Duke University

NIH Research Projects · FY 2026 · 2024-06

PROJECT SUMMARY Chromatin-associated proteins are a major class of genes mutated in autism spectrum disorder (ASD) and intellectual disability (ID). These genetic data indicate the importance of gene regulation for brain development, however a significant challenge for the field is to determine how disruptions of chromatin regulators may converge on specific biological processes in developing neurons. To address this challenge, here we focus in detail on the biochemical and cellular mechanisms by which ID-associated frameshift mutations in the linker histone H1.4 result in impaired neuronal development. As a histone, H1.4 is a direct component of chromatin; thus, studying how mutations in this gene impair neural development offers the opportunity to gain specific biochemical insight into mechanisms of chromatin regulation in neurons. Rahman Syndrome (RMNS) is a rare, genetic form of ID caused by de novo heterozygous mutations in H1-4, which encodes histone H1.4. All RMNS-associated mutations in H1-4 are small insertions or deletions that create a shared C-terminal frameshift. Genetic data indicate that the mutant protein likely functions in a dominant negative or neomorphic manner to lead to RMNS phenotypes, but the biochemical and cellular consequences of expressing RMNS mutant histone H1.4 are poorly understood. We have found that expressing RMNS mutant histone H1.4 in rat hippocampal neurons leads to the disruption of synaptic gene expression and neuronal firing. We hypothesize that RMNS mutant H1.4 disrupts chromatin architecture in differentiating neurons to impair gene expression programs that are required for synapse development and neuronal function. To determine how the RMNS mutation of histone H1.4 leads to aberrant transcription and to evaluate the consequences for brain development, we will use both biochemical and molecular genetic approaches in the developing mouse brain and in human neurons. In Aim 1, we will build a foundation for these studies by using leading edge proteomic and molecular genetic methods to characterize the expression, regulation, and chromatin distribution of histone H1.4 over the course of neuronal differentiation in the mouse. To determine how RMNS mutations disrupt synaptic gene expression in brain development, in Aim 2 we will use a novel in vivo protein tagging strategy to identify proteins that interact with wildtype versus RMNS mutant H1.4. Finally, to determine how RMNS mutations disrupt chromatin regulation, in Aim 3 we will study generate RMNS mutant H1.4 expressing iPSC- derived neurons for biochemical histone H1 proteomics and gene expression analyses. We will then use a novel method for low-input three-dimensional chromatin conformation capture to test the hypothesis that RMNS mutant histone H1.4 disrupts higher level chromatin architecture. These studies will advance knowledge of the causes of brain developmental abnormalities in RMNS, and they will contribute to understanding of the specific mechanisms of linker histone dependent chromatin regulation that are key for neuronal maturation.

NIH Research Projects · FY 2026 · 2024-06

Abstract During embryonic development each body part is programmed to contain an accurate number and arrangement of cells. This accuracy is achieved through precise integration of cell proliferation with other morphogenetic programs. The overarching goal of this proposal is to reveal the physical principles that ensure accurate control of the cell cycle and morphogenesis during Drosophila embryonic development. We will investigate how the integration of the cell cycle oscillator and cytoskeleton leads to generation of the forces that drive nuclear positioning. We will investigate how cytoplasmic flows are generated by cortical actomyosin contractions to reveal novel and quantitative insights on the physical properties of the cytoplasm. Using several biosensors for the main kinases driving the cell cycle, we will study how these activities are integrated to ensure proper cell cycle control. We will also elucidate how transcriptional regulation of cdc25string ensures precise regulation of the timing of mitosis during gastrulation. Specifically, we will dissect the mechanisms controlling mitotic domains on the dorsal side of the embryos, where they are directly linked to the dynamic of the Decapentaplegic (Dpp) morphogen gradients. Collectively, our experiments will define a quantitative framework elucidating how the cell cycle and morphogenesis are regulated accurately during embryonic development.

NIH Research Projects · FY 2025 · 2024-05

Co-occurring substance use and HIV risk among individuals in correctional settings presents a significant public health challenge. Approximately 17% of incarcerated individuals have an opioid use disorder (OUD), and HIV prevalence is notably higher than in the general population. Evidence-based medical interventions—such as pre-exposure prophylaxis (PrEP) and medications for opioid use disorder (MOUD), including buprenorphine—are available but remain underutilized in these environments. Long-acting injectable (LAI) formulations of both PrEP and buprenorphine, recently approved by the FDA, offer new opportunities for improving treatment continuity and outcomes. This project proposes a hybrid implementation-effectiveness type 2 study to evaluate a co-packaged LAI PrEP + buprenorphine (XR-B) intervention in correctional and post-release settings in Maryland and Washington, DC. Specific objectives include: (1) developing a protocol for LAI PrEP + XR-B delivery; (2) evaluating implementation facilitation strategies; and (3) comparing the effectiveness of LAI PrEP + XR-B to oral PrEP + sublingual buprenorphine (SL-B). The long-term goal is to inform scalable, evidence-based practices for improving health outcomes in high-need populations.

NIH Research Projects · FY 2026 · 2024-05

Statement of Work – Sullenger and White MPIs The overall objective of this project is to develop nucleic acid binding polymers that can scavenge extracellular nucleic acid-containing Damage Associated Molecular Patterns (DAMPs) in the setting of pancreatic cancer therapy to mitigate activation of cancer cell migration and metastasis safely and effectively. Our translational hypothesis is that treatment of pancreatic cancer patients with chemotherapy or surgery induces the release of such nucleic acid-containing DAMPs and that administration of a second-generation nucleic acid-containing DAMP scavenger during such therapy will be particularly impactful as it will safely limit the ability of cancer cells to respond to and mobilize as a result of such proinflammatory treatments. The White and Sullenger labs started to collaboratively explore the ability of NASs to neutralize the downstream TLR mediated and pro-invasive effects of extracellular nucleic acids and nucleic acid-containing (NA) DAMPs to improve PC therapy. As we jointly reported, administration of the NAS PAMAM-G3 dramatically reduced metastases of syngeneic KPC pancreatic cancer cells to the liver of mice while also reducing circulating cfDNA levels. Our published data suggest that: 1] inflammation and the presence of activated inflammatory cells and platelets in the tumor and periphery promote cancer invasion and metastasis and 2] modulating inflammation with a NAS controls cancer invasion and metastasis. Though our initial analyses is quite encouraging, it has a major limitation that needs to be addressed to facilitate translation of this NAS strategy to the clinic. Specifically, the NAS PAMAM-G3 we have used has well known dose limiting toxicities. For this reason, the Sullenger lab has developed a new generation NAS, called PAMAM-G3 50:50, that retains the ability to scavenge nucleic acid DAMPs in vitro and in vivo while greatly reducing NAS- associated toxicity. Moreover, the Sullenger laboratory has recently started to evaluate a cationic cyclodextrin polymer (CDP), that has already been translated into the clinic for siRNA delivery in melanoma cancer patients, for its ability to scavenge DAMPs and limit metastasis in the setting of pancreatic cancer. We believe that both of these agents should have wider therapeutic windows than the first generation NAS, PAMAM-G3. The Sullenger and White labs propose to evaluate the potential of these two second generation NASs and the DAMP scavenging therapeutic approach to set the stage for IND-enabling studies for translation of this innovative strategy into the clinic to attempt to improve the outcomes of pancreatic cancer patients. The Sullenger lab will be responsible for all in vitro NAS biochemistry and nucleic acid DAMP analyses as well as all animal models not involving patient derived xenografts. The White lab will be responsible for all studies involving human samples including animal models that utilize patient derived tumor xenografts. Dr. Sullenger and Dr. White will share all findings and meet via Zoom at least twice per month.

NIH Research Projects · FY 2026 · 2024-05

ABSTRACT By harnessing the power of the immune system to attack cancer cells, adoptive T cell therapy (ACT) holds tremendous potential to overcome the limitations of traditional cancer therapies and provide more effective and personalized treatment options for patients. However, the effectiveness of ACT is hindered by T cell exhaustion and the depletion of stem cell memory T cells (TSCM), highlighting the need to improve T cell phenotypes. In several ACT trials, positive responses were closely linked with specific transcriptomic and epigenetic profiles of CD8 T cell subsets in the manufactured T cell product. Epigenetic reprogramming of exhausted cells has been widely postulated as a promising strategy to improve ACT, but we currently lack the knowledge of which genes and epigenetic programs to manipulate. Our long-term goal is to improve cancer treatment outcomes through epigenetic programming of immune cells. To accomplish this goal, we have assembled an experienced and multidisciplinary team with the relevant expertise to address the critical technological challenges and develop next-generation ACT. The overall objective of this project is to translate epigenome editing technologies to enable next-generation ACT through the programming of critical epigenetic nodes that govern complex T cell phenotypes. The central hypothesis is that discovery and targeted perturbation of critical epigenetic nodes can improve T cell function and tumor control. We will accomplish our objective by 1) prioritizing lead targets via high- throughput epigenetic perturbation of primary human T cells, 2) identifying key transcription factors and noncoding regulatory regions driving T cell exhaustion, and 3) investigating the combined effect of master regulatory factors in complex T cell phenotypes. This proposal is innovative because it translates recent developments in epigenome editing technologies and high-throughput targeted epigenetic screening to develop precision interventions that address a fundamental limitation of effective ACT. Collectively, this work will establish a framework for epigenetic engineering in primary human T cells, providing insights into T cell regulation and advancing the development of an enhanced ACT platform.

NIH Research Projects · FY 2026 · 2024-05

PROJECT SUMMARY RATIONALE: Neurological and psychiatric disorders collectively account for approximately 28% of the global burden of disease, with the highest burden being found in low- and middle-income World Bank countries (LMICs) where there are enormous barriers to healthcare. In this setting, there is a dearth of clinicians and researchers trained in the neurosciences, limiting both access to care and research that could impact outcomes. OBJECTIVES: We propose the development of a brain health training program to increase expert brain health research capacity and to develop valid and culturally relevant instrumentation to objectively measure neurocognitive and neurobehavioral capacity. With this focus, we will create a cadre of neuroscientists having the capacity to conduct research across all major disorders impacting the central nervous system. TRAINING PROGRAM: We have brought together leading experts in the neurosciences from Africa (Uganda, Botswana, Congo, South Africa) and the US to create the Neurocognition and Neurobehavior EXperts and Iest Instrumentation (NEXT) program in Rwanda. The proposed training program involves academic instruction, practical laboratory experiences, and research criteria in three areas: brain health (functional neuroanatomy, assessment, neurodiagnostics, neurorehabilitation), research (research design, responsible conduct of research, neuroethics, statistics), and instrumentation (psychometrics). The NEXT program tracks include a 3-year doctoral-level PhD track (8 students), a 2-year Master's degree track (6 students), and a single course track (up to 80 students). Two trainees will be selected for an intensive post doctoral research fellowship at Emory University. RELEVANCE: The overarching intended outcome of this program is to create the foundation of a sustainable brain health program in Rwanda. This includes the introduction of a neuroscience-based academic track at the University of Rwanda and the creation of a cadre of regional experts to conduct independent research, mentor future clinician-scientists, and guide policy having brain health impacts.

NIH Research Projects · FY 2025 · 2024-05

PROJECT SUMMARY Declines in fluid, speed-dependent cognition (e.g., attention and memory) are a hallmark feature of Alzheimer’s disease (AD), but some degree of decline is also seen even in healthy older adults. Based on magnetic resonance imaging (MRI) studies using diffusion-weighted imaging (DWI), the cortical disconnection theory proposes that normal age-related cognitive decline is at least partly explained by the degradation of white matter pathways connecting distributed brain regions, with greater white matter degradation in AD. An important extension of this theory is to understand how white matter disconnection affects brain function in both healthy aging and AD. However, the examination of age- and AD-related differences in white matter connectivity, and its relation to brain function, is constrained by the relatively low resolution of standard DWI data, which cannot accurately resolve fine-grained white matter regions with crossing fiber bundles or high curvature. To address these limitations, this proposal will capitalize on a high-resolution multi-shot DWI protocol that achieves spatial resolutions ≤ 1 mm3 on clinical 3T MRI scanners. Several studies of rodents and younger adults already suggest that high-resolution DWI estimates white matter structural connectivity more accurately than standard resolution DWI. This proposal will translate this earlier work by examining whether standard (1.5 mm3 voxels; 3.375 𝜇l volume) and high-resolution (1 mm3; 1𝜇l volume) measures of white matter structural connectivity differ in their relations to age and ability to explain age-related differences in cognitive performance (Aim 1) or functional connectivity (Aim 2) in healthy adults across the lifespan (n = 140; ages 18- 80 years). This proposal will also examine the sensitivity of high-resolution DWI to white matter disconnection and aberrant structure-function relations in adults with AD (n = 30; Aim 3). Cognition in the healthy adults will be assessed using 12 tests of memory, executive function, and perceptual-motor speed. Structural (DWI) and functional (resting-state functional MRI) connectivity will be assessed using a graph theoretical approach, providing a novel basis for comparison between these different MRI modalities. Results from this clinically feasible high-resolution DWI protocol will help answer fundamental questions about relations between neurocognitive aging, brain structure, and brain function, which is important as the field is moving toward large- scale, multimodal datasets. Ultimately, this project may identify additional age- and AD-related differences in white matter connectivity that cannot be identified using only standard resolution DWI and may inform future interventions targeting white matter connectivity to slow atypical cognitive decline. The wealth of neuroimaging and aging resources available to the applicant at Duke University Medical Center will help her establish an independent research program focused on multiple cognitive domains, graph theory, and advanced DWI methodology, thereby propelling her toward her career goal of becoming a professor at a four-year university.

NIH Research Projects · FY 2025 · 2024-05

PROJECT SUMMARY Chromosomal Instability (CIN) and loss of the short arm of chromosome 17 (17p) are interrelated and relatively common characteristics of many advanced, metastatic cancers. Though these genetic alterations are likely to contribute to synthetic lethalities, the genetic dependencies resulting from CIN and 17p loss have not been extensively identified or explored. A whole-genome CRISPR/Cas9 screen performed in our lab comparing primary and metastatic-derivative cell lines revealed that metastatic tumors, which possess higher rates of chromosomal instability, are more sensitive to inhibition of mitotic regulators associated with chromosome segregation. Knockout of one such regulator, NDE1, selectively inhibited the growth of metastatic, CIN-high cell lines in vitro, and in vivo. Interestingly, dependence on NDE1 is highly correlated with loss of 17p across cell lines in the DepMap database, indicating that both CIN and 17p loss may contribute to NDE1 sensitivity. We hypothesize that inhibiting NDE1 may be an effective way to target cancers harboring 17p loss and increased CIN through a unique, dual action mechanism involving the buildup of errors in faithful chromosome segregation, and synthetic lethality with an NDE1 paralog and binding partner, both of which are located on 17p. To test this central hypothesis, I propose defining the relationship between NDE1 dependence, 17p loss, and chromosomal instability, characterizing the role of NDE1 in proper chromosome segregation, and assessing the therapeutic potential of targeting NDE1 in the context of CIN and 17p loss.

NIH Research Projects · FY 2026 · 2024-05

Project Summary/Abstract Alternative polyadenylation (APA) is a major mechanism of posttranscriptional regulation that generates distinct 3′ untranslated regions (3′UTR) on RNA transcripts from most human genes. Mechanistically, APA is mainly regulated in trans through APA regulators and in cis by single-nucleotide variants (SNVs) enriched in 3′UTRs and downstream gene regions. Several reported examples indicate that APA can regulate the activity of oncogenes and tumor suppressor genes. However, the upstream regulatory mechanism and downstream function of APA in the cancer progression within clinical cohorts, particularly in prostate cancer, remains largely uncharacterized. Prostate cancer presentation is frequently stratified into groups: highly treatable androgen- dependent prostate cancer (ADPC), followed by the lethal castration-resistant prostate adenocarcinoma (CRPC), and ultimately progressing to the most aggressive form, neuroendocrine prostate cancer (NEPC). While significant efforts have been made to characterize gene expression changes during prostate cancer progression, a link between APA and prostate cancer progression has not previously been established. Our preliminary studies found that 3′UTR lengths are significantly shortened in CRPC patients compared with ADPC patients and are significantly lengthened in NEPC patients compared with CRPC patients. We further found that APA factor-regulated genes with altered 3′UTRs can function as novel oncogenes for CRPC or NEPC. Importantly, manipulating 3′UTR lengths of selected novel oncogenes by our developed 3′UTR CRISPR-dCas13 Engineering System (3′UTRCES) generates distinct molecular outcomes, leading to decreased prostate cancer growth in vitro and in vivo. We thus hypothesize that APA drives prostate cancer progression and can potentially be reversed in a clinically meaningful manner. In Aim 1, we will identify APA target genes during prostate cancer progression to castration resistance through multi-omics analyses in prostate clinical samples and cell models. In Aim 2, we will develop a computational model, namely MARS3'aQTL, to infer upstream APA regulators. In Aim 3, we will characterize the molecular regulations, biological functions, and clinical relevance of inferred APA regulators and will use 3′UTRCES to precisely interfere with APA of novel oncogenes. Our proposed studies' successful completion will establish APA as an important targetable posttranscriptional regulatory mechanism, contributing to a more complete understanding of prostate cancer progression.

NIH Research Projects · FY 2026 · 2024-05

ABSTRACT: Malignant primary brain tumors represent the most frequent cause of cancer death in children and young adults and account for more deaths than cancer of the kidney or melanoma. Glioblastoma (GBM) is uniformly lethal, and current therapy is non-specific and produces a median overall survival of <21 months. Moreover, patients with GBM are plagued by reduced systemic T cell counts, reduction in secondary lymph organ cellularity, increased mature T cells sequestered in the bone marrow (BM), and dysfunctional antigen presentation in tumor draining lymph nodes. This combination of BM sequestration of T cells and heterogeneous antigen expression leads to tumor escape and progression. Dendritic cells (DCs) bearing tumor antigen can be delivered as a vaccine and migrate to the draining lymph nodes (DLN) to trigger the formation of potent tumor- specific cytotoxic T lymphocytes (CTLs) capable of eradicating tumor while leaving normal tissue unharmed. However, despite individual cases of remarkable patient responses to antitumor DC vaccination, overall objective responses in early phase clinical trials have remained under 15%. We have found that post dendritic cell (DC) vaccination, both long-term survival and migration of DCs is dependent on the chemokine CCL3. Interestingly, CCL3 can enhance migration of both exogeneous DC vaccine to lymph nodes and endogenous DCs at the tumor site. Separately, but concurrently, T cell sequestration to the BM is also reversed with CCL3 administration. Here, we aim to build on this data preclinically and determine the optimal CCL3 dose for DC migration, determine the mechanism of CCL3 on T cell sequestration, and determine the efficacy of CCL3 in combination with DC vaccines on anti-heterogeneous glioma responses. We hypothesize that systemic administration of CCL3 will increase migration of both exogenous and endogenous DCs and maintain increased levels of activated T cells systemically and within the TME by eliminating sequestration. This proposal will promote the development of CCL3 as a novel anti-cancer drug that enhances the potency and diversity of immune responses generated by DCs will reduce antigen escape and improve the control of diffusely heterogeneous tumors

NIH Research Projects · FY 2025 · 2024-05

ABSTRACT Emotion dysregulation is a transdiagnostic public health problem that is associated with negative effects on psychiatric outcomes and a large global economic burden. The development of novel treatments for emotion dysregulation that are effective across a wide range of psychiatric disorders is therefore an urgent need. A single session of repetitive transcranial magnetic stimulation has been shown to enhance emotion regulation in people with transdiagnostic emotion dysregulation. In these participants we are finding that induction of negative emotion is associated with hyperactivity in the insula, a key salience network node; utilization of emotion regulation skills is associated with downregulation of the insula; and functional connectivity strength between the insula and the dlPFC, a central executive network node, is positively correlated with emotion dysregulation severity. In a comparison group of non-clinical participants, utilization of emotion regulation skills is associated with increased functional connectivity strength between the insula and the mPFC, a default mode network node. These findings may reflect that dynamics between the salience and other large-scale prefrontal networks differ between emotionally dysregulated and non-clinical participants. Therefore, I hypothesize that cortico-insular connections are critically involved in emotion dysregulation and may be a mechanism for the therapeutic response to neuromodulatory perturbation. A connectomic approach will be employed to determine how cortico- insular connectivity is related to emotion dysregulation and affected by brain stimulation. Studies proposed may advance scientific knowledge about the role of functional and structural insular networks in transdiagnostic emotion dysregulation, leading to the refinement of individualized targets in neuromodulatory interventions. The expertise of Sponsor Dr. Kevin LaBar, Ph.D. in emotion regulation neuroscience and Co-Sponsor Dr. Andrada Neacsiu, Ph.D. in neuromodulatory interventions for emotion dysregulation will provide me with high-quality scientific training. The skills and training proposed in this application will allow me to establish the foundation for a physician-scientist career at the intersection of psychiatry and translational neuroscience.

NIH Research Projects · FY 2026 · 2024-05

SUMMARY Dynamic trans-bilayer movement of ions and phospholipids across cell membranes is vital for cellular life and death. In stark contrast to our extensive understanding of membrane ion transport, the mechanisms and consequences of phospholipid flip-flop remain elusive. The TMEM16 transmembrane protein family, implicated in various diseases, such as heart attack, stroke, epilepsy, muscular dystrophy, cancer, and infections like AIDS and COVID-19, comprises both ion channels and lipid scramblases. By closely examining TMEM16 family members, our previous work has eliminated the conceptual barrier between ion channels and lipid scramblases, the two major types of passive transporters on cell membranes cooperating distinct substrates. This conceptual breakthrough has allowed us to determine novel ion and lipid transport principles at the molecular level and uncover new cellular and physiological functions for TMEM16 ion channels and lipid scramblases. In this application, we will investigate TMEM16 lipid scramblases to deepen our understanding of lipid flip-flop and its undiscovered cell signaling roles. We aim to develop innovative methods for monitoring and controlling lipid flip-flop under physiological conditions and to decipher how calcium and voltage activate TMEM16 lipid scramblases at molecular and cellular levels. Additionally, we will employ our novel methods to explore lipid scramblase-mediated cell signaling during cell-cell fusion, an essential yet poorly understood process for human health. Our proposed studies will offer a comprehensive understanding of lipid and ion transport in health and disease, paving the way for developing new therapies to prevent and treat a wide array of diseases, including stroke, heart attack, atherosclerosis, neurological disorders, infectious diseases, and pregnancy complications.

NIH Research Projects · FY 2025 · 2024-04

PROJECT SUMMARY/ABSTRACT Obesity affects 1 in 5 children and adolescents in the United States and prevalence of severe obesity is worsening. Despite advances in treatment for obesity, primary prevention is essential to prevent morbidity and early death. Infants who gain weight rapidly have over 4 times higher odds of developing obesity as a child or adult, and rapid weight gain during infancy also explains a large percentage of the entrenched and worsening racial and ethnic disparities in childhood obesity and subsequent cardiometabolic disease. Infants who are predominantly bottle-fed are at higher risk for excessive infancy weight gain and childhood obesity, yet we do not have effective interventions to reduce excessive weight gain among infants who are bottle-fed. Our preliminary work suggest that two novel intervention strategies are feasible and may reduce excessive infancy weight gain: reducing bottle size; and increasing bottle opacity. Both smaller size and increased opacity attenuate the visual cues that may encourage overfeeding of bottle-fed infants and overriding infants’ hunger and satiation cues. Our past observational work shows the size of the infant bottle is positively associated with total formula fed, weight-for-age, and weight-for-length from 2 months through 12 months of life. Our past experimental work shows that smaller bottles are a feasible intervention from the primary care setting. We have also shown that caregivers using opaque bottles showed greater sensitivity to their infant’s hunger and satiation cues and fed their infants less than when using clear bottles. Opaque bottles were acceptable and feasible in a community-based intervention over 3 months. We aim to test the independent and joint efficacy of these two intervention components among exclusively bottle-fed infants in a randomized, full factorial clinical trial. We hypothesize that smaller and opaque bottles reduce conditional weight gain, decrease volume per feed, and improve caregiver sensitivity to infant hunger and satiation cues. Our objective is to understand the independent and joint efficacy of reduced bottle size and increased bottle opacity and to collect additional feasibility and acceptability data in order to facilitate rapid translation and implementation of optimized bottle feeding strategies. We plan to achieve our objectives by (1) assessing the independent and joint efficacy of reducing bottle size and increasing bottle opacity to reduce excessive weight gain from birth to four months of infant life, (2) determine the effects of reducing bottle size and increasing bottle opacity on changes in volume per feed and on caregiver-infant dyad feeding interactions through video-recorded feeding observations, and (3) quantify and describe factors associated with eligibility, enrollment, and adherence to bottle design interventions. To achieve these aims, we have assembled a team of experts in obesity, primary care-based clinical interventions, infant growth, caregiver-infant dyadic feeding behaviors, factorial designs, and biostatistics. The results from this novel intervention strategy will be used to design a large, multicenter clinical trial of a comprehensive bottle-feeding strategy to reduce the incidence of obesity in early childhood.

NIH Research Projects · FY 2025 · 2024-04

Abstract An estimated 400,000 anterior cruciate ligament (ACL) injuries occur yearly in the United States. Most ACL injuries are non-contact, occurring during rapid deceleration movements such as jumping and cutting. Importantly, the risk of early-onset osteoarthritis (OA) persists regardless of whether ACL reconstruction is performed. Therefore, identifying risk factors for non-contact ACL injury is of great clinical importance. The function of the ACL is to resist tibiofemoral motion during the intense forces associated with dynamic activity. Therefore, measuring the dynamic elongation patterns of the ACL functional bundles will provide imaging biomarkers of elevated tension within the ACL. As the ACL fails under tension, factors associated with elevated ACL bundle elongations reflect increased vulnerability to injury. Non-contact ACL injuries are 2-6x more likely in females as compared to males. Patients with prior ACL injury are up to 15x more likely to sustain injury to their uninjured contralateral knee than uninjured controls. Finally, while controversial, some studies indicate that fatigue increases injury risk. However, there remains limited in vivo data to describe ACL bundle elongations during relevant dynamic activities (such as jumping and cutting) in these groups. Because different studies suggest that different motion patterns are associated with non-contact ACL injury, the kinematic risk factors for this injury remain unclear. Additionally, studies suggest that a number of characteristics of femoral, tibial, and ACL morphology are related to injury risk. However, it remains unclear how these kinematic and morphologic factors interact to influence ACL tension during relevant dynamic activities. Thus, quantifying the influence of these factors on in vivo ACL bundle elongations in these high risk groups addresses a critical barrier to our understanding of what elevates ACL injury risk. Thus, we propose to utilize a novel in vivo multimodal imaging methodology developed by our lab to assess the impact of 6DoF kinematics and characteristics of knee morphology on in vivo ACL bundle elongations (defined as change in length normalized to an unloaded reference length). These relationships will be assessed during jumping and cutting activities in three potentially high risk populations. Specifically, we will investigate these relationships Aim 1.) in both sexes, Aim 2.) in the uninjured knees of patients with prior ACL injury and control knees, and Aim 3.) pre- and post-fatigue. Relevant biological variables including sex, age, BMI, race, isokinetic muscle strength, sport participation, and activity level, will also be considered as covariates that may impact ACL bundle elongations. Furthermore, interactions between covariates will be explored to assess differential effects of one covariate across levels of another. Thus, the proposed studies will have high impact because they will 1.) address a fundamental gap in knowledge regarding the causes of ACL injury 2.) identify those who are at high risk for injury and 3.) identify those who are best targeted for intervention strategies.

NIH Research Projects · FY 2026 · 2024-04

Project Abstract - A Global Syphilis Vaccine Targeting Outer Membrane Proteins of Treponema pallidum The scientific premise of this CRC proposal rests upon our three decades of work defining the molecular architecture of the outer membrane (OM) of Treponema pallidum subsp. pallidum (TPA), coupled with our successes combining bioinformatics, biophysical techniques, and localization methods with live TPA to topologically characterize TPA outer membrane proteins (OMPs) and define the syphilis spirochete’s ‘OMPeome’--its repertoire of OMPs. The central hypothesis is that the principal targets for a syphilis vaccine reside within TPA’s repertoire of rare OMPs. The current application builds upon the work of the current CRC U19 grant awarded to the two mPIs of this proposal (Moody and Radolf). This proposal brings together the expertise in spirochetology (Univ. of Connecticut) and vaccine development (Duke Human Vaccine Institute) to develop vaccines targeting TPA OMPs. The overarching hypothesis is that vaccines targeting TPA OMPs will elicit antibodies that can recognize intact treponemes, provide protection in animal models, and be producible using Good Manufacturing Practices (GMP) at the scale needed to perform a phase 1 clinical trial. This CRC proposal will focus on two main aims: Aim 1) Selection of an ECL vaccinogen panel and production of ECL mAbs with functional activity, with subaim 1.1 of finalizing the ECL vaccinogen panel through robust preclinical animal studies (Univ. of Connecticut) and subaim 1.2 of producing ECL mAbs with functional activity (Duke Human Vaccine Institute), and Aim 2) Selection and optimization of an ECL vaccine platform, with subaim 2.1 of generating functional ECL-specific Abs using mRNA-LNP immunogens (Univ. of Pennsylvania), and subaim 2.2 of generating functional ECL-specific Abs using SpyVLPs (Univ. of Connecticut), to be compared in animal protection studies in subaim2.3. Successful completion of the aims of this CRC U01 proposal will provide the preclinical data needed for an IND submission to the FDA, complete the process development for GMP production, and will result in the full design of a GMP campaign strategy and a plan for toxicology testing.

NIH Research Projects · FY 2026 · 2024-04

ABSTRACT – Genomic Imprinting in the Development of NAFLD By 2030, metabolic dysfunction associated steatotic liver disease (MASLD), formerly called NAFLD, is projected to affect more than 100 million Americans, including 10 million children. MASLD results from overaccumulation of liver fat and can progress to metabolic steatohepatitis and cirrhosis, leading to significant morbidity and mortality even among children and adolescents. Accumulating evidence indicates that early life exposures alter gene expression and influence the risk for MASLD, but regions of the genome targeted by these exposures remain unclear. We hypothesize that aberrant methylation at imprint control regions (ICRs), discrete loci which regulate expression of specific genes and the function of gene networks, contributes substantially to MASLD. Measuring the impact of epigenetic modifications that alter MASLD risk has been challenging due to the lack of tools for comprehensive measurement of epigenetic modifications, longitudinal data, heterogeneous and non-MASLD specific cohorts, accurate non-invasive markers of liver fat and fibrosis, as well as the temporal ambiguity between exposure and outcomes. To overcome these obstacles, we have comprehensively identified DNA methylation-controlled regulatory regions for genomically imprinted genes, many of which are implicated in metabolic disease, growth and development. Once established in the gametes, these ICRs are stably transmitted during mitotic cell division and are thus very similar across tissues. Importantly, imprinted genes play key roles in growth and metabolism, underscoring plausibility in the etiology of MASLD. The overarching goal is to utilize these novel ICRs to identify methylation changes associated with MASLD development. We will leverage the resources of our multi-ethnic, pre-birth, longitudinal cohort, the Newborn Epigenetics STudy (NEST) to evaluate the oldest 500 children (17-21 years). We will identify ICRs at birth associated with increased liver fat, MASLD, and liver fibrosis using magnetic resonance imaging as well as intermediate markers of MASLD and metabolic dysfunction. ICR methylation will be examined in relation to metabolic outcomes, sex, ethnicity and early life exposures. Finally, we will validate the ICR status of MASLD-associated genes in different tissue types to determine their ability to serve as early markers of disease risk. This proposal will contribute significantly to knowledge on the relationship between genomic imprinting and MASLD and clarify risk-mediating regions and exposures that can be used to develop early detection and therapeutic tools to combat the MASLD epidemic.

NIH Research Projects · FY 2026 · 2024-04

Risk of death for Black patients with HPV-negative oropharyngeal cancer (OPC) is significantly worse compared to White, Hispanic, and Asian patients even after adjusting for socioeconomic, demographic and disease related effects. The scientific premise of this proposal is based on the integration of spatial genomics and artificial intelligence (AI) pathomics to conduct a differential expression analysis between Black and White patients with HPV-negative p16+/HPV16+ and p16+/HPV16- (i.e. discordant HPV status) OPC. We have previous experience using these approaches as a basis to understand gene expression changes between Black and White patients with HPV-positive OPC, which is a distinct neoplastic entity when compared to HPV-negative OPC. Therefore, we will identify differences in gene expression and stromal and immune cell topology between Black and White patients to identify biologic mechanism(s) that underlies health disparities with respect to oncologic outcomes. Moreover, within this research proposal, in collaboration with the Duke Cancer Institute, we will create pilot head and neck cancer research funding opportunities to support the career development of young investigators i.e. trainees, post-docs, and early-career faculty <3 years pursuing clinical oncology. We will address these gaps in the field by pursuing the following specific aims: In Aim 1, we will identify ancestry associated driver genes based on the genomic differences associated with OPCs in Black and White patients that effect treatment response, recurrence and overall survival. In Aim 2, we will integrate spatial genomics and AI-guided pathomics between high vs. low pathomic expression within tissue types, controlling for self-reported race. To foster the career development of the head and neck the oncology workforce, in Aim 3 we will work in collaboration with the Duke Cancer Institute to implement a new pilot programs to support young investigators i.e. trainees, post-docs, and early-career faculty <3 across the continuum of head and neck oncology research. We propose to address these important issues and lack of reliable biomarkers for all patients by integrating state-of-the-art techniques in genomics, pathomics, and AI, in a setting fostering the career development of young head and neck cancer investigators. The data generated will be of benefit to all patients with OPCs as we are presently unable to identify poor responders regardless of race.

NIH Research Projects · FY 2026 · 2024-04

Abstract: Adult teleost fish and urodele amphibians can regenerate entire amputated appendages, whereas this ability is restricted to the tip of the distal phalanx in humans and mice. Though modest in comparison to its zebrafish and salamander counterparts, mouse digit tip (MDT) regeneration exhibits similar anatomic stages and proceeds with intramembranous bone regrowth and mostly scarless healing, even in adults. By studying shared principles from organisms with elevated regenerative potential and applying them to mammalian models, we can gain valuable insight into strategies to augment human tissue repair and improve the care of limb loss patients. The catalogue of defined molecular factors in tissue regeneration is expanding and it has become critical to determine how genes involved in regenerative events are engaged upon injury. We previously identified a new class of regulatory sequences we named tissue regeneration enhancer elements (TREEs) that contain sequence information necessary for precise control of regeneration genes. We also reported that TREEs of zebrafish origin can be engineered in viral gene therapy constructs to target expression of pro-regenerative factors to mammalian injury sites and enhance tissue repair. In preliminary collaborative studies between the Poss and Brown groups, we have identified a new set of candidate TREEs from chromatin samples of regenerating MDTs. We have also performed comparative studies guided by single-cell transcriptome analysis of regenerating zebrafish fins and complemented by molecular genetics in mice, to implicate a conserved family of transcription factors in control of digit tip regeneration. Concurrently, we have identified TREEs of zebrafish origin and an effective adeno- associated virus (AAV) capsid variant that can transduce limb tissue and enable selective expression of gene cargoes at an injury site. Our preliminary studies thus unveil a pipeline of TREE control element identification, novel candidate pro-regenerative factor identification, and application of gene therapy methodology. Here, we propose to: 1) identify distal regulatory elements that control key programs in MDT regeneration; 2) define transcriptional mechanisms by conserved factors; and 3) enhance MDT regeneration by spatiotemporal delivery of pro-regenerative factors. This work will increase understanding of transcriptional regulation during mammalian digit tip regeneration and provide important perspective for comprehending, and perhaps changing, existing limitations in the regenerative capacity of human limbs.

NIH Research Projects · FY 2026 · 2024-04

In the United States, 1 in 11 individuals will experience a urinary stone disease (USD) in their lifetime. Laser lithotripsy (LL) via ureteroscopy, in which pulsed laser light is used to progressively break the stones, has become the choice of treatment for USD patients. In particular, “dusting” mode with low pulse energy and high frequency has gained clinical popularity over “fragmenting” mode because of the fine dust produced, which eliminates the need for basket retrieval and ureteral access sheaths, and thus greatly shortens the procedure time. Growing evidence by us and others have discovered that cavitation bubble collapse plays a significant role in stone dusting by Ho:YAG lasers, and maximizing cavitation activities is critically important for improving LL efficiency. Despite the growing enthusiasm, the exact mechanism of cavitation in LL is not well understood and the optimal LL settings for efficient dusting have not been defined. Such a paucity of knowledge is partially due to the lack of imaging technologies for real-time feedback of cavitation activities during the LL treatment. B-mode ultrasound imaging (or active cavitation mapping) is compatible with LL, but cannot image the short-lived transit bubbles that undergo inertial cavitation and may also interfere with the bubble dynamics. Other imaging technologies, such as x-ray CT, fluoroscopy and MRI, cannot detect cavitations due to the lack of contrast. In light of the clinical need for more efficient LL, we propose to develop 3D super-resolution passive cavitation mapping (3D-SR-PCM) that (1) is totally noninvasive and fully compatible with the LL treatment; (2) is capable of real-time monitoring of the 3D cavitation activities at clinically-relevant depths (>10 cm deep) without background interference; and (3) can achieve a cavitation localization resolution of ~50 µm, which is 10-fold better than the B-mode ultrasound imaging. Our long-term objective of this project is to better understand the therapeutic impact of cavitation in LL treatment, and develop more efficient LL with less adverse effects. Our central objective of this proposal is to develop, validate, and optimize a safe, reliable, and precise imaging technology for real-time analysis of 3D cavitation activities during LL treatment. We will pursue three specific aims: (1) develop the first 3D-SR-PCM system with super-resolution bubble localization, automatic fiber tracking, and real-time cavitation analysis; (2) thoroughly validate and optimize the 3D-SR-PCM system on kidney phantoms, and identify the most relevant cavitation characteristics with stone damage under clinically realistic settings; and (3) apply the optimized 3D-SR-PCM system on a swine model in vivo to evaluate the treatment efficiency with cavitation-imaging guidance, and demonstrate its clinical feasibility through a pilot study on USD patients. The proposed 3D-SR-PCM is timely and well justified, considering the growing USD epidemic and the rapid advance in LL technology. We expect the outcome of this project will provide an enabling tool for optimizing the clinical LL strategies, and is anticipated to impact the long-term surgical management of USD and further advance the LL technology.

NIH Research Projects · FY 2026 · 2024-04

Abstract: Infections caused by Mycobacterium tuberculosis (Mtb) have historically been the leading cause of death from a single infectious agent and still result in 1.5 million deaths annually. One of the hallmark characteristics of the disease is an aggregate of immune cells called the granuloma, where macrophages develop epithelioid character, expressing epithelial cadherin (E-cadherin) and other canonical markers of epithelial cells. This structure restrains the bacteria within but is unable to eradicate the infection despite exposing the pathogen to pH changes, hypoxia, and antimicrobial peptides. The unique, lipid-rich cell envelope of mycobacteria protects them from these stressors while simultaneously modulating the host immune response. Thus, targeting bacterial cell envelope components that are specific to the granuloma is a potential avenue for future drugs and therapeutics. Using the zebrafish-Mycobacterium marinum (Mm) model to recapitulate key aspects of TB disease, the proposed aims will uncover novel bacterial elements with importance in mycobacterial granulomas. I will explore how specific cell envelope components impact both mycobacterial survival and host dynamics in mycobacterial granulomas. I will assess how the bacteria sigma factor SigE (σE) impacts survival within the granuloma through cell envelope-mediated interactions with the host immune response. We have identified SigE as a potential modulator of bacterial survival in the granuloma, which will help better understand the host-pathogen interactions that occur in the granuloma. In addition, the knowledge of bacterial factors that allow mycobacterial persistence in granulomas can unveil potential drug targets and therapies to treat tuberculosis.

NIH Research Projects · FY 2026 · 2024-04

Abstract Animal cells achieve and maintain a typical volume through the regulation of ion channels that control osmotic processes. In metazoans, development of properly proportioned bodies also requires volume regulation to be coordinated amongst cells within tissues and across organs and systems. Although ion channels regulating cell volume have been identified in cells in culture, little is known about the mechanisms that regulate cell volume and coordinate volume control throughout whole tissues in vivo. Our proposed studies will investigate in zebrafish a new and conserved pathway controlling cell volume and integrity in vacuolated cells of the notochord. Using genetic, cell biological, physiological, and quantitative in vivo and ex vivo approaches, we will define the physiological mechanisms regulating fluid transport and cell volume in response to mechanical cues.

NIH Research Projects · FY 2026 · 2024-04

ABSTRACT Rift Valley fever virus (RVFV) is an emerging arboviral pathogen and the cause of Rift Valley fever (RVF) in animals and humans. RVF has been reported across much of the African continent and is designated as a priority disease by international organizations due to its epidemic potential and the lack of effective countermeasures. The epidemiology of RVF has particular characteristics, with sudden large outbreaks in animals and humans occurring in spatially and temporally divergent patterns followed by long periods – called the inter-epizootic or inter-epidemic periods (IEP) – where disease activity is purportedly inapparent. Yet enhanced surveillance studies consistently show that the IEP is characterized by continuous disease transmission within susceptible hosts, and this transmission cycle is hypothesized as one mechanism for RVFV persistence during long IEPs. To this end, this project aims to gain insight into the inter-epidemic transmission pathways that result in RVF for livestock and human populations within the Kilimanjaro Region of Tanzania – where prior work by our One Health consortium described an outbreak of RVF among cattle that was contemporaneous with severe cases in humans. This project will: 1) define the prevalence of RVFV for livestock and humans in Kilimanjaro across different socio-ecological zones (e.g. rural, peri-urban, peri-urban, elevation) and exposures (e.g. socio- demographics, local mosquito composition and density); 2) systematically survey the milk value chain in Kilimanjaro to the assess the infectious nature of cow's milk; and 3) leverage data collected during the award period to construct mathematical models that reconstruct the transmission dynamics and epidemiology of RVF in Kilimanjaro. Results from this project will identify the pathways that contributes to RVFV persistence during the IEP and may inform the development of actionable interventions that interrupt this cryptic transmission cycle. This project has direct relevance to the US NIH's continued efforts to conduct foundational research on emerging infectious pathogens and the diseases they cause. This career development award will provide the candidate with expertise in conducting One Health research, and he will acquire advanced skillsets in infectious disease modeling, all of which are necessary for his long-term career goal to become a leader in advancing understanding of zoonotic diseases and implementing mitigation strategies to effectively stop disease transmission. The primary mentor for this award is Dr. Matthew Rubach, an accomplished researcher with substantial experience in conducting clinical research on causes of severe febrile illness and zoonotic diseases in Tanzania. A complementary and diverse group of mentors will provide guidance in One Health (Dr. Jo Halliday), veterinary health (Dr. Gabriel Shirima), and infectious disease modeling (Dr. Paul Johnson). The candidate will use the outstanding resource and interdisciplinary working groups at Duke University, Kilimanjaro Christian Medical Centre, and the University of Glasgow to launch his career as an independent physician-scientist.

NIH Research Projects · FY 2026 · 2024-04

Abstract Oncogene targeted therapies offer a safe and effective approach to suppress cancer since these drugs spare healthy tissues by selectively targeting driver mutations in cancer cells. Despite a strong initial response to these therapies, however, patients eventually progress and relapse within months of treatment due to the survival of residual tumor cells. These residual disease cells are crucial to eradicate since these cells often serve as the reservoir for resistance and relapse. However, the biological and immunological properties of residual tumor cells are poorly understood. We studied the immunological properties of residual cancer cells and the molecular mechanisms governing it. Using a BRAF mutant mouse melanoma model, we found that residual cancer cells that survive in vitro BRAF therapy exhibit immune evasion in vivo. Our preliminary data suggest residual tumor cells evade the immune system by targeting macrophages of the immune system through upregulation of “don’t eat me” markers such as CD47 (cluster of differentiation 47). Our findings are reproducible in human BRAF, ALK and KRAS mutant cancer cell lines that all show CD47 upregulation upon treatment of respective targeted therapies. Importantly, CD47 blockade has been shown to have an antitumor effect in many mouse tumor models and is currently under investigation in clinical trials to determine its efficacy in human patients. Thus, our preliminary data linking oncogene targeted therapies to CD47 upregulation has significant implications for combining these therapies in clinical practice. To further investigate our findings, we propose a series of integrative transcriptomics and functional genomics approaches to uncover potential mechanisms to target and engineer the immune-mediated clearance of residual tumor cells.

NIH Research Projects · FY 2026 · 2024-04

Explicitly or implicitly, there are currently three competing models for the role of the neuromodulator acetylcholine (ACh) in attention. The first asserts that the cholinergic system is spatially imprecise and contributes to a mechanism for arousal but not attention. The second states that the cholinergic system is spatially imprecise and is one component of the mechanism for attention. The third states that the cholinergic system is at the center of the mechanism for attention (implying the system is sufficiently spatially precise to play such a role). In this study, I will test these three competing models, employing electrochemistry and electrophysiology in the visual cortex of macaque monkeys performing a cued orientation-change-detection task. If the release of any neuromodulator is required for the circuit-implementation of attentive effects, the expectation is that the task will drive release of that molecule into V4, and more specifically that release will occur after presentation of a spatial cue, and in the vicinity of neurons whose receptive fields (RFs) represent the cued location. The RF of neurons in visual cortical area V4 will be mapped, then during the task, the cued location will be varied from trial to trial with respect to this RF location. This will be done, first, at a coarse scale (i.e., attend to the recorded quadrant or to one of the other three quadrants) and then at a finer scale, attending to different positions along an iso- eccentricity curve through the RF within the recorded quadrant. Along this iso-eccentricity curve, as the cued location increasingly overlaps the RF location, the prediction is that there will be a corresponding increase in the observed effects of attention on spiking activity (e.g., spike rate increases). A custom dual electrochemistry- electrophysiology recording system will be used to concurrently record both spiking activity and sub-second changes in local ACh concentration. A measure of the spatial extents of attention-dependent spike rate changes and attention-dependent ACh release will be derived by plotting these two metrics over stimulus location. These spatial extent measures (spiking activity changes and ACh concentration changes) will then be compared to rule in or out each of the three competing models for the role of ACh in attention. In addition to offering the first rigorous test of the hypothesis that ACh release supports attention, this study will provide the first measurements of the concentration, timing, and spatial extent of ACh release during an attention task in a primate.

NIH Research Projects · FY 2026 · 2024-04

ABSTRACT One of the great achievements in medical sciences is the improved survival rate among children diagnosed with cancer. Although estimates indicate the 5-year survival rate of children diagnosed with a malignancy is near 80%, most of these individuals prior to the age of 40 demonstrate indices of physical limitation normally associated with the elderly population. Among the treatments received by pediatric cancer patients is multimodal fractionated x-ray irradiation and chemotherapy. Both induce genotoxic stress that eliminates proliferative cancer cells; however, multifarious direct and indirect effects of radiation and chemotherapy can negatively impact normal tissue growth and maintenance especially in actively growing populations. Phenotypes observed earlier in pediatric cancer survivors is the accelerated onset of frailty indices such as neuromuscular decline. In a murine model, we find weeks after image guided pediatric fractionated x-ray irradiation of hindlimbs reduced skeletal muscle fiber size, impaired neuromuscular function, fibrosis, and activation of stress related p53 gene expression. Among the p53 regulated genes elevated weeks after pediatric fractionated radiation is Growth Differentiation Factor 15 (GDF15), a regulator of food intake and body weight in response to stressors such as cancer therapies. Single cell RNA sequencing (scRNASeq) analysis revealed radiation induced GDF15 expression was restricted to muscle resident endothelial cells. Also, we identified a subpopulation of neuromuscular junction associated muscle resident mesenchymal progenitor cells expressing the receptor tyrosine kinase Ret that is implicated in mediating GDF15 activity. We find in our murine model of multimodal pediatric cancer treatment and survivorship, deficits in body weight gain and exacerbation of neuromuscular related phenotypes observed with fractionated radiation alone. Therefore, our long-term goal is to rigorously characterize the mechanisms whereby treatment related GDF15 expression impacts neuromuscular related phenotypes in our murine model of multimodal pediatric cancer therapy and survivorship. Ideally, such insight would be used to uncover interventions to attenuate pediatric cancer treatment related neuromuscular decline that increases morbidity and burdens survivors. To accomplish our objectives, we will utilize, assessment of skeletal muscle integrity, neuromuscular function, imaging analysis, mouse genetics, scRNAseq analysis, flow cytometry, gene expression analysis, pharmacological treatments, and measures of body weight and food intake. The specific aims of this proposal are 1) determine if endothelial cell specific loss of p53 or GDF15 knockout impacts pediatric cancer treatment related fibrosis and neuromuscular deficits, and 2) examine whether Ret activity regulates pediatric cancer treatment related fibrosis and neuromuscular deficits.