University Of Pennsylvania

NSF Awards · FY 2025 · 2025-03

After infancy most mammals stop secreting lactase (the enzyme that breaks down milk’s sugar, lactose) and lose their ability to digest milk. Some humans, however, can digest milk throughout their life because they carry genes that code for lactase persistence (LP). These genes emerged independently, and were selected for, in several populations where milk was a central food staple. This study examines the genetic, microbiome, and cultural factors that allow adult milk digestion among groups of peoples that are milk-dependent but lack LP genes. Additionally, the study examines whether genes that impact lactose tolerance co-evolved with those that permit the metabolization of high levels of milk-derived sugars. The study provides training opportunities in field, genetic, and computational methods and is relevant to human evolutionary history and public health. This study collects novel genetic, microbiome, and phenotypic data to expand current knowledge on genetic and cultural adaptations to milk consumption. Cutting-edge functional genomics approaches enable the identification of novel enhancer regions near the lactase gene and near genes involved with sugar metabolism. Genetic data integration allows for the identification of novel genetic variants influencing lactase persistence. Lactose tolerance tests are applied in tandem with a genome wide association study (GWAS) to identify novel genetic loci linked to milk-induced blood sugar level increases. Gut microbiome sequencing enables the identification of bacteria that aid in digesting lactose and quantifies the extent to which milk preparation practices shape the composition of the gut microbiome. The study creates a large genomic and phenotypic database that informs the understanding of recent human evolutionary history and gene-culture co-evolution. 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 2025 · 2025-03

Nonconvex statistical estimation and learning algorithms are dramatically improving our capacity to efficiently learn from massive datasets, reshaping society through new technological capabilities in healthcare, imaging, transportation, and information processing. Although such learning algorithms have had widespread empirical success, we have yet to find a coherent mathematical foundation that can explain not only why they work and what tasks they provably solve, but also how practitioners can improve their performance either by adjusting the algorithm or even the task itself. The investigator aims to lay this foundation by advancing the design, analysis, and deployment of rigorously justified nonconvex optimization algorithms. This research will create guaranteed procedures for training practical machine learning systems deployed in government and industry, producing more reliable and robust predictive models with fewer data and computational resources. The investigator will incorporate results from this project in education efforts, including course development, local K-12 outreach, and research mentoring of Ph.D. and undergraduate students. In this project, the investigator designs and analyzes nonconvex optimization algorithms. The project focuses on simple iterative methods that compute with data in its ambient form, a class of algorithms that are uniquely scalable to modern high-dimensional statistical estimation and learning tasks. The overarching goal of the project is to understand when these methods converge to local or global optima and to provide efficiency estimates of their performance, measured both in terms of data and computational resources consumed. To achieve this goal, the investigation will draw on the techniques of variational analysis, nonsmooth optimization, machine learning, statistics, and high-dimensional probability. The investigator will leverage these techniques to design and equip simple, scalable iterative methods for nonconvex data fitting problems with strong performance guarantees: generic initialization strategies, rapid local convergence near optima, and seamless adaptation to nonsmooth constraints, models, priors. Such performance guarantees guide the practical implementation of reliable and efficient numerical methods for high-dimensional estimation and learning. 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 2026 · 2025-03

Clinical microbiome studies show that C. albicans, along with elevated levels of S. mutans, in plaque (biofilm) is strongly associated with severe early-childhood caries (S-ECC). Previously, we demonstrated that C. albicans interacts with S. mutans and enhances the accumulation and virulence of plaque-biofilms, amplifying the disease severity in vivo. We found that the presence of C. albicans activates S. mutans Gtf exoenzymes production and promotes extracellular glucan synthesis, cross-feeding/co-metabolism, and acid production, leading to rampant and extensive carious lesions (similar to those found in S-ECC) in a rodent caries model. However, a key question remains unanswered. How C. albicans interacts with S. mutans to initiate the pathogenic process? Recently, we discovered that C. albicans forms highly structured assemblages with S. mutans that are uniquely found in the saliva of S-ECC patients. The C. albicans (Ca)-S. mutans (Sm) assemblage binds avidly to apatite surface through hyphae acting as anchoring points. The attached assemblage displays a new type of cell-group mobility that is mediated by the fungal hyphal formation and elongation in saliva, which promotes rapid surface spreading with enhanced growth and acidogenesis causing more extensive and severe enamel decay than either species alone. Conversely, inactivation of fungal growth or filamentation prevents surface mobility/spreading and cariogenic activity. In this proposal, we will investigate how C. albicans mediates this interkingdom assemblage formation and its emergent properties to initiate pathogenic biofilms. Understanding the early events of Ca-Sm interactions that coordinates effective colonization and rapid biofilm formation will provide new mechanistic insights about the rampant nature and severity of S-ECC carious lesions, which remains poorly understood. Fungal transcription factors (TF) and protein kinase (PK) networks play key roles in Ca filamentation but are highly influenced by the niche environment. Preliminary data show that Ca hyphal morphogenesis in saliva is distinct from previously studied conditions. Thus, we hypothesize that interkingdom assemblage and surface mobility in the unique dental niche depend on novel C. albicans TF and PK networks that respond to saliva and presence of S. mutans, enhancing surface colonization, biofilm spreading and virulence. In Aim 1, we will identify C. albicans TFs and PKs and their target genes that regulate hyphal morphogenesis in saliva with S. mutans mediating interkingdom assemblage formation, colonization, and mobility. In Aim 2, we will investigate how TF and PK networks modulate biofilm initiation/spreading and enamel demineralization. Then, in Aim 3, we will examine the role of TF and PK in biofilm virulence and their impact on oral microbiome and caries severity in vivo. A comprehensive program from laboratory studies to in vivo investigations is offered to understand the underlying mechanisms governing this highly virulent interkingdom interaction and its implications in S-ECC, an unresolved public health problem.

NIH Research Projects · FY 2026 · 2025-03

Project Summary: The outlook for many childhood-onset genetic conditions has markedly improved in the last few decades because of specialized care, supportive treatment, and the introduction of disease-modifying therapies. Thus, increasing numbers of adults with childhood-onset genetic diseases are outliving their historical expectations. These welcome improvements create substantial prognostic uncertainty because the long-term impact of both novel treatments and aging in these diseases are unknown. From a psychosocial perspective, affected adults often grew up with expectations of a shortened lifespan, expectations that may no longer be accurate. Yet, little is known about how they conceptualize their evolving and uncertain prognoses or plan for their futures in light of these changes. Despite NIH’s Inclusion Across the Lifespan policy, these adult cohorts remain under-studied, surviving on an expanding but uncharted frontier of genetic medicine. As outcomes for many childhood-onset genetic conditions evolve with new therapies, there is a critical need to characterize how adults plan their lives given shifting prognoses. If this need remains unmet, genomic medicine risks replicating age biases towards studying younger cohorts, neglecting the needs of adult patients, and missing opportunities to anticipate the psychosocial needs of future adult cohorts with genetic conditions that have shifting prognostic implications. By using a disability studies framework to examine how adults with cystic fibrosis (CF), a prototypical childhood- onset chronic genetic condition, conceptualize their futures and plan their lives, I aim to identify novel clinical opportunities to support patients as they navigate future-oriented decisions amid prognostic uncertainty. The Aims of the proposed research are: Aim 1: Describe how adults with CF plan their futures via qualitative interviews with diverse sample of adult CF patients (N=45) about how their prognostic expectations inform future- oriented decisions. Aim 2: Describe how CF clinicians approach communicating with CF adults about planning their futures via qualitative interviews with clinicians (N=30) about whether and how they incorporate changes in disease trajectory into conversations with CF patients about healthcare and life decisions. Aim 3a: Develop a conceptual model that integrates insights from patient and clinician interviews with theory from disability studies to identify clinical opportunities to support adult CF patients as they plan for their futures. Aim 3b. Explore the relevance of the conceptual model to other contexts via focus groups involving adults with other childhood- onset genetic conditions. The candidate’s career goals are: 1) To produce impactful scholarship that improves the lives of adults with childhood-onset genetic conditions. 2) To pursue a career as an independent ELSI investigator by establishing an externally funded research program. Throughout the K01 Award period, the candidate will undertake coursework and directed readings in qualitative research methods, disability studies, and developmental psychology and will pursue activities focused on building a professional network around disability and genomics.

NIH Research Projects · FY 2026 · 2025-03

Project Summary Infection by many pathogens leads to the formation of structures termed granulomas, organized clusters of macrophages, neutrophils, and lymphocytes, that contain and control the infection, but can also serve as sites for pathogen replication and dissemination. Infection by enteric Yersinia leads to the formation of pyogranulomas (PGs, granulomas enriched in neutrophils) in infected intestinal and systemic tissues. We recently reported that CCR2-dependent inflammatory monocytes are essential for acute control of Yp within infected sites, were required for formation of organized PGs, and were necessary for the activation of neutrophils and production of IL-1 cytokines within the granuloma. We also recently demonstrated for the first time, the existence of intestinal pyogranulomas (PGs), following oral Yersinia pseudotuberculosis (Yp) infection. PGs contain high numbers of viable bacteria relative to adjacent non-PG intestinal tissue, and are enriched in neutrophils and inflammatory monocytes. Intestinal PGs contain similar bacterial burdens as Peyer’s patches, suggesting that PGs represent a previously undescribed site for Yp interaction with the mucosal immune system. Critically, whole-body TNFR1 deficiency, as well as loss of TNFR1 expression on monocytes phenocopied susceptibility of Ccr2-/- mice to Yp infection, and also resulted in failure to form organized PGs or activate neutrophils. Notably, loss of TNFR1 signaling, or loss of monocytes, led to reduced levels of IL-1 production in intestinal PG, and TNFR1 deficiency led to cell-intrinsic loss of IL-1 production. Furthermore, non-hematopoietic IL-1R signaling was needed to mediate control of Yp infection. These findings provoke the hypothesis that a monocyte-driven TNF-IL-1 signaling circuit mediates control of Yp infection. Based on these findings we propose the following specific Aims: First we will define how monocyte-specific TNFR1 signaling contributes to expression of IL-1 in innate immune cells. Second, we will dissect the requirement and sufficiency of IL-1R signaling on intestinal epithelial cells in control of Yp infection. Finally, in collaboration with the Waldor laboratory, we will utilize a newly-developed next- generation chromosomal barcode library together with deep sequencing and a powerful computational toolkit to define the replication and dissemination dynamics of Yp in wild-type and immune-compromised settings.

NIH Research Projects · FY 2026 · 2025-03

Bispecific antibodies have emerged as a promising cancer treatment, with a growing list of encouraging clinical results. Bispecific antibodies enable the binding of two separate targets or the binding of two distinct sites on the same target, simultaneously. This can have important implications when applied as a therapeutic, such as improved specificity and/or unique biological effects. In one common application, bispecific antibodies are designed to physically bring T cells and cancer cells closer together to enhance the immune clearing of cancer cells. Demonstrating the promise of T cell-redirecting bispecific antibodies, blinatumomab, an anti-CD3 x anti- CD19 pair, has produced clinical remission in precursor B cell acute lymphoblastic leukemia at thousand-fold lower dosages than rituximab (anti-CD20 monoclonal antibody), without needing a secondary T cell co- stimulatory signal. In contrast, conventional antibody therapies require cumulative antibody doses ranging from 5-20 g per patient over the course of months to years. Despite the promise of bispecific antibodies (and other immunotherapies), these approaches typically rely on the targeting of a single tumor-associated antigen (TAA) that is over expressed by tumor cells. This can often lead to objective tumor response rates, particularly in solid tumors, since tumor biomarkers are generally not ubiquitously or uniformly expressed by all tumor cells and can be lost during disease progression. To overcome these challenges, we propose to harness a patient’s own anti-tumor autoantibodies, to confer specificity for tumor antigens. Anti-tumor autoantibodies are found within tumors at concentrations that are up to 64-fold higher than normal tissue and evolve with disease progression. Further, there is a rich landscape of recent evidence showing that autoantibodies isolated from tumors are specific to personalized panels of tumor-associated antigens in various cancers, with high clonal and antigen-targeting diversity. This makes autoantibodies well-suited for differentiating between normal and disease pathologies. In our recent work, published in Science Advances, we showed that the transformation of endogenous anti-tumor autoantibodies into highly potent bispecific T cell-redirecting autoantibodies (TRAAbs) can trigger the T cell mediated cytolysis of tumor cells and lead to tumor regression. To our knowledge, this was the first demonstration that increasing the therapeutic efficacy of a patient’s own anti-tumor antibodies offers a viable therapeutic approach; however, a limitation of this approach is the indiscriminate conversion of all serum antibodies into bispecific antibodies, including those with no specificity for the tumor target. The overall goal of this proposal is to engineer a protein therapeutic that limits the transformation of endogenous anti-tumor autoantibodies into highly potent TRAAbs to those antibodies that reside in the tumor microenvironment. The specific aims for this proposal are: Aim 1: Optimize the engineered protein therapeutic to limit TRAAbs to the tumor microenvironment. Aim 2. Determine the distribution and therapeutic efficacy of the engineered protein.

NIH Research Projects · FY 2026 · 2025-03

Project Summary Despite the availability of a variety of intervention approaches for substance use disorders (SUD), many indi- viduals do not benefit from existing interventions either because they are not offered at the time in which they can be most beneficial and/or they are not tailored to the changing needs of individuals. Modern technologies such as mobile and wearable devices provide unprecedented opportunities to deliver just-in-time adaptive in- terventions (JITAIs)–an intervention design that adapts intervention delivery to an individual’s rapidly changing internal state and context, in real-time, real-world settings. However, new research methodologies are needed to capitalize upon these opportunities and inform a new generation of interventions for SUD. Recent advances in Micro-Randomized Trials (MRTs) support the collection of data that can empirically inform the construction of JITAIs. Although the MRT is a major step forward in methodology for building empirically- based JITAIs, new data analytic methods are needed to fully realize the potential of these data. Specifically, there are several important gaps in existing data analytic approaches. First, these methods can be used to evaluate a set of pre-specified JITAIs and assess intervention effects on proximal outcomes (e.g., next-day self-reported substance use) but not to construct an optimal JITAI with respect to a distal outcome (e.g., substance use by a 6-month follow-up). This represents a major gap given that long-term benefits are the primary motivation of SUD treatments. Second, existing methods are limited to intention-to-treat (ITT) analyses, which may provide biased estimates of the effec- tiveness of an intervention when individuals are not fully engaged. The latter is a major challenge in developing effective JITAIs given that engagement in mobile health (mHealth) interventions is often suboptimal and declines over time. When the rate or the pattern of intervention engagement in the study vary from the rate or the pattern of engagement achieved when implementing the intervention in real-world, the results obtained by ITT analyses might not be reproducible. Third, there is no variable selection method in MRT settings to identify the important tailoring variables. These gaps can seriously limit the usefulness and generalizability of the results obtained from MRT data. We propose to close these gaps by developing new methods that can help SUD investigators leverage MRT data to construct a JITAI that is most beneficial in terms of a distal outcome (Aim 1). We further propose to develop new methods that accommodate intervention engagement when estimating the optimal JITAI (Aim 2). We will also develop a method that enables investigators to identify important tailoring variables for inclusion in the optimal JITAI (Aim 3). We will apply these methods to rich data from 3 funded studies, each employing an MRT to inform a JITAI for substance use. These data sets will be used to motivate the new methodologies, as well as to illustrate their utility as we disseminate them to SUD investigators via open-source, easy-to-use statis- tical software (Aim 4). The successful completion of this work will facilitate the development of empirically-based, effective, interpretable and generalizable JITAIs to prevent and treat SUD and potentially other chronic disorders.

NIH Research Projects · FY 2026 · 2025-03

Project Summary: Prostate cancer (PCa) affects nearly 250,000 men in the United States annually, making it the second leading cause of cancer-related deaths in men, with over 30,000 fatalities each year, mainly due to metastatic castration-resistant prostate cancer (mCRPC). The androgen receptor (AR) is a ligand-responsive transcription factor that drives terminal differentiation of the prostatic luminal epithelia. However, AR gets hijacked upon transformation, and the cancer cells become dependent on its activity. This dependency makes androgen/AR-targeted therapies common after initial treatments like surgery or radiation. However, the disease often returns through various mechanisms that restore AR-signaling, driving cancer progression. Despite advancements in antiandrogen therapies, 20-40% of mCRPC patients show resistance to drugs like abiraterone and enzalutamide, and many others develop resistance over time. A major challenge in treating advanced PCa is its reliance on AR-driven oncogenic transcriptional program, which involves multiple cofactors and chromatin proteins. Our long-term goal is to identify, characterize, and therapeutically target these cofactors and chromatin-associated proteins in the context of AR signaling in prostate cancer. We previously demonstrated that NSD2 overexpression is associated with PCa progression. NSD2 (also named MMSET and WHSC1), a histone lysine methyltransferase that catalyzes H3K36me2, is also implicated in the pathogenesis of multiple myeloma and other hematological malignancies. Here, we identify NSD2 as a critical subunit of AR enhanceosome (protein complex that assembles at enhancer regions) required for the oncogenic AR transcriptional program. Our preliminary work indicates that the NSD2 interacts with AR through its HMG-box domain and requires NSD2 catalytic activity to form the functional AR enhanceosome. In Aim 1, we propose to gain deeper molecular mechanistic insights into how NSD2 influences the chromatin landscape to facilitate the assembly of the AR enhanceosome complex and enhance long-range enhancer-promoter looping for a hyperactive AR transcriptional program in PCa cells. In Aim 2, we will evaluate the therapeutic potential of a novel first-in-class small molecule inhibitor of NSD2, which also inhibits its paralog NSD1, in AR-driven prostate cancer in vivo. This includes assessing the on-target activity and anti-tumor effects of the compound in various prostate cancer models, including naïve and Enza-refractory CRPC, primary explant, and organoid models. Together, these studies will provide critical new insights into how epigenetic modifiers regulate oncogenic transcription factors to drive cancer progression.

NIH Research Projects · FY 2026 · 2025-02

Candidate: To achieve her career goal of becoming an independent investigator, K. Jane Muir, PhD, APRN, FNP-BC seeks mentored research training in population health research, machine learning techniques, and causal inference approaches. This career development award identifies modifiable nursing features to improve emergency department (ED) patient outcomes with a focus on reducing differences in clinical outcomes. Research Context: Hospital EDs have the potential to save lives, but pervasive delays and chaotic aspects of emergency care place patient’s health outcomes at risk while widening differences in care among patients with Medicaid or without insurance. Registered nurses direct all processes of care in hospital EDs including triage, throughput, and patient disposition, yet few studies have determined whether they are adequately resourced to do so. This study aims to determine the extent to which hospital nursing models of care are associated with patient outcomes to improve ED care. Specific Aims. 1) To determine which nursing models of care (defined by different combinations of ED and inpatient nursing resources) are associated with ED patient outcomes; 2) To determine the association of nursing models of care on ED patient outcome differences. Research Plan: Datasets include, 1) Penn’s Nurses4All multi-state nurse survey, 2) the American Hospital Association Annual Hospital Survey, 3) AHRQ Healthcare Cost Utilization Project patient database. In Aim 1, machine learning will be used to identify nursing models of care characterized by ED and inpatient nursing resources (nurse staffing levels, nurse work environments, skill mix, nurse practitioners) associated with ED patient outcomes (ED length of stay, ED revisits, disposition against medical advice, 30-day hospital readmissions, in-hospital mortality) in hospitals overall with separate comparisons among hospitals of differing disproportionate share (DSH) status. In Aim 2, differences in ED patient outcomes associated with nursing models of care will be examined across populations. Career Development Plan: With an interdisciplinary and experienced team of mentors, Dr. Muir will pursue didactics, seminars and conferences to complete the training goals, which are to 1) cultivate and apply expertise in the theory, design, and evaluation of research advancing population health; 2) apply advanced machine learning techniques to develop hospital nursing models of care defined by combinations of ED and inpatient nursing resources; 3) expand and apply skills in causal inference approaches with observational data. Environment: The University of Pennsylvania School of Nursing offers an ideal environment to pursue the proposed training and research. Dr. Muir is well-positioned to successfully complete the proposed aims and training because of her experienced mentorship team and extensive resources for career development.

NIH Research Projects · FY 2024 · 2025-02

Project Summary This F99/K00 Transition to Aging Research for Predoctoral Students Award application will facilitate the research training of the applicant to launch an independent career in aging research focused on examining the influence of hearing loss on healthcare engagement and health outcomes. The proposed research will examine how hearing loss influences care engagement, health communication and healthcare use for diverse populations of older adults. In the F99 phase of this award, the applicant will examine the role of hearing loss in self-maintenance practices for chronic illness, termed self-care of chronic illness. This study proposes to (1) describe self-care behaviors in older adults with hearing loss and compare self-care behaviors by hearing loss severity, (2) explore the experience of caring for chronic illness while having hearing loss, and how hearing loss impacts self-care, and (3) describe differences in the experiences of self-care of chronic illness by self-care behavior scores and hearing loss severity. This mixed methods study will use a parallel-convergent design to enroll a single diverse sample of community-dwelling hospitalized older adults with a chronic illness and hearing loss. In the K00 phase of this award, the applicant will expand on F99 work to examine (1) how the intersection of hearing loss and social determinants of health influence healthcare use outcomes and (2) how hearing loss and its intersection with race influence the dynamics of patient-provider interaction. K00 research will be conducted using epidemiologic datasets linked with claims data, prospective data collection, and conversation analysis approaches. Research training will occur in resource rich environments with support from a cross-disciplinary mentorship team with expertise in aging, hearing science, chronic illness, epidemiology, and the required methodologies. The applicant proposes a training plan that includes coursework, practical training, seminars, and engagement in ongoing research. Training will focus on methodological skills, ethical research practices, epidemiology, aging, and career development. This proposal aligns with the priorities of the National Institute on Aging by addressing the burden of age-related diseases and disabilities to improve health and function in aging. Having a greater understanding of how hearing loss influences health in diverse populations aging with chronic illness will inform the development of interventions to improve care delivery and maximize health for the millions of adults aging with hearing loss and a chronic illness.

NIH Research Projects · FY 2026 · 2025-02

Summary/Abstract The objective of this proposal is to develop a molecular toolbox for non-invasive methods to kill cells in situ via specific cell death pathways using safe, penetrant, and tunable temperature stimuli. There is an urgent need for new technology that can engage specific forms of cell death in vivo in a manner that is controlled in space, time, and magnitude. Such technology would enable researchers to systematically interrogate the distinct physiological consequences of programmed cell death through apoptosis, necroptosis, or pyroptosis, and to harness immunogenic types of cell death for cancer therapy. However, current methods to stimulate cell death are either not specific to individual types of death, lack spatiotemporal precision, or are limited by poor tissue penetration. Our proposed molecular tools overcome these limitations by leveraging the unique advantages of temperature as a control signal, including its ability to be modulated non-invasively with high spatial and temporal resolution even deep within tissue. We will generate single protein constructs that can trigger apoptosis, necroptosis, or pyroptosis with either gentle heating or cooling, and validate their efficacy in mouse models of human cancer. The rationale for our work is that this toolset will open powerful new avenues of discovery by enabling the study of programmed cell death pathways in living animals with a high degree of control. Moreover, the ability to selectively engage immunogenic cell death in a dose-dependent manner directly within tumors would establish a novel strategy for targeted, personalized cancer immunotherapy with the potential to be more efficacious and less toxic than current approaches. In preliminary studies, we engineered a temperature-responsive protein called Melt that clusters upon cooling and can induce cell death when fused to caspase-1. We demonstrated that Melt-caspase1 efficiently eliminates cancer cells in vitro and in mouse xenografts with high spatiotemporal precision. Building upon this promising foundation, we will pursue three specific aims: 1) Generate a suite of Melt fusions for cold-induced control of apoptosis, necroptosis and pyroptosis; 2) Test the ability of Melt-induced pyroptosis to stimulate immunogenic cell death and protect against rechallenge in an immune-competent mouse model; and 3) Develop proteins that induce cell death upon gentle heating by fusing effectors to proteins that self-assemble between 37-42C. We will achieve these aims using molecular engineering, live cell imaging, custom devices for feedback-controlled temperature regulation in cells and in mice, and syngeneic mouse models of human cancer. Success in our work will open new horizons for both fundamental discovery and therapeutic translation. Beyond cell death, this technology will enable remote control of a wide array of cell and molecular events directly within living mammals, a capability that promises to broadly impact both basic and applied biomedical research.

NIH Research Projects · FY 2026 · 2025-02

Project Summary Hypertrophic cardiomyopathy (HCM) affects 1 in 500 adults, with treatment limited to managing symptoms rather than addressing the root cause. Around 40% of familial HCM patients have mutations in the gene for myosin- binding protein C (MYBPC3), causing the disease. Theoretically, a single dose of a genomic editor correcting MYBPC3 mutations in the heart would cure the disease for these patients. While the base editors (BE), fusing a nickase-Cas9 (nCas9) to a cytosine deaminase (CBE) or adenine deaminase (ABE), can achieve high-efficiency single nucleotide substitutions, their propensity for off-target editing due to a lack of temporal control over the editing activity poses significant safety concerns. We recently developed split ABE (sABE) that utilizes chemically induced dimerization (CID) to control the TadA deaminase activity. Our sABE retains a high on- target editing activity similar to the canonical intact ABE but displays significantly reduced off-target editing with a narrower activity window and improved precision. When delivered via dual adeno-associated virus (AAV) vectors, sABE has achieved efficient single base conversion on the PCSK9 gene in mouse liver. This gene is a prominent drug target for atherosclerosis and regulating blood cholesterol levels. This achievement marks the first in vivo CID-controlled gene knockdown. We propose addressing the current weaknesses of the sABE system and applying it to HCM treatment. In Aim 1, We will expand CID systems and optimize the splitting strategy in sABE. Our current sABE employs a rapamycin-dependent CID system. Rapamycin has immunosuppressive and autophagic effects in vivo, which can limit its applicability to certain diseases. In Aim 2, we will create sBE variants to achieve versatile base editing with low off-target activity. sABE can only achieve A-to-G editing, while disease-causing mutations can result from other mutation types. We will translate our inducible design to an expanded set of BE systems using sTadA deaminase variants, achieving C-to-T and C- to-G editing, etc. We will then apply sBE variants to knock down PCSK9 in vivo to demonstrate their functionality. In summary, achieving tight spatiotemporal control of the base editing enzyme activity with alternative small molecule inducers will yield versatile split base editors, significantly reducing genomic and transcriptomic off- target editing while maintaining similar levels of on-target editing activity. Correcting MYBPC3-associated HCM and decreasing PCSK9 expression and blood cholesterol levels will validate these sBE systems in vivo, laying the groundwork for their future adoption in treating a broad range of diseases.

NIH Research Projects · FY 2026 · 2025-02

Dermal resident memory CD4+ T cells (dTrm) are uniquely positioned to protect against vector-borne pathogens such as Leishmania major. However, despite their significance, the ontogeny of dTrm cells and the influence of the skin microbiome and T cell intrinsic signals that promote their retention following entry, remain unclear and often controversial. We aim to define how dTrm cells develop and use the information to optimize dTrm cell-mediated immunity in vaccines. In our preliminary studies, we identified protective Th1-biased CD4+ dTrm cells in mice immunized with an experimental peptide vaccine based on an immunodominant leishmanial antigen, phosphoenolpyruvate carboxykinase (PEPCK). Curiously, while a PEPCK-mRNA- lipid nanoparticle (LNP) vaccine promoted the entry of PEPCK-specific T cells into the skin, the T cells were not maintained. We will leverage this novel finding to discover the factors required for developing and maintaining protective dTrm cells globally seeded in the skin. We will: 1) Determine if precursor dTrm cells develop before entry into the skin. This information will provide information on how to expand this population in a vaccine; 2) Identify the role of the skin microbiome in the development and retention of dTrm cells in the skin. The skin microbiome has a large influence on T cells in the skin, and we will test how bacteria influence both the development and retention of dTrm cells; and 3) Expand the pool of protective dTrm cells in the skin. Based on our findings, we will test several approaches to expand the dTrm cell population. These include altering the cytokine milieu and inducing the expression of essential transcription factors for dTrm cell development. These studies will be done in murine models, taking advantage of cutting-edge technology to interrogate T cell responses in immunized mice, including identifying critical genes and proteins associated with dTrm development by cellular indexing of transcriptomes and epitopes, flow cytometry, deletion of genes by CRISPR/Cas9 editing, ectopic expression of genes by retroviral transduction, and in vivo targeted mRNA-LNP delivery of critical factors to promote Trm cells. Leishmaniasis is a neglected disease that causes a wide range of clinical entities. Drugs to cure leishmaniasis are often ineffective and toxic, and there is no vaccine for the disease. The inability to create a protective vaccine is partly because protection is mediated by T cells, rather than antibodies– the target for most vaccines. We have shown that dTrm cells can provide long-lasting protection in mice that have healed a primary infection, and we hypothesize that vaccines that expand a dTrm cell population will lead to an effective vaccine. Thus, these studies have direct relevance to developing a vaccine for leishmaniasis. Additionally, there are many vector-borne diseases with no vaccines, and we believe that expanding dTrm cells in vaccines for other vector-borne diseases will be essential for optimal protection against a wide range of infections.

NIH Research Projects · FY 2025 · 2025-02

Project Summary Bone marrow is a highly complex and heterogeneous environment, composed of hematopoietic, mesenchymal, nervous and endothelial tissues. It is the site of a wide range of diseases including blood and immune disease (leukemia, myelodysplastic syndromes, aplastic anemia, myelofibrosis), skeletal disease (osteoporosis, osteopetrosis, Paget’s disease), vascular disease (avascular necrosis, ischemic and hypertension-related bone disease), and cancer metastasis (breast, prostate, thyroid). There is a great appreciation for the role of mesenchymal cells in orchestrating bone remodeling, angiogenesis and hematopoiesis – functions that are critical to maintaining health and that deteriorate with aging – but there is little knowledge on how this population is maintained or how it helps regulate the cellular diversity in bone marrow. Transcriptomic data has started to shed light on bone marrow cell regulation, but since bone marrow is primarily composed of hematopoietic cells, current data, particularly from humans, lack a holistic representation of all bone marrow cell types. Additionally, there is a lack of human data regarding how mesenchymal stem and progenitor cells are maintained, how they repopulate the bone marrow stroma and which mesenchymal cells are critical to each stage of hematopoiesis. This project seeks to achieve two interconnected goals: generate a complete single-cell transcriptomic and regulomic map of human bone marrow cells and experimentally establish the mesenchymal hierarchy in human bone. In order to accomplish this, we developed a protocol to isolate single-cell suspensions from human femoral head tissue, enrich for rare or difficult to isolate cell populations (i.e. mesenchymal cells, adipocytes, hematopoietic stem and progenitor cells) and isolate single-nuclei for simultaneous transcriptomic and epigenomic sequencing. This information will provide insight into how cell fates are regulated and what cell-cell interactions support cell fate decisions. Of particular interest is what epigenomic changes determine mesenchymal cell fate and what epigenomic states of hematopoietic cells make them more amenable cell regulation be one mesenchymal cell over another based on transcriptomic signatures. Transcriptomic and epigenomic studies will complement in vivo explant and in vitro differentiation studies to determine mesenchymal lineage differentiation patterns. By performing flow sorting for four progenitor cells, the studies will define the differentiation potential and position each cell on the step-wise differentiation pathway to osteogenesis and adipogenesis. This will provide insight into how mesenchymal cell fate is regulated and what cell types are required for hematopoietic recruitment and maintenance. By unraveling the complex bone marrow cellular signaling networks, the long-term goal is to develop novel therapies for osteoporosis and blood disease and provide the research community with a rich dataset to catalyze further discoveries.

NIH Research Projects · FY 2026 · 2025-02

Project Summary/Abstract The goal of this proposal is to understand how replication and transcription are spatially and temporally coordinated at molecular scales during embryonic development. A long-standing assertion is that RNA Pol II (Pol II) is completely evicted from DNA during replication to avoid steric conflict between transcription and replication machinery. This eviction hypothesis rests largely on in vitro or population and time-averaged genomics or biochemical experiments. These either may not recapitulate endogenous contexts or are unable to capture the interactions kinetics of Pol II’s association with chromatin with adequate temporal resolution (of seconds to minutes). In fact, recent in vivo experiments suggest that Pol II is not completely evicted from chromatin but remains in proximity (~40nm) to promoters and gene bodies during replication. This relationship is facilitated by Pol II interacting with PCNA, a sliding clamp protein associated with actively replicating domains in the nucleus that grow and disappear as replication completes. These contradictory findings call into question the molecular mechanisms that govern the coordination of transcription and replication machinery. I propose to use advanced live microscopy and single molecule tracking to directly measure the interaction kinetics of transcription and replication proteins in real time in developing Drosophila embryos. This work will provide molecular scale insights on the coordination of replication and transcription in an in vivo context. Our lab has established high resolution light-sheet microscopy which enables tracking of single protein molecules within the nuclei of live Drosophila embryos. Single molecule tracking reveals how individual proteins move within the nucleus and the kinetics of protein-protein and protein-chromatin interactions. Using these approaches along with perturbations to replication and transcription, I will investigate the chromatin binding kinetics of Pol II and PCNA as transcription is activated post-replication. To further understand how transcription is reactivated post-replication I will similarly investigate the distributions and kinetics of the pioneer transcription factor Zelda that is a ubiquitous activator in Drosophila embryos. Preliminary data from our lab shows that Zelda is excluded from domains of active replication marked by PCNA yet is detected near these sites. I hypothesize that i) Pol II is retained within actively replicating domains to avoid complete eviction from chromatin to enable rapid re-engagement and that ii) Zelda swiftly transitions from its excluded state post-replication to engage at these sites to facilitate transcriptional activation.

NIH Research Projects · FY 2026 · 2025-02

Project Summary/Abstract Stress and stressful environments often lead to maladaptive behaviors that can increase vulnerability to substance use disorders. The paraventricular nucleus of the thalamus (PVT) has been implicated in both stress and drug reward with a population of mu-opioid receptor-expressing neurons within the PVT (PVT- MOR+ neurons) potentially important for the maladaptive behaviors associated with stress. Individuals’ response to stress varies with some showing resilience, while others show susceptibility and are prone to the detrimental effects. While the relationship between stress and substance use disorders is well documented, there exists a significant gap in knowledge regarding the role of individual differences in stress susceptibility in mediating opioid reward. Furthermore, the neural pathways and cell types mediating the development of resilience or susceptibility phenotypes is unknown. The overall goals of this proposal are to A) determine how susceptible and resilient phenotypes influence subsequent opioid-taking and B) if a population of mu-opioid receptor (MOR) positive neurons in the PVT are necessary for the development of stress phenotypes. Our hypothesis is a susceptibility phenotype following social defeat stress will result in increases in opioid intake. We further hypothesize that activation of PVT-MOR containing neurons is necessary for establishing susceptibility following chronic social stress. Aim 1 will investigate how chronic stress response phenotypes influence opioid self-administration. Aim 2 will examine the role of PVT-MOR+ neurons in the development of stress phenotypes using chemogenomic tools to manipulate PVT-MOR+ neurons during chronic social defeat stress. Successful completion of these aims will establish if stress responsive subtypes differentially influence opioid reinforcement and will clarify the role of PVT-MOR+ neurons in behavioral responses to chronic social stress. Through this fellowship, Ms. Tyner will accomplish the defined Training Goals, including 1) technical training and mastery of neural circuit manipulation and viral imaging techniques, 2) expertise in the neurobiology of stress, 3) refined written and oral scientific communication, and 4) professional development towards a career in academic research. This fellowship will also facilitate progress towards achieving her current and future research goals to enable success as an independent researcher.

NIH Research Projects · FY 2026 · 2025-02

The human virome is vast, differs among human individuals, and changes over the human lifespan. While the virome influences human health in many ways, true understanding of its impact is limited by incomplete characterization of the human virome composition. The goal of the Penn Virome Characterization Center (VCC), titled “The Oro-Respiratory-Gut Virome Axis Over Space and Time”, is to define the human virome and its dynamics in priority body sites in generally healthy individuals without acute illnesses, reflecting typical US populations across the lifespan among community-living participants. Our program will provide key insights into virome composition, richness and complexity, and produce extensive data and methodologies for future studies of disease associations. We will focus on key body sites identified by the NIH Human Virome Program as priority targets, including Oral/Dental, Gut (fecal), Respiratory Tract (upper and lower) and Blood. Our collaborative team has published extensively on the microbiome and virome, and in the course of these studies assembled unique cohorts with biobanked specimen repositories suitable for rapid initiation of the Penn VCC program. Most of our sampling is from an urban setting in the mid-Atlantic region. To achieve our goals, the Penn VCC will: (a) Take advantage of rich in hand biobanks of human samples for rapid efficient analysis, and enroll and phenotype new subjects to obtain longitudinal oro-respiratory-gut-blood specimens, from existing cohorts of healthy children, adolescents, adults and elderly individuals; (b) Comprehensively describe and quantify virome populations from distinct oro-respiratory-gut-blood sites, including viruses of humans, bacteria, and human eukaryotic commensals, and implement methods for molecular identification of host cells for novel viruses and characterization of viral DNA modification; (e) Define the commonalities and differences of virome communities across oro-respiratory-gut biogeography and blood over time within individuals and between different individuals and age groups; (f) Protect participants, investigators, and NIH by ensuring adherence to ethical norms and regulatory requirements, while conducting rigorous legal and qualitative research that addresses unanswered questions raised by human virome research that will inform future policy and research practice via our Ethical, Legal, and Social Implications Core; (g) Ensure harmonization and integration with other VCCs of the Human Virome Program, and establish open sharing of data to maximize research value; (h) Support collaborative research among HVP investigators using specimens, data, and novel insights generated through the VCC; and (i) Ensure appropriate representation of community populations as research participants.

NIH Research Projects · FY 2026 · 2025-01

A core goal in biomedical research is to understand signaling pathways that regulate cellular decisions in complex three-dimensional environments such as cells in developing or regenerating tissue. Directly manipulating these signaling patterns in vivo is often complicated or provides little control. This is especially the case for questions including the complex and heterogeneous extracellular matrix. Excitingly, recent advances in engineered in vitro systems have made complex three-dimensional environments – including organoids and organ-on-chip systems – more controlled. However, while these systems are powerful, they have largely ignored the dynamic reciprocity between a cell and its newly deposited (nascent) environment. The key challenge is to implement this dynamic nascent extracellular matrix into the existing model of cell-matrix dynamic reciprocity – the Bissell model – to identify the mechanisms of cellular decisions within complex tissue environments. If successful, this improved model will unlock the potential of engineered model systems to better represent target tissue environments and as a testing bed for novel drug testing. The overarching theme of my research group is to develop and apply engineering tools to drive the understanding of cell-cell and cell-ECM interactions. Developed tools in my group include dynamic chemistries and engineering approaches to mimic and manipulate organoids and their nascent matrix, and magnetic actuation to induce dynamic changes in a cell’s environment. We apply these tools to further develop complex cellular systems that implement dynamic changes of the nascent matrix to better guide translatable discoveries. Project one: Manipulating the local nascent environment to guide cellular decisions in complex cellular systems. Controlled modulation of matrix components and their physical properties enables spatiotemporal control over stem/progenitor cell differentiation pathways and their decision making, and towards controlling complex tissue development. We will develop new chemistries to directly manipulate the physical properties of the nascent matrix in situ and within engineered nascent matrix hydrogels. Our goal is to develop improved in vitro models of dynamic reciprocity, including cell-nascent matrix interactions that will be extendable to other cellular systems and disease models. Project two: Remote control of cell-cell and cell-matrix interactions in complex cellular systems. To better control and manipulate cellular systems dynamically, we will take advantage of magnetic actuation of cell-cell contacts, including cadherins, and cell-matrix contacts, including integrins. We propose to develop cadherin- mimetic magnetic microgels to exert forces onto cells within organoids and matrix-binding magnetics to introduce forces to the nascent matrix. These models will enable characterizing how physical forces exerted onto cells and their nascent matrix guide cellular decisions within complex three-dimension environments.

NIH Research Projects · FY 2026 · 2025-01

Project Summary/Abstract Psychiatric disorders carry a significant public health burden and are among the most debilitating health conditions that individuals can face. Despite tremendous efforts to collect and analyze genetic and clinical data from millions of participants across various psychiatric disorders, understanding the intricate pathophysiological pathways and identifying objective biomarkers for psychiatric risk management and patient treatment remain formidable tasks. The rapid evolution of large- scale imaging genetic data resources unlocks new possibilities to uncover shared neural foundations and genomic pathways pertinent to psychiatric disorders. Nevertheless, current research on imaging biomarkers in psychiatric genetics predominantly centers on mapping genetic effects of common variants within a single imaging modality, such as brain structural magnetic resonance imaging (MRI). These approaches often miss more extensive biological mechanisms that span across multiple imaging modalities and organs, and neglecting rare variants, which may have larger biological effects. In response to NOT-MH-21-175 (further analysis of human connectome data), we aim to is to conduct thorough secondary data analyses to enhance the integration of multi-organ imaging data of the brain and body, psychiatric genetic studies, and functional genomic data resources. These novel approaches will incorporate multi-organ multi-modal imaging data resources and will encompass a comprehensive range of genetic variants, from common to rare, across the full allele frequency spectrum. Our specific goals are as follows. Aim 1 involves integrating common and rare variations to understand the genetic co-architecture of psychiatric disorders and multi-organ imaging phenotypes of the brain and body. Leveraging our expertise in medical imaging data analysis, we will harness the latest open-access imaging genetic datasets from 9 studies involving more than 100,000 subjects, linked to extensive brain MRI, cardiac MRI, abdominal MRI, and retinal imaging data. In Aim 2, we will link imaging data to the tissue and cell type-specific functional landscape of psychiatric disorders and identify involved imaging traits within gnomically supported brain and body regions and cell types. By the convergence of evidence from genomic and imaging data integration, we aim to decode the complex genetic underpinnings of psychiatric disorders, leading to improved biological targets. Aim 3 will evaluate the putative causal mechanisms between imaging phenotypes and psychiatric disorders. This research project aims to substantially advance the integration of both population-scale imaging resources and curated functional genomic databases into psychiatric genetic studies, providing a novel multi-organ perspective that broadens the scope for identifying biomarkers and understanding the etiology of psychiatric diseases.

NIH Research Projects · FY 2026 · 2025-01

I am a Neurocritical Care Neurologist at the University of Pennsylvania (Penn), aiming to become an independent investigator with expertise in functional neuroimaging and disorders of consciousness (DoC) after brain injury. I have previously used functional magnetic resonance imaging to identify a novel brain network potentially associated with consciousness in humans – termed the arousal-awareness integration network (AAIN) – and have collected preliminary data to suggest that AAIN dysfunction may serve as a final common pathway in the pathophysiology of DoC after brain injury. My long-term career goal is to better understand the network dysfunction that characterizes DoC across etiologies of brain injury, and to target this pathophysiology with novel diagnostic, prognostic, and therapeutic strategies. The goals of this project are to use novel approaches to clarify the physiologic relevance of network connectivity in DoC, and to evaluate the AAIN's association with consciousness and consciousness recovery. In patients with acute brain injury in the intensive care unit, the proposed research will prospectively investigate: (1) the extent to which network connectivity reflects the dynamic level of consciousness (`state'), versus recovery potential (`fate') by determining how connectivity changes with alterations in consciousness; (2) whether AAIN connectivity, relative to other networks, is associated with state of consciousness, which will help clarify which network is most fundamental to consciousness; and (3) whether AAIN connectivity, relative to other networks, is associated with the future recovery of consciousness, which may help guide families in high-stakes decisions about continuing or withdrawing life-sustaining treatment. This project could advance our understanding of consciousness while facilitating the development of a novel clinical biomarker for consciousness detection and prognostication. My career development plan leverages the expertise of leaders in the field to acquire skills in advanced neuroimaging (advised by primary mentor, Dr. John Detre), clinical research in DoC (advised by co-mentor, Dr. Brian Edlow), and rigorous statistical approaches to the prognostication of consciousness recovery (advised by co-mentor, Dr. Jonathan Elmer). At Penn, I will benefit from a unique environment and unparalleled resources, with access to a robust infrastructure for neuroimaging research, a state-of-the-art MRI scanner that is dedicated to research and embedded within the neurocritical care unit, and an innovative clinical program that specializes in DoC. This project, along with the associated mentorship and training plan, will provide me with the knowledge and expertise necessary to develop an independent career in DoC research.

NIH Research Projects · FY 2026 · 2025-01

Innovative scan protocols with combined long axial FOV PET and spectral CT for improved quantification in oncology: The goal of this project is to i) develop methods and protocols to obtain high-quality dynamic information from both PET and CT modalities and, ii) use dynamic Spectral CT for PET kinetic modeling to reduce overall scan time to match existing clinical PET protocols (~20 minutes). This technological advancement will enhance diagnostic capabilities in oncology, with broad applications in other diseases, while also reducing the burden on patients with shorter imaging protocols. We will achieve these goals through 3 specific aims: (1) Develop spectral CT protocol and processing pipeline for IDIF and perfusion maps; (2) Develop advanced modeling to combine Spectral CT and PET data for kinetic modeling; (3) Evaluate PET-Spectral CT to quantify blood flow and glucose metabolism in breast cancer. In Aim 1 we will implement an ultra-high-pitch spectral CT protocol to generate dynamic iodine maps with extensive axial coverage, including the blood pool and targeted tissue/organ. Our deep learning (DL) based approach utilizes spectral CT to differentiate anatomical background from iodine enhancement, preserving image quality and minimizing radiation exposure. We aim to improve iodine contrast bolus delivery and timing, assessing concentration and injection/acquisition timing. In Aim 2 we will utilize a custom flow phantom for testing and validation of the CT methods in Aim 1 methods as well as pre- and post-reconstruction data corrections in PET. Initial assessments of the relationship between CT, PET, and CT-informed PET IDIFs and resultant kinetic perfusion parameters will be made with the flow phantom. In Aim 3 we will test the CT-dose reduction, bolus optimizations and PET data correction methods developed in Aims 1 and 2 in a translational porcine model (N=4) followed by a cohort of breast cancer patients (N=10). Patlak graphical analysis will also be implemented to compare the net influx rate of FDG when the IF is estimated with and without CT information. These metrics will be compared across modalities and estimation methods with the goal of developing a clinically translatable hybrid Spectral CT-PET protocol for dynamic imaging. Our overall outcomes will be solutions that facilitate the acquisition of high-quality dynamic information from both PET and CT, to be leveraged for PET kinetic modeling purposes, and used for clinically practical protocols to enhance the accuracy of oncological diagnostics.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY Determining how neurons are assembled into functional circuits will provide insight into developmental disorders of the nervous system and may suggest therapeutic approaches to promote nerve regeneration. To navigate to their correct targets, axons must precisely modulate their responses to extracellular cues. For example, to cross the midline commissural axons must first respond to attractive cues while preventing the response to repulsive cues. After entering the midline, axons switch their response to allow them to be repelled away from the midline to establish connections that are essential for coordinated sensory and motor behavior. Netrin attracts commissural axons to the midline in both invertebrates and vertebrates through receptors of the Deleted in Colorectal Carcinoma (DCC) family (encoded by frazzled (fra) in the fly). We have shown that Fra (and likely DCC) signals in two ways to direct axon guidance across the midline. First, Fra responds to its canonical Netrin ligand to promote local cytoskeletal rearrangements and second, Fra acts independently of Netrin to directly regulate gene transcription. There are three major knowledge gaps in our understanding of Fra and Dcc signaling mechanisms that this proposal will address: 1) the molecules that coordinate Netrin- dependent regulation of the actin cytoskeleton during axon attraction in vivo are incompletely defined, 2) many of the factors that control the transcriptional activity of Fra (and Dcc) are unknown, 3) the downstream target genes that are regulated by Fra's transcription factor function and how they contribute to Fra and Dcc- dependent processes are largely unknown. Three complementary sets of preliminary findings underpin our proposed research. First, analysis of pathogenic variants of human DCC (hDCC) have led to the discovery of an essential and conserved role of the Wave Regulatory Complex (WRC) in Netrin-dependent axon attraction. Second, we have completed an in vivo proteomic screen for Fra interacting proteins and have identified and validated several novel interacting proteins and pathways that are likely to contribute to Fra-dependent signaling. Third, we have performed RNA sequencing in embryonic neurons isolated from Fra loss of function and gain of function conditions and have identified a set of 15 reciprocally regulated transcriptional targets. This preliminary data provides the foundation and rationale for the experiments that we are proposing to 1) define how Fra/Dcc signaling recruits and regulates the WRC during axon guidance, 2) define how the COP9 complex works with Fra to regulate transcription and 3) define the contribution of Fra's transcriptional targets to axon and dendrite targeting. Our research will identify new molecular components of both Netrin-dependent and Netrin-independent Fra/Dcc signaling and may inform future studies of these signaling pathways in both neuronal and non-neuronal contexts and may offer insights into pathological conditions in humans that are caused by aberrant DCC function.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY High-grade serous ovarian cancer (HGSOC) and triple-negative breast cancer (TNBC) exhibit shared clinical and genomic characteristics, including poor prognosis, homologous recombination deficiencies, and potential immunoreactivity. These diseases present a pressing and unmet medical need for the identification of new therapeutic targets. Histone acetylation enzymes have emerged as compelling drug targets due to the demonstrated clinical success of histone deacetylase inhibitors in hematological malignancies, affirming the feasibility of this therapeutic approach. Imbalanced histone acetylation, a hallmark of epigenetic alteration in cancer, disrupts genome organization and gene transcription, thereby promoting tumorigenesis. While both histone acetyltransferases (HATs) and histone deacetylases (HDACs) are involved in regulated histone acetylation, HATs are generally associated with accessible chromatin and increased transcription activity, essential for hyperproliferating tumor cells. Consequently, targeting HATs holds promise as a more efficient strategy for cancer treatment. However, the development of HAT-targeting therapy in the clinic has lagged significantly behind HDACi. Although potent and selective HATis with in vivo efficacy in cancer models have been developed at the preclinical stage, the identification and prioritization of specific HAT targets among the 37 distinct HATs in humans have posed challenges. Utilizing a novel systems biology approach developed by our team, we conducted a comprehensive characterization of genes encoding HATs in cancers, leading to the identification of KAT6A as a promising clinical actionable drug target. Preclinical studies have shown the potential of KAT6i as a monotherapy and in combination with FDA-approved drugs in KAT6A-dependent tumors. Notably, potent and selective KAT6is have been successfully developed and are currently undergoing evaluation in phase 1 clinical trial. We hypothesize that KAT6A hyperactivation disrupts the delicate balance of histone acetylation, leading to aberrant genome accessibility and dysregulated transcriptional programs (KAT6A addiction). Therefore, KAT6A serves as a novel therapeutic target in a subset of tumors primarily driven by the recurrent amplification of the KAT6A gene. In this application, we aim to assess the therapeutic potential of newly developed KAT6is as monotherapy and in combination with targeted therapy drugs in preclinical models of HGSOC and TNBC. We have assembled a team of collaborators with added expertise and resources to address the following specific aims: Specific Aim 1: Characterizing the epigenetic changes induced by KAT6i treatment in KAT6A-dependent cancer. Specific Aim 2: Defining the mechanism of action of KAT6i treatment in KAT6A- dependent cancer. Specific Aim 3: Evaluating the therapeutic potential of KAT6i as mono- and combination therapy for cancer. Through these proposed studies, we anticipate providing a strong rationale for KAT6A as a novel drug target for the treatment of cancer.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY Highly processed foods have become increasingly prevalent in our diets over the past several decades. These foods often contain high levels of high fructose corn syrup (HFCS), and excess intake of HFCS is associated with obesity and a host of associated metabolic diseases, such as type II diabetes and cardiovascular disease. However, the gut-brain mechanisms through which fructose is sensed, and how it influences homeostatic feeding circuits in the brain, are largely unknown. My preliminary data suggest that fructose engages the vagus nerve to inhibit activity in hunger-promoting agouti-related protein (AgRP)-expressing neurons in the brain. Building on this finding, this proposal will test the hypothesis that Y2-expressing vagal afferents sense fructose in the gut and are sufficient and necessary for gut fructose-induced inhibition of AgRP neuron activity. Aim 1 will use single- cell resolution two-photon calcium imaging to determine the real-time activity dynamics of fructose-activated vagal afferents. In a complementary experiment, we will combine retrograde tracing to label gut-innervating vagal afferents, with activity-dependent gene labeling via in situ hybridization in mice, to determine the molecular identity of gut-innervating fructose-activated vagal afferents. Aim 2 will leverage an activity-dependent technique to gain genetic access to fructose-responsive neurons to determine their role in modulating AgRP neuron activity Through these experiments, we expect to reveal the gut-hypothalamic nutrient sensing pathway for fructose. These results will not only provide new insight for the field of gut-brain signaling, but will also have clinical applications by uncovering new potential drug targets for obesity treatment. My Sponsor, Dr. Amber Alhadeff, is an expert in the gut-to-AgRP neural pathways involved in nutrient sensing, and my Co-sponsor, Dr. Guillaume de Lartigue, has extensive experience with anatomical tracing and two-photon microscopy of vagal afferents. My prior research experience, combined with the expertise of both Sponsors, make me uniquely positioned to successfully complete all proposed experiments during my graduate training. Therefore, funding of this NRSA proposal to support my dissertation studies will set me on the trajectory toward my ultimate goal of becoming an independent investigator that explores gut-brain circuits that regulate feeding.

NIH Research Projects · FY 2026 · 2025-01

Project Summary/Abstract Chronic pain is a pervasive burden affecting over 50 million US citizens. The experience of pain is subject to modulation based on context; however, the mechanism of this modulation is unknown. This project aims to unravel the intricate dynamics of prefrontal cortical microcircuits during expectation-driven endogenous pain relief, or placebo analgesia. Specifically, this research focuses on mu opioid receptor (MOR) expressing anterior cingulate cortex (ACC) neurons. This proposal integrates a preclinical placebo analgesia model in mice with cutting-edge techniques such as in vivo single-neuron resolution calcium imaging and the application of a novel opioid biosensor (δLight), to investigate the nuanced interplay between nociceptive signaling and the opioid system within the ACC during placebo analgesia. In Aim 1, viral expression of GCaMP8m under a MOR promoter (mMORp-GCaMP8m) enables visualization of nociceptive MOR+ neurons during placebo conditioning. In Aim 2, the role of enkephalin in the ACC during placebo analgesia will be investigated through CRISPR-mediated excision and biosensor-based monitoring. These aims strive not only to elucidate the neural correlates and potential mechanisms of placebo analgesia, but also to contribute to a broader understanding of how pain is endogenously regulated in a context-dependent manner by opioid systems in prefrontal cortex. As a pivotal component of this research endeavor, a training plan is outlined to foster the development of Ms. Oswell’s skills, preparing her for an independent research career. The successful execution of these aims not only contributes to the scientific understanding of placebo analgesia but also provides significant training opportunities, ensuring she is well-equipped with the requisite skills for a future independent research career. Through hands-on experience in state-of-the-art methodologies, she will acquire expertise that extends beyond the immediate scope of this project, cultivating a foundation for continued contributions to the broader field of pain neuroscience. The interdisciplinary nature of this research, combining molecular biology, neuroimaging, and behavioral analysis, provides a unique platform for skill diversification and the cultivation of a holistic research perspective. This project holds promise not only for advancing our understanding of pain modulation but also for shaping Ms. Oswell into a proficient and independent researcher poised to make substantial contributions to the scientific community.