University Of Pennsylvania

NIH Research Projects · FY 2026 · 2025-07

Project Summary The opioid epidemic remains a major public health crisis in the U.S. Despite the effectiveness of current FDA- approved medications to treat opioid use disorder (OUD), there is still a high rate of relapse following detoxification. Thus, there is a critical need for conceptually new research investigating the neurobiological mechanisms underlying opioid taking and seeking that could lead to novel druggable targets. Our exciting preliminary studies indicate that systemic infusions of amylin, a satiation factor and neuropeptide, attenuate oxycodone self-administration and reinstatement at doses that do not alter food intake or compromise the antinociceptive effects of oxycodone. Consistent with these findings, activation of amylin receptors in the mesolimbic reward system is sufficient to reduce oxycodone self-administration and reinstatement. While our pilot studies highlight a novel neuropeptide system that could be targeted to reduce opioid taking and seeking, there are significant gaps in our understanding of central amylin receptors and their role in addiction-like behaviors. We recently began to identify cell type-specific patterns of amylin receptor expression in the brain and showed that amylin receptors are expressed on dopamine and GABA neurons in the ventral tegmental area (VTA), as well as dopamine d1 receptor (D1R) and dopamine d2 receptor (D2R) -expressing medium spiny neurons (MSNs) in the nucleus accumbens (NAc). One goal of this proposal is to build upon and expand our pilot studies by comprehensively defining the functional roles of cell type-specific VTA and NAc amylin receptors in opioid taking and seeking using viral-mediated methods in transgenic rats (Aim 1). We will also determine the individual contributions of these amylin receptor-expressing cell types to amylin's efficacy (Aim 1). Very little is known about the cellular and molecular mechanisms underlying the suppressive effects of amylin pharmacotherapy on voluntary drug taking and seeking. Therefore, we will combine viral-mediated gene delivery approaches with in vivo fiber photometry in transgenic rats to determine how amylin pharmacotherapy alters intracellular calcium dynamics in specific VTA and NAc cell types of oxycodone-experienced and drug- naïve rats in order to understand how activation of central amylin receptors alters midbrain and striatal circuits to suppress opioid taking and seeking (Aim 2). Finally, we will use an unbiased, single nuclei transcriptomics approach to characterize the effects of oxycodone alone and in combination with amylin pharmacotherapy on amylin receptor expression and differently expressed genes (DEGs) in all VTA and NAc cell populations (Aim 3). Findings from these studies will provide our first insights into how oxycodone alters amylin receptor expression and signaling in specific midbrain and striatal cell populations and identify opioid dysregulated genes that are putatively restored by amylin pharmacotherapy. Overall, the research proposed in this application will advance a novel framework for the development of pharmacotherapies aimed at targeting central amylin receptor-expressing cells to treat OUD.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY Improved understanding of B cell biology may uncover therapies that will restore immune tolerance in autoimmune disease with minimal safety risk. Tumor necrosis factor (TNF) superfamily members are key regulators of B cell biology, and blockade of several of these pathways has shown significant clinical benefit in autoimmunity with minimal infectious risk. For example, B cell activating factor (BAFF)-blockade provides benefit in lupus, a proliferation-inducing ligand (APRIL)-blockade provides benefit in IgA nephropathy, CD40L-blockade shows promise in Sjogren’s disease, and all these therapies are well-tolerated. TNF superfamily member receptor activator of NF-κB (RANK), and its ligand RANKL, are highly expressed by memory B cell subsets. Human genetic variation in RANK is associated with the autoantibody-mediated disease myasthenia gravis. In mice, transient blockade of RANKL increases the quantity of antigen-specific vaccine titers. Yet, the impact of RANKL on human humoral immunity remains unknown. Understanding RANK-RANKL B cell control in humans may identify a new pathway to augment antibody responses to vaccination or dampen autoantibody responses in autoimmunity with limited safety risk. To address this gap in knowledge, I captured the in vivo human experiment by collecting plasma and peripheral blood mononuclear cells from individuals receiving denosumab, an FDA approved RANKL-blocking antibody commonly used in the treatment of osteoporosis, and matched controls following Covid-19 booster vaccination. I observed a significant increase in Covid-specific IgG responses, total IgG1, and autoantibody formation to numerous self-antigens in individuals receiving RANKL- blockade. As RANK signaling can induce FAS in osteoclasts and FAS expression in B cells is a critical negative regulator of T cell-dependent B cell responses, I hypothesize that blockade of RANKL broadly increases human B cell responses to vaccination and self-antigen by impairing B cell FAS-mediated apoptosis. This hypothesis will be tested through the following 3 aims: 1) Determine the impact of RANKL-blockade on humoral and cellular immunity to recent Covid and influenza vaccination versus remote tetanus vaccination, 2) Define the site of impaired B cell tolerance and the pattern of autoantibodies following RANKL-blockade, and 3) Test the impact of RANKL-blockade on FAS-mediated B cell survival in vitro and in vivo. Collectively, these studies will address the existing knowledge gap of RANKL control of human B cell responses and may provide the foundation to target RANK-RANKL to tune human humoral immunity.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY Childhood disruptive behavior disorders (DBDs) are one of the most common forms of psychopathology in children, with a worldwide pooled prevalence of nearly 6%. Much research has linked the presence of callous- unemotional (CU) traits (i.e., low empathy, guilt, and prosociality) to increased risk for severe DBDs across development. Moreover, children with DBDs and CU traits show poorer treatment outcomes when compared to children with DBDs alone. Heterogeneity within the clinical phenotype of CU traits is thought to compound this treatment gap. In particular, children with CU traits can be further differentiated into primary (i.e., low fear, low anxiety) and secondary (i.e., anxiety and trauma) subtypes. Theoretical models suggest that primary and secondary CU traits develop through distinct etiological pathways. However, the physiological mechanisms which underlie the development of these subtypes are not well understood and studies have yet to use advanced machine learning techniques to derive biologically informed subtypes. Here, I propose that the development of CU traits occurs via two distinct etiological pathways underpinned by unique profiles of physiological reactivity and exposure to adversity. As part of an ongoing, longitudinal study, preschool aged children (n=500) complete a variety of behavioral tasks while parasympathetic nervous system (PNS) function, as indexed by respiratory sinus arrythmia (RSA), is measured. Parent-reported questionnaires assessing CU traits, DBDs, anxiety, and exposure to adversity are also collected. I hypothesize that RSA will differentially predict CU trait at baseline and 2-year follow-up, such that higher RSA will be related to CU traits for children exposed to more adversity, and lower RSA will be related to CU traits for children regardless of exposure to adversity (Aim 1). Additionally, I hypothesize that a novel semi-supervised machine learning algorithm will identify “biotypes” of CU traits based on patterns of PNS function. Specifically, I expect to find a primary profile characterized by low levels of RSA across tasks, associated with lower anxiety and fearlessness, and a secondary profile characterized by high levels of RSA, associated with anxiety and emotion dysregulation (Aim 2). The overarching goal of this proposal is to gain a better understanding of how subtypes of CU traits develop based on distinct patterns of physiological reactivity in the context of differential exposure to adversity. These findings can inform etiological models of CU traits, while simultaneously informing early identification and precision treatment of childhood DBDs. With guidance from a strong mentorship team with relevant expertise (Drs. Waller, Wagner, Kimonis, Davatzikos, and Zisser), my proposal and the accompanying training plan provide an ideal foundation for my planned career as an independently funded, leading clinical scientist studying biomarkers of externalizing psychopathology.

NIH Research Projects · FY 2025 · 2025-07

Enter the text here that is the new abstract information for your application. This section must be no longer than 30 lines of text. This is a new application to support the Predoctoral Training Program in Genetics (GEN-TG) at the Perelman School of Medicine (PSOM), University of Pennsylvania. The primary goal of the GEN-TG is to recruit, train and mentor outstanding individuals who will become future leaders in genetic research as well as effective educators and mentors. To accomplish this goal, we have established a flexible, evidence-based training plan that integrates technical and professional skills in preparation for research intensive careers in academia, industry and government. The program brings together a select group of 53 faculty from 17 departments or schools with demonstrated expertise and commitment to the training and mentoring of students. The combination of a research-oriented medical school and a strong university base provides an outstanding environment for graduate education. The Genetics and Epigenetics (GE) graduate program was established in 1995 and is the major training entity in the discipline of Genetics at PSOM. GE is one of six subgroups of the multidisciplinary Cell and Molecular Biology Graduate Group, that administers the broad core curriculum for our predoctoral trainees, including advanced coursework in Genetics. Each student's individualized curriculum emphasizes the integration of research training, responsible conduct of research, scientific rigor and reproducibility, and mentorship. After coursework is completed, the student must pass an oral preliminary exam that consists of a defense of a written NRSA-style thesis research proposal. The student then carries out a significant genetics research project under the direction of a laboratory mentor and the advice of a thesis committee, with additional oversight and mentoring from GEN-TG faculty. Students are appointed to the GEN-TG for two years of support after they have completed coursework, selected a thesis lab and passed their Preliminary exam. The GEN-TG provides added value to the graduate program through the following training activities: honing presentation skills at the weekly Genetics Research in Progress talks; guidance in the preparation of individual fellowship applications; establishing best practices in peer review at mock-study sections; building a strong cohort identity at the annual Penn Genetics retreat; networking with Distinguished Seminar speakers; supporting travel to attend scientific conferences; exploring career options in Philadelphia Biotech; and hands on training in near-peer mentorship. GEN-TG trainees have outstanding publication records, retention rates and career outcomes. Our objectives over the next five years include: 1) foster the next generation of geneticists by preparing our trainees to traverse their chosen career path successfully; 2) assist trainees to explore career options within and beyond academia; 3) manage upward pressure on total training time; 4) improve mentorship skills for faculty and trainees.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY Genetic bi-allelic deficiency of LCAT (LCAT-D) is an ultra-rare, autosomal recessive condition that manifests as two different syndromes, familial LCAT deficiency (FLD) and fish-eye disease (FED). Both syndromes are characterized by excess of unesterified cholesterol (UC), significantly reduced HDL-C levels and corneal opacities. In addition, FLD patients develop chronic progressive renal disease, which represents the main cause of morbidity and mortality in this condition. Additional manifestations of FLD include anemia, increased levels of triglycerides and the presence of an abnormal lipoprotein called LpX. While the consequences of LCAT-D on lipoprotein metabolism have been extensively characterized, much less in known about the natural history of LCAT-D. Clinical presentation and progression of disease appear to be heterogeneous across individuals, and we still lack a clear understanding of the factors contributing to such variability, not only between FED and FLD, but also within each syndrome. Current treatment for FLD is limited to symptomatic management of its sequelae. Novel therapeutics targeting LCAT are at different stages of development, including an AAV-hLCAT gene therapy approach at the University of Pennsylvania. Unfortunately, their further development is limited by inadequate understanding of the natural history of LCAT-D. Identification of suitable biomarkers to determine eligibility and assess efficacy is urgently needed. The overarching goal of this proposal is to: 1. Identify biomarkers that can guide enrollment criteria, including diagnostic biomarkers to distinguish FLD from FED, and prognostic biomarkers to enrich enrollment with FLD patients that have a higher risk of rapid CKD progression (Aim 1). 2. Identify pharmacodynamic and efficacy biomarkers to aid the identification of target response and the efficacy of the intervention tested (Aim 2). 3. Identify comorbidities and concomitant medications that may impact pharmacodynamic measurements (Aim 3). We will achieve these goals by performing unbiased retrospective long-term deep data extraction from medical records of FED and FLD patients that will enroll in our ongoing natural history study (NCT06217588). These data will be complemented by laboratory assays assessing LCAT mass and activities, and other markers of esterification, including %CE, done in samples obtained from the patients enrolled in the study and from pre-existing samples and data from selected cohorts of LCAT-D patients and patients with secondary LCAT deficiency. The successful completion of this study will provide key diagnostic biomarkers and needed information on the rate of decline of key markers of disease, as well as the effect of concomitant medications or co-morbidity that may affect the primary pharmacodynamic parameters. Overall, these data will be pivotal for the refinement of the inclusion and exclusion criteria and be instrumental to identify the primary clinical efficacy endpoints and establish the duration of the clinical trial.

NIH Research Projects · FY 2025 · 2025-07

PROJECT SUMMARY Hepatocellular carcinoma (HCC) is the predominant subtype of liver cancer and arises almost exclusively in the context of chronic inflammation. The healthy liver is a modulator of systemic immune tolerance, recognizing harmless dietary and environmental antigens from the gut and degrading them without inducing inflammation. Dysregulation of this tightly controlled response can lead to chronic inflammation, compensatory hepatocyte regeneration, and eventually fibrosis and hepatocarcinogenesis. While viral hepatitis is the predominant risk factor for HCC, metabolic disorders such as metabolic-associated fatty liver disease (MAFLD) are expected to increase the global incidence of HCC in the coming decades. With treatment options limited due to the mutational heterogeneity of HCC and poor immunotherapeutic responses in patients with MAFLD-associated HCC, there is a clinical need to identify the metabolic vulnerabilities of cancer and immune cells in the tumor micro- environment to exploit specific dependencies of HCC and improve cancer therapy. To this end, we have identified one of the most downregulated metabolic pathways in HCC to be catabolism of the branched chain amino acids (BCAAs). The BCAAs are leucine, isoleucine, and valine, all of which are essential amino acids that contribute to protein synthesis or undergo catabolism to support anaplerotic processes. The majority of BCAA catabolic enzymes are often downregulated in HCC, apart from two enzymes responsible for the first step in the pathway: BCAT1 and BCAT2. Both enzymes catalyze the reversible transamination of BCAAs by transferring the amino group to α-ketoglutarate to generate glutamate and the respective branched chain ketoacids. Loss of BCAA catabolism downstream of the BCATs may represent a unique vulnerability of HCC and confer dependence on BCAT1 or BCAT2 expression for tumor growth. Here, I demonstrate that high BCAT1 expression, but not BCAT2, correlates with worse overall survival in patients according to data from The Cancer Genome Atlas. Moreover, BCAT1 protein expression is high in many well-characterized human cell lines of HCC, and BCAT1 knockdown inhibits growth in a high BCAT1-expressing line, which can be rescued by BCAT1 re-expression. This suggests that tumors with high BCAT1 still utilize the BCAA pathway to support their growth, and this could impact BCAA availability within the tumor microenvironment as a result. Thus, I hypothesize that BCAT1 promotes HCC cell growth through a tumor-intrinsic mechanism, and this suppresses anti-tumor immunity by reducing the amount of BCAAs available to immune cells in the tumor microenvironment. For my first aim, I will determine how BCAT1 knockdown leads to growth inhibition with metabolite supplementation and will confirm the relevance of BCAT1 to HCC growth in vivo. In my second aim, I will investigate whether BCAT1 expression regulates BCAA uptake and how changes in BCAA levels impact immune cell populations in the liver. Collectively, these findings will provide insight into BCAA utilization in the context of BCAT1 and establish a critical crosstalk between tumor and immune cell metabolism in HCC.

NIH Research Projects · FY 2026 · 2025-07

The symbiotic relationship between microbiota (MB) and the human host is a key component of health and its dysregulation is associated with many disease states, including inflammatory, cardiovascular, and autoimmune diseases. Homeostatic regulation of gut microbial communities and their metabolic products is achieved by several host mucosal immune mechanisms, most critically, the production of secretory IgA (sIgA) that directly coat large fractions of the MB. Until recently it has been believed that sIgA is the only secretory immunoglobulin (sIg) involved in the maintenance of MB homeostasis. However, this paradigm is now being challenged by findings showing that sIgM also coats a large proportion of the human MB, often in association with sIgA. The goal of this proposal is to determine the unique and synergistic physical and functional consequences derived from the targeting of MB by different sIg isotypes. Upon sIgA binding, bacteria undergo agglutination, motility arrest, and enchained growth (EGrowth), processes that ultimately dictate whether bacteria are physically excluded from the mucosal barrier or colonize it. In addition, sIgA binding to MB regulates bacterial gene expression and metabolite production, thus further contributing to the sIgA functional properties. While sIgA effects on MB are beginning to be deciphered, the effects of sIgM coating and the potential synergies between sIgM and sIgA have never been investigated. In contrast to humans and fish, laboratory mice lack sIgM-coated MB thus precluding the use of this animal model in studies involving sIgM-MB interactions. Using our highly tractable fish model, we have established that both sIgT (a functional homolog of sIgA) and sIgM coat their gut MB (either alone or in combination). Moreover, we have generated new preliminary data that support our central hypothesis that MB coating by sIgT and/or sIgM has unique, complementary, and synergistic functional consequences on the modulation of several key processes critical for MB homeostasis, including MB agglutination, EG, motility, gene expression and metabolism. To address this hypothesis, we will first evaluate the differential and synergistic effects of sIgT and sIgM binding to MB on agglutination, EGrowth, motility and colonization followed by evaluation of the effect of sIgM and sIgT binding on MB metatranscriptome, gene expression and metabolism. We will then explore sIgs-mediated modulation of bacterial gene expression through novel superoxide-dependent and independent mechanisms, evaluate regulation of bacterial antibiotic production upon sIgs coating of MB, and explore where are MB-specific sIgM generated in the host while assessing whether sIgM and sIgT recognize the same or different MB molecules. The proposed work has the potential to uncover novel mechanisms by which sIg-coating of MB play unique and synergistic roles in physically regulating MB colonization or clearance, and in modulating gene expression as well as enhancing or limiting the metabolic influence of MB on the host. Moreover, the knowledge derived from this paradigm shifting research will be used in future projects to identify, design and evaluate MB- and sIg-targeted interventions for critical human diseases associated with dysregulation of the microbiome such as Chron’s disease, inflammatory bowel disease, and colorectal cancer.

NSF Awards · FY 2025 · 2025-07

In the rapidly expanding field of data science, the ability to understand group actions in data analysis is pivotal for a broad spectrum of scientific tasks. In mathematical terms, a “group” is a collection of elements combined with an operation that links any two elements to form a third, adhering to closure, associativity, identity, and invertibility principles. A “group action” involves applying elements of a group to another set’s elements, transforming them in structured ways, such as through rotations or reflections. These transformations are crucial in many data processing applications, including cryo-electron microscopy (cryo-EM), image registration, and multi-reference alignment. Each observation in these problems involves a common, unknown signal and an unknown group element, with the primary goal being to infer both the signal and the group elements accurately. This project aims to significantly advance statistical understanding and develop effective methodologies for handling data influenced by group actions. The wide existence of such data ensures that the progress we make towards our objectives will have a great impact not only on the statistics and machine learning community but also on a much broader scientific community, including fields such as structural biology, computer vision, and signal processing. This project will have educational outcomes that result in curriculum development, teaching, and outreach activities, including activities to K-12 students through the University of Pennsylvania Data Science Academy. The project will advance applications in image recognition and time series alignment, which have broad application in areas like medical imaging. This project is structured around three main aims, each designed to tackle distinct aspects of group actions. First, the PI will improve the accuracy of orbit recovery in scenarios where the prior distributions of group elements are non-uniform, developing computationally efficient procedures that are effective under realistic conditions. Second, the PI will develop theories and methods for group synchronization problems, particularly under high noise levels and in situations with incomplete data, aiming to reduce the error of group recovery and provide entrywise inference. Third, the PI will address theoretical and computational challenges in the multi-reference alignment problem, developing procedures specifically designed for the cyclic structural nature of data, thereby enabling more precise uncertainty quantification. Together, these aims will not only enhance the theoretical understanding of and the ability to analyze group actions but also lead to the development of accurate and computationally efficient algorithms designed to tackle real-world challenges in data analysis where group actions are integral. This research project will have impacts more broadly, in that it will result in software development and in the education of technical experts. These experts will use this software to advance applications in image recognition and time series alignment, which have broad application in areas like medical imaging. These activities will then advance applications in image recognition and time series alignment, which have broad application in areas like medical imaging. 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 · 2025-07

The Algebraic Geometry Northeastern Series (AGNES) is a series of biannual conferences in the field of algebraic geometry. The conference is hosted on a rotating basis by an association of universities in the Northeast region. This award supports six AGNES conferences, which will be held at Dartmouth College on November 8-10, 2024, at Rutgers University in Spring 2025, at the University of Massachusetts, Amherst in Fall 2025, at Stony Brook University in Spring 2026, at Brown University in Fall 2026, and at the University of Pennsylvania in Spring 2027. Each AGNES conference has two goals. First, each conference promotes the dissemination of cutting-edge research in mathematics. The centerpiece of each conference is a series of research lectures by top mathematicians; there are also educational talks for graduate students and events which promote new collaborations or development of peer relationships. Algebraic geometry is a field in the mathematical sciences concerned with solution sets of polynomial equations. It has deep connections to many other areas of pure mathematics, such as topology, arithmetic, number theory, differential geometry, dynamical systems, and homological algebra. At the same time algebraic geometry has found important applications in many subdisciplines of applied mathematics, including cryptography, complexity theory, mathematical biology, and computer vision. The scientific scope of AGNES is greatly enriched by lectures from neighboring mathematical subjects, such as arithmetic geometry, dynamics, complex geometry, and computational geometry. Further information about conference events can be found at the website: http://www.agneshome.org/ 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 · 2025-07

ABSTRACT Pulmonary fibrosis (PF) is a devastating and progressive lung disease with a rising incidence and prevalence and minimally effective therapies. PF pathology is characterized by aberrant mesenchymal and epithelial populations that disrupt the normal alveolar lung architecture. In the alveolar epithelium in human PF and murine PF models, a subset of Alveolar Type 2 (AT2) cells lose their quintessential transcriptional program, entering but failing to exit an “aberrant transitional” state between AT2 and Alveolar Type 1 (AT1) cells. These cells are found in proximity to pathological collagen producing fibroblasts and they are transcriptionally enriched in fibrotic mediators, suggesting they may pathogenically activate alveolar fibroblasts. Why these aberrant transitional AT2s arise, why they fail to differentiate to AT1s, and how they contribute to fibrosis is not understood. We have identified that the aberrant transitional AT2s are enriched in the PKR-like ER kinase (PERK) cell stress pathway and its downstream Integrated Stress Response (ISR). How PERK-ISR signaling relates to the emergence and persistence of the aberrant AT2s and to the epithelial contribution to fibrogenesis is also not known. In this proposal we will answer these questions by utilizing complimentary in vivo models of lung fibrosis with a human model system. For our murine models, we use mice that express a human PF-associated mutation in the AT2 restricted Surfactant Protein C gene (SftpcMut mice) where intrinsic AT2 stress pathways initiate spontaneous lung fibrosis without a “second hit” lung injury and the established Bleomycin (Bleo) fibrosis model of extrinsic AT2 activation and lung fibrosis. Our human system is an induced pluripotent stem cell (iPSC)-based AT2 (iAT2s) model from a PF patient harboring a SFTPCMut. In both murine models we have found that AT2s are enriched in PERK-ISR signaling as they enter and persist in the aberrant transitional state. We have developed a strategy to sort the most fibrogenic aberrant transitional AT2s from the mouse lung and found that in ex vivo co-culture systems these cells induce a fibrotic response in alveolar fibroblasts. We also discovered that SFTPCMut iAT2s develop an aberrant transitional gene signature and are enriched in PERK-ISR signaling and profibrotic mediators. We will now test our hypothesis that PERK-ISR signaling promotes AT2s to enter and restrains their exit from an aberrant transitional state that contributes to lung fibrosis by pathogenically activating alveolar fibroblasts. We will [Specific Aim 1] determine how aberrant transitional epithelial cells directly contribute to fibrosis and test our hypothesis that an altered alveolar niche signaling circuit initiated by the aberrant transitional epithelial cells promotes fibrotic tissue remodeling. We will [Specific Aim 2] define how PERK-ISR signaling impacts epithelial differentiation and promotes fibrosis and test our hypothesis that PERK-ISR signaling contributes to AT2 persistence in the aberrant transitional state. When completed these studies will fill critical knowledge gaps in PF biology by defining the direct contribution of the aberrant transitional cell to fibrosis and the mechanistic impact of the targetable PERK-ISR pathway in PF pathogenesis.

NSF Awards · FY 2025 · 2025-07

This project is about using recent advances in mathematics to improve lattice based cryptography. Lattice based cryptography is a foundation for advanced cryptographic schemes that are resistant to attacks by quantum computers. Secure cryptography is essential for digital communication, for example for ensuring the safe transfer of sensitive financial data. The new mathematical advance behind this project is the efficient construction of lattices that have both addition and multiplication operations and that are more densely packed than the ones now typically used in cryptography. The central goals of the project are to improve these constructions, to develop faster algorithms to operate on these lattices, and to build more efficient cryptographic applications using them. The technical advances in this project concern number fields with small root discriminants. This project will study the computational complexity of constructing such number fields and of performing arithmetic operations in them. Prior work on infinite families of number fields with small root discriminants has focused on existence theorems. This project will build on work of two of the P.I.s on efficient explicit constructions of such families. The goal is to improve these constructions using a variety of mathematical techniques including Galois cohomology, explicit Chebotarev theorems and recent advances on Hilbert's 12th problem via p-adic methods and modular forms. Another goal is to develop fast Fourier methods for performing arithmetic operations of the kind needed in cryptography. The relevance of number fields with small root discriminants was noted by Peikert and Rosen in 2006. They showed that such fields lead to very small connection factors relating the difficulty of solving the worst case of the short vector problem to the difficulty of solving the average case of the short integer solution problem. The cryptographic protocols to be studied using the rings of integers of the above fields include collision resistant hash functions, homomorphic commitment schemes, streaming authenticated data structures, zero-knowledge proof systems, and some types of digital signature schemes. 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 · 2025-07

PROJECT SUMMARY This proposal aims to elucidate the mechanisms by which TET family enzymes act on 5-methylcytosine in genomic DNA and to exploit these insights to devise controllable epigenome editors. Modifications to cytosine bases play an important role in diverse processes, including development, pluripotency, and oncogenesis. The best-studied cytosine modification is methylation at the 5-position (5mC), typically occurring at cytosine-guanine dinucleotides (CpGs) which are most often found in clusters, including CpG islands (CGIs). The methylation status of CpGs over regions of DNA plays a key role in dictating whether associated genes are actively transcribed or silenced. While methylation can be stably maintained as part of the epigenetic code governing cell identity, changes in methylation are equally important, allowing the genome to be dynamically responsive to the cellular environment or during development. This critical process of active DNA demethylation is mediated by TET family enzymes, Fe(II)/α-ketoglutarate-dependent dioxygenases that can oxidize 5mC to generate 5- hydroxymethylcytosine (5hmC). Importantly, 5hmC is itself a substrate for further oxidation, generating 5- formylcytosine (5fC) and 5-carboxylcytosine (5caC). The different oxidized 5mC bases (ox-mCs) open up distinct pathways for DNA demethylation, which differ in their obligate dependence on DNA replication versus DNA repair, resulting in major different implications for the timing and consequences of gene activation. Despite the critical importance of 5mC oxidation in regulating gene expression, the mechanisms by which TET enzymes reactivate silenced genes has remained enigmatic. This gap is due in part to limitations in the tools used to sequence and track dynamics of 5mC and ox-mCs, and the fact that most studies employ all-or-none approaches with TET deletion or inactivation. The urgency for better understanding demethylation dynamics is fueled by the discovery that TET enzymes can also be harnessed as powerful epigenome editing tools when targeted to specific genomic loci by CRISPR-Cas proteins. This proposal aims to apply a series of innovative enzymatic and sequencing tools to reveal and exploit the mechanisms of TET enzymes. Specifically, the mode of TET action on CpG clusters will be revealed by the combined application of novel sequencing methods that can localize multiple different ox-mCs in single DNA molecules and engineered TET variants that can either stall or accelerate through multiple oxidation states of 5mC. These tools will be applied to decipher the footprint of TET on model substrates in vitro and in cells, and these observations will be applied to dissect T cell differentiation as biological model. Building on these mechanistic insights into targeted DNA demethylation, we will devise novel controllable epigenome editors that can allow for efficient and targeted DNA demethylation and mimic physiological gene activation. Together, our aims will thus newly reveal the mechanisms by which TET enzymes act on CpG clusters and advance novel biotechnological tools that leverage TET enzymes to reshape the epigenome.

NIH Research Projects · FY 2025 · 2025-07

Support for an interdisciplinary Graduate Pharmacology Training Program, is proposed to prepare students for productive careers in disciplines where fundamental knowledge of quantitative and systems pharmacology is required. Alumni from a similar training program at Penn have gone on to academia, industry, regulatory science and consulting, to name a few of the variety of opportunities training in pharmacology provides. Our Predoctoral Training Program trains Pharmacology Graduate Group (PGG) students in classic principles of pharmacology including pharmacokinetics, dynamics, drug-receptor interactions as well as in emerging fields of proteomics and metabalomics, biologics and cell therapies. The PGG is supported by an interdepartmental group of 50 PTP faculty from 22 departments in 5 Schools of the University of Pennsylvania. The Office of Biomedical Graduate Studies (BGS) ensures curricular development, quality control and uniform admission standards across all Graduate Groups, including PGG. Direct management of the Pharmacology Training Program is done by a three-person Leadership Committee that, in corrdination with the Executive Committee of the PGG, defines and reviews policy and selects trainees. PTP Faculty membership is governed by three criteria: (1) expertise in a relevant field of study, (2) significant contribution to training, and (3) extramural funding to support trainees. Admission of students to Graduate Programs is a tiered decision, first by the PGG admissions committee, then by a BGS-wide admissions committee. Subsequent apppointment to the pharmacology training grant is decided by its Leadership Committee. Support for each trainee will encompass their second and third years of graduate school. The first year is fully supported by BGS. Trainees must re- apply after the first 12 months and while most students are re-appointed for a second 12-month period, this intermediate evaluation enhances progress and productivity. Prior to appointment , students must successfully complete required courses in Cell and Molecular Biology, Fundamentals of Pharmacology and Medical pharmacology and Human physiology and Statistics. Coursework extends over 1.0 year with lab rotations ending after 1.5 years. All students will take a yearly course on the responsible conduct of scientific research. Students will also receive training through seminars, journal clubs, annual retreats, scientific meetings, oral and poster presentations, and social events that encourage interactions. Successful completion of a comprehensive "Candidacy" examination marks the start of independent research toward the dissertation. Thesis research is conducted under the supervision of a faculty advisor and is monitored by a thesis committee and the PGG Academic Review Committee. The dissertation defense takes place when the thesis advisor and committee concer that the body of work is complete. Based on the number of potential trainees, and the uniqueness of this training program at Penn, we request support for 12 predoctoral trainees/year for the next 5 years.

NIH Research Projects · FY 2023 · 2025-07

Summary Over 600 human proteins have been recently prioritized as key cancer targets, with nearly half being considered ‘intractable’ by standard small-molecule inhibition approaches, due to target instability and active site accessibility constraints. By redirecting the ubiquitin-proteasomal pathway (UPS) for targeted protein degradation, the proteolysis-targeting chimera (PROTAC) technology provides a potential solution, enabling rapid and continuous target consumption as well as the stronger pharmacological effects than small molecule inhibition. Nonetheless, PROTACs suffer from similar developmental hurdles as small molecules and cannot be easily designed for motif or post-translational modification-specific targeting. To address these hurdles, research efforts have shifted toward gene therapy approaches by introducing the concept of protein-mediated protein degradation. Here, E3 ubiquitin ligases are redirected by replacing their natural substrate binding domains with “off-the-shelf” binding domains, including nanobodies, antibodies, and DARPins, to generate target-specific ubiquibodies. To augment this platform, we recently exploited natural protein-protein interaction information to develop algorithmic pipelines that prioritize target-selective peptides which can be fused to the E3 ubiquitin ligase conjugation domains to induce target protein degradation. In this project, we will augment our current methods to enable the development of these ubiquibodies (uAbs) for any protein, including those deemed ‘intractable’ by small molecule-based means. To do this, we will automate a bipartite algorithmic pipeline that leverages recent advancements in protein language modeling as well as existing co-complex databases to design peptide binders to diverse protein targets, including those with solved co-crystals as well as those with minimal structural information. Specifically, our pipeline will take user-specified target proteins as inputs, and generate prioritized lists of candidate peptide binders as outputs, enabling subsequent generation of uAbs for target degradation. Through library-on-library fluorescence-based assays in human cells and subsequent encapsulation of uAb mRNA in lipid nanoparticles (LNPs), we will develop a scalable method to test and translate our degraders for downstream in vivo validation. In total, this work will generate a robust peptide design tool that will enhance targeted protein degradation efforts and lay the foundation for programmable proteome editing.

NIH Research Projects · FY 2025 · 2025-06

Project Summary Immune thrombotic thrombocytopenic purpura (iTTP) is a life-threatening thrombotic microangiopathy characterized by thrombosis due to dysregulated platelet activation. iTTP results from autoantibodies directed against ADAMTS13, a plasma protease that regulates platelet activation via cleavage of von Willebrand Factor (vWF) multimers. Autoantibodies bind to and reduce ADAMTS13 activity resulting in ultralarge-vWF multimers that induce inappropriate platelet activation and microvascular thrombi leading to anemia and tissue ischemia with end-organ damage. Current therapeutic approaches oJer transient benefit and comprise daily plasma exchange therapy with or without the addition of anti-CD20 (rituximab) or anti-vWF monoclonal antibodies (caplacizumab). However, relapses are frequent and often accompanied with significant patient morbidity. Moreover, subclinical organ injury is increasing appreciated as an important aspect of iTTP, underscoring the need for curative therapy. Chimeric Antigen Receptor (CAR) engineered T cells targeting CD19 (CART-19) have recently been shown to be eJective in patient with other antibody-mediated autoimmune diseases, including systemic lupus erythematosus, which were refractory to traditional therapies. A significant downside of this approach is that CD19 is a pan-B cell antigen and, as such, all B cells, including protective memory B cells acquired over a lifetime of infections and immunizations, are eliminated. Therefore, to develop a robust curative therapy for iTTP, we have developed two complementary CAR T cell strategies to selectively deplete ADAMTS13-secreting B cells. One strategy targets IGVH1-69, the antibody variable gene most commonly found on anti-ADAMTS13 B cells while the second utilizes ADAMTS13 domains on the CAR construct to also engage anti-ADAMTS13 B cells. Our preliminary work suggests these two novel CAR T platforms would target anti-ADAMTS13 B cells but spare the majority of protective B cells. We propose an in depth pre-clinical evaluation of each platform, and their combination, to assess their eJicacy and safety profiles in comparison to the pan-B cell ablative CART-19 standard. We will also conduct a detailed analysis of engagement anti-ADAMTS13 BCRs by these novel CARs that will provide mechanistic insights into their function and could help optimize their design. Our team has extensive experience in taking CAR T therapies through concept, pre-clinical, and clinical phases of development. This pre-clinical work serves as critical go/no-go studies to guide their clinical- phase development with the aim of providing a curative therapy for patients with iTTP.

NIH Research Projects · FY 2025 · 2025-06

ABSTRACT This proposal aims to define the role of p53 in regulating chaperone-mediate autophagy (CMA) and the consequences of this regulation on tumorigenesis. p53 holds the distinction of being the most frequently mutated gene in human tumors. Inactivation of p53 is essential not only for the initiation of a remarkably wide range of tumors but also for their continued survival and proliferation. Thus, understanding the mechanism by which p53 suppresses tumorigenesis is a central objective in cancer biology. p53 is known to induce apoptosis, cell cycle arrest, and senescence. However, emerging evidence indicates that these canonical activities may not fully account for p53-mediated tumor suppression, underscoring the need to identify and characterize additional cellular processes that are regulated by p53. CMA is a highly selective form of autophagy. Unlike macroautophagy, which delivers proteins and organelles in bulk to the lysosome for degradation, CMA targets a subset of cytoplasmic proteins individually. This selectivity permits CMA to regulate intracellular processes. CMA is demonstrated only in vertebrates, and it might have evolved later during evolution to modulate increasingly elaborate biological processes. However, both the regulation and functions of CMA remain unclear. We recently found that CMA governs the balance between self-renewal and differentiation of embryonic stem cells. Our preliminary results further suggested that p53 may be an important activator for CMA. Through this activation, p53 inhibits key enzymes in the tricarboxylic acid (TCA) cycle, a central metabolic hub essential for oncogenic growth. These findings reveal previously unrecognized connections among p53, CMA, and metabolism that impact tumorigenesis. Our central hypothesis is that p53 is an important regulator of CMA, and p53-mediated CMA activation and metabolic regulation contribute to tumor suppression. We propose three specific aims: (1) Elucidate the function of p53 in regulating CMA, (2) Define the molecular mechanisms by which p53 regulates CMA, and (3) Determine the role of p53-mediated CMA activation and metabolic regulation in tumor suppression. The proposed studies will reveal a fundamental component of p53 biological function. They will also likely suggest new targets for cancer therapy.

NSF Awards · FY 2025 · 2025-06

This award will provide support for three Annual Geometry Festivals, the first taking place at Duke University in the Spring of 2026, and the second and third taking place at the University of Maryland and at Stonybrook University. The conferences will cover major recent accomplishments and outstanding problems in geometry to a wide audience of interested students and faculty. This conference will be beneficial to graduate students, early career mathematicians, and advanced undergraduate math majors. The conference will bring together geometers covering a wide swatch of fields. Topics covered will include differential geometry, partial differential equations and numerical analysis, algebraic geometry, spectral geometry, geometric and harmonic analysis, dynamical systems and mathematical physics. Speakers were selected not only for their subject expertise, but also for their ability help engage younger geometers and bring them up to speed in current areas of research, and to encourage interaction and collaboration amongst researchers at a variety of career stages. Here is a URL for the initial iteration of the conferences: https://sites.google.com/view/geometry-festival-duke-2026. 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 · 2025-06

Anyone using the internet today is exposed to dozens of targeted digital advertisements online. These advertisements represent an enormous number of different advertisers, and are displayed to internet users in dozens of different ways. As a result, many questions about targeted advertising online arise, including: What and how many ads are people exposed to online? How do these ads differ between different people or groups? How do they change over time in response to people’s online activities? And, most importantly, what effects do they have on people? Addressing these questions can help improve public policy and regulation for the advertising industry, and provide insights and guidance to Americans, whose online experiences are being shaped by targeted advertisements. The research team will build software systems to collect and analyze the content and impacts of digital targeted advertisements on real users. The project also includes initiatives to promote STEM engagement among local high school students in the Philadelphia area. Targeted digital advertisements are both highly personalized and executed through dozens of intermediaries. As a result, understanding the targeting mechanisms, content, and impacts of targeted digital advertising requires an end-user-centered approach that focuses on what content people actually see, and what effect it has on those real people. To do this, the project will make advances in algorithmic auditing methods, which allow people without access to systems’ internal workings to study their outputs and impacts. The research team will develop two systems of auditing infrastructure to enable multi-platform auditing and real-time ad analysis, deploying them with live participants to run experiments that causally measure the impacts of current targeted ads and user-centered alternatives. The project will contribute innovations in sociotechnical AI auditing methods, an open-access Targeted Ad Observatory for analysis by other research teams, and educational materials to engage local STEM students. 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 · 2025-06

Project Summary/Abstract The overall goal of my research program is to mechanistically understand how circadian and metabolic rhythms shape how fully differentiated cells become functionally specialized, or “mature”. Cycles in energy availability, due to earth’s 24-hour rotation, are the oldest, most consistent environmental input for life on earth. Critical cellular functions—from gene expression to protein synthesis—are thus optimized to 24-hour rhythms as a key adaptation to daily life, from bacteria to most cells in our body. How such rhythmic physiology determines maturity is central to cell and developmental biology, and can be harnessed to build fully functional tissues for regenerative medicine. In the past decade, chronobiology studies have not only revealed novel insights into mammalian cell biology, but also identified several uncharted areas with abundant opportunities for further investigation. For the next five years, we will focus on two such areas: 1) How do circadian and metabolic rhythms determine cell maturation? 2) How do circadian and metabolic rhythms maintain cell maturity? We will address these questions in defined human in vitro and mouse in vivo model systems through the following projects: a) using in situ multimodal mapping of single-cell gene expression and functional states, we will determine how several circadian clock transcription factors program, synchronize, and entrain maturing cellular activities; b) using in vivo multiplexed gene editing, we will elucidate the molecular mechanisms by which clock components and feeding sustain mature tissue chronophysiology. Since chronic misalignment between endogenous and external rhythms triggers multi-system dysfunction (e.g., metabolic, cardiovascular, and neural syndromes), we believe that cracking the mechanisms of circadian and metabolic rhythms in the acquisition and maintenance of mature cell phenotypes will not only reveal critical knowledge to chronobiology, but may also result in actionable insights for diagnosing and treating disease.

NIH Research Projects · FY 2026 · 2025-06

Abstract CHALLENGE: Pathogen persistence within a host results in lifelong infection and causes significant morbidity and mortality. Viruses that endure, like HIV-1, evade clearance by establishing latent infection and escaping immune responses. Latency is a reversible form of nonproductive infection in which virus transcription is suppressed. As such, latent viruses fly under the radar of the host immune surveillance. Previously tested latency reversal strategies have failed due to inadequate potency and off-target toxicity. Further, HIV-1 evades host immune responses through its extreme genetic diversity and direct immune dysregulation. While passive antibody and cellular immunotherapy have shown promise in clearing virus, efficient, cost-effective delivery of these interventions has hampered their use. “Kick and kill” cure strategies, which have not been effective to date, seek to combine latency reactivation (“kick”) with interventions that clear reactivated virus (“kill”). APPROACH: mRNA gene therapy has revolutionized vaccine development and is poised to address shortcomings described above. As opposed to other technologies, mRNA does not come with risk of insertional mutagenesis or require serologic prescreening. Importantly, its production uses cell-free procedures, remains inexpensive at scale, and is both flexible and rapid. mRNAs encoding any gene of interest are delivered in lipid nanoparticles (LNPs) that can be engineered to target specific cell types for timed, transient protein expression. To address shortcomings in existing HIV Cure strategies, I have recently developed three novel mRNA-based approaches: CD4-targeted delivery of the viral protein Tat, which drives HIV-1 transcription; hepatic delivery of mRNA encoding monoclonal antibodies that are secreted into circulation; and chemokine-targeted mRNA delivery to effector immune subsets. Here, I propose to develop these approaches as novel platforms that can be combined into the first mRNA-only kick and kill cure strategy. INNOVATION: In this application, I outline an interdisciplinary research program in which I design, study, and apply cutting edge mRNA gene therapy to tackle previously insurmountable obstacles to an HIV cure. Using all aspects of my clinical and scientific training, I have designed interventions grounded in an understanding of both HIV-1 and human biology and tailored my delivery vehicle to suit this application. But, these approaches will find application elsewhere; as they are developed, I envision applying these tools to disrupt viral transcriptional programs, deliver pathogen-specific antibodies, and modify host effector cells as interventions against other diseases. These novel mRNA-based approaches have the chance to translate the success of the mRNA-LNP platform from vaccines to HIV eradication with the potential for worldwide use.

NIH Research Projects · FY 2026 · 2025-06

Cell differentiation is a complex process in which bioenergetic metabolism, specifically glycolysis and oxidative phosphorylation (OxPhos), plays a pivotal role. This role extends beyond ATP production and encompasses factors like the accumulation of intermediate metabolites such as 2-hydroxyglutarate (2HG). In fetal growth plates, hypertrophic chondrocytes are central to bone development. Despite their importance, our understanding of their regulation by bioenergetic metabolism is still in its infancy. The fetal growth plate is a hypoxic and high glycolytic structure. Hypoxia-inducible factor 1α (HIF1) is crucial for the survival of growth plate chondrocytes. HIF1 promotes glycolysis and lactate fermentation while reducing OxPhos activity. The survival role of HIF1 relies on its partial inhibition of OxPhos. Recently, we generated a mutant mouse lacking Mitochondrial Transcription Factor A (TFAM) in limb bud mesenchyme. TFAM, encoded by the nuclear genome, primarily regulates the transcription of mitochondrial-encoded electron transport chain subunits. Loss of TFAM did not impact the differentiation of mesenchymal progenitor cells into chondrocytes, chondrocyte proliferation, survival, or physiological death. However, it significantly delayed chondrocyte hypertrophy, leading to shorter bones. Thus, despite the high glycolytic activity of the fetal growth plate, its proper development still relies on OxPhos. While TFAM is not a known nuclear transcription factor, single-cell RNA sequencing (scRNA seq) of mutant and control growth plates at birth revealed significant changes in the expression of numerous genes involved in chondrocyte biology. These changes included reduced expression of HIF1 target genes and increased expression of genes involved in chondrocyte biology. Intriguingly, enhancing HIF1 activity appeared to slow down endochondral bone development, suggesting that decreased HIF1 signaling does not mediate the delayed hypertrophy observed in TFAM mutant growth plates. Untargeted metabolomics analysis revealed that levels of intermediate metabolites like 2HG were significantly elevated in mutant cells. Additionally, exposing control cells to hypoxia, thus activating the HIF1 signaling pathway, resulted in a similar increase in 2HG and other intermediate metabolites. 2HG plays a role in controlling cell differentiation by modulating epigenetic events and chromatin accessibility. Based on our preliminary data, we will test the following hypothesis: 1. loss of TFAM in growth plate chondrocytes delays hypertrophy by suppressing OxPhos, resulting in elevated levels of intermediate metabolites, such as 2HG, which modulate chromatin remodeling, ultimately affecting the nuclear transcriptome; 2. Augmented HIF1 activity replicates the effects of TFAM loss by delaying chondrocyte hypertrophy, reducing OxPhos activity, and increasing 2HG levels or other intermediate metabolites.

NSF Awards · FY 2025 · 2025-06

This I-Corps project is based on the development of a system that converts salty water into concentrated acid and base using only electricity. Acid is widely used in metal refining processes from primary extraction to product finishing across metals including nickel, lithium, rare earths, and steel. However, current acid production and recycling methods are often associated with the generation of hazardous gases that must be managed. The transportation of acids and precursor chemicals adds further logistical complexity and costs to metals processing plants, especially where primary extraction occurs at remote mining sites. Spent acid is often wholly or partially neutralized on site to form salts of calcium, magnesium, sodium, or iron, which can pose environmental risks and space constraints to mine or refinery sites and surrounding ecosystems. Acid procurement and waste management can account for 30% to 80% of operating expenditures in metals leaching processes. This technology may offer a safer, cleaner, and more energy-efficient alternative for metal extraction with no direct emissions or hazardous byproducts. In addition, this technology may enable sustainable domestic supply chains for critical materials. This I-Corps project utilizes experiential learning coupled with first-hand investigation of the industry ecosystem to assess the translation potential of a bipolar membrane electrodialysis platform for acid and base production. The adoption of bipolar membrane electrodialysis in metallurgy has been limited due to the low concentrations of acid electrodialysis can produce with current technology. Metals leaching typically requires more concentrated acids and bases than in common applications of electrodialysis such as in the beverage or desalination industries. This technology allows for the doubling of acid concentrations versus traditional electrodialysis by optimizing membrane compositions, reactor component sizing, and operational parameters. Unlike conventional acid generation technologies, this approach avoids combustion, phase changes, or hazardous byproducts. The research builds on recent advances in membrane materials and stack design to improve efficiency and durability, allowing for high-throughput operation with minimal downtime. This technology may enable production of concentrated acid and thus, has important applications in metallurgy, where concentrated acids are widely used to extract metals from ores. This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.

NIH Research Projects · FY 2025 · 2025-06

Project Summary A limited understanding of clinically relevant signaling pathways has limited the development of therapeutic agents for human glomerular disease. Our long-term goal is to identify novel therapeutic targets for the treatment of glomerular disease by elucidating the details and functional significance of key signaling pathways that regulate podocyte injury and survival. Our preliminary data have shown that YAP silencing disrupts podocyte focal adhesion architecture and actin cytoskeletal integrity. Using RNA-Seq to compare the transcriptomic profile of control and YAP knockdown mouse podocytes, the most significantly enriched gene ontology molecular function term among the differentially upregulated genes is “potassium channel regulator activity”. YAP silencing significantly increases KCNN4 gene and encoded KCa3.1 (intermediate-conductance calcium-activated potassium channel) protein expression. KCa3.1 inhibitors decrease podocyte actin cytoskeletal disruption, pathogenic intracellular calcium signaling and potassium efflux along with decreasing podocyte injury in an acute in vivo model of glomerular disease. The overall objective of this application is to define the role of YAP in podocyte survival and as a potential therapeutic target in proteinuric kidney disease. Our hypothesis is that YAP nuclear expression is essential for maintaining the integrity of the podocyte actin cytoskeleton, including through repression of KCNN4 gene expression. The loss or inactivation of YAP enhances pathogenic KCa3.1 signaling in podocytes. Our hypothesis will be tested by pursuing two specific aims: Aim 1 will define the mechanism of Hippo-YAP regulation of podocyte KCa3.1 function. We will define the mechanism by which Hippo-YAP regulates KCa3.1 channel function in podocytes and the mechanism of podocyte injury induced by calcium-activated potassium channel activity. We will also test the in vivo interaction and functional role of YAP inhibition of KCNN4/KCa3.1 function. In Aim 2, we will develop and test the efficacy of KCa3.1 inhibitors for podocyte protection. We will use rational design and medicinal chemistry to optimize KCa3.1 inhibitors for podocyte protection, test the ability of novel KCa3.1 inhibitors to protect podocytes from injury while decreasing pathogenic potassium efflux and intracellular calcium signaling, and test the ability of KCa3.1 inhibitors to decrease albuminuria and slow kidney disease progression in relevant in vivo glomerular disease models. Currently, there are no KCa3.1 inhibitors in clinical development for podocytopathies, FSGS or proteinuric kidney disease. The work proposed is expected to characterize the role of Hippo-YAP signaling in regulating KCa3.1 function in podocytes while developing a novel therapeutic strategy for proteinuric kidney disease.

NIH Research Projects · FY 2025 · 2025-06

Irreversible blindness affects over one million people nationally and over 40 million globally, with most cases of acquired blindness leaving the visual cortex intact. A cortical visual prosthesis (CVP) is a promising therapeutic strategy for these individuals. However, the production of perceived visual sensations through stimulation of long-range cortico-cortical connections has proven to be variable and inconsistent. There is a lack in understanding of the biophysical mechanisms of cortical stimulation and its effects on neural dynamics and behavior. Investigating these mechanisms has been limited by the model system and the neuro-technologies available to interrogate that system. Projections from the posterior parietal cortex (PPC) to the prefrontal cortex (PFC) in non-human primates (NHPs) are involved with saccades in visual-spatial attention and provide an ideal and generalizable system to study long-range cortical dynamics during behavior. Retrograde adeno- associated viruses (retroAAVs) can specifically target PPC-PFC projection neurons for optogenetic stimulation. In Aim 1, a multimodal optoelectronic device with microLED and transparent microelectrodes will be developed to enable colocalized optogenetic stimulation and electrical recording of the projection neurons with high spatiotemporal resolution. In Aim 2, the retroAAV injection protocol will be optimized to achieve the highest transduction efficacy. In Aim 3, the optoelectronic device and retroAAV strategy will be implemented to introduce microstimulation of the PPC-PFC neurons in NHPs during a visual-spatial task. Our preliminary data on successful optoelectronic device development, retroAAV expression, and optogenetic stimulation and single-unit recording of PPC-PFC projection neurons reduce risk and validate feasibility for our proposal. The results will improve our understanding of the biophysical mechanisms of how stimulation affects multiregional cortical dynamics and visual behavior, informing the future development of effective CVPs.

NIH Research Projects · FY 2026 · 2025-06

PROJECT SUMMARY/ABSTRACT Numerous new PET radiotracers introduced in the past decade have improved our ability to non-invasively detect disease and characterize tumor biology. These new tracers complement imaging with FDG, a marker of glucose metabolism widely used in the clinic. The integration of these tracers into clinical practice has transformed the treatment of many diseases, ushering in a new era of precision medicine guided by molecular imaging. Modern PET protocols, though, limit our ability to fully exploit this new tracer technology. As a PET scanner cannot distinguish between tracers, different tracers must be imaged on separate days to allow for decay of the first prior to imaging the second. This necessary delay hampers the radiologist’s ability to render a timely unified diagnosis of the two image datasets, inconveniences the patient, needlessly consumes resources, and stresses our already strained healthcare system. In this project, we will develop novel methods to image two PET tracers with different molecular targets back-to-back, in single imaging session, using a novel whole-body PET scanner, the PennPET Explorer. Using this whole-body scanner, we will study protocols where a first tracer is injected at a lower dose and a second tracer at higher dose, without the patient leaving the scanner, with images obtained in the same imaging session. We will study two pairs of tracers as representative models for this new class of protocols for dual-tracer PET. In Aim 1, we will develop protocols to image Fluoroglutamine, a research radiotracer of glutamine metabolism, in combination with FDG in women with breast cancer. This will serve as a prototype for quantification with a pair of research radiotracers, where kinetic analysis is implemented. In Aim 2, we will develop protocols to image two FDA approve radiotracers with dual-tracer PET. Aim 2 will image FDG paired with Fluoroestradiol (FES), a clinically approved radiotracer that measures estrogen receptor (ER) expression in breast cancer. In this aim, we will develop a delayed (static) dual-tracer FES/FDG image protocol ready for immediate clinical translation, noting the clinical need to interpret FES-PET in the contact of FDG-PET. We will also develop novel 4D radiomic image analysis techniques to analyze the robust dual-tracer data collected in this aim to predict response to ER-targeted therapy, a currently unmet clinical need. In the Aim 3, we will translate the dual-tracer protocols developed in the first two aims to scanners with shorter axial field-of-views, including long-axial field-of-view scanners with varying lengths that are now clinically available as well as standard-axial field-of-view scanners that currently populate image centers across the country. As a result, the protocols developed will apply to the entire range of available scanners, increasing the overall impact of this proposal. The outcomes of this proposal will advance the capabilities of PET—optimizing the utilization of new radiotracers—to characterize and guide treatment in oncology.