Duke University

NSF Awards · FY 2024 · 2024-08

When executing a program, a computer maintains a value for each variable in the program. This project instead considers a type of execution called "symbolic execution", wherein the computer maintains a description of the value of each variable; this description might be its actual value, but it might instead be characteristics of its value. For example, at a certain point in the program's execution, the computer might know only that the variable contains a value larger than five. Symbolic execution has found myriad applications for security analysis and defense, software testing, and debugging. However, it is also much slower than regular execution, which precludes its use for several important purposes. This project’s novelties are in a variety of innovations to make symbolic execution faster. This project's impacts are to enable new applications for symbolic execution in the areas of computer security, debugging, and diagnostics. The investigator seeks to involve students from underrepresented groups in the research and to release software embodying these improvements. The innovations explored in this project to accelerate symbolic execution of a program include methods to reduce instrumentation of the program needed to execute it symbolically; to more quickly find a control-flow path to reach a desired point in the program; to limit the paths explored to a subset allowed by policies on (otherwise unknown) inputs; and to otherwise limit symbolic execution costs. One application that serves as a proving ground for progress in the project is a method by which a verifier determines (using symbolic execution) that the messages received from a remote component are consistent with the claimed or sanctioned component software, e.g., as a means of defending the receiver from malicious inputs. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-08

Project Summary/Abstract There is a critical need to develop innovative strategies in early adolescents with type 1 diabetes mellitus (T1DM) that develop self-efficacy and facilitate positive parental involvement. In the absence of accessible interventions, we can expect adherence to diabetes self-management tasks (like blood glucose monitoring [BGM] and BG review) to decline and glycemic control (hemoglobin A1c [HbA1c]) to deteriorate, as typically seen during adolescence. The objective of this pilot RCT is to assess the feasibility, acceptability and preliminary effects of incorporating the structured care of a pet fish into a family-based diabetes self-care routine combined with communication skills training (Routine+CC) on HbA1c, BGM frequency and BG review in early adolescents with suboptimal control of T1DM. The rationale for the intervention is based on three established principles: Social Cognitive Theory, Habit Formation Theory, and Family Systems Theory. Participants in the Routine+CC group will be given a Betta fish with supplies. Participants will be told to feed their fish in the morning/evening/bedtime and perform BGM at that time; and perform fish tank maintenance activities once a week in collaboration with their parent(s) and review their blood glucose (BG) trends with the parent at that time utilizing a collaborative communication framework. The hypothesis is Routine+CC will have high recruitment, fidelity and retention rates; and will provide activity-based cues to perform BGM and BG review, a positive competence experience that enhances self-efficacy in performing diabetes self-care tasks, and an opportunity for positive parental engagement in the youth’s diabetes care. The hypotheses will be tested by pursuing these specific aims: Aim 1: Conduct a rigorous feasibility and acceptability pilot RCT of (a) Pet Fish + Collaborative Communication (Routine+CC) (n=20) which combines routine (pairing fish care with family-based diabetes self-care tasks) and collaborative communication (family will receive training on collaborative communication skills); (b) Pet Fish (Routine) (n=20) which only has the routine; as compared to (c) Control (n=20) which is usual care. Obtain quantitative and qualitative feedback on feasibility (recruitment, fidelity, retention) and adolescent/caregiver acceptability; and Aim 2: Assess the preliminary effects of the Routine+CC intervention on HbA1c, BGM frequency and adherence to weekly BG review at intervention completion (12 weeks) plus a follow-up period (12 weeks). Examine whether Routine+CC changes the hypothesized target mechanisms (parental involvement, diabetes self-efficacy, habit strength) based on survey measures. This approach is innovative, because it proposes a novel approach for youth with TIDM that is resource-efficient and home-based, offers new concepts in habit-formation research and presents a potential benefit of pet care for children with chronic conditions. The proposed research is significant because it is expected to provide the necessary quantitative and qualitative data related to intervention feasibility, acceptability and preliminary efficacy data that will directly inform and ensure the success of a subsequent larger fully powered multi-site randomized efficacy trial.

NIH Research Projects · FY 2025 · 2024-08

Ovarian cancer accounts for more deaths than any other cancer of the female reproductive system. In 2021, there were 21,410 new cases of OC and 13,770 deaths. Remarkable progress has been made in ovarian cancer treatment, resulting in a 33% decline in mortality in the past few decades; unfortunately, equitable access to these therapies remains a challenge. While survival rates improved from 40% to 47% among non- Hispanic (NH) White women, survival has stagnated at 35% for NH-Black women. A well-established predictor of the ovarian cancer survival disparity is lack of access to quality treatment. In analysis of the SEER-Medicare database between 2008-2015, only 14% of NH-Black ovarian cancer patients received guideline-concordant surgery and full cycles of recommended chemotherapy, contributing to poor survival. There is also growing recognition of the enduring impact of societal stressors on health outcomes. Yet only a handful of studies have examined these factors in relation to OC disparities, and none have evaluated its contribution via healthcare access (HCA) domains, or via pathways that involve chronic stress associated with discrimination. In this R37 extension, we propose to build on the well-established ORCHiD (Ovarian Cancer Epidemiology, Healthcare Access and Disparities) research infrastructure to examine longitudinal trajectories of HCA and investigate the individual and joint associations of societal stressors with HCA domains and ovarian cancer treatment and survival outcomes in diverse patients. Our proposed extension substantially moves us towards translational impact by addressing key gaps in the literature regarding the mechanisms through which healthcare access domains impact OC disparities.

NSF Awards · FY 2024 · 2024-08

Non-technical abstract: The research focuses on the development and understanding of a novel type of quantum devices, the “multiterminal Josephson junctions”. Conventionally, Josephson junctions are made of two superconductors – materials in which electric current can flow without resistance – connected through a non-superconducting region. The principal investigator’s team has recently made Josephson junctions with several superconducting regions connected together. The breakthrough became possible by making the non-superconducting region from graphene – a single atomic monolayer of graphite. Graphene is known to be ballistic – in can conduct electrons over large distances without scattering – enabling efficient coupling between multiple superconducting terminals. The project enhances understanding of the multiterminal Josephson junctions, which are both expected to have fascinating physical properties and may eventually find applications in future quantum devices. The research also contributes to training of students on all levels in quantum nanotechnology. The project includes outreach in the local community, with a special focus on broadening the participation of underrepresented groups in STEM. Technical abstract: This project builds on the recent progress achieved by this research team and other groups in making multi-terminal Josephson junctions. One of the central themes of this work is the analogy between the Andreev bound states in a multiterminal Josephson junction and the band structure of a crystal. In particular, it has been predicted that multiterminal Josephson junction could be used to emulate topologically nontrivial bands containing Weyl points. These bands can be explored to search for the predicted topological signatures, such as the quantized transconductance. The project further aims to look for the predicted topological contributions to the multiplet resonances. Finally, the team plans to explore the physics of the “Josephson triode” – a tunable superconducting diode recently realized in this type of samples – with a particular focus on its topological properties. Multi-terminal Josephson junctions have been touted as “synthetic topological matter” which should offer new insights into the role of topology in condensed matter systems. The project intends to enhance our understanding of these structures, which is relevant to a whole class of hybrid superconductors samples. The multi-terminal Josephson junctions are both expected to have fascinating physical properties and may eventually find applications in quantum microwave devices. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-08

DART.3―Revolutionizing Neuropsychiatric Treatment through Noninvasive, Programmable Cell-Type-Specific Neuropharmacology ABSTRACT Neuropharmacology is central to the treatment of brain disorders, and new tools are needed to untangle how canonical drug-receptor interactions are transformed by brain circuits to impact complex behavioral states. DART (drugs acutely restricted by tethering) offers a new path forward, by making it possible to deliver clinical drugs to one cell type at a time, observe ensuing behaviors and cellular dynamics, and reconstruct mechanisms from parts to the whole. Since its debut, we have worked closely with a growing user base to refine and deploy the technology. In particular, the second-generation DART.2 offers thousandfold cellular specificity, histology tracers of target engagement, and a catalog of antagonists, agonists, and allosteric modulators of several receptors. Here, we propose to advance the technology in three new domains of innovation with DART.3. Aim 1 enables noninvasive, whole-body delivery for precise drug engagement. Aim 2 broadens the therapeutic range to Nicotinic and 5-HT2A receptors and cellular locales. Aim 3 provides two kinds of programmable rapid reversibility. Our priorities in technical innovation reflect the most pressing gaps identified by our community of collaborators. Understanding complex animal behavior is our standard. Thus, we characterize tools in behaving animals, develop robust systems of rigor and reproducibility, and invest in our user base by providing reagent distribution, training and support through frequent interactions. These priorities are empowering a thriving and intellectually diverse neuroscience community to transform how we study the brain.

NIH Research Projects · FY 2025 · 2024-08

Project Summary Copper (Cu) is an essential micronutrient for nearly all eukaryotic organisms. Cells must maintain a careful Cu homeostasis to balance cellular needs while minimizing toxicity due to excess Cu accumulation. Fungi, such as Candida albicans, must acquire, route, store, use, and continually monitor levels of Cu to thrive in human hosts. To survive as a human pathogen, C. albicans must be able to withstand the manipulation of Cu metal concentrations at the host-pathogen interface, forcing the fungi to rely on complicated Cu homeostasis pathways to adapt to Cu-replete and Cu-depleted environments. Although commonly used as a treatment, fluconazole is fungistatic, merely inhibiting growth of C. albicans and increasing the risk of developing resistant strains. Fluconazole induces a Cu-deficient response in C. albicans, despite an increase in total Cu levels compared to untreated cells. The Cu-deficient response includes the induction of the Cu-import gene CTR1, repression of the Cu exporter CRP1, and a switch in expression of cytosolic superoxide dismutase from CuZnSod1 to MnSod3, controlled by the transcription factor Mac1. Mac1 is regulated by Cu levels in the cell, binding Cu ions under Cu- replete conditions and binding DNA under Cu-deficient conditions. Despite understanding the metalloproteins involved in the Cu-deficient response, there are still numerous gaps in our understanding of the response. It is known that Mac1 must lose its Cu ions to bind DNA; however, the molecular basis for activation, including the discrete entity involved, is unknown. The work described within this proposal seeks to further illuminate our understanding of the Cu-deficient pathway of Cu homeostasis in C. albicans by investigating the role of CuZnSod1. CuZnSod1 has been primarily characterized its role as global antioxidant, removing reactive oxygen species; however, there are novel roles proposed for Sod1 in sensing the Cu levels of the cell and participating in the Cu homeostasis response by directly activating Mac1. Here, I aim to probe the location and metalation state of CuZnSod1 under fluconazole-induced Cu-deficient conditions and characterize the conditions for a direct interaction between CuZnSod1 and Mac1, using biophysical techniques. Collectively, this work will allow us further insight into the Cu-deficient homeostasis mechanisms of C. albicans and will allow us in the long-term to apply this understanding to design more effective antifungal treatments exploiting metal-specific weakness to increase potency and decrease changes of developing resistant strains.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY/ABSTRACT Biotechnology advances in research settings are rapidly growing, but the gap between translational medicine to clinical utility is lacking. In addition, there is a disconnect between undergraduate bioengineering education and workforce skills development that reduces the bioeconomy human capital needed to address critical biotechnology and biomanufacturing infrastructure in the United States. The proposed project aims to bridge the gap between biotechnology research and clinical applications by developing a bench-to-industry-to-bedside pipeline for undergraduate and graduate biomedical engineering students. Through team-based learning experiences, the project emphasizes collaboration and feedback cycles among critical stakeholders in biotechnology and biomanufacturing. The objectives include augmenting existing biotechnology design courses with emerging technologies such as adeno-associated viruses, recombinant protein production, and chimeric antigen receptor T cells that reinforce common innovation and manufacturing pipelines for student teams. We will also integrate industry and clinical mentorship from local biotechnology companies and medical practice and incorporate ethical frameworks in biotechnology design cycles. To address healthcare equity with rising costs of novel biotechnologies and information access, student designs will emphasize human-centered universal design and education to diverse populations as part of comprehensive needs assessment and stakeholder analyses. By enhancing the Duke Biomedical Engineering curriculum with advances in gene, cell, and molecular therapy, the project seeks to cultivate essential bioengineering skills for a future in molecular and cellular medicine. The intended educational outcomes include fostering skill development in molecular, cellular, and genetic engineering, promoting successful transitions into careers in the biomedical research workforce, and equipping students with the knowledge and experience to navigate the evolving biotechnology landscape and needs of all biotechnology stakeholders. With an estimated impact on approximately 100 undergraduate students and 50 graduate students over the five-year duration of the project, this program aims to significantly contribute to the bioeconomy and advance the translation of biotechnology advancements into clinical applications.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY Hutchinson-Gilford Progeria Syndrome (HGPS) is a rare, accelerated aging disease caused by a single point mutation (SNP) in the LMNA gene. This mutation results in the production of an aberrant protein called progerin which disrupts proper nuclear function within cells. Patients with HGPS die at an early age most often due to complications from severe atherosclerosis. The progression of this atherosclerosis is not fully understood. Animal models that exclusively express progerin in either vascular smooth muscle (vSMC) or endothelial (EC) cells still exhibit thickening of the vessel adventitia despite no progerin-positive cells in this region. This suggests multiple vascular cell types can influence the same pathological characteristics, possibly through paracrine signaling. Having a better understanding of which cell types within the vasculature contribute most to the fibrosis, calcification, and stiffening that occur during atherosclerosis would be useful in generating more effective, targeted therapies. Treating HGPS has proven difficult, with only one therapy approved for clinical use. Other treatment options have been effective in animal models but have shown only modest benefits in patients. A new genetic engineering therapy using an adenine base editor (ABE) can correct the LMNA point mutation and significantly improved survival in an HGPS mouse model. Understanding what level of mutation correction is required in the vasculature to alleviate pathological features would aid in translating this new technology to the clinic. The goal of this proposal is to use an in vitro human vascular model of HGPS to better understand the influence of different vascular cells on the progression of HGPS-induced atherosclerosis and to estimate what level of mutation correction would be needed to improve the vascular pathology. We hypothesize the adventitial fibroblasts are the primary contributors to vascular fibrosis, calcification, and stiffening in HGPS, and that mutation correction within these cells will effectively reduce these disease characteristics. To test this hypothesis, we will generate a tissue-engineered blood vessel (TEBV) disease model using cells from HGPS patients. In Aim 1, we will determine the relative contributions of fibroblasts, vSMCs, and ECs on HGPS vascular disease by testing different combinations of progerin-expressing cells in the TEBV model and evaluating disease progression. For Aim 2, we will treat HGPS TEBVs with different doses of ABE, delivered by adeno-associated virus, and evaluate editing efficiency in each cell type and progression of atherosclerosis. When completed, these aims will provide a more physiologically relevant human model of HGPS vascular disease for testing new treatments. Additionally, they will improve our understanding of how different vascular cells contribute to HGPS disease progression and to what extent correcting the LMNA mutation within these cells will abate or reverse atherosclerosis. Broadly, this work will also show the utility of using human microphysiological systems to model disease and test the efficacy of new therapeutics.

NIH Research Projects · FY 2025 · 2024-08

ABSTRACT The force-gated ion channel Piezo2 plays a crucial role in various mechanosensory functions, including light touch detection, proprioception, urination, blood pressure regulation, and lung inflation. Additionally, gain-of- function and loss-of function mutations have been implicated in Gordon syndrome, distal arthrogryposis, and overall deficits in proprioception. Given the crucial role of Piezo2 in physiologically significant mechanosensory processes and its direct association with human diseases, it is imperative to gain a mechanistic understanding of how Piezo2 functions as a sensor of mechanical stimuli. Although the physiological roles of Piezo2 are well understood, its fundamental properties—specifically, the tension-response relationship and gating kinetics— and how they both are affected by tissue-specific alternative splicing, are unknown. The overall objective of this proposal is to determine the fundamental biophysical properties of the human force-gated ion channel Piezo2 and systematically investigate six domains that undergo tissue-specific alternative splicing to determine their necessity and sufficiency in modifying function. My rationale is that knowing the biophysical properties of Piezo2 will enable us to understand its physiological roles in distinct tissues and cell types. I hypothesize that human Piezo2 and its splice variants exhibit distinct sensitivities and dynamic ranges in membrane tension sensing and differ in their gating kinetics. The scientific premise for this hypothesis is based on 1) the fact that Piezo2 is alternatively spliced in a tissue-specific manner, which implies an underlying difference in function, and 2) my preliminary data demonstrating distinct tension-response relationships in two Piezo2 splice variants. My research plan will determine the tension-response relationship (Aim1) and gating kinetics (Aim2) of human Piezo2 and the effects of its six spliced domains on these properties. The proposed research plan is innovative because it develops two innovative tools: Piezo2 constructs that allow for a structure-function investigation of spliced domains and a rigorous and unbiased image analysis program. It then applies these tools to explore two innovative concepts: quantification of the Piezo2 tension- response relationship and the hypothesis that spliced domains affect both this property and gating kinetics. The significance of this work is that it provides an understanding of the function and physiology of human Piezo2. This is essential for rationalizing the purpose of Piezo2 tissue-specific splicing, interpreting structural data of Piezo proteins, and comprehending the physiological forces, both in intensity and temporal dynamics, Piezo2 can sense or not. Its positive impact is that it will identify domains that selectively modulate Piezo2 function, which may become targets for treating Piezo-related diseases, such as itch, inflammatory pain, and chronic pain.

NSF Awards · FY 2024 · 2024-08

This project aims at developing algorithms for the analysis of mechanical systems with imperfect or uncertain geometries, essential for Digital Twins (DTs). DTs of geometrically complex systems may require months to train because the generation of computational grids is time-consuming and labor intense. These difficulties will be bypassed by combining the Shifted Boundary Method (SBM), an immersed geometry computational method, with probabilistic Subdivision Surfaces (SSs), for geometric representation. Probabilistic geometry representations are needed because the exact geometry of the system in operation is usually only partly known. This project aims to transform the current design and analysis cycle, by making Computer-Aided Design (CAD) and mesh generation more flexible, automated, and better integrated with the analysis phase. This research will enable an ecosystem of computational methods that can robustly and efficiently interact with the meta-algorithms for DTs (e.g., reduced-order modeling, machine learning, uncertainty quantification, and optimization) and will benefit the “democratization” of computing to professionals who are non-expert in this field. Computations in complex geometries pose at least two major challenges: (1) the representation of geometries with imperfect/uncertain CAD models or imaging-based data; and (2) the quantification of the effect of geometric uncertainties on the performance of systems. Unfitted Finite Element Methods (UFEMs; e.g., cutFEM, the Finite Cell Method, Immerso-Geometric Analysis, etc.) simplify mesh generation by immersing geometries in a pre-existing simple grid. However, UFEMs suffer from numerical instabilities or poor matrix conditioning, whenever small cut cells are present. UFEMs require more involved data structures and cut-cell integration may become extremely complex, or even unfeasible for geometry representations with gaps and overlaps. The as-is geometry of parts may be different from design models due to manufacturing uncertainties and wear caused by operation. The Shifted Boundary Method (SBM) shifts both the location of boundary conditions from the true boundary to an approximate boundary (with no cut cells) and their value by means of Taylor expansions. This yields a simple, robust, accurate and efficient method for very complex geometries, which may include gaps/overlaps. The SBM will be applied to high-order hierarchical B-spline discretizations, which have superior monotonicity and regularity properties. The SBM can be adapted to uncertain geometries using probabilistic SS, which combine standard (deterministic) subdivision surfaces with stochastic SPDE representation of random fields. Uncertainties will then be propagated to the output quantities of interest. Observation data from the product in operation will be probabilistically synthesized with the uncertain simulation data obtained by forward propagation. This approach will be extended to a variational Bayesian framework for the inference of geometry and other model parameters using the observation data. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NSF Awards · FY 2024 · 2024-08

This award provides travel support for 14 early career researchers to attend the 2024 Thematic Conference on Uncertainty Quantification for Machine Learning Integrated Physics Modeling (UQ-MLIP 2024), to be held in Crystal City, Arlington, Virginia, 12-14 August 2024. This thematic conference will provide an interdisciplinary coverage of uncertainty quantification for scientific machine learning and physics modeling. It will bring together leading experts, scientists, and young researchers from both academia and industry, with the goal of exchanging the latest developments on these topics and identifying challenges and opportunities to push this interdisciplinary research effort forward. The conference will feature technical presentations by invited speakers, a poster contest, and two panel sessions addressing challenges and future directions. This award will broaden the participation of a diverse set of participants, including women and underrepresented minorities, early career researchers and students, mid-career and senior faculty, as well as representatives from federal agencies and private companies. Dissemination will be achieved through workshop proceedings. A detailed summary about challenges and opportunities will be made available to the community at large. Computational models of real-world systems are increasingly integrating data-driven models from the field of machine learning with physics-based models derived on, or informed by, first-principles. It is thus of greatest importance to carefully characterize and quantify the uncertainties associated with each model class under realistic scenarios where data can be scarce and limited. Furthermore, the propagation of parametric and model-form uncertainties to the outcomes of the integrated models demands for the construction of novel approaches or extensions of existing methodologies. Other topics that would benefit from such developments include digital twinning, model reduction, large scale integrated computations, and decision making in computational science and engineering. Applications of these methods hold the promise to push the boundaries of modeling, inverse identification, and simulation and experimental characterization in mechanics of materials and structures across scales. This thematic conference will facilitate the exchange of information on these topics, providing interdisciplinary collaboration and networking opportunities to a broad and diverse audience including early career researchers, faculty, students, stakeholders, and industrial partners. This project is jointly funded by the Division of Civil, Mechanical and Manufacturing Innovation (CMMI) in the Engineering (ENG) directorate and the Office of Advanced Cyberinfrastructure (OAC) in the Computer and Information Science and Engineering (CISE) directorate. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-08

Latino children in the United States (US), who represent one quarter of all children, are more than 50% more likely to die of their cancer diagnosis than their Non-Latino counterparts. Latino families in the US navigate several adverse healthcare, community, and population health-level indicators that limit their access to quality healthcare and community and population-level resources and in turn may shape their child’s risk of developing and dying from cancer and their ability to advance resilient grief outcomes. Despite higher cancer mortality and limited interventions to address these health disparities, there remains a striking lack of research focused on generating targeted solutions for Latino populations who are living through the loss of a child. Further, the identification of resiliency factors has remained undiscovered because the experience of cancer loss in Latino populations has not yet been explored from a strength-based and multi-level perspective. The overall purpose of this study is to explore the experiences and grief outcomes of Latino families in the US who are living through the loss of a child to cancer. This study will utilize a modified multi-level version of the Center for Latino Adolescent and Family Health Framework, that has been integrated with empirically driven grief concepts in a mixed methods design that integrates data from validated measures representing the framework’s domains across healthcare, community, and population-levels (n=60), with qualitative descriptive data from individual interviews (n=~20) with the deceased child’s primary caregiver. The aims are to: Aim 1. Examine the relationships among healthcare-level (healthcare access), community-level (grief conventions), population health-level (stress), multi-level resiliency factors (community dynamics) and crucial grief outcomes (coping, mental health, post-traumatic growth, meaning-making) of Latino families in the US who are living through the loss of a child through self-reported measures and questions. Aim 2. Elucidate the experiences of Latino families in the US who are living through the loss of a child to cancer in the context of the mechanisms of healthcare, community, and population health-level factors through in-depth, individual semi-structured qualitative interviews. Aim 3. Develop a comprehensive understanding of how multi-level resiliency factors across the domains of healthcare, community, and population health influence adaptive grief outcomes among Latino families in the US after the loss of a child to cancer by integrating self-reported measures and interview data. The findings will enhance the understanding of the mechanisms that may produce adverse outcomes in pediatric oncology bereavement among bereaved Latino families and inform the development of tailored and targeted interventions to address them.

NIH Research Projects · FY 2025 · 2024-08

PROJECT ABSTRACT Fatigue, the most distressing symptom, affects between 60-90% of all individuals with breast cancer. The prevalence of fatigue increases to 80-96% when these individuals are receiving chemotherapy, a common treatment modality. This high prevalence of fatigue is a concern for clinicians, patients, and their families as fatigue is a debilitating symptom that interferes with the individual’s physical wellbeing, including their ability to perform daily activities, and decreases social functioning. However, a lack of understanding of what causes fatigue makes tailoring interventions to reduce fatigue difficult. Yet, while investigators have researched many biological pathways in relation to fatigue, one mechanism that deserves further investigation is the oral microbiome. Compelling evidence suggests that the microbiome influences symptoms such as fatigue and physical well-being. Importantly, individuals with breast cancer have been found to have dissimilar oral microbiomes to health controls, which are then further altered during chemotherapy. Additionally, studies on individuals with chronic fatigue syndrome have suggested that the oral microbiome might play a role in the development of fatigue. Given the high prevalence of fatigue experienced by individuals with breast cancer, exploring this relationship between oral microbial changes, fatigue, and physical well-being is urgently needed. Investigating the oral microbiome of individuals with breast cancer has the potential to inform our understanding of the biological process of fatigue in a cancer that affects millions of individuals annually. The objective of this longitudinal study is to inform our understanding of a biological mechanism associated with fatigue and physical well-being and clarify the impact of chemotherapy on the oral microbiome in individuals with breast cancer receiving chemotherapy. This study will use data and biospecimens that have been collected through the Duke University 1000 Patient Project (1KP). The 1KP project is an ongoing clinical data repository in which patients, including individuals with breast cancer undergoing a mastectomy, are consented for comprehensive data (demographics, disease, fatigue, and physical well-being) and biospecimen collection, such as oral microbiome samples, at multiple timepoints pre and post operatively over the course of 6 months. The aims are to: Aim 1. Characterize oral microbial changes (relative abundance, alpha diversity) prior to chemotherapy and up to 6 subsequent months in 25 individuals with breast cancer. Aim 2. Explore whether oral microbial changes (relative abundance, alpha diversity) are associated with fatigue and physical well-being measured by the Patient-Reported Outcomes Measurement Information System in 25 individuals with breast cancer receiving chemotherapy. Results from the proposed study will enhance the understanding of the relationship between the oral microbiome and fatigue in individuals with breast cancer receiving chemotherapy and inform future research on the creation of tailored intervention to ameliorate fatigue and enhance physical well-being in individuals with breast cancer.

NSF Awards · FY 2024 · 2024-08

The geographical ranges of most species across the Earth are made up of multiple populations (all individuals of that species in a defined area) and these populations often experience quite different conditions. In addition, how the environment will change in the future will also differ among populations and will ultimately determine whether each population persists, which in turn will determine whether the entire range of a species will contract, expand, or shift in space. In the face of environmental change, such shifts in geographical ranges can have important implications for ecosystem health and human well-being. Given the variation in climate across time and space for populations of even a single species, to understand and predict how geographical ranges will shift with changing climate requires both long-term and multi-population studies. This project will synthesize data collected over decades (up to 29 years to date) for a total of 40 populations of two species of tundra plants that are widely distributed across western North America. Tundra plants are especially suited to studies of geographical range shifts driven by climate warming, because they are adapted to cold climates but can also benefit to some degree from warming. The researchers will use the long-term data to examine how survival, growth, and reproduction of the tundra plants respond to variation in multiple, measured environmental factors (both climatic and non-climatic) at multiple spatial scales (from the scale of individual plants to the scale of entire regions), and will assess how year-to-year variation and long-term trends in those factors are likely to either enhance abundance or increase extinction risk of those plants within populations and thus increase or decrease the geographical ranges of the species. All the data on both performance of individual plants and environmental factors affecting that performance will be made publicly available for any future studies of how environmental changes affect geographical range shifts. The three questions this research aims to address are, first, how does demographic buffering – the evolution of reduced temporal variance in demographic rates (survival, growth, reproduction, and recruitment), especially those that most influence population growth – arise from responses of those rates to specific climatic drivers? Second, what role do differences in demographic responses to environmental variation between individuals within populations, between populations in a region, and between regions play in reducing temporal variance in population growth and thus enhancing species persistence locally, regionally, and across entire geographical ranges? Third, do long-term demographic data reveal evidence of adaptive demographic lability – an increase in the population growth rate due to temporal variation in vital rates caused by nonlinear responses to climate, the opposite of demographic buffering - and if so, does its presence depend on the type of environmental driver (e.g., abiotic vs. biotic drivers)? This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2024-08

ABSTRACT Pregnant patients sometimes face impossible choices between earning necessary income and adherence to prenatal care; these issues were highlighted in our formative qualitative work with Black pregnant patients. Increased prenatal care adherence and positive maternal and infant health outcomes may be promoted by addressing work-related pregnancy care barriers. Obstetric clinicians are uniquely positioned to ensure pregnant patients are educated about federally mandated employment protections during their pregnancy, however very few clinicians have formal training about employment laws that govern accommodations during pregnancy and postpartum. To fill this unmet need, a patient- and community-engaged multidisciplinary team and I developed, and pilot tested PROvider ReMote ObsTetric-Related Employment Education (PROMOTE). Our research aims will use a provider-level randomized trial comparing PROMOTE vs usual care. During Aim 1, we will determine the efficacy of PROMOTE to increase the likelihood that obstetric clinical teams counsel patients about work and pregnancy. In Aim 2, we will determine the effectiveness of PROMOTE to reduce undesired wage or advancement reduction, increase access to accommodations, and improve prenatal care adherence and pregnancy health outcomes. The rich dataset that we will collect, including electronic medical record review, patient survey and qualitative interviews will be leveraged as we develop and pilot test ObsTetric HEAlth Outcomes ResearCH Mentoring (TEACH), a year-long mentored career development and obstetric health disparities research program tailored to medical trainees. A patient- and community-engaged multidisciplinary stakeholder group and I will develop the TEACH mentorship program including career and research mentorship. We will leverage existing institutional resources at Duke and integrate TEACH seamlessly into the unique Duke medical school curriculum which provides a year for mentored research. We will pilot test TEACH with six Duke medical trainees to determine feasibility and acceptability of TEACH. The proposed work is led by an early-stage investigator clinically trained as a Maternal Fetal Medicine subspecialist with master’s level research training, formal health disparities research training, and experience mentoring trainees in medicine. The proposed project integrates research that will yield an intervention that empowers obstetric clinicians with practical skills to address employment-related barriers to prenatal care and work that will develop and pilot testing an obstetric health disparities mentored research experience tailored for medical students.

NIH Research Projects · FY 2024 · 2024-08

Abstract: Heterochronic parabiosis and serum transfer models have given evidence for the rejuvenating properties of young blood in old mice and equally potent deleterious effects of old blood in young mice. The critical factors or even tissues responsible for such effects remained elusive due, in part, to the complexity of the parabiosis models. Taking an organ-specific approach may lead to better understanding of how individual tissues contribute to the effects we have seen with parabiosis. Since the liver is a critical organ regulating whole-body metabolism and is the primary source of secreted proteins in circulation, it is an intriguing target to manipulate its biological age and determine resulting functional and molecular changes. To alter the biological age of the liver, we will utilize the heterochronic hepatocyte transplantation technique to incorporate young cells in the aged liver, and vice versa. Isochronic hepatocyte transplantation and non-surgical groups will serve as controls. We have generated preliminary data showing successful transplantation into both young and old livers and see whole-body effects on adiposity to validate efficacy of the model. We hypothesize the donor cells will engraft and influence the host liver to take on either, a more youthful or accelerated aging state based on the donor cell age. Moreover, we hypothesize that transplanted cells will contribute to the liver secretome and mimic profiles of the young and old plasma proteome. We will test these hypotheses through the following aims. Aim1 will test the hypothesis that transplanted cells will alter the host liver function, cellular landscape and biological age. We will measure clinical markers of liver function, including AST, ALT and BUN after heterochronic transplantation. In addition, we will use the recently developed 10xGenomics Xenium Spatial transcriptomic platform to determine how this procedure affects the cellular landscape of the liver, while using epigenetic clocks as an indices of biological age of the tissue. Aim2 will test the hypothesis that donor hepatocytes will alter host liver secretome dependent on the donor cell age. Hepatocytes-derived secreted proteins will be measured using the MetRS mouse model to label proteins in the plasma and compare the plasma proteome across groups. The proposed studies will fully test the hypothesis that donor cells will engraft into the host liver and influence tissue function, cellular phenotype and secretome, which may mimic the effects of heterochronic blood sharing.

NIH Research Projects · FY 2025 · 2024-08

Pain, fatigue, and distress are highly prevalent co-occurring symptoms in patients with advanced cancer. When these symptoms interfere with patients’ daily lives, it can diminish their ability to live congruently with their values—greatly reducing quality of life (QoL). An exclusively medical approach to managing pain, fatigue, and distress can have intolerable side effects and limited responsiveness. There is a need for psychosocial symptom management interventions designed specifically for patients living with advanced cancer. Emerging evidence suggests patients may benefit from Acceptance and Commitment Therapy (ACT), a third-wave Cognitive- Behavioral Therapy (CBT) approach that emphasizes acceptance, mindfulness, and engagement in values- guided activity. Though pilot results in patients with advanced cancer have been promising, ACT-based interventions have rarely been tested in well-powered efficacy trials. Building on our team’s extensive prior work, we developed a psychosocial intervention called ENGAGE. ENGAGE includes training in traditional CBT skills (e.g., activity pacing) to decrease symptom severity, when possible, and ACT skills (e.g., mindfulness) to promote acceptance of experiences that cannot be fully controlled (e.g., prognosis), with the goal of decreasing symptom interference and improving QoL. Our two pilot trials of ENGAGE provide strong support for its feasibility, acceptability, and promise for improving patient-reported outcomes. This proposal progresses our work to an NIH Phase II randomized controlled trial (RCT) to evaluate ENGAGE's efficacy for reducing symptom interference in patients receiving cancer care in medically underserved areas. Delivered through videoconferencing, this brief (4 weekly, 45-minute sessions) intervention has been designed to be acceptable for patients residing in medically underserved areas by including graphical displays of concepts, videos modeling skills, and audio content summaries. In this RCT, we will randomize 190 patients with Stage IV cancer (breast, prostate, lung, or colorectal) and moderate-to-severe symptom interference to ENGAGE or Supportive Care control. Patient-reported outcomes will be assessed at baseline, 2 months, and 4 months. Aim 1 is to determine ENGAGE's efficacy for reducing symptom interference (primary outcome) at 2 months (primary endpoint). Aim 2 is to determine ENGAGE’s efficacy for improving secondary outcomes (i.e., self-efficacy for symptom management, acceptance, mindfulness, values-based activity, symptom severity, and QoL) at 2 months. Aim 3 is to test the maintenance of ENGAGE’s effects on primary and secondary outcomes at 4 months. An exploratory aim seeks insights for future implementation efforts using mixed-methods data collection from patients, oncology providers, and clinic leaders. This trial will be one of the first to test the efficacy of an ACT-based intervention for patients with advanced cancer who are receiving care in medically underserved areas. ENGAGE is readily implementable and, if shown to be efficacious, our team is well positioned to move rapidly into implementation work, with the long-term aim of reducing suffering in patients with life-threatening illnesses.

NIH Research Projects · FY 2025 · 2024-08

ABSTRACT Nutrient absorption in the neonatal mammalian gut relies on intracellular digestion by a population of enterocytes known as vacuolated or neonatal enterocytes. Recently, we showed these cells are conserved between zebrafish and mammals and are specialized in the uptake and digestion of dietary proteins. As these cells possess a prominent lysosomal vacuole and are present in non-mammalian vertebrates such as fish, we refer to them as Lysosome Rich Enterocytes (LREs). In our previous studies, we found that protein uptake in zebrafish and mouse LREs is mediated by a scavenger receptor complex composed of cubilin (Cubn), its transmembrane partner amnionless (Amn), and the endocytic adaptor Dab2. However, the cellular mechanisms that allow LREs to internalize cargo from the intestinal lumen at an astounding rate are unknown. Here, we will leverage the experimental advantages of the zebrafish system and the conserved biology of LREs to investigate the endocytic machinery that allows these cells to support dietary protein absorption in the intestine. Specifically, we will elucidate the mechanisms regulating a specialized form of clathrin mediated endocytosis that confers LREs a high capacity for luminal protein uptake.

NIH Research Projects · FY 2026 · 2024-08

PROJECT SUMMARY / ABSTRACT Technical advances in magnetic resonance imaging (MRI) have led to a wide range of imaging techniques, contrast mechanisms, and clinical applications. However, despite marked progress in radio-frequency (RF) and shim coil technologies, the traditional MRI scanner architecture currently used on virtually all scanners still has major limitations. RF coil arrays require wired connections to the bulky receiver chain in the scanner and the machine room via bulky cable assemblies, which can result in long setup times, patient discomfort and motion, lower signal-to-noise ratio (SNR) from crosstalk, loss of transmit power from power dissipation, and RF burns from induced currents. These issues are further exacerbated with modern high-channel-count or flexible RF coil arrays. In addition, conventional low-order spherical harmonic shim coils require wired connections to amplifiers in the machine room and cannot effectively shim localized static magnetic field inhomogeneities (∆B0) in the human body, leaving artifacts that severely degrade the image quality in many applications. We previously proposed two coil designs to address some of these limitations: 1) Our novel integrated RF/wireless (iRFW) coil design enables MR imaging and the wireless transfer of data from/to peripheral devices with a single coil array for low-throughput applications such as wireless physiological monitoring, but not yet for the wireless transfer of MRI data, which requires further development; 2) Our integrated parallel reception, excitation, and shimming (iPRES) coil design enables MR imaging and an effective shimming of localized B0 inhomogeneities with a single integrated RF/shim coil array. However, such iRFW and iPRES coil arrays remain limited by the bulky wired connections and receiver chain required to transfer the MRI data. Our goal is to address these limitations by developing a highly innovative wireless MRI scanner architecture based on a stand-alone, platform-independent high-channel-count wireless integrated RF/shim coil array with on-board received chain and cloud-based data processing workflow that will enable wireless MRI and localized B0 shimming with a single coil array. This paradigm shift in MRI scanner architecture will eliminate all cables from the coil array and the bulky receiver chain embedded in the scanner, thus drastically reducing the system complexity, footprint, and cost, while making the entire receiver chain and data processing workflow (including with third-party advanced reconstruction methods) compatible with scanners from different manufacturers, and improving the freedom of positioning, patient comfort, safety, SNR, spatial fidelity, image quality, diagnostic accuracy, and clinical utility for a wide range of MRI applications throughout the human body. Specifically, we will develop the technology to enable this novel wireless MRI scanner architecture and we will integrate it with a 48-channel wireless integrated RF/shim head/neck coil array to demonstrate its feasibility and advantages for human brain imaging, which will open up exciting new avenues for MRI.

NIH Research Projects · FY 2025 · 2024-08

ABSTRACT A small number of conserved signaling pathways are key players in cell fate decisions that govern embryonic development, maintenance and health of adult tissues, and progression of disease states. For example, embryonic development in numerous species hinges on the dynamics and regulation of the BMP signaling pathway. Our current knowledge of BMP signaling pathway specificity is based upon foundational genetic and in vitro experiments, but we have limited understanding as to how cells produce diverse but specific responses to similar signaling inputs in vivo. In the Drosophila embryo, a steep gradient of BMP signaling is dynamically established prior to gastrulation. This gradient is interpreted by different populations of cells to establish the dorsal-ventral axis of the embryo, with cells at the dorsal midline turning on a unique set of transcripts compared to more lateral cells. We do not know how the dynamics of gradient formation and the final gradient pattern are interpreted by cohorts of cells in the embryo to produce the correct spatiotemporal transcriptional response. This study aims to shed light on how cells perceive and respond to varying BMP signaling inputs, with the goal that these discoveries will be broadly applicable to other biological contexts. The proposed research program employs cutting-edge quantitative live imaging techniques, the creation of predictive mathematical models, and the adaptation of tools for studying BMP signaling in diverse insect species. Specifically, I will develop a predictive model for BMP target gene transcription that can correlate BMP signaling dynamics with timing and spatial patterns of gene expression. Second, I will examine crosstalk between BMP signaling and EGFR/ERK signaling by developing live imaging approaches and applying quantitative methods. Finally, I will expand the research scope by building tools to examine BMP signaling dynamics in the red flour beetle, Tribolium casteneum. By leveraging the unique characteristics of Tribolium, such as slower development and distinct embryonic tissue architecture, I will elucidate how the conserved BMP signaling pathway adapts and functions across species. The proposed multi-faceted approach, ranging from mathematical modeling to cross-species comparisons, promises to unveil fundamental principles of cell signaling and provide a foundation for further advancements in developmental biology and translational applications. To accomplish these goals, I have formed an exceptional committee of advisors who can aid in my diverse approach to understanding signaling dynamics in vivo. Together with my advisor, Dr. Stefano Di Talia, I have designed a training plan that will provide me with the skills required to run a productive, independent laboratory that tackles complex questions about the role of conserved signaling in developmental biology.

NIH Research Projects · FY 2024 · 2024-08

ABSTRACT Sickle cell disease (SCD) results in premature aging and early mortality. Although we (using >15 years follow up [f/u]) and others have shown that life expectancy in SCD has improved, adults with SCD still have a life expectancy of only 58 years. Early mortality is associated with SCD-related end-organ damage, especially to the heart, lung, kidneys and central nervous system, as well as with vaso-occlusive event frequency and biomarkers such as sVCAM-1. Most remarkable, however, is that adults with SCD >50 years (now 13% of adults with SCD) have physical function similar to non-SCD adults over the age of 80. While frequency of death peaks in the 5th decade, a subpopulation of patients live into their 70's. Yet little is currently known about how best to evaluate and care for SCD patients over age 50 or predict which patients will achieve that milestone. We hypothesize that multi-omics, by representing several SCD-related and unrelated physiologic processes, strongly determine-by direct and indirect effects-both SCD organ severity and survival. Analysis of these relationships can inform our understanding of the variability in survival and the pace of aging in SCD. Elucidating the multi-omic contributions to SCD severity and mortality will also be critical for developing models of assessment and care for this population. NHLBl's TOPMed program is an extraordinary opportunity to facilitate personalized medicine for SCD, including improving our understanding of factors affecting severity and mortality. Our cohort (OMG-SCD), together with other TOPMed SCD cohorts, total >4000 samples with whole genome sequence (WGS) results and rich clinical data, including organ-function phenotypes and clinical laboratory data; several studies also have survival data. The OMG-SCD cohort also has stored plasma samples, some of which have previously been used to identify proteins associated with kidney damage. Plasma protein activity can be modulated by N-linked glycosylation and thus contributes to health and aging. Yet the role of proteomics and N-linked glycosylation is unexplored in the context of aging in SCD. Discovery of proteomic biomarkers could illuminate the underlying biologic mechanisms of accelerated aging and mortality in SCD. Here, we propose to: (1) Identify novel clinical and 'omic risk factors, including telomere length, mitochondrial copy number, and clonal hematopoiesis, for organ dysfunction and early mortality and (2) Identify proteomic and glycoproteomic biomarkers for premature mortality. Our work is poised to yield significant discoveries regarding the nature of aging and to identify 'omic risk factors of poor prognosis in SCD, thus facilitating precision medicine-guided care models in SCD.

NIH Research Projects · FY 2025 · 2024-08

Title: Mitochondrial Regulation of Brain Network Dynamics in Stress Project Summary Stress-related neuropsychiatric disorders, such as bipolar disorder, schizophrenia, major depressive disorder (MDD), and post-traumatic stress disorder (PTSD), often exhibit mitochondrial dysfunction. Mitochondria play a critical role in responding to environmental stressors and influencing brain health throughout an individual's life. However, the specific impact of mitochondrial dysfunction on brain network activity under stress conditions and its subsequent effect on cognitive and behavioral changes remain poorly understood. The proposed research will use a multi-method approach that combines molecular, cellular, and circuit-level analyses to understand how mitochondrial bioenergetics and quality control mechanisms contribute to the synchronized brain network activity underlying cognition in normal and pathological states. The research strategy will focus on understanding basic mechanisms that regulate mitochondrial functions and how these processes impact brain networks in stress conditions. A comprehensive understanding of how key mitochondrial processes regulate the spatiotemporal dynamics of neural activity will provide fundamental insights into how these critical cellular processes contribute to brain function. In the present proposal, Dr. Quigley will test the hypothesis that the properties of mitochondria in distinct brain regions confer vulnerability to cellular stress that can be detected at the network level, contributing to behavioral outcomes. To do so, Aim 1 aims to investigate region and cell type specific mitochondrial phenotypes throughout the brain associated with stress. The approach will use a combination of imaging techniques, biochemistry, and physiological assays to examine subcellular differences in mitochondrial dynamics and protein expression under conditions of stress. Aim 2 aims to identify joint molecular and electrophysiological markers for changes in allostatic load induced by mitochondrial dysfunction. This will be achieved by first examining the relationship between in vivo mitochondrial functions, brain circuit activity, and stress by characterizing a method of light-activated induction of oxidative stress, followed by measurements of electrical activity and the use of statistical methods to make comparisons to behavioral stress. Subsequently, candidate markers will be identified from proteomic experiments to inform the measuring of joint neurophysiological and molecular biomarkers associated with stress and the application of advanced statistical methods to identify associated patterns that correlate with behavioral states. The proposed research will pave the way for further independent studies and may aid in developing new therapeutic avenues that span diagnostic categories. Combined with the applicant’s prior graduate training in acquiring and analyzing in vivo electrophysiology data, this work will facilitate Dr. Quigley’s transition to an independent academic research career by incorporating the use of molecular genetic techniques, imaging, and advanced computational analyses.

NIH Research Projects · FY 2025 · 2024-08

ABSTRACT This application is for a K01 Mentored Research Scientist Development Award to provide dedicated career development training for Dr. David Lee to launch his own independent research program. Dr. Lee has conducted basic science research in the areas of muscle cell biology, aging and metabolism. This K01 will enhance Dr. Lee’s ability to 1) become an expert in biology of Fibro/Adipogenic progenitor cells (FAPs) and muscle fibrosis, 2) conduct mass spectrometry-based science with state-of-the-art equipment, 3) use novel computational approaches to perform informatics analysis on large scale metabolomics data (to go form spectral peaks to specific pathways), 4) develop and implement strategies to manipulate Wnt driven glycolysis in FAPs to counteract muscle fibrosis and 5) develop high-quality independent research program that will allow collaborative and inclusive opportunities for science. To achieve the goals, Dr. Lee has devised a clear and focused training plan with clearly assigned individuals who will each contribute unique expertise to the aforementioned areas of training. Dr. Lee’s primary mentor is Dr. Christopher Newgard (career development, metabolomics interpretation). Dr. Lee’s Co-mentor is Dr. James Bain (mass spectrometry, metabolomics). Dr. Lee has also enlisted three Research advisors: Dr. Jianhong Ou (Bioinformatics, -omics data reduction/interpretation), Dr. Matt Hilton (career development, FAPs biology), Dr. James White (muscle, aging, sarcopenia). There exists a heavy social and economic cost associated with lost independence and mobility due to muscle weakness and injury. There are currently no effective strategies to fully counteract sarcopenia to restore mobility and strength. No rigorous research has investigated the fibrogenic development of FAPs in age-related muscle fibrosis, and I hypothesize that FAPs could be a cellular target for anti-fibrosis therapies to delay or prevent the loss of muscular strength in the context of sarcopenia. The first objectives (Aim 1) of the proposed studies are to determine the onset of FAP fibrogenesis through a time course study of sarcopenia development to inform therapeutic strategy. Second (Aim 2), I will determine if greater FAP glycolysis is a driver of FAP fibrogenesis using orthogonal metabolomics and glycolytic flux analysis. Finally (Aim 3), informed by my preliminary data, I will mechanistically determine if Wnt signaling drives FAP glycolysis and fibrogenesis making it a target for anti- fibrosis therapy to delay muscle weakness in sarcopenia. Together, the K01 training and mentorship will enable Dr. Lee to transition into an independent research career and become a leader in developing therapeutic strategies to combat age-related muscle weakness and fibrosis for translation therapeutics.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY/ABSTRACT Duchenne muscular dystrophy (DMD) is a devastating X-linked disease with no current cure. DMD is caused by mutations in the gene encoding dystrophin. Dystrophin is a protein necessary for the maintenance of muscle structure and is essential for skeletal and cardiac muscle integrity. As muscle damage progresses, the respiratory muscles become weak and fibrotic leading to hypoventilation and respiratory insufficiency. Sadly, most patients die from respiratory failure. Further, 1/3 of DMD patients also have neurological manifestations and central nervous system (CNS) pathology. However, the impact of the CNS pathology in the respiratory- related morbidity in DMD is unknown. Defining the respiratory muscle and neuropathology is essential as novel gene therapies using AAV-microdystrophin (AAV-µDys) for DMD enter clinical trials and become FDA-approved. Several pre-clinical trials with AAV-μDys reveal promising results with dystrophin production resulting in improved survival and ambulation; however, in these studies the diaphragm was not adequately transduced, and respiratory outcome was not assessed. Thus, there remains a critical need for a therapy that will halt or reverse respiratory disease. In our first aim, we will comprehensively examine breathing and the respiratory motor units in novel humanized mouse models and compare these to the established mdx DMD mouse model. We will then utilize these DMD mouse models to elucidate the impact of dystrophin deficiency on respiratory neuro-pathology. Finally, we will examine the efficacy of a novel AAV capsid carrying μDys in treating respiratory pathology and neuro-pathology in the DMD mouse models. The fundamental hypothesis driving this proposal is that dystrophin deficiency in both the respiratory muscles and CNS leads to breathing impairments, and AAV-μDys will correct both the respiratory myopathy and neuropathology. In Aim 1 we will identify the impact of dystrophin deficiency on respiratory function and histopathology in the humanized mouse models of DMD. In Aim 2, we will perform physiological, histological, transcriptional, and molecular studies to define the impact of dystrophin loss on the respiratory centers and motor neurons of the medulla and cervical spinal cord. Then, we will examine the ability of a novel AAVcc47-µDys to effectively transduce and correct respiratory muscle and neuro-pathology (Aim 3). The proposed experiments are innovative because the impact of dystrophin deficiency on breathing and neuropathology in humanized mouse models has not been previously examined. Defining this pathology will provide clinically relevant outcome measures for future therapeutic studies. Finally, the use of the novel AAVcc47 to deliver µDys and target respiratory pathophysiology and neuropathology will provide a much needed therapeutic option for respiratory insufficiency in DMD. Since AAV- µDys gene therapy is already in clinical trials, this work has the strong potential to quickly translate to clinic and inform and impact the clinical treatment of boys and young men with DMD.

NIH Research Projects · FY 2024 · 2024-07

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. Visceral adiposity is a key risk factor for the development of type 2 diabetes and metabolic syndrome, owing to its role in promoting a chronic inflammatory state. While behavioral traits are a key driver for the accumulation of visceral adipose tissue (VAT), it is now well recognized that the microbiota is critically involved in regulating VAT development and remodeling. However, there is a gap in knowledge regarding the specific mechanisms underlying this process. We recently identified Clostridium immunis, a new human commensal bacterium, that reduces serum triglycerides, body weight, and VAT in mice, and we have purified and characterized an exopolysaccharide (EPS) secreted by C. immunis that recapitulates these activities. Moreover, we have demonstrated group 3 innate lymphoid cells (ILC3s) and IL-22 are both critically required for C. immunis and its exopolysaccharide to protect against metabolic diseases. Collectively, our preliminary data establish that C. immunis is the first human commensal bacterium that negatively regulates the development of VAT in an immune-dependent manner. The overarching goal of this proposal is to determine the mechanism by which C. immunis EPS decreases VAT. We hypothesize the C. immunis EPS results in decreased VAT by reducing ILC3-derived IL-22, thereby increasing energy expenditure. In the first Aim, we will analyze energy balance under a range of relevant environmental conditions. In the second Aim, we will determine the immunomodulatory effects of C. immunis EPS. Taken together, the proposed experiments will provide foundational insight into microbiome–immune–metabolism interactions. More broadly, these studies will highlight new microbiome-derived therapeutic opportunities for treating visceral adiposity and its associated complications.