Univ Of North Carolina Chapel Hill

NIH Research Projects · FY 2026 · 2023-05

Technologies that can closely monitor surgical recovery and wound healing for timely, proactive treatments represent an essential keystone to developing next-generation personalized medicine that can further reduce patient pain, prevent morbidity and death, and improve individual wellbeing. Microsurgical tissue transfer entails surgical elevation of a portion of tissue (or flap) based upon its defined vascular supply in the form of a single artery and vein. While this reconstructive strategy is well-accepted, failures do occur and almost always result from early microvascular thrombosis. This flap-threatening event occurs in 6-14% of cases, and if untreated flap necrosis and reconstructive failure are inevitable. The most common flap monitoring strategies is serial physical examination and external doppler examination. However, these strartegies are limited by its inherently subjective nature and the requirement for skilled bedside personnel to check the flap frequently. And the intermittent assessment is subject to delay in the diagnosis of malperfusion, since clear signs of malperfusion may take several hours to become obvious. Recent developments in wearable electronic sensors with built-in systems on chip enable opportunities for real-time monitoring of physiological conditions of targeted tissues. However, wearable biosensors that feature skin-interface pose a challenge: to sense physiological parameters such as oxygenation of tissue microenvironments at depth. In the case of flap monitoring, existing devices such as ViOptix are only able to monitor flaps which bear a cutaneous skin. This deficiency means that muscle flaps must be monitored with indirect sensing technology through neighboring skin, which is predisposed to delay recognition of muscle malperfusion. This absence of direct, real-time monitoring technology for muscle-only flaps gives rise to the fundamental and overarching unmet clinical need: to advance technological platforms for deep-tissue monitoring. We propose a soft wearable intelligent patch (SWIP) that incorporates microneedle waveguides to enable deep-tissue sensing of oxygenation without implantation procedures for continuous monitoring of recovery after microsurgical tissue transfer. We aim for the proposed device to enable physiological measurements from 4 different locations of skin to yield both local (tissue oxygenation, pulsation intensity, and blood flow rate) and global (pulsation rate and respiration rate) physiological information continuously and simultaneously. The sensing interface will rely on biocompatible, optical waveguides in the form of microneedles to enable light-matter interaction at deep tissue (~ 2 cm below the skin surface). The device will be equipped with a control module that provides a series of signal pre-processing and a Bluetooth Low Energy (BLE) interface to advertise the data for further processing by a cloud-based computing device. We envision that the proposed SWIP will advance diagnostic technology for reconstructive surgery and beyond, and offer real-time monitoring to facilitate precise customization and personalization in surgical recovery and rehabilitation.

NIH Research Projects · FY 2026 · 2023-04

The North Carolina Translational and Clinical Sciences Institute (TraCS) is a dynamic regional network of universities, research institutes, healthcare providers, and >250 community collaborators across our state. Based at UNC Chapel Hill, TraCS has strategically fostered partnerships with NC A&T, which brings expertise in workforce development, engineering, and data science, and NC State, with a nationally ranked school of veterinary medicine and additional expertise in engineering and computer science. Over the next seven years, we will catalyze development, testing, implementation, and dissemination of translational science and research to improve human health, by completing our Overall Aims. Aim 1: Perform and support research that advances the science of translation with the goal of clearing barriers to rapid translation of research towards implementation of best evidence to our patients and their communities. Aim 2: Maximize the informativeness of translational research, to drive its impact to improve clinical benefits and reduce harms. Aim 3: Train and maintain an expert, multidisciplinary translational research workforce at all professional levels, prepared to meet the challenges of current and future health crises. Aim 4: Implement evidence-based strategies to improve human health using our learning health care system as our laboratory. We have intensified our efforts in dissemination and implementation, recognizing that this is a critical barrier to effective translation of innovation to health. We will capitalize on (i) the substantial resources of our learning health care system and (ii) the collective expertise of TraCS, honed over 14 years of supporting world-class translational research, to realize our vision of “A healthier North Carolina through innovation”. We will innovate methods and processes, engender trust, and recruit research participants to allow participation in research for all residents of our state. We will implement our advances to ensure benefits for our participants, their communities and patients everywhere. With broad stakeholder engagement and rigorous evaluation of our efforts, we will maximize the relevance of our research to those we serve. TraCS will lead and support efforts across the CTSA consortium to create a more efficient research environment.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY Breast cancer is now the most prevalent cancer, with 2.3 million women diagnosed and 685,000 deaths globally. Although, the death rate has declined by about 1% per year over the last decade due in large part to earlier detection through screening, the current screening standard, digital mammography, lacks sensitivity and is challenged in radiographically dense breasts. Sensitivity in dense breasts is improved by magnetic resonance imaging (MRI), but required contrast enhancement and substantial additional cost are drawbacks. A non-contrast, low-cost alternative, ultrasound strain elastography (SE) has been shown to achieve high sensitivity and specificity for breast cancer diagnosis. However, while SE diagnoses suspicious breast massed by interrogating tissue stiffness, the underlying cause of stiffening is not discerned. Among the many possible causes, stromal collagen organization is newly a focus of intense interest due to its recently proven association with breast cancer stage, prognosis, treatment response, and other clinical features. Therefore, delineating stromal collagen organization in vivo would not only improve breast cancer diagnosis and treatment management but also help to elucidate the complex and poorly understood pathophysiology of breast cancer development, progression, and aggressiveness. One approach to noninvasively evaluating stromal collagen organization in the breast is ultrasound shear wave elastography (SWE), but shear wave propagation is inhibited in 63% of malignant breast masses, leading to low-quality measurements. The lack of a robust ultrasound approach to interrogating stromal collagen organization represents a major gap in exploiting a rapidly emerging and highly promising biomarker for breast cancer. To fill this gap, our group has developed Viscoelastic Response (VisR) ultrasound, an on-axis approach to interrogating collagen fiber organization via stiffness anisotropy in breast masses and surrounding stroma without observing shear wave propagation. Our preliminary in vivo data, acquired in 30 women with BIRADS-4 or -5 masses, demonstrate that VisR-derived ratio of mass-to-surrounding tissue stiffness anisotropy (RMSA) differentiated biopsy-confirmed malignant (n=9) from benign (n=21) breast masses with a sensitivity of 95% and a specificity of 89%. Further, in our pilot feasibility study of four women monitored serially while undergoing neoadjuvant chemotherapy (NAC), VisR RMSA trended from malignant to benign indication with response to treatment. The success of our investigations motivates further advancement of VisR RMSA measurement technology, its extension to revealing the relationship between VisR RMSA outcomes and stromal collagen organization, and broader application to cancer detection and treatment monitoring. We hypothesize: Advanced VisR RMSA correlates to stromal collagen fiber organization, which is relevant to differentiating malignant from benign breast masses and predicting pathologic complete response (pCR) to NAC in women, in vivo.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY/ABSTRACT Reducing polypharmacy by discontinuing medications with reduced benefits and increased risks is a priority in older adults (OAs) with Alzheimer’s Disease and Related Dementias (ADRD). Low-dose aspirin for primary or secondary prevention of atherosclerotic cardiovascular disease (ASCVD) has been proposed as a target for discontinuation, with limited real-world data suggesting high variation in prescribing and discontinuation. Guidelines recommend against aspirin for primary prevention in OAs, while supporting its use for secondary prevention; however, applicability to OAs with ADRD is questionable. Reduced life expectancy for OAs with ADRD may translate into lower long-term ASCVD benefits, while increased potential for drug interactions may increase short-term bleeding risk. On the other hand, higher ASCVD risk among OAs with ADRD may position them for greater risk of ASCVD events previously observed in other populations in weeks after discontinuing aspirin, and the anti-inflammatory and anti-thrombotic effects of aspirin could also protect against further progression of cognitive or functional decline. The exclusion of OAs with ADRD from randomized trials of aspirin and limited availability of observational data on aspirin use (a non-prescription drug) leaves patients, providers, and caregivers with little evidence about benefits and harms to guide informed decisions about aspirin discontinuation. Our long-term goal is to improve decision-making, care quality, and outcomes for OAs with ADRD, through improved evidence and treatment guidelines about medications optimization as ADRD progresses. The proposed retrospective cohort study will use records on daily aspirin use uniquely available for a national cohort of Veterans Affairs (VA) nursing home (NH) residents with ADRD, linked to Minimum Data Set (MDS) assessments, electronic health records, and VA and Medicare utilization data over 2016-2023. Specific aims are to (1) Identify clinical and socio-environmental factors predicting aspirin discontinuation in OAs with ADRD after NH admission, stratified by ASCVD status; (2) Examine effects of discontinuing aspirin on ASCVD events, major bleeding, emergency department/hospital admissions, and mortality, stratified by ASCVD status; and (3) Examine effects of discontinuing aspirin on cognitive function, functional dependence, and behavioral/psychological symptoms of dementia. To focus our aims on generating robust, clinically- and policy-relevant evidence, we will use pharmacoepidemiologic methods to reduce selection bias and confounding. Aim 2 is a prevalent new-user study applying covariate balancing methods and competing risk models, with supplementary analyses to address time-varying aspirin exposure and confounders. Aim 3 will assess time-varying exposures and confounding using inverse-probability-weighted marginal structural models. This study will inform future practice guidelines to address if aspirin can be safely discontinued in OAs with ADRD and empower patients, caregivers, and providers to make informed decisions.

NIH Research Projects · FY 2026 · 2023-04

Abstract Osteoarthritis (OA) is the most prevalent degenerative disease in older adults with the incidence rising rapidly after age 50 and leveling off after age 70. OA is also one of the common causes of chronic pain and the leading cause of physical disability in older adults. Currently, there is an unmet need for therapeutic strategies to improve the outcome for patients with OA. Our latest work identifies a list of microRNAs (miRNAs) in human cartilage and demonstrates a strong association with a robust anabolic effect. This effect is joint-specific and follows a distal-proximal axis gradient (high in ankle and low in hip). Studies show that a joint's identity is maintained by synovial cells and that there is a distinct miRNA profile in different joints. Together, this suggests that the miRNAs we identified in cartilage may originate from synovium and be involved in maintaining joint homeostasis. In Aim 1, I will determine the synovial cell types that express these regenerative miRNAs within human joints and the effects of age on the expression of these miRNAs. In Aim 2, I will determine the signaling pathways responsible for the miRNA-mediated anabolic effects in cartilage and the effects of age on these pathways. I will conduct gene set enrichment analysis to determine miRNA-mediated pathways and then use proteomics to validate these pathways. Through this project, I will determine the miRNA-mediated mechanisms by which synovial cells promote endogenous anabolic effects in the human joint. The key career enhancement of this award will be the training in computational bioinformatics to analyze the complex datasets generated by the project, and further training in aging biology to understand how aging impacts the regeneration process. To facilitate progress toward independence, the training plan will include the coursework/workshops in computational bioinformatics and aging biology, extensive internal and external scientific meetings, and career professional development activities and mentorship. The research and career development plan detailed in this proposal will be conducted with a team of outstanding mentors. Dr. Yi-Ju Li, a professor of Biostatistics & Bioinformatics and an expert in statistics and bioinformatics, will serve as the primary mentor and focus on the training in bioinformatics, statistics, and professional skill development. Drs. Cathleen Colón -Emeric, Virginia Kraus (Duke), and Patrik Önnerfjord (Lund University, Sweden) will serve as co-mentors; they will facilitate training in translational aging research, OA research, and proteomics, respectively. The environment at the Duke University and Duke Molecular Physiology Institute, where the main research activities are located, are ideal for the research and training activities outlined in this proposal. This award will enable me to elucidate the novel contributions of miRNAs to joint tissue homeostasis. Advancements in this area of research have the potential to develop as new therapeutic strategies aimed at improving the quality of life for patients with OA.

NIH Research Projects · FY 2026 · 2023-04

Lung cancer is the leading cause of cancer-related deaths in the U.S., accounting for about 132,000 deaths per year. While targeted therapies of lung adenocarcinoma have improved overall survival, similar advances in lung squamous carcinoma (LUSC) have been stagnant. Extensive molecular profiling through the Cancer Genome Atlas (TCGA) effort revealed that LUSC tumors are highly idiosyncratic and rarely driven by solitary actionable pathways. Also, metastasis, rather than primary tumors, is responsible for the majority of cancer-related deaths. However, the mechanistic underpinnings of how LUSC spreads are very poorly understood. In this proposal, we will investigate the roles of a circle RNA (circRNA), CDR1as, on its regulation of LUSC metastasis. To investigate the regulatory role of CDR1as on LUSC metastasis, we have recently developed sophisticated LUSC models that metastasize to sites common to human disease. By integrating clinical LUSC TCGA data with our mouse models, we have identified CDR1as as a key driver of LUSC metastasis. We have found that CDR1as plays a key role in stabilizing the coding mRNA transcript for cerebellar degeneration-related protein 1 (CDR1). We found that CDR1as and CDR1 are each necessary for LUSC metastasis, and CDR1 over-expression alone is sufficient to promote LUSC metastasis. We found CDR1 interacts with several specific Golgi trafficking proteins, and CDR1 expression corresponds with poor LUSC survival and tightly couples with an epithelial-mesenchymal transition (EMT) program. Additionally, we have found key structural elements of CDR1as that promote its RNA stability and expression levels. Taken together, key questions arise, such as: 1) How does CDR1 trafficking in the Golgi vesicles promote migration and metastasis? 2) Can oligo-mediated targeting of CDR1as block LUSC metastasis and prolong survival? The objectives of this proposal are to define how CDR1 promotes LUSC metastasis through Golgi trafficking. We will also determine the biologic and therapeutic implications of interrupting structural elements of CDR1as.

NIH Research Projects · FY 2026 · 2023-04

Much of the basic research on cardiac biology has been focused on cardiomyocytes (CMs), aiming to unravel the basic principles underlying cardiac physiology and pathophysiology for future development of therapeutic interventions to treat cardiac diseases. Besides CMs, the heart contains many other cell types including endothelial cells, fibroblasts, and a wide variety of immune cells. During heart development and homeostasis, non-myocytes (nonCMs) have been increasingly recognized to play active roles in regulating various CM behaviors. Yet a lack of detailed information on the cellular identities and cell states of the nonCMs associated development and homeostasis is a major hurdle to precisely delineating the biological events in heart development and homeostasis. In our recently published study, we delineated nonCMs cellular and transcriptomic dynamics during cardiac regeneration. Through scRNA-seq, we identified major nonCM cell types, including multiple macrophage (MC), FB and EC subpopulations with unique tempo-spatial distribution. Prticularly, we found that MC exists in multiple definable states that exhibit dynamic functional changes from acute inflammatory response to inflammation resolution. Interestingly, perturbing MC function resulted in defective cardiac regeneration. Combining Topologizer and RNA velocity analyses, we uncovered dynamic transition between MC functional states and identified factors involved in mRNA processing and transcriptional regulation associated with the transition. However, whether and how nonCMs interact at the subpopulation level, and MC dynamic functional change affects nonCMs interactions and hence heart regeneration remains largely unexplored. In this research proposal, we hypothesize that heart regeneration is a highly orchestrated process involving temporally regulated MC function executed by the distinct subtypes and their interactions with other nonCM cell types. To test this hypothesis, we aim to 1) define the role of the inflammatory MCs (iMCs) subtype in nonCM interaction and heart regeneration, 2) delineate the role of the immune surveillance MC (isMCs) subtype in nonCM interaction and heart regeneration, and 3) study the molecular mechanism governing transition of MC functional states.

NIH Research Projects · FY 2026 · 2023-04

Project Summary Alcohol (ethanol) dependence and relapse in abstinent alcoholics are major health problems throughout the world and neurochemical pathways that modulate these disorders are currently under investigation. However, the neurobiology underlying binge drinking, a dangerous pattern of behavior that proceeds and contributes to dependence, has received far less attention. Thus, it is of paramount importance to identify the neurocircuitry in the brain that modulates binge drinking as such knowledge will provide insight into the initial stages of alcohol use disorders (AUDs). In the last funding period of this grant we showed that NPY signaling in regions of the extended amygdala modulate binge-like ethanol drinking via mechanisms that overlap with NPY mechanisms involved with dependence-like intake. The goal of the present application is to extend our understanding of the NPY neurocircuitry that modulates binge-like ethanol drinking with a novel focus on NPY neurocircuitry originating in the medial prefrontal cortex (mPFC). The mPFC provides top-down regulation of the extended amygdala, in part through a glutamatergic mPFC  basolateral amygdala (BLA) circuit that is modulated by Y1 receptor (Y1R) signaling. Knowledge obtained from the proposed studies will greatly expand our understanding of the NPY neurocircuitry that modulates binge-like ethanol intake by linking together cortical and extended amygdala circuitry. The mPFC integrates information from limbic and cortical regions and has been implicated in modulating goal-directed (via prelimbic (PL) circuits) and habitual (via infralimbic (IL) circuits) seeking of ethanol and drugs of abuse, as well as anxiety and fear learning. Recent evidence has revealed a critical role for NPY signaling in the mPFC, and we have provided strong pilot evidence showing that infusion of an Y1R agonist into the PL, but not IL, region of the mPFC blunts early-experience binge drinking, and that chemogenetic silencing of Y1R+ neurons originating from the PL and projecting to the BLA blunt early-experience binge intake. The proposed aims will use powerful and innovative chemogenetic and transgenic tools, electrophysiology, and histological approaches to determine if a history of binge-like ethanol drinking induces alterations of NPY and NPY receptor signaling in the mPFC (Aim 1), if site-directed infusion of a Y1R agonist or Y2R antagonist, or viral-mediated overexpression of NPY in the mPFC blunts binge-like ethanol drinking (Aim 2), and if chemogenetic silencing of Y1R+ pyramidal neurons of the mPFC and which project to the BLA blunt binge-like ethanol intake (Aim 3). Together results from these proposed studies will address a critical gap in the literature regarding the role of the mPFC, NPY signaling in this region, and the functional interaction between the mPFC and extended amygdala in the modulation of binge-like ethanol intake in mice.

NIH Research Projects · FY 2026 · 2023-04

PROJECT ABSTRACT/SUMMARY Survivors of adolescent and young adult cancers (AYAs) are a vulnerable and underserved subgroup of survivors at increased risk for long-term health effects, including obesity, diabetes, cardiovascular disease, additional cancers, and frailty. Furthermore, over half of AYAs already have overweight or obesity and obesogenic lifestyle behaviors are common among AYAs, which exacerbates their cardiometabolic risk. Given that obesity is associated with poorer outcomes in cancer survivors, there is a critical need for weight management interventions—yet, no weight loss interventions have been developed to meet the unique needs of AYAs. Extant research, including our own preliminary data, indicate that AYAs prefer tailored interventions (both with respect to developmental stage and cancer context) that are delivered remotely via website or app. Thus, we propose to develop a 6-month mHealth weight management intervention designed specifically for AYA cancer survivors and test its efficacy in a randomized controlled trial. The intervention will be rooted in self-determination theory in order to bolster intrinsic motivation for health behavior change by enhancing perceived competence, relatedness, and autonomy. It will include evidence-based behavioral weight loss strategies adapted for AYA survivors (e.g., simplified dietary self-monitoring, adaptive goal-setting, tailored feedback on progress) as well as personalized dietary and physical activity goals. The intervention will be paired with digital monitoring tools (wireless scale and activity tracker), as well as access to a closed Facebook group to foster peer support. AYA survivors nationwide (N=240, diagnosed between ages 15-39 [current age 18-39], posttreatment, body mass index [BMI] 25-50kg/m2) will be randomized to one of two arms: 1) mHealth intervention as described above, or 2) self-guided (digital tools + health education + Facebook). Randomization will be stratified by BMI, sex, and race/ethnicity. Assessments will occur at 0, 3, 6, and 12 months. Percent weight change at 6 months (primary outcome) will be assessed using a remote collection protocol via video weigh-in and wireless scale to facilitate enhanced reach across the US. Secondary outcomes in the full sample include frailty (frailty index), objectively measured physical activity (ActiGraph GT9X Link), dietary intake (ASA24), and quality of life (SF-36), as well as questionnaires assessing hypothesized psychosocial mediators targeted by the intervention. A subsample of participants (n=80) will complete in-person visits at each clinical site at 0, 6, and 12 months to assess changes in body composition, waist circumference, frailty, and biomarkers of aging and cardiometabolic disease. We hypothesize that compared to the self-guided arm, AYAs in the intervention arm will manifest greater percent weight loss at 6 months and better maintenance of weight loss from 6 to 12 months. We also will examine the psychological and behavioral mechanisms of action to inform future optimization efforts, and explore demographic and clinical- related moderators of intervention response. Our findings will accelerate the development of effective remotely- delivered mHealth weight loss interventions to improve outcomes and reduce the burden of morbidity in AYAs.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY/ABSTRACT The Inflammatory bowel diseases (IBD), Crohn’s disease (CD) and ulcerative colitis (UC), are chronic inflammatory diseases of the gastrointestinal tract with no cure. Genome-wide association studies (GWAS) have found >250 genomic loci associated with IBD, but variant contributions to mechanisms driving IBD pathogenesis and disease prognosis remain unclear. Each locus typically contains tens to hundreds of variants, the vast majority of which are in non-coding regions suggesting a role in gene regulation. For most loci, the causal variant, the affected regulatory element, and the target gene being regulated are unknown. We hypothesize that regulatory variants contribute to IBD phenotypes by altering gene transcriptional programs driving phenotypic heterogeneity. We propose to identify putative casual regulatory variants using two, orthogonal, high-throughput analyses. Aim 1: Regulatory quantitative trait loci (QTL) for chromatin accessibility (caQTL) and transcription factor binding (tfQTL) associate genetic variation with alterations in regulatory activity. For variants in GWAS loci, these analyses will identify regulatory variants with potential contributions to regulation in disease-relevant cell types and tissues. Aim 2: Alternatively, massively parallel reporter assays (MPRA) systematically interrogate allelic effects on transcriptional regulation of thousands of genetic variants. MPRA using vectors of human DNA regulatory elements containing IBD associated variants of interest can be performed in mouse cells or organs due to the well-established conservation of transcription factor motifs between human and mouse. We will use MPRA to determine variants that alter regulatory activity in colon, ileum, and mesenteric lymph nodes under both normal and LPS-stimulated inflammatory states. Aim 3: Integrating results from QTL and MPRA assays, we will select high confidence putative IBD regulatory variants for intestinal epithelial cell focused functional validation in patient derived 2D intestinal monolayer systems. The long-term goals of this project are: 1) To fill the gap between our ability to detect genetic, gene regulatory, and gene expression variation linked to IBD and our ability to explain how that variation ultimately contributes to IBD; and 2) To provide a unique data resource for IBD investigators to access for their own studies.

NIH Research Projects · FY 2026 · 2023-04

PROJECT SUMMARY/ABSTRACT Physical activity is a protective factor of Alzheimer's disease and related dementias. While there are many benefits to physical activity, adults with intellectual disability (ID) often do not engage in healthy physical activity levels. With higher rates of early-onset dementias and age-related conditions in adults with ID, developing improved strategies and programs that promote physical activity in this population is critical. Barriers to physical activity include a lack of inclusive physical activity opportunities, access to community exercise facilities, and reliance on external supports. The purpose of this NIH Stage 1 mixed methods study is to test the feasibility, acceptability, fidelity, and prepare for a Stage 2 efficacy study of a multi-level physical activity intervention. Step It Up+ is a 16-week multi-level program that utilizes self-management strategies, including goal setting, self-monitoring, and visual supports to help adults with ID engage in the program more independently through an interactive web-based dashboard. Adults with ID work with a support coach to increase daily step counts and time spent in moderate-to-vigorous physical activity by engaging in individualized aerobic and strength training activities. Aim 1 is to modify, adapt, and refine our existing Step It Up home-based program at the individual level of the socioecological model to include the community level through a weekly inclusive group fitness class (Stage 1A). This phase will include usability testing, pilot testing of the RCT assessment battery, and semi-structured interviews with adults with ID and surveys and focus groups with additional stakeholders (parents, caregivers, and exercise professionals) to demonstrate the features and functionality of the interactive web-based dashboard. Aim 2 is to conduct a pilot randomized controlled trial (Stage 1B). This phase will use a waitlist control experimental design with 120 adults with ID in two sites (North Carolina and Texas). Feasibility will be assessed quantitatively by measuring recruitment, attendance, and completion rates. Qualitative data from focus groups and interviews will augment the feasibility quantitative findings and provide feedback on consumer acceptability. Content experts will evaluate treatment fidelity by reviewing a subsample of video-taped sessions. We will measure the impact of Step It Up+ on increasing physical activity and explore hypotheses regarding the potential efficacy of the intervention's effect on physical activity. We will compare primary outcomes of physical activity using daily step counts, activity intensity minutes, body composition, and secondary outcomes of wellbeing, functioning, and social belonging. The findings from this project will lay the essential groundwork for a large-scale, for an R01 to conduct a Stage 2 primary prevention trial in adults with ID who are pre-symptomatic of AD/ADRD. Our long- term goal is to promote healthy physical activity behaviors through inclusive community settings, examine the long-term impact of these programs in preventing Alzheimer's disease and related dementia in aging adults with ID, and improve wellbeing in adults with ID.

NIH Research Projects · FY 2026 · 2023-04

Project Summary/Abstract Many individuals with monogenic and idiopathic forms of autism spectrum disorder (ASD) exhibit brain enlargement early in life. However, the underlying cellular and molecular mechanisms leading to early brain overgrowth in ASD are unknown. To identify the mechanisms leading to brain overgrowth, we will use an appropriate model system, iPSC-derived organoids, from a well-powered, deeply phenotyped cohort with multiple control groups, the Infant Brain Imaging Study (IBIS). IBIS is the largest longitudinal neuroimaging study of infants (>250 participants) at high familial risk for autism by virtue of having an older sibling/proband with ASD. Importantly, IBIS participants have previously undergone longitudinal neuroimaging at multiple time points in infancy (between 6-24 months of age) and school age, extensive behavioral assessments at these time points, as well as rare and common variant genotyping. The extensive phenotypic data generated in this cohort make it an ideal population from which to generate iPSC-derived organoid models and relate in vitro phenotypes to in vivo brain growth and behavioral trajectories. Our study also represents a unique opportunity to evaluate how well organoid phenotypes model the in vivo brain growth trajectories of the individual from whom they were derived. In this proposal, we will derive and validate iPSCs from blood for participants from high risk families who developed ASD (HR+), high risk participants who did not develop ASD (HR-), and low risk individuals without ASD (LR-) totaling 99 participants. We will differentiate the iPSC lines to cortical organoids to model inter- individual differences in brain development. We will use single cell (sc)RNA-seq to identify cell types, cell cycle states, and differentiation trajectories in each participant-derived organoid across two time points modeling the period of cortical neurogenesis, totaling 2.38M sequenced cells. We will validate cell type counts and states using tissue clearing followed by lightsheet microscopy of the cortical organoids. We will identify cell types, fate decisions, and cell cycle states that correlate with both cross-sectional and longitudinal cortical surface area growth and ASD symptoms and cognitive ability over time. Leveraging this unique, deeply characterized clinical cohort, we will determine both the in vivo relevance of cortical organoids and the cellular and molecular mechanisms underlying brain overgrowth in ASD.

NIH Research Projects · FY 2026 · 2023-04

Project Summary One of the underexplored aspects of neuronal biology is that as postmitotic neurons become mature, they undergo dynamic changes to ensure that the mature nervous system is capable of long-term survival and function. Understanding these mechanisms that are critical for the long-term homeostasis of the adult brain is important as their dysfunction could increase the vulnerability of neurons to age-related neurodegeneration, such as Alzheimer’s disease (AD). We have identified miR-29 as a microRNA that is strikingly induced during brain maturation. In contrast to the high miR-29 levels that are maintained in the normal adult brains, miR-29 levels are reduced in Alzheimer’s Disease patients. miR-29 is recognized to target many of the genes in the AD pathways including BACE1, NAV3, and IFITM3. To evaluate the functional importance of miR-29, we generated mice in which miR-29 can be conditionally deleted. Mice deficient for miR-29 in the brain are born normal but then progressively decline, exhibiting neurological defects and early lethality. These results show that miR-29 is physiologically important for the maintenance of long-term homeostasis in the adult brain. Reduction in miR-29 levels could therefore increase the vulnerability of mature neurons to become dysfunctional in the context of AD. We have recently generated mice in which miR-29 levels can be conditionally reduced in the adult brain. Thus, the overall focus of our proposal is to define the consequences of miR-29 reduction in the adult brain and to evaluate the therapeutic potential of miR-29 for AD. Specifically, in Aim 1, we will conduct single-cell RNA- seq analysis to identify the specific cell types that are most impacted by miR-29 reduction in the adult brain. We will also examine if miR-29 reduction causes changes in dendritic spine morphology and neuronal arborization in the adult brain. Our results have revealed that an essential function of miR-29 is to regulate non-canonical (non-CG), CH methylation in the mature brain via its targeting of the methyltransferase Dnmt3a, where the loss of miR-29 results in CH hypermethylation and reprogramming of gene expression. In Aim 2, we will examine if CH hypermethylation is a common feature of AD in three distinct mouse models of AD and AD patient brain samples. Our hypothesis is that restoring miR-29 levels would have therapeutic benefit for AD. Thus, in Aim 3, we will examine whether AAV-mediated delivery of miR-29 confers benefit in human stem cell and preclinical mouse models of AD. Overall, we are excited to be working on a molecule, miR-29, that has a unique and essential function in the mature brain. Our studies will help define its mechanisms of action as well as evaluate its therapeutic potential in the context of Alzheimer’s Disease.

NIH Research Projects · FY 2026 · 2023-03

ABSTRACT Type III interferons (IFN-λ) play a key role eliciting antiviral immunity at epithelial surfaces, controlling infections locally without the inflammatory immune pathology triggered by the more potent systemic type I IFN (IFN-αβ) response. However, the effects of IFN-λ in the skin have not been extensively investigated. Herpes simplex virus type 1 (HSV-1) infects epithelial cells and establishes a life-long infection in sensory neurons. Using a mouse skin infection model, we found that multiple lines of mice lacking IFN-λ signaling developed more severe HSV-1 skin lesions compared to wild-type mice. However, disease severity was uncoupled from viral loads in the skin, suggesting a role for IFN-λ in suppressing inflammatory immune pathology. We found that IFN-λ signaling in both keratinocytes and leukocytes was necessary for protection from severe skin disease and that Ifnlr1-/- mice exhibited greater neutrophil infiltration into HSV-1 skin lesions compared to WT mice. We hypothesize that IFN- λ protects against inflammatory pathology during HSV-1 infection by suppressing neutrophil recruitment and activation. The greatest burden of HSV-1 disease in humans results from reactivation of lifelong latent infections, rather than new infections. We have developed a simple and tractable mouse model to induce reactivation of latent HSV-1 from dorsal root ganglia which produces recurrent HSV-1 skin lesions. Using this model, we found that Ifnlr1-/- mice develop more severe reactivation disease compared to WT mice. Flaviviruses and other arboviruses are inoculated into the skin by blood-feeding mosquito or tick vectors. We found that IFN-λ signaling reduced viremia caused by Zika virus and Langat virus only when the viruses were inoculated in the skin, not when the skin barrier was bypassed by subcutaneous inoculation. Vector saliva is known to enhance arbovirus pathogenesis in part by recruiting neutrophils, a key IFN-λ responsive cell type. We hypothesize that IFN-λ signaling in the skin restricts flavivirus dissemination and that this effect is greater in the context of vector feeding. Aim 1: Define the mechanism by which IFN-λ restricts acute HSV-1 skin disease. 1A: define the IFN-λ responsive leukocyte populations that limit skin lesion severity. 1B: define the pathogenic immune responses suppressed by IFN-λ in the skin. 1C: investigate how IFN-λ signaling in keratinocytes limits HSV-1 skin disease. Aim 2: Define the mechanism by which IFN-λ restricts recurrent HSV-1 skin disease. 2A: define the role of IFN-λ specifically during HSV-1 recurrent disease. 2B: assess the kinetics of viral replication and skin lesion formation using live animal luminescence imaging. Aim 3: Define skin-specific effects of IFN-λ against mosquito-borne and tick-borne flaviviruses. 3A: define the IFN-λ responsive cell types that restrict replication of mosquito-borne flaviviruses and tick-borne flaviviruses in the skin. 3B: investigate the effect of vector saliva on IFN-λ mediated antiviral immunity in the skin.

NIH Research Projects · FY 2026 · 2023-03

Deborah Stephens, DO is an Associate Professor at the Lineberger Comprehensive Cancer Center (LCCC) at the University of North Carolina (UNC) who focuses on evaluation of novel targeted agents and combination therapies in patients with chronic lymphocytic leukemia (CLL) and B-cell lymphomas with the ultimate goal of improving clinical care and outcomes. The scope of her clinical trials program [including National Cancer Institute (NCI)-sponsored, investigator-initiated, and industry-sponsored trials] is significant as it improves the care not only for local patients but has also produced research outcomes leading to changes in the standard of care for patients. Through several national and local leadership roles, Dr. Stephens dedicates 75% of her time to research. At UNC, she serves in the roles of Director of the CLL and Lymphoma Research Program and as a Co-Program Leader for the UNC LCCC Cancer Therapeutics Program. Specifically, 40% of her total time will continue to focus on NCI-sponsored clinical trials, which is solely supported by the R50 Research Specialist Award. Her R50-related activities include: (1) her role as international principal investigator for the S1925 EVOLVE CLL Trial (NCT04269902; 15% effort); (2) her role as CLL Lead for the SWOG Leukemia Working Group (10% effort), where she organizes a task force of other CLL specialists to develop SWOG CLL clinical trials. In this role, she will also serve as a mentor and co-investigator on S2504 PIRAMID Trial for Richter Transformation (NCT07220187) and is mentoring a junior faculty to open an additional SWOG CLL trial for patients with relapsed disease. She will be author on the recently completed ECOG and Alliance CLL clinical trials (NCT03701282 and NCT03737981). She meets monthly with CLL leaders from ECOG and Alliance to discuss trial issues and plan next studies and participates in bi-annual SWOG meetings; (3) her role as an active member of the SWOG Lymphoma Working Group (10% effort), where she continues to perform research on data from the S1001 (NCT01359592) and S0816 (NCT00822120) clinical trials and her role mentoring Dr. Boyu Hu who is the adult chair for AHOD2131 and will chair S2506; (4) and her role as Experimental Therapeutics Clinical Trials Network (ETCTN) hematology expert for LCCC’s UM1 grant with The Ohio State University as Lead Academic Organization UM1-CA186712 (Carson), where she works alongside Dr. Claire Dees (Medical Oncology) to represent UNC. (Her effort related to UNC’s ETCTN Group is 10%, with this R50 grant to continue funding 5% of this effort.) As such, continued support from this R50 Grant, allows Dr. Stephens to continue her work and plans for meeting the objectives of the NCI-funded clinical trials research program.

NIH Research Projects · FY 2026 · 2023-03

PROJECT ABSTRACT The HIV-1 reservoir is thought to contain cells that vary in their susceptibility to clearance by the host immune system. Identifying mechanisms that allow infected cells to avoid clearance could inform the development of cure strategies that are able to more effectively eliminate HIV-infected cells in the long-lived reservoir. Persistence during long periods of untreated infection, when immune surveillance for HIV-infected cells is highest and the immune environment is highly inflammatory, may select for infected cells with an enhanced ability to evade detection and clearance by the host immune system. This study will focus on two groups of reservoir cells that carry intact proviral genomes and likely differ in susceptibly to clearance. The first are "early" reservoir cells that were infected (or are a clone of a cell infected) early in untreated infection. The second are "late" reservoir cells that were infected (or are a clone of a cell infected) near the time of ART initiation. These "late" reservoir cells have persisted in the reservoir for less time and have primarily persisted during antiretroviral therapy when HIV- specific immune responses are blunted and HIV-infected cells typically have very long half-lives. In contrast, "early" reservoir cells persisted for long periods of untreated infection when most HIV-infected cells have a short half-life. We hypothesize that early proviruses will have epigenetic features that make them resistant to T cell stimulation and less susceptible to clearance. In this study we will identify intact "early" and "late" proviruses in CD4+ T cells isolated from the blood of 15 participants on suppressive ART (Aim 1a). We will then examine phenotypes predicted to impact reservoir clearance. Specifically, we will examine whether "early" and "late" proviruses differ in their sensitivity to T cell stimulation (Aim 1b) and/or have different genetic (proviral sequence and integration site) or epigenetic features (Aim 1c). To test the hypothesis that "early" proviruses are more difficult to clear, we will measure the susceptibility of "early" and "late" proviruses to killing by autologous CD8 T cells (Aim 2a) and assess whether disruption of epigenetic regulators alters their susceptibility to killing by autologous CD8 T cells (Aim 2b). Our focus on "early" and "late" viruses allows us to explore potential mechanisms associated with clearance in populations that likely differ in their susceptibility to clearance, however, the proposal work will also reveal if features other than proviral age are predictive of susceptibility to clearance. We will then explore strategies to improve antigen presentation in infected cells (Aim 3a) and assess whether improved antigen presentation can enhance killing by autologous CD8 T cells in populations typically refractory to clearance (Aim 3b). The overarching goal of this project is to define a population of cells with an enhanced ability to evade clearance, identify host and proviral features associated with clearance evasion and test whether better antigen presentation can make HIV-infected cells more susceptible to clearance. Successful completion of these aims will connect proviral genetics and epigenetics to virus reactivation and killing to inform the development of improved clearance strategies.

NIH Research Projects · FY 2024 · 2023-03

Abstract The majority of validated social-communication measures for children with autism spectrum disorder (ASD) were designed to behaviorally phenotype or diagnose this core symptom domain and not to detect change over time. The lack of validated outcome measures has somewhat stifled intervention research efforts as well as early interventionists’ ability to monitor the effects of their programs on children’s social-communicative outcomes. It has become difficult to sort out whether non-significant treatment effects are a result of truly ineffective interventions, or rather poorly validated measures for this population. We know that the early, foundational social-communication and language skills of children with ASD predict their functioning into adolescence and even adulthood, with functional use of language by age 5 being one of the best predictors of long-term prognosis. As such, clinicians have begun to focus their efforts on improving children’s social- communication and language skills using both developmental and behavioral interventions as well as pharmaceutical treatments. In reviewing these treatment trials, it is clear there is a range of intervention success and even more clear that we lack a consistent set of outcome measures for the social-communication symptom domain. Our group has been working to validate measures of key social-communication and language skills and are able to draw from a wealth of prior psychometric data to adapt and validate a brief observational measure that can be easily used within clinical trials as well as within routine care and practice. We propose to adapt and psychometrically validate a measure of social-communication and language skills for young children, ages 12 – 60 months, preliminarily or formally diagnosed with ASD. Specifically, we plan to validate the Early Communication Indicator for Autism Spectrum Disorder (ECI-ASD) using a robust and representative multisite sample of well-characterized children with ASD (n = 400). The current version of the ECI is norm-referenced and allows for progress monitoring, which means the measure can be used in a formative, data-driven fashion to monitor children’s intervention progress and make changes if children are not improving, but it also can provide summative outcome data. In addition, the current measure can be scored live using a mobile app. Currently, no other current social-communication measure specifically designed for children with ASD easily allows for ongoing intervention progress monitoring, data visualization, and live scoring, which will make the adapted ECI-ASD a unique outcome measurement tool. Further, the ECI-ASD will provide the ability to enter data into an online platform to compare the progress of children with ASD to normative data and this represents a sorely needed and clear innovation above current measurement approaches. The expected deliverable is a novel outcome measure of key social-communication and language skills that is psychometrically sound, minimally burdensome to administer and score, and sensitive to the incremental change expected for young children with ASD.

NIH Research Projects · FY 2026 · 2023-03

PROJECT SUMMARY Maternal duplication of the chromosome 15q11-q13 locus (Dup15q syndrome) is a major genetic cause of autism spectrum disorder. UBE3A is contained within this genetic locus; compelling evidence suggests it is the main driver of Dup15q syndrome pathophysiology. Accordingly, the normalization of UBE3A levels might effectively treat the syndrome, but this has been difficult to test. To allow us to test this idea, we have generated conditional Ube3a-overexpression mice that have construct validity for the principal cytogenetic abnormalities underlying Dup15q syndrome: interstitial chromosome 15 duplication (1 extra copy of Ube3a) and isodicentric chromosome 15 (2 extra copies of Ube3a). Thus, our models will be valuable for determining the extent to which increases in UBE3A protein levels are necessary and sufficient to drive Dup15q-relevant pathophysiology. With these mice, we are also able to genetically normalize Ube3a expression at different ages, allowing us to identify the window during which normalization provides substantial therapeutic benefit. To advance an informed intervention strategy for Dup15q syndrome, we will use our new mouse models to (1) establish the behavioral and physiological consequences of UBE3A overexpression, (2) determine when normalization of UBE3A rescues pathophysiology, and (3) optimize an approach to appropriately knock down UBE3A and correct pathophysiology in Ube3a-overexpression mice.

NIH Research Projects · FY 2025 · 2023-03

ABSTRACT A conserved long non-coding RNA (lncRNA) called Kcnq1ot1 is essential for proper mammalian development. Defects in proper gene silencing by Kcnq1ot1 result in Beckwith-Wiedemann syndrome, a birth defect with a high mortality rate whose prevalence is increased in children conceived by in vitro fertilization. Despite its critical role in development, the mechanisms that Kcnq1ot1 uses to silence genes remain poorly understood. While it is known that Kcnq1ot1 silences genes by recruiting histone-modifying enzymes called the Polycomb Repressive Complexes and G9a, it is not clear how sequence elements in Kcnq1ot1 facilitate these interactions. As one possible clue, another repressive lncRNA that is essential for development, Xist, has been shown to interact with RNA-binding proteins (RBPs), that in turn, recruit the Polycomb silencing enzymes. In my own studies, I have found that many of these same RBPs associate with Kcnq1ot1 at levels significantly higher than with other RNAs in the transcriptome, implying that they too are important for the function of Kcnq1ot1. The objective of my study is to further develop the mechanism of Kcnq1ot1 by identifying functional sequence domains in the transcript, identify the RBPs that bind these regions, and determine their involvement in recruiting the Polycomb and G9a silencing enzymes. If successful, I hope to not only elucidate the mechanism of Kcnq1ot1, which is conserved in mice and humans, but further our general understanding of lncRNA function in early development. Long term, this information could eventually contribute to optimization of in vitro fertilization and discovery of genes implicated in infertility. The activities I propose will provide me with critical training in RNA biology and genomics, molecular biology, developmental biology, teaching and mentorship, oral communication skills and networking, and scientific writing, each of which will be critical for my future career as a principal investigator at a primarily undergraduate institution.

NIH Research Projects · FY 2025 · 2023-03

PROJECT SUMMARY (ABSTRACT) The mu opioid receptor (MOR) is widely expressed throughout the nervous system and mediates both analgesic and addictive effects of opioids. Despite the current opioid crisis in the United States, opioid drugs offer unparalleled analgesic efficacy and are prescribed for a variety of pain conditions. To dissociate opioid analgesic and addictive effects, it is essential to determine which neural circuits and cell types mediate each of these effects. MOR signaling in medial prefrontal cortex (mPFC) is of particular significance due to the mPFC function as a key integrative node in ascending and descending pain circuits and its strong connectivity to subcortical reward circuitry. However, how opioids eventually alter the activity of glutamatergic mPFC output neurons to modulate opioid addiction and pain behaviors is unknown. The main outputs of the cortex include two distinct populations of layer 5 glutamatergic projection neurons: intratelencephalic (IT) and pyramidal tract (PT) neurons. IT neurons predominantly project intracortically and to striatum, while PT neurons project broadly and are poised to directly modulate neural activity throughout the brain. Despite evidence supporting a MOR-dependent role for mPFC in pain and addiction, preliminary data suggest that MOR is not expressed directly on IT or PT cells. This project aims to (1) determine how opioid exposure impacts mPFC IT and PT cell activity in mice and (2) delineate the contribution of mPFC IT and PT cells to opioid addiction and analgesia. In order to determine how opioids modulate IT and PT cell activity, dual-color calcium imaging will be used as a proxy for neuronal activity. Neural dynamics of each population will be characterized at baseline, during acute and chronic morphine exposure through subcutaneously implanted pumps, and during naloxone-precipitated withdrawal. To assess the contribution of IT and PT cells to opioid addiction, mPFC IT or PT cell activity will be chemogenetically suppressed in addiction models including morphine conditioned place preference and oxycodone self- administration. Finally, the contribution of IT and PT cells to morphine analgesia and opioid-induced tolerance and hyperalgesia will be assessed through chemogenetic suppression of mPFC IT or PT cells after acute and chronic morphine exposure in assays for thermal (hotplate) and mechanical (von Frey) pain. This project will establish foundational knowledge in the dissociation of the addictive and analgesic effects of opioids, illuminating targets for nonaddictive pain therapies. Through this research proposal and associated training plan, I will gain excellent training in neuroanatomy, addiction and pain behaviors, and neural dynamics in a supportive training environment at the University of North Carolina MD/PhD Program. This training will provide me with the technical and professional skills necessary to become a leader at an academic medical center and pursue my goals of practicing pain medicine and researching innovative non-addictive therapies for pain as a physician-scientist.

NIH Research Projects · FY 2026 · 2023-03

Psychedelic drugs continue to be used and abused by large numbers of individuals within the US and recently there has been a resurgence of interest in the actions of psychedelic drugs like lysergic acid diethylamide (LSD). It is currently unknown how LSD and related psychedelic drugs exert their actions at the molecular and atomic levels. In this MERIT extension we will elucidate the molecular mechanisms by which LSD and related drugs interact with multiple G protein coupled receptors. We will also determine the potential relevance of differences between human and rodent receptors for the actions of LSD and related drugs in vitro and in vivo. Finally we will examine the effects of LSD and related drugs on the gene transcriptional changes which occur specifically in neurons which express receptor essential for the actions of LSD. Taken together, these new findings will provide molecular and atomic details regading the actions of psychedelic drugs in vitro and in vivo.

NIH Research Projects · FY 2026 · 2023-03

PROJECT SUMMARY Heart Failure has resisted the downward trend in mortality seen in other diseases and new therapies are needed. Heart failure is driven by the hypertrophy and loss of cardiomyocytes, which comprise the bulk of the heart by volume and provide the main contractile force necessary to move blood throughout the body. Over 90% of cardiomyocytes in mammalian species are polyploidal and/or multinucleated. These extra copies of DNA have been studied in the context of the low regenerative capacity of the heart, yet comparatively little effort has focused on other potential adaptive or maladaptive effects of this increased nuclearity. Historically, the large size of the mature cardiomyocyte has precluded its analysis using unbiased single cell RNA sequencing. Research has focused on RNAseq in isolated nuclei, which by its very nature lacks information about nuclearity within the studied hearts. Recently, new technology has allowed for unbiased, high-throughput single cell RNAseq of whole, mature cardiomyocytes. When the results of these single cell studies are compared to the isolated nuclei studies, researchers find that, using the same clustering algorithms, there are approximately twice as many distinct cellular transcriptome clusters as there are distinct nuclear transcriptome clusters. This result suggests the existence of a form of ‘nuclear code’ in which a multinucleated cell’s transcriptome is ‘encoded’ by nuclei drawn from two or more nuclear clusters, a result supported by observations from other multinucleated cell populations. These encoded cells could display differences in contractility, resistance to apoptosis, or even proliferative potential, with significant implications for cardiac function and the development of heart failure. The goal of this Steven I. Katz proposal is to 1) Utilize cutting edge single cell and single nucleus sequencing technology to identify and confirm this nuclear encoding, 2) Explore the spatial distribution of mono and multi- nucleated cells across the heart, and 3) Identify how this encoding responds to genetic and environmental perturbations in healthy and failing hearts. As requested by the mechanism, in this proposal we set forth a plan for an ambitious new direction for our laboratory’s research, built upon rigorous work documented in the literature and supported by collaborators and subject matter experts. The proposal combines bioinformatic, imaging, and next generation sequencing approaches to identify how changes in the nuclear code of multinucleated cardiomyocytes leads to differences in response to cardiac injury and stress. Upon completion, this grant will result in a more complete understanding of the role of multinuclearity and polyploidy in the mammalian heart, with subsequent revelation of numerous new avenues of research for diagnostic and therapeutic approaches to combat heart failure.

NIH Research Projects · FY 2026 · 2023-03

Project summary The incidence of HPV-associated (HPV+) head and neck squamous cell carcinoma (HNSCC) has dramatically increased over the last few decades and continues to rise. Despite the magnitude of this epidemic, mechanisms of HPV-driven carcinogenesis in HPV+ HNSCC have not been thoroughly investigated. Compared to patients with tobacco-associated HNSCC, those with HPV+ HNSCC have increased overall survival and higher response to treatment, which usually consists of chemo- and radiation therapy; however, survivors frequently suffer from treatment’s toxic side effects, such as swallowing and speech dysfunction. In addition, approximately 25% of HPV+ HNSCC patients develop recurrent or metastatic disease, for which there are limited treatment options. A pressing goal in head and neck oncology is to decrease the morbidity of therapy for HPV+ HNSCC through treatment de-escalation. However, biomarkers that identify HPV+ patients with good prognosis, who may be appropriate for de-escalation therapy, are lacking. Using three independent cohorts, we found that constitutively active NF-κB (usually arising from genetic defects in NF-κB regulators, including TRAF3 and CYLD) correlates with survival and should be explored as a prognostic biomarker in HPV+ HNSCC. Our preliminary data suggest that survival benefits of patients, whose tumors harbor overactive NF-κB, are attributed to better tumor response to therapy and that both, inherent NF-κB-driven tumor characteristics (e.g. downregulated expression of oxidative stress response, NRF2 target genes), as well as a distinct tumor microenvironment (e.g. elevated number of tumor infiltrating CD4+ T cells), may contribute to increased sensitivity of NF-κB active tumors to radiation. We previously reported that mutations in TRAF3 and CYLD were associated with a lack of HPV integration, leading us to hypothesize that NF-κB activation may enable cells to maintain HPV episomes. Since the canonical HPV carcinogenesis model depends on HPV integration, we also hypothesize that activation of NF-κB may be critical for an alternative mechanism of HPV carcinogenesis driven by HPV episomal maintenance. To explore our hypothesis, in Specific Aim 1, we will investigate the impact of NF-κB signaling on HPV gene expression and episomal maintenance. In Specific Aim 2, we will explore the significance of NF-κB pathway on cellular proliferation, survival, and cellular transformation in response to HPV. Finally, Specific Aim 3 will explore mechanisms of NF-κB mediated radiation sensitivity in HPV+ HNSCC.

NIH Research Projects · FY 2026 · 2023-03

ABSTRACT Inhibition of oncogenic KRAS is a highly pursued goal in drug discovery efforts, as RAS mutations are found in ~25% of human cancers. One key oncogenic KRAS mutation (KRASG12C) contains a reactive cysteine at a hotspot location, is the fourth most prevalent mutation in KRAS-driven tumors and is found at particularly high frequency (40% of RAS mutations, 13% overall) in non-small cell lung cancer (NSCLC). Excitement has accelerated rapidly around the discovery and application of covalent, Cys12-specific inhibitors of KRASG12C in recent years as these compounds have shown efficacy in advanced clinical trials, with Sotorasib (AMG510, LUMAKRAS™) recently receiving FDA approval for treatment of locally advanced or metastatic NSCLC. However, the kinetic mechanisms underlying the activity of this class of inhibitors remain poorly understood impeding rational drug design efforts. To address this gap in knowledge, we developed a new fluorescence- based approach to kinetically characterize the reactions of the proteins with these acrylamide-based inhibitors. Intriguingly, we find that the two clinical compounds, AMG510 (Amgen) and MRTX849/Adagrasib (Mirati Therapeutics) possess distinctly different kinetic properties, which we propose to investigate in further detail here. Recognizing the oxidative environment promoted by oncogenic KRAS signaling and tumorigenesis, we also evaluated the redox sensitivity and oxidative status in cells and found that Cys12 of KRASG12C is prone to oxidation. Moreover, oxidation at this site prevents inhibitor attachment, suggesting that redox modification of KRASG12C may constitute a mechanism of resistance to AMG510 and other covalent inhibitors. Other unanswered questions remain in the field, including the propensity of the protein-drug adducts to undergo chemical reversibility of the Michael addition reactions through which they bind, and how this could be affected by altered acrylamide “warheads”. Aim 1 proposes structural and kinetic studies combined with computational modeling and molecular dynamics simulations to elucidate the differential mechanisms of KRASG12C inhibitor engagement, inactivation and reversal. As development of treatment resistance remains a significant hurdle for targeted inhibition strategies, Aims 2 and 3 propose to investigate the linkage between the redox sensitivity of KRASG12C and inhibitor efficacy by measuring functional, signaling-relevant outputs for recombinant proteins and biological samples (lung cancer cell lines and patient-derived organoids). For Aim 2, lung cancer cells under variably oxidizing conditions, with and without inhibitor present, will be assessed for KRASG12C modifications and downstream signaling outputs; use of HyPer-DAAO genetic constructs with targeting to the plasma membrane will allow spatiotemporal and dosage control over hydrogen peroxide production within cells, near KRASG12C. In Aim 3, the relationship between tumor redox properties and inhibitor efficacy will be investigated using fresh NSCLC tumor specimens carrying KRASG12C mutations. Cumulatively, the results will provide improved under- standing of the drugs and will serve as a platform for characterizing and developing future direct KRAS inhibitors.

NIH Research Projects · FY 2026 · 2023-02

PROJECT SUMMARY Consistent with the research priorities of the National Institute on Aging, this R01 application will investigate the optimal intervention sequence to achieve weight loss in older adults with obesity and ≥ 2 Medicare-defined multiple chronic conditions (MCC). The growing prevalence of obesity in older adults, particularly in those with common chronic conditions such as diabetes, hypertension, or arthritis, increases the risk of functional decline, nursing home placement, and early mortality. Weight loss interventions can mitigate such adverse outcomes; however, differential response to treatment is often observed due to a patient’s clinical heterogeneity. Clinicians lack guidance on the most effective lifestyle-based intervention, and which intervention to try if the first one fails. An innovative Sequential, Multiple Assignment, Randomized Trial (SMART) design will be conducted to identify optimal intervention approaches for weight loss in older adults with MCC, tailoring strategies for non-responders to weight loss. During the 52-week, two-stage trial, 180 older adults with obesity and MCC will be enrolled to compare two weight loss interventions: a prescriptively focused, medically tailored, weight loss intervention (prescriptive), and a behaviorally focused, health coaching intervention (behavioral). Consistent with a SMART design, at 8-weeks, early non-responders (weight loss of <2.5%) will be randomized to: (a) more sessions of the original assignment; (b) a combination of prescriptive and behavioral interventions; or (c) a switch to a prescriptive, medically tailored strategy (initial, first-line behavioral arm participants) or to a behaviorally focused health coach-delivered strategy (initial prescriptive arm participants). The SMART will enable the identification of the treatment combinations that maximize weight loss at 52-weeks. To this end, the proposal aims to: 1) test the superiority of an initial (first-line) prescriptive or behavioral intervention using an adaptive strategy for early non-responders; 2) assess the patterns of initial weight loss and compare strategies for non-responders; and 3) examine the cost-effectiveness from a societal perspective for maintaining weight loss of the proposed treatment sequences at 78-weeks (26-weeks post-intervention completion). The primary outcome is percent weight loss at 52-weeks; secondary outcomes include global health and physical function, anthropometry, behavioral treatment targets and risk factors, and clinical indices. Based on preliminary data, it is hypothesized that older adults with obesity and MCC will achieve greater weight loss with a prescriptive, medically tailored intervention, and the estimated adaptive intervention strategy tailored to a patient’s characteristics will lead to better outcomes than a fixed intervention. If the trial is successful, the adaptive strategy will be compared to a fixed prescriptive or behavioral strategy in a future comparative effectiveness trial. The proposed approach should benefit patients facing competing and complex medical issues who are underrepresented in clinical trials. This study aligns with the NIH Strategic Plans for Obesity, Nutrition Research, and Precision Health, and is responsive to the Institute of Medicine’s call for telehealth research that may influence policy by advancing health delivery science.