55% of non- preferred method users reported a ‘medical reason’ for nonuse. Further, qualitative data on medical reasons for non-use revealed IMC application by providers. Non-preferred method use is undesirable, as it can lead to dissatisfaction, discontinuation, and unplanned pregnancies. Identifying interventions that effectively reduce non-preferred method use and IMCs is an important contribution to global public health. The applicant’s long- term objective is to identify effective and scalable interventions for reducing medical barriers to contraceptive care for women living in low-resource settings. The proposed project will 1) estimate the impact of two social accountability interventions on non-preferred method use at the population level; 2) determine the frequency and elucidate the nature of non-preferred method use due to IMCs using innovative mystery client data collected among 137 public-sector Kenyan facilities, and 3) use qualitative methods to investigate provider perspectives on non-preferred method use and IMCs to explore key factors. The applicant hypothesizes that social accountability interventions, in which community oversight motivates providers to improve their performance, could increase patient-centeredness of care and therefore reduce non-preferred method use. To test this hypothesis, Aim 1 will use difference-in-difference methods to analyze pre- and post-intervention data from a randomized controlled experiment assessing two social accountability interventions in Kisumu, Kenya. Aim 2 will use mixed methods to analyze mystery client data collected from all public facilities in Kisumu, Kenya. Aim 3 proposes in-depth interviews with family planning providers in Kisumu, where the applicant will build a new skill – standardized vignettes – to understand and contextualize provider decision-making around IMCs. These data collection methods will overcome major methodological challenges that have prevented research into IMCs in the past 20 years. Results will contribute important new information for improving contraceptive care in low-resource settings. Additionally, the proposed rigorous training and research plans will support the applicant in developing specialized subject knowledge, building mixed methods expertise, and advancing in their development as an independent researcher.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY Understanding what shapes patterns of genetic diversity in natural populations is an inherently challenging problem. Despite the challenges, studying variation in DNA sequences within individuals in a population has the power to yield insights into selective pressures operating in the natural environment of a species, uncover historical events like the migration of humans and their pathogens across the world, and identify the genetic basis of traits like human diseases. One major challenge facing population-genomic inference is that most current state-of-the-art approaches have been developed to study human-like genomes, that are sparsely populated with functionally important elements and thus effects of selection on nearby sites can be ignored. These assumptions however do not apply to compact (gene-dense) genomes where direct and indirect effects of selection are pervasive. My research program is geared towards understanding how the joint effects of selection with other evolutionary processes operating simultaneously in a population, shape patterns of variation across the genome. As many pathogenic species tend to have highly compact genomes, experience strong bouts of selection as well as drastic repeated bottlenecks, and undergo asexual reproduction or self- fertilization often (which further increases the effects of selection), our methods would be absolutely essential to perform inference in such species. We will employ computational, statistical, and theoretical approaches to broach these questions, utilizing the development of new methods and their applications to publicly available whole-genome sequence variation data. The strength of selection against new mutations, a crucial piece of information for modeling how selection shapes variation, has been estimated predominantly for coding regions, despite the fact that in many species the majority of functional DNA that impacts fitness is non-coding. My first goal will therefore be to generate a better estimate of the shape of the genome-wide fitness effects of new mutations. As lower rates of recombination result in stronger effects of selection, my second goal is to better understand how selection against deleterious mutations affects genome-wide patterns of variation in species that undergo high rates of self-fertilization and to develop methods that account for the effects of selection. My third goal is to apply our methods to perform inference of demography and identification of recent selective sweeps in species with compact genomes, like those of Plasmodium falciparum and vivax. My work will result in a better understanding of how natural selection shapes genomic variation, as well as the development and application of methods that jointly account for multiple evolutionary processes. This will be crucial to perform accurate evolutionary inference in compact genomes. My long-term vision is to apply our methodological advances to human pathogens to learn about their population history and adaptation to their hosts.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY The retinoblastoma (Rb) protein is a prototypical tumor suppressor due to its role in restricting proliferation. In its active form, Rb functions as the key regulator of the G1/S cell cycle transition by physically binding to E2F transcription factors, an interaction that inhibits activation of genes necessary for DNA replication. To relieve transcriptional repression of pro-proliferative genes and promote cell cycle progression, Rb is inactivated through its phosphorylation by the cyclin-dependent kinases 4 and 6 (Cdk4/6). This inactivation is the direct target of the Cdk4/6 inhibitors that are currently used in combination with anti-hormone therapy for treatment of hormone- receptor positive, HER2-negative metastatic breast cancer. Despite the clinical success of Cdk4/6 inhibitors, primary and acquired resistance in patients remain a major challenge, and these drugs are ineffective against other tumor types. Identifying other mechanisms that regulate Rb and mediate sensitivity to Cdk4/6 inhibitors should provide avenues for addressing some of these clinical challenges. I have preliminary data that suggests Rb is subject to rapid turnover throughout the cell cycle, and I have found that Rb protein levels are decreased following Cdk4/6 inhibition. Finally, I have found that Rb protein levels can be stabilized and rescued by disrupting components of the ubiquitin-proteasome system, indicating that the Rb protein is regulated by this proteolytic pathway. Collectively, these findings challenge our current understanding of Rb as a stable protein and are clinically relevant because a change in Rb protein abundance following Cdk4/6 inhibition can potentially compromise the ability of Rb to restrict proliferation and maintain proper cell cycle arrest. Thus, the overall goal of this proposal is to understand how ubiquitin signaling affects Rb’s tumor suppressive function. Based on my preliminary findings and current understanding about the role of Rb in cell cycle control, my central hypothesis is that the regulation of Rb protein by the ubiquitin-proteasome system contributes to proper cell cycle progression and mediates response to Cdk4/6 inhibition. In Aim 1, I will use mass-spectrometry and a CRISPR/Cas9 approach to identify the ubiquitin ligase (E3) that degrades Rb and live-cell imaging to determine the timing of Rb degradation in the cell cycle. In Aim 2, I will investigate how the degradation of Rb affects cell cycle progression using various biochemical, genetic and fluorescent imaging approaches. By identifying a non- degradable Rb mutant and engineering cells lines that express an inducible form of this mutant, I will be able to determine the extent to which disrupting Rb degradation affects sensitivity to Cdk4/6 inhibition. Completion of these aims will provide me with diverse training experiences in cell, systems and cancer biology and shed light on how Rb is regulated beyond phosphorylation-dependent inactivation.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY/ABSTRACT This proposal represents a five-year research career development program focused on understanding intestinal epithelial cell (IEC) barrier dysfunction in peanut allergy and on providing a strong foundation for the applicant, Dr. Erin Steinbach, to build an independent basic and translational research career. The candidate is currently a tenure track Assistant Professor of Medicine at the University of North Carolina (UNC) School of Medicine in the Division of Rheumatology, Allergy, and Immunology. She is a member of the Thurston Arthritis Research Center and Center for Gastrointestinal Biology and Disease. The proposal builds on the candidate’s previous mucosal immunology research and clinical allergy training by integrating five new realms of expertise represented by her mentor team of Drs. Shehzad Sheikh and Wesley Burks and her advisors’ expertise in IEC biology, gene regulation, metabolomics and integration of large datasets, and prospective human studies. This work will add to the existing knowledge of a diverse group of diseases associated with increased IEC permeability and will foster the development of novel therapies targeting the intestines for severe allergic reactions. Through formal coursework, seminars, and guidance from her team, the candidate will develop the skills needed to run a successful research program. She will develop a unique skillset for competing successfully for R01-level grants that will support her transition to an independent physician scientist career. Peanut is the most common cause of death from food-related anaphylaxis. Severity of a food allergic reaction is partly determined by IEC barrier dysfunction, suggesting that peanut is adept at affecting the IEC barrier. No one has worked out how or why people with peanut allergy have increased IEC barrier permeability. Our preliminary data show that barrier permeability after the development of peanut allergy persists even in isolated, cultured IECs. We will use a novel Collaborative Cross mouse model (CC027 strain) and primary human 2D “enteroids” (primary human-derived IEC monolayers) to broaden the understanding of increased IEC permeability in peanut allergy. CC027 mice develop peanut allergy through the oral route without adjuvant, which is normally needed in animal models to establish peanut allergy. The CC027 peanut-allergic mouse develops increased intestinal permeability. Peanut-allergic CC027 mouse-derived IECs have altered proliferation and cholesterol metabolism, gut dysbiosis, and produce very high levels of Angiopoietin-like 4 (ANGPTL4). Serum ANGPTL4 levels are higher in peanut-allergic, compared to non-allergic, pediatric patients. Alterations in the gut microbiota are present in food allergy and affect the IEC barrier, but its effects in IEC barrier function in peanut allergy remain elusive. We hypothesize that in established peanut allergy, more local IEC ANGPTL4 production and an altered microbial metabolome disrupts intestinal epithelial stem cell metabolism and ultimately IEC barrier integrity.

NIH Research Projects · FY 2025 · 2024-08

Project Summary/Abstract Necrotizing enterocolitis (NEC) is a severe, often fatal intestinal disease of prematurity. Mortality rates approach 50% if surgery is required to resect necrotic and irreversibly damaged intestinal tissue. Extensive preclinical research supports a central role for deleterious unrestrained inflammation in intestinal injury and destruction of the intestinal epithelium. Despite the insight gained from decades of research, treatment options for infants with NEC are non-specific and often ineffective in the most severe cases. Factors that have impeded the development of novel effective therapies for this disease include an incomplete understanding of disease mechanisms in human infants, a lack of biomarkers, and the fragility of critically ill preterm infants. In addition, NEC is a developmental disease associated with intestinal immaturity, which poses a further challenge in that interruption of these developmental processes can have lifelong consequences. To overcome these obstacles, future NEC research must be patient-focused, innovative, and highly collaborative. I have spent the last 14 years dedicated to studying NEC and the immunologic mechanisms of disease. I created the largest biorepository in the world for NEC, which contains tens of thousands of samples obtained prospectively from premature infants that developed NEC and age-matched control infants. The NEC Biorepository is a multi-institutional collaborative of leading investigators in NEC research from across the country. The development of this biorepository was motivated by an urgent need to study larger patient cohorts and the logistical difficulties of obtaining intestinal samples from critically ill neonates undergoing emergent surgery. In addition, my laboratory is pioneering efforts in biotherapeutic design and implementation for NEC. We recently identified the polyfunctional cytokine interleukin-22 as a potentially powerful new immunotherapy for this disease, and we currently hold a patent for its use in the treatment of NEC. This proposal will utilize the invaluable samples in the NEC Biorepository, our collaborative network, and our extensive expertise in this specialized field to accelerate the growth and development of a new era of NEC research. We will define new mechanisms of immunopathogenesis and employ innovative interventions to interrupt the exaggerated injurious response that leads to NEC. In this effort to reprogram intestinal immunity, we will redefine the paradigms that currently exist in our vague understanding of NEC pathogenesis. This NIDDK Catalyst Award provides our research program with the opportunity to use groundbreaking techniques to answer the elusive questions regarding the central disease mechanisms in NEC. We are determined to cure this devastating disease by interrupting the deleterious inflammatory response that results in intestinal injury in preterm neonates.

NIH Research Projects · FY 2026 · 2024-08

ABSTRACT Most neurodegenerative diseases are sporadic with unknown etiology that likely stems from the interaction between genetics and the environment, the latter of which continues to remain poorly understood. Epidemiological studies have linked environmental exposures to the development of TDP-43 pathology, widely considered a hallmark of frontotemporal dementia (FTD) and amyotrophic lateral sclerosis (ALS) but has now been expanded to a host of other age-related neurodegenerative disorders including Alzheimer's disease (AD) (collectively referred to as TDP-43 proteinopathies). To begin addressing some of the major gaps in our understanding of how environmental risk factors are linked to neurodegeneration, we performed a high-content microscopy screen to identify environmental toxicants that mediate TDP-43 dysfunction and identified pathology “promoters” that have never been linked to TDP-43 or dementia, thus opening the possibility that we can 1) uncover new risk factors that impact the onset or progression of neurodegeneration, 2) identify the cellular pathways and disease contexts through which they operate, and 3) provide a new paradigm for therapeutic interventions that are based on environmental susceptibility. We assembled a team of experts covering TDP-43 pathogenesis, genetic modeling, high-content screening, and computational modeling to address our overarching hypothesis that distinct classes of at-risk environmental toxicants drive TDP- 43 dysfunction, neurodegeneration, and memory & motor decline. To address this hypothesis, we will evaluate a high priority subset of environmental toxicants identified in our screen based on their prevalence and levels of human exposure. In Aim-1, we will examine the pathophysiological relevance of toxicant exposure using in vivo and neuronal models that allow us to interrogate their functional impact. In Aim-2, we will evaluate exactly how toxicants mediate TDP-43 dysfunction and will examine direct cysteine-dependent and indirect stress-responsive mechanisms that would link environmental toxicants to enhanced TDP-43 aggregation. In Aim-3, we will evaluate the extent to which toxicants affect TDP-43-dependent pathways vs. global disruption of other key FTD/ALS-associated signaling pathways. This study is innovative because it is the first systematic survey of at-risk chemicals affecting TDP-43, enabling us for the first time to connect environmental agents to neurodegenerative disease. Our study is also significant because it could radically transform our understanding of how an environmental agent may drive or perhaps increase susceptibility to disease, which could logically inform strategies to alleviate or mitigate disease burden.

NIH Research Projects · FY 2026 · 2024-08

Prenatal Alcohol Exposure (PAE) is a leading cause of cognitive disabilities known as Fetal Alcohol Spectrum Disorders (FASD). Interventions that ameliorate PAE’s impact are a high priority. One such intervention is the micronutrient choline, a one-carbon (1C) donor essential for healthy brain development; choline improves cognitive outcomes in both FASD and its preclinical models. However, its benefits are not always clear, and an important modulator of these responses are the known polymorphisms in choline- and 1C-metabolizing genes that affect the metabolism, transport, and utilization of choline and its methyl groups and thus choline needs. We recently reported that for one such gene, the choline transporter SLC44A1, minor alleles known to increase choline needs are associated with greater memory improvement in alcohol-exposed children who received supplemental choline; that is, those who carry the effect alleles have a higher choline need that is met by the supplement. Moreover, in a separate study of control and alcohol-exposed children who never received choline supplements, those same alleles are again associated with greater cognitive impairment, indicating that alleles that increase choline needs also increase the vulnerability to PAE, likely because their greater needs went unmet by current dietary practice. Here, we further explore the choline genetic landscape of those with PAE to identify additional choline-related polymorphisms that modulate cognition and behavior. Specifically, we hypothesize that select polymorphisms in additional genes that affect choline needs significantly influence behavioral outcomes in FASD. Aim 1 tests the hypothesis that there exist polymorphisms in additional choline-related genes that influence behavioral outcomes in those with PAE who consume normal dietary choline (Gene x Exposure). This aim utilizes an existing dataset from those with PAE who were deeply genotyped and phenotyped with respect to neurobehavior. Aim 2 directly tests the influence of a strong candidate gene identified in our preliminary data, ALDH1L1, on cognition and behavior. ALDH1L1 controls the size of choline-derived 1C pools and therefore choline levels. Using an established haploinsufficient mouse model, we will establish 1L1’s role in affecting behavior and vulnerability to PAE. These findings (i) provide novel insight into how alleles affecting choline needs impact behavior in any population; (ii) further informs how choline improves outcomes in FASD; and (iii) informs the optimization of future choline interventions for at-risk pregnancies and individuals. This study represents an early application of Personalized Medicine to FASD.

NIH Research Projects · FY 2025 · 2024-08

Project Summary Despite public health efforts to reduce prenatal alcohol exposure (PAE), 11.7% of pregnant women in the U.S. consume alcohol and 3.9% binge drink. PAE can lead to growth retardation, facial dysmorphology and neurobehavioral disabilities, the key manifestations of Fetal Alcohol Spectrum Disorders (FASD). Although the growth deficits correlate with the cognitive and behavioral impairments, we lack mechanistic insight into the basis for these growth deficits. A major driver of fetal growth deficits is maternal metabolic disruption, and although alcohol is well-known to alter metabolism, surprisingly, this has been minimally investigated in PAE. My preliminary data address this knowledge gap, and show that alcohol reduces maternal-fetal glucose pools, with commensurate rise in fetal amino acid catabolites including urea. Although alcohol stimulates skeletal muscle catabolism, I find no sign that this happens in alcohol-exposed dams. Instead, my preliminary studies find that alcohol-exposed dams fail to undergo an expected adaptation to an insulin-resistant state. Moreover, I find that alcohol-driven changes in maternal fasting glucose and insulin correlate with the alcohol-mediated reductions in fetal body and brain weight. Launching from these data, this proposal uses an established mouse model of PAE and tests the hypothesis that alcohol prevents the dam from entering an insulin-resistant state, thus limiting fetal glucose availability, and thereby increasing fetal reliance on amino acids to support gluconeogenesis instead of growth. The first phase of the K99 portion quantifies maternal-fetal fed/fasted glucose levels, gluconeogenesis, and their related metabolite pools to understand how alcohol alters maternal-fetal energetics to reduce fetal glucose availability. The second phase of the K99 portion quantifies insulin signaling, and glucose storage and utilization in maternal liver, to test the hypothesis that alcohol prevents maternal adaptation to insulin resistance. Finally, the R00 phase will identify the underlying mechanism(s) that mediates this adaptive failure in alcohol-exposed dams, focusing on major candidate drivers including leptin-resistance, and prolactin and placental lactogen activity. Future studies will test these candidates functionally using organ-targeted transgenic and knockout mouse models. I will be trained in the methodologies of whole-animal and cellular metabolic assessment and an in-depth understanding of insulin/leptin signaling in pregnancy. I will also be trained in mouse behavioral assessment, looking toward future studies of how these metabolic changes may impact cognitive function. This work provides novel, mechanistic insight into the basis for the fetal growth deficits that typify FASD, focusing on key, heretofore overlooked metabolic processes. These data may also inform the development of a metabolite fingerprint or hormone test to identify at-risk pregnancies, and they will offer insights into nutrient-based interventions that could improve gestational outcomes in those pregnancies.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY The pathophysiology of cognitive dysfunction in prepubertal children with Type 1 Diabetes Mellitus (T1DM) is not understood. It is likely that hyperglycemia, hypoglycemia, vascular disease, insulin resistance, and diabetic ketoacidosis (DKA) play important roles. However, the absence of longitudinal studies in T1DM children and our lack of understanding of the pathogenesis of neuronal defects limit our ability to prevent cognitive disability and identify and treat those at highest risk. Despite preliminary evidence that T1DM is associated with alterations in brain structure and function, major gaps remain in our knowledge about how T1DM impacts the developing brain. The overall premise of this project is that T1DM in children is associated with significant neurocognitive and brain alterations and the risk of developing neurocognitive deficits increases with both biological and psychosocial stress that ultimately delay/impede the normal maturation of associative brain systems supporting attention, learning and memory, executive and social cognition. Our hypothesis is that children with greater net exposure to biological and psychosocial stress will have greater deficits in trajectory of brain development underlying complex cognition. Our multidisciplinary team of neuroimaging experts and endocrinologists will examine brain and cognitive alterations in children with T1DM across three aims: Aim 1 will measure the differential impact of age of T1DM diagnosis on brain structure, micro-organization, metabolism and function and mental health. Aim 2 will Identify biological and psychosocial moderators and Risk and Resilience factors. We will evaluate the impact of both biological stressors and social determinants of health (e.g., SES, Education, Adversity exposure, rurality, access to healthcare services, race, etc.) on neurocognitive function and maturation in children with T1DM. In Aim 3 we will develop a recruitment, retention and dissemination strategy to prepare for future clinical trials that build a precision risk- and resilience-prediction model. The findings from this observational study will bridge gaps in our understanding of the development of neurocognitive dysfunction in T1DM. It will further inform key clinical targets to address to ensure the optimal care of children with T1DM. The power of a larger network will further allow the exploration of complex 2-way or even 3-way interactions between biological and social determinants of health and neurofunctional outcomes in children with T1DM.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY/ABSTRACT People living with HIV (PLWH) are at heightened risk for chronic lung diseases. The mechanisms underlying this risk are poorly understood. Viral infections are major causes of acute respiratory events including pneumonia as well as asthma and COPD exacerbations, all of which can increase the risk of chronic lung impairments. The nasal mucosal “secretome”, composed of cytokines/chemokines and other soluble mediators, limits the initial spread of viral infections through controlling viral entry, releasing cytokines/chemokines, as well as numerous immune modulating mediators. The nasal microbiome, including bacterial microbiome and fungal mycobiome, is a critical component of the respiratory immune system, acting as a barrier against harmful pathogens Our research group has established approaches to rigorously characterize and quantify the nasal mucosal secretome and microbiome. Our overall objective is to determine nasal mucosal immune and microbiome alterations and clinical implications associated with these changes among PLWH compared to seronegative individuals. We hypothesize that alteration of the nasal mucosal milieu among PLWH is associated with adverse pulmonary outcomes. We will characterize the nasal immune environment (Aim 1) and microbiome (Aim 2) of PLWH compared to seronegative individuals. Through longitudinal assessments embedded in these Aims we will define the variability in these measures. We will then define nasal mucosal endotypes associated with pulmonary consequences among PLWH compared with seronegative MWCCS participants (Aim 3). Successful completion of these aims will lead to a rigorous characterization of the nasal mucosal and microbiome changes among PLWH, identifying potential endotypes associated with adverse pulmonary outcomes in this population. These results will provide valuable insights to guide therapeutic strategies and interventions to modify nasal mucosal inflammation and nasal microbiome and its associated complications in this vulnerable population.

NIH Research Projects · FY 2025 · 2024-08

Chronic conditions impose significant challenges on workers in low-wage work environments, often worsening health outcomes. Chronic pain-related conditions contribute to disability and work limitations. Work accommodations, which enable individuals with disabilities to perform job duties, have the potential to mitigate such limitations. However, a significant percentage of workers in the United States, particularly those in low-wage work environments who face additional challenges, do not receive work accommodations. The lack of accommodations denies potential benefits to both employees and employers. Despite the high prevalence of chronic pain-related conditions among workers in low-wage work environments, little research has explored the role of work accommodations for advancing health outcomes in this population. This study aims to address this gap using explanatory sequential mixed-methods research. The first phase will quantitatively examine the use of work accommodations by frequency and type, as well as the extent to which employer-level factors influence workers receiving work accommodations. The second phase will qualitatively explore the experiences of workers in low-wage work environments with chronic pain-related conditions and their process of obtaining work accommodations. The third and final phase will integrate the quantitative and qualitative data, using the qualitative results to inform interpretation of the quantitative findings. By examining work accommodations for workers in low-wage work environments with chronic pain-related conditions, this study will inform future policies and promote workplace health. The findings will contribute to a deeper understanding of the experiences and challenges faced by workers in low wage work environments and guide interventions to improve workplace health. This project is essential for developing the applicant’s skills in designing, conducting, and analyzing mixed-method studies, as well as quantitative and qualitative data. This study aligns with the applicant’s goal of becoming an independent researcher focused on reducing workplace challenges and identifying strategies to improve health and quality of life by addressing workplace conditions.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY/ABSTRACT Cough hypersensitivity syndrome (CHS) is a disorder characterized by chronic coughing, which leads to serious health sequelae such broken ribs, unintended urine leakage, and fainting. Because coughing is a respiratory symptom across a variety of chronic lung diseases, the investigation of CHS aligns with the research priorities identified in the Strategic Vision of the National Heart, Lung, and Blood Institute. A critical barrier to progress is that clinical practice presents two distinct treatment approaches for CHS. Pharmacologic options include neuromodulating medications and nerve block injections, while nonpharmacologic options rely on cognitive behavioral principles via speech therapy. The lack of empirical evidence to guide which of these extremely different treatments to recommend for an individual patient results in a trial-and-error approach, resulting in increased misdiagnosis and unnecessary, risky, and expensive treatments. The exact components of nonpharmacologic speech therapy that target underlying mechanisms of change in CHS remain unknown, presenting a barrier to future treatment optimization. The central rationale for the proposed research is that cough symptomology can identify CHS patient subgroups, which will more accurately predict which patients respond to different CHS treatments. Preliminary work demonstrates that the symptom of the urge-to-cough using the general Labeled Magnitude Scale successfully detects changes in healthy adults, which needs to be trialed as a metric of cough symptomology in patients with CHS. The current project addresses these critical gaps by (Aim 1) investigating whether cough symptoms predict responses to treatments for CHS, and (Aim 2) determining the feasibility of a pilot randomized controlled trial of a nonpharmacologic treatment, acceptance and commitment therapy, in patients with CHS. Aim 1 will provide essential clinical guidance by establishing a scientifically informed approach to predict which patients with CHS are candidates for pharmacologic versus nonpharmacologic treatments. Aim 2 will demonstrate feasibility, acceptability, and preliminary data for pursuing larger randomized controlled trials in specific nonpharmacologic interventions for CHS. The proposed K23 award will address these research needs by focusing on essential career development training in the areas of clinical trials, cough neurobiology, respiratory psychology, and scientific leadership. Taken together, completion of the proposed project will logically lead to an independent patient-oriented research program focused on improving symptom burden and quality of life in patients with CHS across the spectrum of chronic lung disease.

NIH Research Projects · FY 2025 · 2024-08

Project Summary/Abstract Post-cesarean surgical site infections (SSIs) contribute to maternal morbidity and mortality globally; as rates of cesarean delivery increase, so will the number of SSIs. Timely SSI diagnosis and treatment can improve maternal outcomes. However, in many settings, particularly in rural areas, postoperative wound monitoring is challenging due to physical and financial barriers. The overall goal of this proposal is to improve strategies for post-cesarean SSI monitoring by validating and updating two existing image-based diagnostic algorithms. The original algorithms were trained on image-SSI diagnosis dyads collected on women delivering via cesarean in rural Rwanda. One algorithm, using visible images (photographs), had a 83% sensitivity and 75% specificity. The second algorithm, using thermal images, had 95% sensitivity and 84% specificity. In this proposed research, we will prospectively follow 6,000 women in Rwanda, Ghana and Mexico (2,000 per site) and collect wound images and SSI diagnoses at postoperative day (POD) 10. These sites were chosen because of: a) high SSI rates; b) the potential to integrate an accurate SSI diagnostic algorithm into existing community health worker follow-up programs; and c) the diversity in skin tones across the study sites. Using this data, we will assess the generalizability of the existing visible image and thermal image algorithms, evaluating the sensitivity and specificity overall and by country (Aim 1). We will then retrain the algorithm to improve predictive properties across diverse populations, and we will explore adding clinical data to the algorithms to improve accuracy (Aim 2). Finally, for a subset of 1,200 women who are SSI negative at POD10, we will reevaluate at POD20 and POD30, and use these image-SSI diagnosis dyads to explore the need for later SSI monitoring and the ability to predict delayed SSIs with images captured at POD10 (Aim 3). The culmination of this research will provide strategies for home-based monitoring of cesarean-related SSIs that can accommodate a range of skin tones.

NIH Research Projects · FY 2024 · 2024-08

PROJECT SUMMARY/ABSTRACT Significance: Autism spectrum disorder (ASD) is an increasingly prevalent neurodevelopmental condition in the United States, for which the etiology largely unknown. ASD is a spectrum, characterized by restrictive and repetitive behaviors, and difficulties and differences in communication and learning. More severe and profound forms of ASD require lifelong support. Early diagnosis of ASD is critical as it allows for the opportunity to develop strengths and build skills, increasing independence later in life. It is important to explore the possible causes of ASD that occur in the perinatal period to target potential cases for screening and intervention. Previous research has shown that common indicators of hormonal variability including thyroid dysfunction and metabolic conditions among pregnant people are associated with ASD in their children. Research on other prevalent exogenous and endogenous hormonal exposures and their association with ASD, is inconclusive. Specific Aims: The overarching goal of this proposal is to evaluate how variability in the perinatal hormone environment is associated with ASD. My specific aims are 1) to investigate if there is an increased risk of ASD in children of pregnant people who used hormonal contraception (HC) in the 3 months prior to conception or during pregnancy, compared to children of pregnant people who did not use HC in this timeframe, and if this risk increases with progestin-only contraceptives, 2) to investigate if there is an increased risk of ASD in children of pregnant people who reported diagnosis of endometriosis, polycystic ovary syndrome (PCOS), or uterine fibroids, compared to children of pregnant people who did not report diagnoses of these conditions, and 3) to investigate if, among a subgroup of pregnant people with reported subfertility, the risk of ASD in their children is altered by certain fertility treatments compared to those who did not receive fertility treatment. Approach: These aims will be assessed using data from The Study to Explore Early Development (SEED), a multi-site case control study of ASD in the United States. Logistic regression models will be used to estimate the odds of the exposure in cases compared to controls, for each aim. Inverse probability of exposure weights (IPEW) and inverse probability of observations weights (IPOW) will be calculated and implemented into logistic regression models to control for bias from confounding and missing data, respectively. Fellowship Information: The applicant is a PhD student in the Department of Epidemiology at the University of North Carolina at Chapel Hill (UNC), and a predoctoral trainee in the NICHD funded T32 training program in reproductive, perinatal, and pediatric epidemiology. The proposed training plan will provide the applicant with the necessary resources to build on her rigorous education at UNC, successfully complete her doctoral dissertation, and pursue the professional skills necessary to prepare her for a career as an independent investigator in perinatal epidemiology.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY/ABSTRACT Despite major advances over recent decades, cardiovascular disease (CVD) remains a leading cause of death in the USA and globally and risk for factors that predispose to CVD remain incompletely understood. Although symptomatic food allergy is relatively uncommon in adults, nearly 1 in 5 adults make IgE antibodies to food that can be detected in circulation; the majority of those with IgE antibodies are unaware of the IgE, do not have symptoms related to food and routinely consume the foods to which they make IgE. We recently published findings from a study of 4996 adults in the National Health and Examination Survey (NHANES) with replication in 960 adults in the Multi-Ethnic Study of Atherosclerosis (MESA) that showed a significantly increased risk of CVD-related mortality associated with IgE antibodies specific for cow’s milk. Although a novel observation, these findings fit within an increasing body of evidence that inflammation, including Th2-related allergic pathways, contribute to CVD development. The goal of this proposal is to expand our research in order to address several important questions: i) do IgE antibodies specific for other food allergens also confer CVD risk? ii) which specific subtypes or markers of CVD are associated with IgE to foods? iii) does consumption of a food to which an individual is sensitized increase risk of CVD? To address these questions, we will comprehensively assess the relationship between food antibodies and CVD mortality in the discovery sub- cohort of MESA that we have previously studied (Aim 1), validate the findings of our preliminary data and Aim 1 in the entire MESA cohort, examining which CVD phenotypes are associated with food specific IgE and whether consumption of allergen increases risk (Aim 2), and take advantage of an ongoing cohort of those undergoing clinically indicated coronary CT angiography (CAVA-CCTA) to understand the relationship between food specific IgE and atherosclerotic plaque. Together, these aims are designed to provide strong evidence in support of our hypothesis that food sensitization, in a manner dependent on consumption, confers CVD risk. Confirmation of our recently published findings would have high potential for clinical impact because they suggest an opportunity for precision nutritional prevention of CVD, could change food allergy guidelines which currently encourage those who have asymptomatic IgE to foods to consume the food, question whether oral immunotherapy for food allergy could have unintended long-term consequences, and suggest a potential use of anti-IgE therapies for CVD.

NIH Research Projects · FY 2024 · 2024-08

Abstract Recombinant adeno-associated virus (rAAV) is a non-replicative DNA viral vector that is used in three FDA approved gene therapies and many ongoing clinical trials. Despite the popularity of rAAV as a therapeutic vehicle, there exists no generalizable method of tightly controlling rAAV transgene dose. After transduction, most rAAV genomes persist as concatemeric episomes that are retained over time in non-proliferating cells. Recent evidence suggests that rAAV episomes are chromatinized and epigenetically regulated, although further inquiry is needed to clarify the relationship between epigenetic modifications and rAAV transgene expression. The Hathaway Laboratory has developed a novel, titratable, and reversible system of epigenetic editing that uses bifunctional small molecules, called Chemical Epigenetic Modifiers (CEMs), to recruit endogenous epigenetic machinery to targeted genetic loci. We hypothesize that potent and specific next-generation CEM compounds (SLF* CEMs) could be applied to chemically regulate rAAV transgene expression. In Aim 1, SLF* CEM-based systems of rAAV regulation will be constructed and screened for efficacy in human cell lines. In Aim 2, the putative epigenetic mechanism of CEM-mediated rAAV regulation will be interrogated through knock down/replacement and CUT&RUN-Sequencing experiments. The results of this study could provide a foundation for the development of a system of epigenetically tuning the dose of rAAV- based gene therapies without the need for vector readministration.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY Wildfire smoke increasingly contributes to ambient air pollution and more specifically PM2.5 and has been associated with increased respiratory virus morbidity and mortality. We have previously shown that exposure of human volunteers to woodsmoke (WS) significantly modifies acute antiviral host defense responses to influenza. Similarly, we have preliminary evidence indicating that simulation of wildfire smoke affects viral infections in human nasal epithelial cells (HNECs) in vitro. Since concurrent exposures to wildfires and respiratory viruses will likely increase worldwide due to climate change, understanding the clinical manifestation, cellular mechanisms, and chemical components causing modified antiviral defense presents an important research gap critical to public health. We have already demonstrated that computational modeling approaches can establish the presence of harmful chemicals in different wildfire smoke conditions as drivers of toxicity. Integrating these translational research and computational analysis models, the central hypothesis of this proposal is that wildfire smoke affects multiple steps of antiviral defense pathways in the respiratory mucosa and that chemical signatures within the complex pollutant mixtures can be associated as drivers of biological responses. Specific Aim 1 will use in vitro exposure and infection of HNECs to determine the mechanisms of wildfire smoke-induced changes in antiviral defense function and identify the chemical signatures mediating these responses. HNECs will be exposed to a variety of different emission mixtures simulating different wildfire scenarios. Changes in viral infection and multi-omic assessment of immune and inflammatory responses will be assessed. Using computational modeling, chemical and molecular signatures driving the responses in HNECs in vitro will be determined. Specific Aim 2 will determine the effects of WS exposures and influenza infection on nasal immunity in humans in vivo, using inoculation with the live attenuated influenza virus (LAIV) vaccine as a model to elicit antiviral defense responses in the respiratory mucosa. Volunteers will be randomized for exposures to WS/Filtered Air followed by inoculation with LAIV/Placebo. Using multi-omic platforms, samples will be analyzed for immune biomarkers, as well as markers of viral replication and sinonasal symptoms, followed by computational analysis associating biomarkers with infection outcomes. This application aims to comprehensively assess the impact of wildfires on respiratory mucosal antiviral defenses by examining viral entry and replication, antibody production, as well as mucosal metabolomics, transcriptomics, and proteomics mediating cellular defenses. The parallel investigations of in vitro (SA1) and clinical (SA2) outcomes will yield both immediate translational impact and in-depth mechanistic insights into the effects of wildfires on respiratory antiviral defenses. Ultimately, this application will elucidate the mechanisms and chemical signatures through which wildfire exposures disrupt respiratory innate immune defenses and correlate these changes with alterations of nasal mucosal host defense functions in humans in vivo.

NIH Research Projects · FY 2026 · 2024-08

Every year 400,000 people with Alzheimer’s disease and related dementias (ADRD) are hospitalized, transfer to a skilled nursing facility (SNF), and transition to home, assisted living, or long term care. After discharge, one in five patients are re-hospitalized within 30 days, and their caregivers experience high levels of strain and psychological harm. To promote effective transitions from SNF to home and other destinations, we designed the Connect-Home ADRD transitional care intervention (C-H ADRD), an adaptation of Connect-Home transitional care. Our recent trial of Connect-Home generated evidence for intervention efficacy in a population of 327 patient and caregiver dyads. In a supplemental award, we identified unique needs of SNF patients with ADRD and their caregivers. We designed the C-H ADRD intervention to address those needs: (1) before discharge, SNF staff develop an ADRD transition plan with patient and caregiver dyads and (2) after discharge, a Dementia Caregiving Specialist provides phone-based support for the ADRD transition plan. In a pilot study, we demonstrated C-H ADRD’s feasibility, acceptability, and potential to reduce caregiver strain and patient neuropsychiatric symptoms (NPS). The next logical step is an efficacy test of C-H ADRD. Our Objective is to conduct a parallel arms, cluster randomized trial (CRT) of C-H ADRD, enrolling 360 ADRD patient and caregiver dyads in 12 SNFs. The Specific Aims are: Aim 1: Conduct an efficacy CRT of C-H ADRD (intervention) vs. usual discharge planning (control) and compare caregiver outcomes 30 days after discharge: (1) caregiver strain (primary) and (2) depression and preparedness for care transitions (secondary). Aim 2: Compare patient outcomes between intervention and control arms 30 days after discharge: (1) patient NPS (primary) and (2) quality of life and acute care use (secondary). Exploratory: Examine whether caregiver strain modifies intervention effect on patient outcomes. Aim 3: Examine potential variability in Connect-Home ADRD outcomes and implementation. (3a) Determine intervention impact by patient discharge destination and other factors for caregiver and patient outcomes. (3b) Identify factors at the dyad-level (e.g., patient discharge destination) and SNF-level (e.g., quality rating) contributing to fidelity to the C-H ADRD protocol. IMPACT: C-H ADRD has potential to improve health outcomes of people with ADRD and their caregivers.

NIH Research Projects · FY 2025 · 2024-08

Project Summary This developmental R21 study proposes a secondary data analysis addressing specific questions regarding the mediating pathways by which childhood maltreatment (CM) experienced by mothers may place their children at greater risk for externalizing and internalizing symptoms in adolescence. Using innovative models from developmental neuroscience, this study explores novel frameworks for understanding associations between maternal CM and offspring psychopathology. The subsample is drawn from the ongoing Family Life Project (FLP) and is comprised of mothers who reported that they did or did not experience CM at or before the age of 14 (N=1122). There is growing interest in the long-term consequences of CM and its association with the socioemotional adjustment of survivors and their children. Prior research links CM history across numerous domains of adult interpersonal functioning, including elevated depression, intimate partner violence (IPV), and problems in the parental role. Children of mothers with trauma histories are reported to have significant adjustment difficulties when compared to children of women without self-reported CM. Despite the strength of evidence linking maternal CM history and offspring psychopathology, there is heterogeneity in outcomes for survivors of trauma and their children. Our goal is to identify the underlying mechanisms by which maternal CM may be related to psychopathology among offspring in adolescence. We propose a dimensional model of adversity focused on examining how different types of risk factors may be differentially related to the development of children whose mothers report CM. Using novel methods to achieve a precise and systematic identification of the processes that account for the heterogeneity of outcomes among offspring of women with CM histories, will not only open new avenues for research identifying factors that may protect or interrupt potential links between maternal CM and offspring psychopathology, but also provide more clear road maps for future intervention development. A major area of innovation includes the use of a rich longitudinal dataset that has a comprehensive array of biological, neurocognitive, behavioral, caregiving, and ecological measures from age 6-months – age 16, allowing us perhaps for the first time, to examine the intergenerational effects of CM. To our knowledge this is the first study of its kind to test complex longitudinal relations among early adversity, deprivation, fear learning, stress physiology, hypervigilance, and key outcomes in a racially and economically diverse sample. The high prevalence rates for CM in the United States, suggests a significant number of children are living in homes with mothers reporting trauma histories making this an important direction for further inquiry.

NIH Research Projects · FY 2025 · 2024-08

Project Summary Heart attacks, or myocardial infarctions (MI), remain a leading cause of death worldwide. As adult human hearts have limited regenerative capacity, cardiomyocytes (CMs) lost after MI are replaced by fibrotic scar tissue, leading to cardiac dysfunction and often heart failure. A promising approach to replace lost CMs is direct reprogramming of cardiac fibroblasts (CFs) into induced cardiomyocyte-like cells (iCMs) using the specific cardiac lineage transcription factors Mef2c, Gata4 and Tbx5 (MGT). Over the last decade, considerable progress has been made in understanding the transcriptional and epigenetic regulation of iCM generation. However, post- transcriptional processes, including translation and mRNA modification, are also important to regulate gene expression. Post-transcriptional regulators have recently been shown to play critical roles in cell fate determination, but their roles in direct cardiac reprogramming are largely unknown. Recently, Dr. Xie identified the RNA binding protein Ybx1 and the m6a reader Igf2bp1 as critical post-transcriptional barriers to iCM induction. Excitingly, upon removing Ybx1, a single factor, Tbx5, could successfully reprogram fibroblasts into iCMs. Thus, in Aim 1, Dr. Xie will use ChIP-seq and ATAC-seq to characterize the mechanisms by which the innovative cocktail Tbx5+siYbx1 generates iCMs. Then, taking advantage of the reduced cargo size of this cocktail, Dr. Xie will develop a clinically relevant lipid nanoparticle-mRNA delivery system to achieve in vivo iCM conversion in a mouse MI model. Post-transcriptional mRNA m6a methylation is another regulator of gene expression important in cell fate maintenance. In addition to serving as a barrier to cardiac reprogramming, Dr. Xie found that the m6a binding protein, Igf2bp1, formed cellular granules in CFs. She, therefore, hypothesizes that Igf2bp1 sequesters or otherwise regulates the stability of m6a mRNAs in CFs by regulating cellular RNP granule dynamics and that loss of Igf2bp1 leads to a shift in mRNA metabolism promoting cardiac fate conversion. To test these hypotheses, aim 2 is to identify m6a alternations associated with iCM generation, map precise RNA binding sites of Igf2bp1 and elucidate the underlying mechanism through which Igf2bp1-mediated cellular granule formation regulates the induction of iCMs. The proposed work will not only fill critical gaps in our mechanistic understanding of how post-transcriptional regulation impacts cardiac fate conversion, but also advance iCM reprogramming closer to therapeutic application through the development of an LNP-mRNA delivery system. With an excellent mentoring team and a stimulating research environment at UNC Chapel Hill, Dr. Xie will learn cutting-edge techniques and hone her professional skills, which will provide a solid foundation for her future research in studying the mechanisms of cardiovascular disease and to use insights from this work to develop novel therapies to improve patients’ lives. This K99/R00 award will support the development of her unique and comprehensive research program and catalyze the transition to independence.