University Of Pennsylvania

NIH Research Projects · FY 2025 · 2022-02

ABSTRACT / SUMMARY Acute critical illnesses rapidly lead to severe organ damage and loss of life. These illnesses include sepsis, stroke, and acute respiratory distress syndrome (ARDS). Here we focus on ARDS, which is acute inflammation of the lungs’ air sacs, and the cause of death in COVID-19. For ARDS and these other diseases, we have developed ligand-targeted nanocarriers that localize drugs to the inflamed microvasculature of affected organs. As we moved towards clinical translation, we found the key step is gaining control of complement, a set of plasma proteins that bind microbes and aid their clearance. But we found complement-nanoparticle interactions are a “double-edged sword”, with both benefits to optimize, and deleterious features to resolve. First, we found that complement protein C3 rapidly opsonizes particular nanoparticles, and that such C3- opsonized nanoparticles then act as “decoys” to ameliorate ARDS mouse models (e.g., nebulized LPS) by ~75%. The C3-coated nanoparticles accumulate in marginated leukocytes, which are key to ARDS pathophysiology, and cause those cells to leave the lungs. However, C3 opsonization induces an anaphylaxis-like reaction called CARPA (complement-activation-related pseudo-allergy). Therefore, in Aim 1, we will engineer nanoparticles that can function like C3-coated decoys to ameliorate ARDS, but without CARPA. We will also investigate the mechanism underlying nanoparticle decoy therapy. Then we will test the translational potential of these optimized decoy nanoparticles by testing them in fresh, perfused, ex vivo human lungs. Second, we found that the ligand-targeted nanoparticles we have been developing for drug delivery for years also induce CARPA. Therefore, in Aim 2, we will re-engineer our ligand-targeted nanoparticles to prevent CARPA. We will test a drug carrier we have previously used to concentrate drugs in the alveolar microvasculature of the lungs: liposomes conjugated to anti-PECAM antibodies that bind endothelial cells. We will test in vitro and in vivo in mice whether various engineered versions of anti-PECAM liposomes can evade C3 opsonization and CARPA, and thereby achieve more specific delivery to the lungs. Lastly, we will test these CARPA-avoiding nanoparticles with plasma from ARDS patients, as such patients have perturbed complement. Upon completion of these two Aims, we will have developed two technologies that may aid therapy of ARDS: 1) Decoy nanoparticles that safely cause marginated leukocytes to leave the lungs, and thereby ameliorate ARDS-like phenotypes; 2) A technology for preventing complement side effects such as CARPA when delivering ligand-targeted nanoparticles. As marginated leukocytes play pivotal roles in most acute critical illnesses, and CARPA sensitivity is common to those as well, the technologies developed here may impact not only ARDS, but also sepsis, stroke, and more.

NIH Research Projects · FY 2026 · 2022-02

Project Summary Periodontal disease (PD) is a prevalent oral inflammatory condition that is epidemiologically associated with systemic disorders (comorbidities), such as, cardiovascular disease, rheumatoid arthritis, and type-2 diabetes. An independent association between PD and comorbidities remains even after adjustment for confounders. A possible factor contributing to this independent association is that PD can cause low-grade systemic inflammation, which may in turn influence comorbidities. The relationship between PD and systemic comorbidities is bidirectional in that systemic diseases can also promote susceptibility to PD. However, there is no known unifying causal mechanism that can explain how PD affects and is affected by comorbidities. To mechanistically explain the reciprocal association between PD and comorbid conditions, a novel hypothesis is proposed based on the recent concept that systemic inflammatory stimuli can cause epigenetic rewiring of hematopoietic stem and progenitor cells (HSPCs) in the bone marrow, which enables these cells to give rise to ‘trained’ myeloid cells that can respond more strongly to future stimuli. This concept represents a form of innate immune memory and is known as ‘trained innate immunity’ (TII). TII can be protective in infections but potentially detrimental, hence maladaptive, in inflammatory disorders. Thus, given that chronic inflammatory diseases are – in large part – driven by the action of inflammatory myeloid cells, inflammation-driven transcriptomic and epigenetic alterations in their bone marrow progenitors are likely to influence the initiation or the progression of different chronic inflammatory disorders that emerge as comorbidities. This project involves investigation of the comorbidity of PD with another inflammatory bone loss disorder, rheumatoid arthritis. The overarching hypothesis is that maladaptive TII constitutes a mechanistic basis for the comorbid connection of PD and arthritis, which are studied using validated preclinical models, ligature-induced PD (LIP) and collagen antibody-induced arthritis (CAIA), respectively. The objective of Aim 1 is to show that inflammation-adapted HSPCs in the bone marrow mediates the comorbid association of PD and inflammatory arthritis. That the proposed maladaptive effect is mediated by inflammation-adapted (‘trained’) HSPCs will be tested by bone marrow transplantation experiments. Aim 2 was designed to investigate whether experimental PD induces transmissible epigenetic modifications in bone marrow progenitors towards a maladaptive inflammatory phenotype that underlies the development of inflammatory comorbidities. Further studies are proposed to show that interleukin-1β acts on HSPCs to mediate LIP-induced trained myelopoiesis and increased disease activity (Aim 3). If successful, this project will provide a unifying network for and mechanistic insights into the interconnection of inflammatory comorbidities and maladaptive TII in the bone marrow. Such conceptual framework could also provide a platform for novel therapeutic interventions targeting inflammatory comorbidities via pharmacological modulation of TII.

NIH Research Projects · FY 2026 · 2022-02

PROJECT SUMMARY Accumulating evidence strongly supports the idea that sleep is a crucial variable in neurodegenerative disease: disease progression disrupts sleep, and disrupted sleep worsens brain degeneration. Sleep is thought to represent a powerful untapped therapeutic modality through which neurodegeneration can be modified. Yet how sleep and neurodegeneration are coupled at a mechanistic level is poorly understood. Defining cellular and molecular links between sleep and neurodegeneration has been difficult, limiting the ability to pursue sleep modification as a therapeutic avenue. We propose leveraging a neurodegeneration model in Drosophila to dissect mechanisms of disrupted sleep in detail, including use of high throughput genetic screens available in simple systems, with the goal of defining molecular pathways linking sleep and brain integrity. We have found that expression of the human neurodegenerative disease protein TDP43 (linked to Alzheimer’s, frontotemporal dementia, and motor neuron disease) causes a robust sleep impairment. Our initial data suggest that the Drosophila sleep phenotype results from dysfunction in glia, which are known to be critically involved in sleep regulation. Importantly, expression of TDP43 in glia also causes brain degeneration and shortened lifespan in flies, providing a strong rationale for investigation of TDP43 as a key link between sleep and neurodegeneration. Because TDP43 pathology has been described in many human neurodegenerative diseases including Alzheimer’s, focused study of the mechanisms linking TDP43 with sleep are likely to be broadly relevant. Here we will investigate the molecular mechanisms linking TDP43-associated brain degeneration and sleep. In Aim 1, we propose to define the glial subtype critical for the sleep effect and examine how sleep loss affects the subcellular localization and accumulation of TDP43. A preliminary genetic screen for modifiers of TDP43-induced sleep dysfunction has already defined several suppressors, including Ataxin-2, a known human disease gene that interacts with TDP43 in neurons. In Aim 2, we will examine this suppressor and others in detail to define molecular and cellular mechanisms of the interaction. Finally, our preliminary data indicate that restriction of sleep opportunity (Sleep Restriction Therapy, SRT) can reverse sleep defects in TDP43 flies. In Aim 3 we will examine SRT in TDP43 flies, and conduct a genetic screen to define the molecular pathways through which SRT improves sleep in this brain degeneration model. Taken together these aims will shed new light on the molecular and genetic links between sleep dysfunction and brain degeneration, and provide the foundation for novel therapeutic targets that leverage sleep to promote brain integrity.

NIH Research Projects · FY 2026 · 2022-01

PROJECT SUMMARY T2R bitter taste receptors are G-protein coupled receptors (GPCRs) originally identified on the tongue, but which also serve diverse roles in other tissues. “Extraoral” (outside the tongue) taste receptors are likely important in airway infection, asthma, pre-term labor, and even cancer. In airway epithelial cells, T2Rs 4, 14, 16, and 38 recognize quorum-sensing molecules produced by gram-negative bacteria, including the opportunistic pathogen Pseudomonas aeruginosa. Stimulation of these receptors modifies production of antibacterial molecules, including nitric oxide and defensins. Some polymorphisms reducing the functionality of T2Rs increase patient susceptibility to upper respiratory infection and chronic rhinosinusitis. Targeting T2Rs as therapies requires better elucidation of their expression and downstream effects. This must be studied in primary human cells, as mouse T2Rs differ in number and agonists that activate them. We identified several T2R receptors in primary human monocytes and macrophages. Macrophages are important innate immune cells that phagocytose and kill bacteria as well as secrete pro- or anti- inflammatory cytokines to modify immune responses. Stimulation of these receptors activates calcium release and nitric oxide production that acutely (within 5-15 min) enhances phagocytosis of bacteria. Preliminary data suggest that T2Rs may be targets for stimulation of innate immune responses to kill bacteria without the use of conventional antibiotics, reducing pressures for antibiotic resistance. This may be especially useful in upper respiratory diseases. Acute and chronic rhinosinusitis account for >20% of all antibiotic prescriptions in the US, and thus are important drives of antibiotic resistant micro-organisms. Chronic rhinosinusitis and associated nasal polyps also cause dysosmia, hyposmia, and/or anosmia in many patients. We will further define the signaling pathway that increases macrophage phagocytosis and test how it affects bacterial killing (Aim 1). As bacteria such as P. aeruginosa can invade cells and live as intracellular pathogens to evade further immune detection, we will test how T2Rs affect macrophage bactericidal activity. This work will also be among the first to use live cell imaging and fluorescent biosensors to study macrophage GPCR signaling in real time. Next (Aim 2), we will examine how T2R activation reduces inflammatory responses to toll-like receptors (TLRs). Our preliminary data suggest novel pathways of T2R-TLR crosstalk possibly involving Akt. Finally (Aim 3) we will compare phenotype, T2R expression, and T2R function in macrophages from inflamed chronic rhinosinusitis nasal polyps with naïve serum-derived macrophages from the same patients. We will examine if macrophage infiltration of polyps is altered with T2R polymorphisms or bitter taste perception. We also examine if increased macrophage numbers in nasal polyps affects chronic rhinosinusitis disease phenotypes.

NIH Research Projects · FY 2026 · 2022-01

Project Summary/Abstract Callous-unemotional (CU) traits, defined by low empathy, guilt, and prosociality, predict very high risk for childhood disruptive behavior disorders (DBD) and adverse adult outcomes, including violence, psychopathy, and crime. Standard treatments for DBD are not as effective for children with CU traits. To inform personalized treatments for DBD, a better understanding is needed of specific risk factors for CU traits beginning early in childhood. The overall objectives of this proposal are to identify specific child-level and parenting-level risk factors that predict CU traits across early childhood. The central hypothesis is that reduced sensitivity to cues of threat and affiliation will specifically predict CU traits. High parental harshness, low warmth, and low emotion scaffolding are also hypothesized to predict increases in CU traits, including via interactions with low threat sensitivity and affiliation among both parents and children. The rationale for the proposal is that by successfully isolating specific child- and parenting-level risk factors that predict CU traits, novel treatment strategies can be developed to reduce CU traits, and in turn, disrupt pathways to DBDs, including via multi-level treatment modules that target behavior, physiology, and attention. The hypotheses will be tested by pursuing the following specific aims: (1) Establish phenotypic markers of CU traits in early childhood; (2) Establish phenotypic markers of parenting practices; and (3) Illuminate shared temperament and parenting factors that predict CU traits. All three aims will be pursued within a prospective longitudinal study. Participants (N=500) will be recruited from community and clinical settings at the University of Pennsylvania (n=250) and Boston University (n=250) and assessed at time 1 (aged 3–4) and time 2 (aged 5-6). Under the first aim, low threat sensitivity and affiliation will be tested as child- level risk factors for CU traits across multiple levels of analysis, including parent-report and multiple new computer and observational tasks that allow simultaneous measurement of attentional (i.e., eye-tracking) and physiological (i.e., respiratory sinus arrythmia) processes. Under the second aim, low threat sensitivity and affiliation of parents, assessed again across multiple levels of analysis, will be tested as predictors of different parenting dimensions of harshness, low warmth, and low emotion scaffolding. Under the third aim, the interplay of parenting behaviors and shared temperament features of parents and children will be tested as dyadic risk factors for CU traits. The proposal is innovative because it will generate new measures of precision risk factors that predict CU traits, is guided by extensive preliminary data, hypotheses are tested during a critical developmental period for understanding CU traits, and the sophisticated quantitative analysis incorporates a rich multi-method measurement framework that will generate new knowledge about the biological, cognitive, social, and emotional processes underlying CU traits. The proposed research is significant because it will identify biomarkers and behavioral indicators of illness (NIMH Priority 2.2) and lead to improved capacity to operationalize specific risk factors to target in interventions to mitigate risky pathways to CU traits and DBD.

NIH Research Projects · FY 2026 · 2022-01

PROJECT SUMMARY The coronavirus disease 2019 (COVID-19) pandemic is a public health crisis, characterized by pneumonia and multiorgan dysfunction. We previously demonstrated that community-acquired pneumonia increases the long- term risk of cardiovascular disease. There is an urgent need to investigate the incidence and mechanisms of cardiovascular disease in COVID-19 survivors. Thus, we propose a novel investigation of the intermediate and long-term cardiac, vascular, and renal consequences of COVID-19. Acutely, COVID-19 is associated with microvascular and macrovascular thrombotic events and inflammatory- and stress-related injury in the heart, kidneys, and vasculature that may put COVID-19 survivors at particularly elevated risk of chronic complications. Our study team has combined expertise in the study of post-pneumonia cardiovascular risk, vascular and kidney pathophysiology, epidemiologic outcomes research, and implementation of longitudinal prospective cohort studies. Our goal is to examine the natural history of cardiac, vascular, and kidney disease in COVID-19 survivors, and to identify risk factors for adverse longitudinal outcomes in these patients. We propose a prospective cohort study evaluating 1) cardiovascular events in a large, electronic health record-based cohort of survivors of COVID-19 in our health system compared with matched controls (“MACE cohort”) and 2) detailed vascular and renal phenotyping in a smaller cohort of COVID- 19 survivors compared with matched controls (“deep phenotyping cohort”). In the MACE cohort, we will collect detailed hospitalization, demographic, and clinical data as well as records for post-COVID-19 hospitalizations. An expert physician panel will prospectively adjudicate hospitalization records to evaluate for post-COVID-19 MACE (heart failure hospitalization, acute coronary syndrome, serious arrhythmia, stroke, peripheral artery disease, and death). In the deep phenotyping cohort, we will perform serial quantitative measurements of vascular health in large, medium-sized, and small arteries, specifically: (1) pulse wave velocity (the reference standard measure of large artery stiffness), (2) flow-mediated dilation (a measure of endothelial function), and (3) microvascular structure assessed by sublingual imaging. We will also perform serial measurements of kidney function (estimated glomerular filtration rate, albuminuria, markers of tubular injury, and exploratory ultrasound images to estimate fibrosis). We aim to assess the long-term incidence of and risk factors for MACE in COVID- 19 survivors, and to evaluate the trajectory of microvascular and macrovascular health and kidney function over time in these patients. Our mechanism-driven approach will provide critical guidance on longitudinal cardiovascular risk and vascular and kidney damage following COVID-19 infection. The results of this study will enhance our understanding of the long-term target organ effects of COVID-19 and identify risk factors that can be targeted by future interventions to ultimately reduce the risk of adverse outcomes in COVID-19 survivors.

NIH Research Projects · FY 2026 · 2022-01

Metabolic Imaging of Targeted Therapies in Cancer PROJECT SUMMARY/ABSTRACT Given the paradigm shift in cancer therapy including the ever-growing increase in the use of targeted therapies, foremost small-molecule kinase inhibitors in cancer therapy, there is an urgent need to develop reliable imaging techniques to detect and monitor the efficacy of such inhibitors in cancer patients. Because direct evaluation of cell signaling is practically not feasible and changes in tumor volume occur late after treatment initiation given the predominantly cytostatic effect of the inhibitors, we are proposing an alternative approach to monitor changes in tumor metabolism induced by kinase inhibition. This will be achieved in three stages: 1) analysis of gene expression/proteomic/phosphoproteomic to identify metabolic pathways perturbed by inhibition of the signaling pathway performed both in vitro and in vivo in the mouse xenotransplant models using patient derived cultured and primary cells (PDX), 2) metabolomic and metabolic fluxomic analysis of effect of kinase inhibition on metabolic pathways, also done in in vitro and in vivo settings, 3) analysis of biomarkers of inhibitor response validated by the above “-omics” studies by imaging techniques, preferably non-invasive, such as 1H MRS or chemical exchange saturation-transfer (CEST) with standard FDG PET imaging serving as control. In these proof-of-principle studies, we will focus on mTOR, the serine/threonine kinase hyperactive in the majority of cancer types, and employ direct and indirect inhibitors of mTOR, rapamycin/rapalog and Torin2, respectively, as index kinase/kinase inhibitor system. We will use diffuse large B-cell lymphoma (DLBCL) as experimental cancer model. In preliminary studies, we have demonstrated that rapamycin decreased concentrations of lactic acid in patient-derived lymphoma cell lines, both cultured in vitro and xenotransplanted into mice, as detected by unique 1H MRS imaging-based detection pulse sequences developed by us and our collaborators. The rapamycin-induced decrease in glycolytic metabolism correlated with and, importantly, markedly preceded inhibition of tumor cell growth, strongly supporting the notion that image-based evaluation of the key metabolic response is predictive of biological tumor cell response to the inhibition. The response also correlated with and, hence, was at least in part attributable to decreased expression of hexokinase II, other glycolytic enzymes and enzymes from other key metabolic pathways including phosphoribosyl-amidotransferase and other enzymes involved in glutaminolysis. Utilizing 13C MRS and 13C LC-MS, we have confirmed mTOR control of glycolysis and also noted decreases in fatty acid and sterol metabolism as well as inhibition of the pentose phosphate shunt and the TCA cycle. We anticipate that the proposed studies will extend our knowledge of the impact of mTOR inhibition on malignant cell metabolism and, ultimately, set the stage for future clinical evaluation of MRS or other imaging method(s) for monitoring response to inhibitors of mTOR and other cell-signaling kinases in DLBCL and other types of cancer.

NIH Research Projects · FY 2026 · 2022-01

Project Summary Efforts to accurately estimate the prevalence of pain and quality of pain management in the population of patients with Alzheimer’s Disease and Related Dementias (ADRD) have met with varied success. Communication about pain can be extremely challenging for patients with advanced dementia, and the etiology of pain difficult to determine. In addition, patients may resist pain treatments due to their inability to understand the purpose of analgesia and may receive decreased benefit from analgesia due to the disruption of the placebo effect. Family caregivers (family members, spouses, friends or others who assume the critical caregiving role) are at high risk for chronic stress, deteriorating physical health, financial difficulties, and premature death. They suffer from high rates of depression, anxiety and grief; pain management for their patients has been one of the most commonly expressed concerns. Based on preliminary work whereby we examined pain management challenges and needs of caregivers of patients with ADRD, we designed a behavioral intervention entitled ENCODE to assist caregivers in effectively identifying and communicating their pain management challenges and needs. We propose a 5- year randomized clinical trial in which caregivers of patients with ADRD will be randomly assigned to a group receiving standard care with the addition of “friendly calls” (attention control group) or a group receiving standard care with the addition of the ENCODE intervention (intervention group). The specific aims are to assess the impact of the intervention on caregiver outcomes including quality of life, health, anxiety and depression, and their perceptions of the intervention. Additionally, in order to facilitate the adoption of the intervention in practice we will conduct a cost analysis demonstrating the costs associated with its delivery and identify barriers and facilitators to adoption.

NIH Research Projects · FY 2026 · 2022-01

Hypertensive disorders of pregnancy (HDP) are a leading cause of maternal mortality and morbidity in the US. In addition to contributions to maternal mortality, HDP increase the risk of chronic hypertension (cHTN), which can lead to cardiovascular (CV) disease. Neighborhood greenspace has been associated with improved CV outcomes in the general population, but few studies have evaluated the relationship between greenspace and the development of HDP and subsequent cHTN. The broad objective of this proposal is to fill this gap by evaluating the impact of greenspace and a greening intervention on HDP and cHTN. We will merge GeoBirth, a cohort that will include 92,000 pregnant patients in Philadelphia, PA from 2008 to 2021, with several neighborhood greenspace datasets, including data from a randomized controlled trial of a greening intervention recently conducted by our team. Our overarching hypothesis is that living near greenspace lowers risk of HDP and cHTN. We propose the following aims: 1) Observational: To determine if neighborhood greenspace is associated with HDP and cHTN after pregnancy. We will analyze associations of residential greenspace with risk of HDP using multilevel logistic regression adjusted for individual and neighborhood-level confounders (1a). We will determine if greenspace exposure is associated with risk of cHTN within 5 years after pregnancy using survival analysis methods (1b). 2) Quasi-experimental: To measure the impact of a neighborhood greening intervention on HDP and cHTN after pregnancy, using a quasi-experimental design. We will analyze the impact of vacant lot greening on risks of HDP and cHTN in the 5 years after pregnancy, using a difference-indifferences analysis of women living near lots that were greened compared to control lots between 2008-2017. 3) Experimental: To examine the impact of a greening intervention on changes in blood pressure during pregnancy, using a previously conducted RCT. We will use data from a completed cluster RCT (2011-2014) vacant lot greening to examine the impact of greening on blood pressure changes (trajectories are associated with development of HDP) from 20 weeks’ gestation to delivery, among GeoBirth participants living near 541 vacant lots in three trial conditions: greened lots, trash clean-up only lots, and control lots. The proposed study will be the largest investigation into whether neighborhood greenspace can mitigate HDP risk and cHTN risk after pregnancy.

NIH Research Projects · FY 2026 · 2022-01

Hepatocellular carcinoma (HCC) is the most rapidly rising cause of cancer mortality in the United States. The majority of patients with HCC present with incurable disease at diagnosis and, despite the approval of targeted therapies, life expectancy remains less than 20 months. The diagnosis of HCC as well as its response to treatment rely primarily on imaging biomarkers which have replaced tissue-based methods. Recent studies demonstrate that the dismal prognosis for these patients issues, at least in part, from deficiencies of current clinical imaging paradigms in diagnosing HCC as well as in identifying residual or recurrent HCC after treatment. Clinical imaging paradigms for HCC diagnosis and the assessment of treatment response are based on anatomic imaging features that often fail to identify HCCs or provide functional measures of response to targeted therapies. Indeed, the sensitivity of standard-of-care (SOC) contrast-enhanced (CE) MRI for small HCCs can be as low as 20%. Similarly, SOC imaging provides inadequate assessments of response to therapies. Addressing this deficiency requires the development of new imaging paradigms that provide functional measures of HCC biology to improve accuracy, sensitivity and specificity as well as inform the application of therapeutics. The development of novel functional imaging strategies for HCC has been limited by the absence of methodologies that can tailor imaging probe selection to the relevant HCC biology as well as a dearth of representative animal models. Using genome editing and metabolomics, our laboratory has demonstrated the fundamental dependence of HCC cells on lactate dehydrogenase and NADPH-dependent reductases to be promising imaging targets for Dynamic Nuclear Polarization 13Carbon Magnetic Resonance Spectroscopic Imaging (DNP-13C-MRSI), an emerging imaging technology. The proposed project will build on this prior work to study the ability of DNP-13C-MRSI to: 1) improve the accuracy of diagnosis and treatment response assessment of HCC following SOC therapies as compared to conventional imaging and 2) inform treatment selection. We hypothesize that DNP-13C-MRSI provides a unique technology through which to leverage fundamental enzymatic dependencies of HCC cells and enable functional molecular imaging for diagnosis and treatment response assessment. To test this hypothesis the proposed project will use unique animal models of HCC developed in our lab to pursue three aims: (1) to optimize a DNP-13C-MRSI pulse sequence that enables sensitive, accurate and reproducible measurements of regional pyruvate metabolism in autochthonous HCCs at high spatial resolution; (2) to determine the sensitivity, specificity and accuracy of DNP-13C-MRSI of HP 1-13C- pyruvate uptake and metabolism for identifying HCCs; and (3) to determine the accuracy of DNP-13C-MRSI of HP 1-13C-pyruvate and/or 1-13C-dehydroascorbic acid (DHA) for identifying and characterizing residual disease/local recurrence following TAE as compared to SOC imaging. The achievement of the proposed aims holds the potential to transform the imaging and treatment of patients with HCC, a devastating disease.

NIH Research Projects · FY 2026 · 2022-01

SUMMARY Despite the emergence novel therapeutic modalities, including BRAF inhibitors and immunotherapies, over 7,100 people are expected to die each year from malignant melanoma, primarily from metastatic dissemination and therapy resistance. The proposed studies leverage the expertise of the two co-PIs in tumor microenvironment, the Integrated Stress Response (ISR) and melanoma progression to test the overall hypothesis that as part of the adaptive response to tumor microenvironmental (TME) stress, bi-directional interactions between melanoma cells and fibroblasts, endothelial cells and adipocytes promote survival, metastatic dissemination, and therapy resistance. Preliminary results and published reports from the two PIs indicate that the ISR is activated in human melanomas and that genetic or pharmacological disruption of the ISR severely impairs primary tumor growth and metastasis in multiple experimental tumors, including melanomas. The studies will delineate the salient roles of the transcription factor ATF4, a major transcriptional effector of the Integrated Stress Response, in a pro-survival and pro-metastatic program mediated by the non- canonical tumor suppressor BRN2, a transcription factor effector of multiple melanoma-associated signaling pathways. To test the central hypothesis, we will pursue three specific aims: In Aim 1 we will use a novel, conditional global ATF4 knockout mouse model, as well as Fibroblast (FB)-specific and Endothelial (EC)- specific ATF4 knockout mice and cells to determine the role of ATF4 expression in each TME component on melanoma progression and metastasis. Preliminary results with global or FB-specific deletion of ATF4 results in a severe deficiency in tumor growth of flank melanoma tumors. In Aim 2 we will study how SRC promotes melanoma progression via ATF4-BRN2 cooperativity. Both BRN2 and ATF4 can repress anoikis/apoptosis, and in preliminary studies we reveal that BRN2 interacts with ATF4. Therefore, we will test the hypothesis that Extracellular Matrix (ECM)-driven integrin signaling, and monounsaturated fatty acid (MUFA) uptake from adipocytes and lymph converge on SRC to impose a cooperative BRN2-ATF4 anti-apoptotic and pro- metastasis gene expression program, driven in part by hippo signaling. Finally, under Aim 3, we will determine how MUFAs dictate melanoma phenotype. Here we will dissect a novel mechanism underpinning MUFA- mediated nuclear localization of -catenin and test the hypothesis that the resulting SRC-driven nuclear CAV1- -catenin complex promotes a pro-metastasis gene expression program via BRN2-ATF4, and the contribution of ATF4 and BRN2 to melanoma phenotypic heterogeneity and tumor immune infiltration. By delineating how the ISR uses ATF4 to coordinate the output of the TME to shape melanoma progression, we will identify therapeutically exploitable pathways for anti-melanoma approaches.

NIH Research Projects · FY 2025 · 2022-01

ABSTRACT Sensory systems have evolved to help meet the behavioral needs of organisms to ensure survival. Across the animal kingdom, two sensory functions are paramount: first, to identify specific objects in the environment and endow them with salience; and second, to move closer to objects of desire and away from objects that are best avoided. These properties – identification and localization – are the “what” and “where” questions of sensory information processing, respectively, and each of the senses provides a unique snapshot of the world that complements the other senses. In the unique case of olfaction, orthonasal and retronasal olfactory channels ensure that odor sources can be identified and tracked with fidelity at distal, proximate, and intraoral distances. This proposed project will focus on the “where” question of information processing in the human olfactory system. In particular, we aim to understand the capacities, constraints, and mechanisms by which odor cues orient and steer a navigator in the right direction. In this regard, a singular aspect of odors is their ability to travel through the air over long distances, such that the olfactory system can gather valuable predictive information not only about the physical location of an odorous source, but also about the navigator’s position within a physical landscape. Critically, while elegant neurobiological studies on odor navigation have been conducted in insects and birds, basic research on this topic in mammals, including humans, is sparse. Our planned studies are inspired by groundbreaking experiments showing that different types of neurons can encode and map physical spaces, including “place cells” (representing specific locations in space) and “grid cells” (representing an internal coordinate system to self-orient in this space). Here we will use functional magnetic resonance imaging (fMRI), virtual reality (VR) techniques, and computational methods to determine whether we can infer the presence of “grid-like” fMRI responses when human subjects navigate through an odor-rich two-dimensional landscape. In Aim 1, subjects will be asked to navigate a VR arena in which the only informative sensory cues are olfactory, enabling us to test whether subjects can learn the spatial relational positions among a set of odors, and whether grid-like responses arise during odor navigation. Aim 2 will test the stability of olfactory grid-like responses by assessing whether contextual changes in the VR arena induce remapping of olfactory cognitive maps. Aim 3 will test the behavioral limits of human olfactory navigation by progressively peeling away all remaining visual cues in the arena. Together these studies should bring fundamental understanding to the capacities and constraints of human olfactory navigation, and should highlight neural mechanisms underlying the “where” question of human olfaction, and more broadly, how the olfactory system tracks and locates odor sources in odiferous environments.

NIH Research Projects · FY 2026 · 2022-01

Under inflammatory conditions, bone destruction can be linked to excessive activity of bone-resorbing osteoclasts (OCs), which results not only from the differentiation of too many OCs, but also from over- maturation of OCs. While most current bone loss treatments prevent bone loss by reducing OC numbers, it may be better if future therapeutic strategies focus on targeting OC maturation rather than early OC differentiation to avoid inhibiting coupled bone formation that depends on interactions between bone-forming osteoblasts and OCs. In an effort to target maturation, we previously identified immunoglobulin superfamily member 11 (IgSF11) as a novel cell surface receptor that regulates OC differentiation but not new bone formation. To characterize IgSF11 signaling, we analyzed, by mass spectrometry, proteins phosphorylated after IgSF11 activation and identified Pyruvate kinase M2 (PKM2), the enzyme that catalyzes the last step of glycolysis, as a downstream target. This finding highlights a potentially greater than previously known determinative role for metabolic regulation during OC differentiation and inflammatory bone loss. We therefore propose the following specific aims: 1. Examine the role of IgSF11-PKM2 signaling in inflammatory bone loss. We will investigate OC-expressed IgSF11 in the context of inflammatory bone loss by using an LPS-induced model of bone loss. To test the contribution of PKM2-dependent effects, we will treat LPS-induced IgSF11-/- mice with small molecule modulators (TEPP-46, shikonin) of PKM2. Our preliminary data suggests that TEPP- 46 activation of PKM2 reduces DSS-induced bone loss. To examine whether IgSF11 expression affects colitis- associated bone loss, we will perform DSS-induced colitis experiments using IgSF11-deficient mice. We will also perform DSS-induced colitis experiments using IgSF11-/- mice treated with TEPP-46 or shikonin. These studies will be critical to establishing the intersection of IgSF11 and PKM2 contributions to clinically relevant inflammatory bone loss. 2. Characterization of IgSF11-PKM2 signaling mechanisms in osteoclast differentiation. We have formulated a four-step model of IgSF11-PKM2 function during OC differentiation, which we will test with the aid of hCD3-iFL, a retroviral (RV) construct to directly activate intracellular IgSF11 in differentiating OCs. We will first investigate possible crosstalk between RANK and IgSF11-PKM2, which we speculate is mediated by TRAF6-dependent K63-linked polyubiquitination of the IgSF11 scaffold protein PSD- 95. Second, we aim to identify kinases proximal to the IgSF11-PSD-95 complex that phosphorylate PKM2. Third, we will use RV mutants to confirm the importance of various PKM2 modifications, PKM2 allosteric confirmation, and PKM2 subcellular localization to OC differentiation. Finally, PKM2 is a well-characterized enzymatic regulator of glycolysis, so we will employ metabolic assays and inhibitors to confirm the significance of this aspect of PKM2 function to OC differentiation. These studies will be critically important to initial validation and characterization of a putative IgSF11-PKM2 pathway and its function during OC differentiation.

NIH Research Projects · FY 2026 · 2021-12

PROJECT SUMMARY/ ABSTRACT Despite effective strategies for prevention, early detection, and treatment, colorectal cancer (CRC) remains the second leading cause of cancer death in the United States. While there have been considerable policy and system-level efforts to increase CRC screening rates, uptake remains well below national goals of 80% participation, and racial/ethnic disparities remain. Colonoscopy and FIT are considered evidence-based and top-tier tests for CRC screening, but most screening programs rely heavily on one of these tests. The overall goal of this proposal is to evaluate a multi-level intervention that could durably increase CRC screening rates by incorporating principles of behavioral economics, such as opt-out framing, simplifying choice, and effort reduction. Through partnership with the University of Pennsylvania Health System (UPHS), we will develop a centralized program that includes clinician-directed nudges facilitated by the electronic health record (EHR) and direct outreach to patients. We will also evaluate the effectiveness of sequential choice (colonoscopy, then FIT to those who defer or decline) compared to offering colonoscopy only. This is a 3-year pragmatic clinical trial with 2 x 3 factorial design at 30 diverse primary care practices with 20,000 average-risk patients who are overdue for screening. Clinicians will be cluster randomized in a 1:1 ratio (at the practice level) to A) colonoscopy only, or B) sequential choice (colonoscopy, then FIT if no colonoscopy is completed) nudges in the EHR during clinic visits (Aim 1). Concurrently, patients will be individually randomized in a 1:2:2 ratio to 1) no outreach, 2) colonoscopy only, or 3) sequential choice outreach of colonoscopy, then FIT (Aim 2). The primary outcome is completion of guideline-recommended colorectal cancer screening. Through surveys and qualitative interviews, we will explore patient and clinician factors impacting the effectiveness of the intervention.

NIH Research Projects · FY 2026 · 2021-12

Project Summary / Abstract My long-term career goal is to become a leading independent investigator developing and evaluating surveillance, preparedness, and operations response strategies to combat the public health burdens from respiratory viral surge events. Respiratory viral surge events, in which hospitals face capacity strain from an influx of infected patients, range from annual respiratory viral seasons dominated by seasonal influenza to rarer and more severe epidemics such as due to novel influenzas (e.g., H1N1) and coronaviruses (e.g., COVID-19, SARS, MERS). Optimizing outcomes for both infected patients and uninfected patients admitted during viral surges (i.e., “bystander patients”), requires that hospitals display: (1) adaptation—the ability to improve care and outcomes for infected patients by implementing new care processes based on accumulated experience, and (2) resiliency—the ability to continue to deliver high quality care to uninfected patients despite the presence of a surge event. However, it is unknown what enables hospitals to display adaptation and resiliency, thereby threatening care quality for all patients during viral surges. I am an Instructor of Medicine in the Division of Pulmonary, Allergy, and Critical Care at the University of Pennsylvania Perelman School of Medicine. My preparations for this career path include masters degrees in clinical epidemiology and biomedical ethics, mentored research training resulting in high-impact first-author publications serving as preliminary data, national invited talks at universities and academic conferences, and clinical work as a pulmonologist and medical intensivist at a major academic referral center. This grant application seeks to combine my and my mentorship team’s experience in defining and studying healthcare capacity strain with purposefully selected career development activities to achieve my complementary training and research goals including methodologic training in advanced statistical modeling, qualitative research methods, implementation science, and cost-effectiveness analysis. The specific aims of this grant are to: (1) Quantify adaptation by determining how hospitals’ cumulative seasonal experiences with influenza affect processes of care and clinical outcomes among high acuity patients with influenza. (2) Measure resiliency by determining how hospitals’ daily capacity strain and cumulative experience during respiratory viral surges affect processes of care and clinical outcomes among bystander patients (i.e., without infection) at risk for acute respiratory failure. (3) Identify organizational characteristics that may influence how hospitals achieve, or struggle to achieve, adaptation and resiliency in the face of a respiratory viral surge event. At the end of the proposed K23 award, I expect to understand how care delivery and outcomes change over the course of a respiratory viral surge event and what organizational factors may account for observed differences in hospital adaptation and resiliency. These findings will have substantial positive impact by facilitating testing organizational interventions to improve hospital adaptation and resiliency, which will be the focus of my initial R01 applications at the conclusion of the K23 award.

NIH Research Projects · FY 2025 · 2021-12

ABSTRACT Current treatment strategies for major depression, the leading cause of disability worldwide, leave more than half of patients with no meaningful treatment benefit. This R61/R33 proposal was developed in keeping with NIMH Strategic Objective 3.1 to “Develop new interventions based on discoveries in genomics, neuroscience and behavioral science.” We will test the efficacy of a new psychotherapeutic strategy, the first real-time fMRI neurofeedback therapy to use cloud-based pattern classification to decode the patient’s attentional state and dynamically modulate task stimuli (in a “closed loop”) based on this state, rather than the standard approach of conveying feedback through a separate gauge that tracks activation in a localized region of interest. The overall objective of this R61/R33 is to test whether closed-loop real-time fMRI neurofeedback that specifically targets our hypothesized attentional mechanism of depression (i.e., neural perseveration of negative states) reduces depression severity. This study will be the first dose-finding test of real-time fMRI effect on negative attention bias. During the R61 Phase 60 participants with MDD ages 18-65 years will be randomly assigned to treatment with active neurofeedback (n=30) vs sham feedback (n=30). We have three aims: 1) Establish neural target engagement using pre-post change in neural perseveration of negative attentional states in active vs sham neurofeedback; 2) Determine the lowest “dose” of training necessary to reduce neural perseveration of negative states; 3) (Exploratory) Establish behavioral target engagement using perseveration on negative images in an eye gaze task comparing active vs sham neurofeedback. Go/No-Go Criteria for R61: There will be at least a medium effect size (Cohen’s d ≥ 0.4) in neural perseveration reduction comparing real to sham NF. Our pilot study (Mennen et al 20) found an effect size of 0.83. Therefore this criterion is both clinically meaningful and realistic and will demonstrate target engagement. During the R33 Phase 80 participants with MDD ages 18-65 will be randomly assigned to treatment with active neurofeedback (n=40) vs sham feedback (n=40). We have three aims: 1) To conduct a randomized controlled trial (RCT) to compare the effect of real-time neurofeedback vs. sham on depression outcome; 2) To determine the relationship between the markers of neural perseveration established in the R61 phase and the reduction in depressive symptoms; and 3) To determine the durability of the treatment effect by comparing MADRS scores at 3 months followup between those who received real as compared to sham NF. Impact. This project will establish real-time fMRI neurofeedback as a means of reducing neural perseveration of negative states as a treatment for MDD. Results from this line of research will inform feedback strategies and improve understanding of neural mechanisms underlying negative attention and MDD. Our cloud based platform would be readily scalable and allow dissemination to thousands of hospital and research facilities.

NIH Research Projects · FY 2026 · 2021-12

Project Summary Ameloblastoma accounts for 14% of all odontogenic tumors and African-Americans are five times more likely to develop ameloblastoma compared to Caucasians. Despite radical surgery, 10% of ameloblastomas recur and 25% of recurrent ameloblastomas occur in the black racial group. The biological determinants of ameloblastoma racial disparity are unclear and there are no specific biological markers to predict recurrence. Most ameloblastomas display genetic mutations of BRAF that encodes the serine/threonine protein kinase B-Raf, an activator of MAPK/ERK-signaling pathway. BRAF oncogenes induce the expression of key autophagic markers that include LC3, p62 and BECLIN1. High expressions of p62, ATG7 and LC3 have been identified in all variants of ameloblastoma and our in vivo mouse ameloblastoma tumor model displayed elevated LC3 and p62 levels. These suggest ameloblastoma recurrence can be attributed to autophagic cell survival mechanisms of residual invasive neoplastic odontogenic epithelium. Interplay of autophagic regulator BECNLIN1 with RUBICON [Run domain Beclin-1-interacting and cysteine-rich domain-containing protein], a component of LC3-associated phagocytosis (LAP) dysregulates autophagosomal maturation and endocytic trafficking to promote tumor migration and invasiveness. Our hypothesis is that autophagy reactivates residual invasive odontogenic epithelium by LAP-mediated entosis and recycling of bioenergetic cellular components. Our collaborative group has a relatively large cohort of ameloblastoma tissues and have generated epithelial-derived (EP-AMCs) and mesenchymal-derived (MS-AMCs) ameloblastoma cell lines from BRAF V600E+ multicystic/follicular ameloblastomas. To elucidate biological mechanisms contributing to racial disparity in Black versus White racial groups, we will determine prognostic biomarkers of ameloblastoma recurrence and assess how LC3-mediated autophagic ‘cargo’ processing orchestrate recurrence disparity. In Aim 1 we will determine whether autophagic proteins are pro-oncogenic adaptors associated with ameloblastoma racial disparity, aggressive phenotype and propensity for recurrence. In Aim 2, we will assess whether residual invasive ameloblastic epithelium survive using LAP-mediated entosis and recycling of bioenergetic cellular components. While ameloblastoma is relatively rare, understanding the interplay of two converging cytoprotective pathways in ameloblastoma growth pattern and recurrence has the potential to lead to new prognostic biomarkers and precision-guided therapies to alleviate racial disparities in BRAF+ tumors like ameloblastoma.

NIH Research Projects · FY 2026 · 2021-12

SUMMARY Our long-term objective is to develop highly translatable animal models for testing cancer immunotherapies. Animal models have been essential in cancer research. However, mouse tumor transplantation or tumor genetic models lack critical components for studying human anti-tumor responses such as high mutational load, tumor microenvironment (TME) and tumor heterogeneity or mice have major differences from humans in innate and adaptive immune responses. In response to PAR-20-131, we will use our novel humanized mouse models (HuMice) to test gamma-delta T (γδT) cell- based immunotherapies. Since Vγ9Vδ2+ T cells, a subtype of γδT cells that are most commonly used in adoptive immunotherapy, are unique to primates, traditional mouse models are not ideal to study human γδT cells. This proposal will maximize translational potential of mammalian models by studying Vγ9Vδ2+ T cells-based therapy in HuMice with HLA-matched human melanomas from cell lines or patient-derived xenografts (PDX). Our laboratories have established >500 melanoma PDX and >300 melanoma cell lines, which represent all clinical, genetic and biologic groups of the disease. In Aim 1, we will study adoptive transfer of enhanced γδT cells. We will test a new Vγ9Vδ2+ T cell expansion method in HuMice. We will then equip γδT cells with DR5-CAR that targets both myeloid derived suppressive cells and melanoma cells. We will study alterations in the TME after treatment. To avoid potential toxicity of targeting DR5, we will develop a novel combinatorial CAR that targets PD-L1 and DR5. In Aim 2, we will stimulate endogenous γδT cells for cancer therapy. We will use bromohydrin pyrophosphate (BrHPP) and resiquimod to expand endogenous human γδT cells to treat melanoma bearing HuMice. We will study whether BTN3A1 mAbs expand human γδT cells and govern antitumor functions of both γδ and CD8+ T cells in HuMice. We will then combine expansion of endogenous γδT cells with anti-PD-1 mAbs in melanoma-bearing HuMice. We expect that HuMice will allow us to test expansion of endogenous Vγ9Vδ2+ for cancer therapy for the first time in a model system and our new expansion methods are effective and may be optimized and tested in future clinical trials.

NIH Research Projects · FY 2026 · 2021-12

Project Summary/Abstract Acute myeloid leukemia (AML) is a hematopoietic malignancy characterized by aberrant self-renewal and blocked differentiation of myeloid progenitor cells. Many of the oncogenic drivers of AML converge in dysregulation of epigenetic and transcriptional regulation pathways, generating de novo dependencies on these regulators. A handful of epigenetic dependencies have been identified in AML, however, single-agent inhibitors against epigenetic regulators have shown limited therapeutic efficacy in patients with AML. To improve our limited understanding of epigenetic-related synergistic genetic interactions in AML, we developed a highly efficient CRISPR-Cas12a-based method enabling us to perform double deletion genetic screening. Our preliminary studies identified and validated two pairs of novel interacting synthetic sick combinations of epigenetic regulators in AML: bromodomain containing protein 9 (BRD9) and Jumonji domain-containing protein 6 (JMJD6) as well as the lysine acetyltransferase 6 (KAT6) and JMJD6. JMJD6 is a bi-functional arginine demethylase and lysyl-hydroxylase regulating transcription enhancer activation and was identified in both synergistic pairs of epigenetic factor deletions. BRD9 is a component of chromatin remodeling SWI/SNF complex and was previously identified as an AML specific dependency. KAT6A is a histone acetyltransferase and transcriptional co-activator. We hypothesize that BRD9/JMJD6 and KAT6A/JMJD6 interact synergistically at the level of transcription and chromatin. The presence of JMJD6 in both interactions suggests that JMJD6- deficiency sets up a unique transcription and chromatin state that sensitizes AML cells to distinct epigenetic stresses. In Aim 1, we will investigate the synthetic sick interactions of BRD9/JMJD6 and KAT6A/JMJD6 in vitro and in vivo and in Aim 2, we will dissect the AML-specific synthetic sick interactions of BRD9/JMJD6 and KAT6A/JMJD6 at the molecular level. Our proposed studies will offer basic mechanistic insight into how these novel AML synthetic sick epigenetic interactions sustain AML pathogenesis. Successful completion of the proposed studies holds significant promise towards developing innovative epigenetic pathway-directed therapies and revealing fundamental biological insights into the pathogenesis of AML. 1

NIH Research Projects · FY 2026 · 2021-12

PROJECT SUMMARY SETD2 is mutationally inactivated in many cancer types including lung adenocarcinoma. In published work, we demonstrated that Setd2 inactivation has potent tumor promoting effects in an autochthonous mouse model of KRASG12D-driven lung adenocarcinoma. SETD2 uniquely catalyzes histone H3 lysine 36 trimethylation (H3K36me3), which marks actively transcribed gene bodies, facilitating chromatin resetting after gene transcription. In the current project, we aim to understand the provocative observation that SETD2 inactivation potently drives tumor cell proliferation due to a defect in one-carbon metabolism and activation of mTORC1 signaling. We demonstrate that SETD2 loss is associated with an enrichment in the abundance of S- adenosyl methionine (SAM) and multiple other metabolites that are part of SAM-adjacent metabolic pathways. We will test the hypothesis that the disuse of SAM that results from the lost activity of the SETD2 methyltransferase leads to SAM accumulation, enhanced one-carbon metabolism, and activation of mTORC1 signaling, all supporting cell growth and proliferation. Consistent with this hypothesis, we demonstrate that limiting dietary intake of methionine reduces KRAS-driven lung adenocarcinoma growth and reverses the effects of SETD2 inactivation. Thus, we will assess the efficacy of clinical and pre-clinical drugs that target the methionine cycle for potential synthetic lethal interactions with SETD2 deficiency. Finally, downstream of activated mTORC1 signaling we observe prominent transcriptional programs of hypoxia inducible factors (HIFs) and peroxisome proliferator-activated receptor γ (PPARγ) coactivator 1 α (PGC1α). Consistently, we observe multiple pathophysiological changes that are associated with increased activation of HIF and PGC1α transcription, such as alterations in mitochondrial biogenesis and the co-enhancement of oxidative phosphorylation and glycolytic pathways. Thus, we will test the requirement of these master transcription factors for effectuating phenotypes downstream of SETD2 inactivation.

NIH Research Projects · FY 2026 · 2021-12

Philadelphia Regional Center for Children’s Environmental Health (PRCCEH) is a new children’s center which will provide the infrastructure to integrate CEH research expertise at University of Pennsylvania (Penn) Perelman School of Medicine (PSOM), Children’s Hospital of Philadelphia (CHOP), Drexel University, Temple University, Thomas Jefferson University, Lehigh University, Franklin & Marshall College, Villanova University and University of Delaware to improve children’s health by reducing environmental exposures in early life across our region. The Center will be led by Directors Rebecca Simmons, M.D., and Aimin Chen, M.D., Ph.D., and Deputy Director Marilyn Howarth, M.D. The mission of the PRCCEH is to disseminate children’s environmental health knowledge to health care providers, community members, and policy makers, to develop, test and implement new translational products, and to engage researchers and community partners to make policy, practice, and behavioral changes to reduce environmental exposures in early life. The vision and the mission of the PRCCEH are oriented to the Philadelphia region to address pressing CEH issues. The Center consists of 27 experts in pediatrics, epidemiology, occupational and environmental medicine, toxicology, social ecology, implementation science, community engagement, risk communication, nursing, bioinformatics, and other areas. The Center is comprised of an Administrative Core and Executive Committee, a Development Core, and a Translation Core, and is advised by Internal Advisory Committee and Community Stakeholder Advisory Committee. The Center will focus on four primary research and translation areas: a) Asthma prevention, motivated by increased rates of asthma hospitalization in Philadelphia and a Community Asthma Prevention Program (CAPP) with more than 20 years of experience; b) Lead exposure and harm reduction, motivated by high rate of elevated blood lead levels in children <6 years of age at the population level; c) Air pollution, motivated by high annual particulate matter and ozone pollution in Philadelphia-Reading-Camden Metropolitan Statistical Area (among the 25 worst polluted areas in the U.S.); and d) Endocrine disrupting chemicals (EDCs), motivated by increasing disease burden from EDC-related preterm birth, obesity and diabetes, and neurodevelopmental disorders. The Center members are experienced in these translational areas and will achieve the following Specific Aims in the next 5 years. Aim 1. Build the PRCCEH as a regional infrastructure center for CEH research and translation; Aim 2. Expand PRCCEH membership and leverage institutional resources to promote CEH translation; Aim 3. Nurture and mentor early-stage investigators (ESI) in CEH research and attract established environmental health (EH) scientists into CEH; Aim 4. Translate CEH research to community members and stakeholders to improve children’s health; Aim 5. Establish two pilot programs for CEH translation; Aim 6. Provide efficient administrative services to facilitate research translation.

NIH Research Projects · FY 2026 · 2021-12

PROJECT SUMMARY Incisional hernia (IH) is a common, overlooked surgical health problem spanning a broad range of patients and stakeholders. In the U.S., over 153,000 IHs are repaired per year with expenditures exceeding $7 billion. Evidence-based interventions, including preoperative optimization, surgical techniques, and prophylactic mesh, can reduce risk; however, multi-level factors impede clinical translation. One critical barrier is the need for accurate, generalizable risk prediction to link risk recognition, behavior change, and outcomes. Pre-operative risk assessment enables providers to leverage risk information to guide decision-making, surgical planning, and informed consent. Current limitations of IH prediction have created barriers to IH prevention. Our proposal addresses the need for patient-specific, clearly presented risk information to enhance health care, enable individualized risk assessment, and close the gap between optimal practice and actual clinical care in hernia prevention. Our preliminary research has defined the clinical and economic burden of IH, characterized inefficiencies in treatment-oriented paradigms, defined key patient populations for prevention, and demonstrated effective risk reductive surgical techniques. We also show the benefit of using electronic health record-based prediction over administrative claims datasets and the power of machine learning to maximize model performance. Most recently, we created a pilot, portable, clinical decision support-mobile user interface for prediction, setting the stage for this proposal. Our approach is hallmarked by use of a unique multi-source database, innovative applications of machine learning, stake-holder engagement, and inter-disciplinary collaboration. In this proposal, we will identify and discover factors associated with IH using data from >130,000 patients with longitudinal follow-up and characterize intra-operative risk factors using natural language processing. Machine learning will enable improved predictive performance (Aim 1). Models will be tested on a geo-temporally diverse data source and end-user input will guide and prioritize features, format, and functionality, leading to creation of a provider-adapted Hernia Calc housing the predictive models (Aim 2). Hernia Calc will be evaluated in real-world practice to assess contextual determinants and to create a stakeholder-driven implementation protocol to identify strategies to support widespread dissemination (Aim 3). Our approach addresses barriers to IH prevention through development of optimized, validated, specialty- specific IH risk models integrated within a provider-informed interface and implementation strategies for clinical use. This work will lead to a broad, significant, and sustained impact on the field, catalyzing a major pivot towards hernia prevention, enabling precise risk prediction for abdominal surgery patients. Completion of our aims will augment knowledge of hernia and improve health outcomes in surgery allowing a pivot in practice towards prevention and aligning our proposal with Core Missions of the NIH.

NIH Research Projects · FY 2026 · 2021-12

PROJECT SUMMARY Surgical outcomes for patients with intractable epilepsy are modest, in large part due to mis-localization of seizure generators. Currently, epileptologists localize seizure generators by manually identifying the seizure onset zone on intracranial EEG. There is also abundant interictal EEG data (between seizures) that is largely ignored because, until now, clinicians lacked rigorous approaches to interpret this data. Recent work by Dr. Conrad, her co-authors, and others, validated two quantitative methods to analyze interictal EEG: interictal epileptiform discharges (IEDs) and functional connectivity (FC). This work demonstrates that quantitative analysis of IEDs and FC reveals regions — referred to here as hubs — that help identify seizure generators (Conrad et al., Brain, 2020; Conrad et al., Network Neuroscience, 2020). This proposed project will evaluate IED and FC hubs in order to elucidate their mechanisms and their potential utility in surgical planning. The central hypothesis of this proposal is that epilepsy measurably alters the interictal network, allowing us to probe seizure generators with interictal data. The objectives of the proposal are: to determine the temporal stability and spatial correspondence between IED and FC hubs (Aim 1), to determine if these hubs localize seizure generators (Aim 2), and to actively probe the interictal network with cortical stimulation to verify hypothesized FC hubs (Aim 3). These studies will expand our understanding of epileptic networks. If successful, quantitative analysis of interictal EEG will enhance localization of seizure generators and improve surgical outcomes. This proposed project will also provide critical career development training to Dr. Conrad, an Epilepsy Instructor at University of Pennsylvania. The proposal builds on Dr. Conrad's background in quantitative EEG analysis and clinical epilepsy. Dr. Conrad will be mentored by Brian Litt, a world-renowned expert in computational epilepsy; Danielle Bassett, a leading expert in network theory; and Eric Marsh, a leading expert in quantitative IED analysis. The training plan in this proposal includes mentored completion of the proposed research, as well as a rigorous program of didactics, workshops, lab meetings, and directly relevant clinical work. Together, these experiences will provide essential training in time-series statistics, machine learning, subject enrollment, cortical stimulation, and EEG data acquisition. This proposed project and training plan will launch Dr. Conrad on an independent research career focused on using a multimodal approach to evaluating interictal data in order to improve our understanding and treatment of epilepsy.

NIH Research Projects · FY 2025 · 2021-09

SUMMARY Alzheimer's disorder (AD) is a devastating neurodegenerative disease and the most common cause of dementia. There are approximately six million Americans with AD and 29.8 million worldwide, making AD one of the most pressing public health issues as the world's population continues to age. Presently, there is no known effective prevention or cure exists, and current AD medications only alleviate symptoms or slow decline rates. The landscape of AD drug trials is gloomy. One possible reason is that AD is a heterogeneous disorder but trials are designed treating it as a monolithic disease. Although lifestyle and environmental risk factors clearly affect AD, the primacy of genetic influences suggests that categorization by genetic basis should be prioritized in developing effective interventions. Genetics can offer insights on risk prediction, disease mechanism, and new therapeutic targets. Heritability of AD estimates range from 49-79%, but the conventional single nucleotide variants (SNVs) identified to date only account for <50% of AD heritability. Multiple studies have highlighted the roles of copy number variants (CNVs) in AD. We hypothesize that a systematic investigation of genome-wide CNVs at the full spectrum (i.e. small and large in size, common and rare in frequency, and coding and no-coding in genomic regions) from whole-genome sequencing (WGS) can further enhance the knowledge of AD etiology and risk. Leveraging the rich resources from the Alzheimer's Disease Sequencing Project (ADSP), we propose to focus on a large multi-ethnic WGS sample (n>17,000) composed of AD cases and normal healthy elderly controls, and to (1) detect and genotype CNVs from WGS for ADSP case-control samples; (2) perform association analysis to identify genome regions of CNVs contributing to AD; and (3) conduct cross-ethnic association studies to find ethnic-shared or ethnic-unique AD-associated CNVs. Successful completion of our aims will provide (i) the first large-scale CNV investigation of AD genetics using WGS data; (ii) new CNV calling method for WGS based on the current best practices; (iii) new CNV association strategies to address issue of breakpoint non-alignment and enhance association power; (iv) multi-ethnic characterization of shared and unique CNV risk factors for AD; and (v) optimized computational pipelines with open- source code and released standardized images (e.g., Docker images and Bioconductor packages) that are easily deployable in other large-scale WGS association projects.

NIH Research Projects · FY 2025 · 2021-09

The overarching goal of our proposed Penn Artificial Intelligence and Technology (PennAITech) Collaboratory is to identify, develop, evaluate, commercialize, and disseminate innovative technology for monitoring aging adults and those with Alzheimer’s Disease (AD) and Alzheimer’s Disease Related Dementias (ADRD) in their home environment and the artificial intelligence (AI) methods and software for analyzing data generated by those technologies. The collaboratory is motivated by the need for a comprehensive pipeline from technology-based monitoring of aging adults in the home, collection and processing monitoring data, integration of those data with clinical data from electronic health records, analysis with cutting-edge AI methods and software, and deployment of validated AI models at point of care for decision support. We have assembled a team of experts with experience building and implementing each component of this pipeline and will promote this vision and existing tools and software (AIM 1). A central focus of the PennAITech Collaboratory is to advance this vision through the solicitation, review, and funding of pilot grants focused on technology and AI development to advance the science of care management and aging in place for vulnerable older adults or those with AD/ADRD receiving skilled home and community-based services (AIM 2). Funded pilot projects will be supported in AIM 3 through cores focused on administration (Core A), stakeholder engagement (Core B), technology identification and training (Core C), clinical translation and validation (Core D), networking (Core E), and ethical and policy issues (Core H). Finally, the collaboratory will work closely with the planned coordinating center to facilitate the success and translation of all collaboratory projects (AIM 4). We are committed to improving the health of aging adults and those with AD/ADRD through these specific aims.