University Of Pennsylvania

NIH Research Projects · FY 2026 · 2022-04

PROJECT SUMMARY The complexity of microbial communities inhabiting the healthy human skin have been illuminated in recent years, as well as signatures associated with inflammatory skin diseases such as atopic dermatitis. However, the molecular and biochemical mechanisms that mediate microbial symbiosis with the skin are not well understood. We recently found that homeostatic skin barrier function, epidermal differentiation, and recovery from injury are dependent on the microbiota in mice. These effects were mediated by the keratinocyte aryl hydrocarbon receptor (AHR). The AHR is a ligand-dependent xenobiotic receptor of foreign/toxic substances that is also known to regulate epidermal differentiation, tight junction/adhesion, and antimicrobial innate immune responses. The objective of this project is to identify and characterize microbial ligands of the AHR, and to test their utility in treating skin barrier dysfunction and infection. Epidermal barrier dysfunction is a key feature of atopic dermatitis, a common skin disorder characterized by chronic and relapsing, itchy, inflamed, skin lesions and dysbiotic microbiota. To test the hypothesis that commensal bacteria mediate AHR signaling to promote skin barrier integrity and defense, we propose three aims. 1) Define microbial mechanisms of keratinocyte AHR regulation that promote barrier repair; 2) Establish pharmacodynamic actions of microbial AHR ligands to promote skin barrier function and antimicrobial defense; and 3) Evaluate microbial AHR ligands in treatment of epidermal barrier dysfunction and S. aureus infection in murine models. Findings from these studies will provide novel, accessible targets to promote skin barrier function and repair, while advancing fundamental understanding of cutaneous host-microbiota interactions.

NIH Research Projects · FY 2025 · 2022-04

PROJECT SUMMARY/ABSTRACT This proposal describes a five-year training plan for the development of an independent research career focused on the role of the distal airway epithelium in human lung injury and regeneration. Specifically, the applicant strives to understand the regulation of a novel human specific secretory population called respiratory airway secretory cells (RASCs), which can act as progenitors for alveolar type 2 (AT2) cells, and understand how this population is altered in Chronic Obstructive Pulmonary Disease (COPD). The applicant is in her final year of Pulmonary and Critical Care fellowship at the Hospital of the University of Pennsylvania with previous PhD training in cellular and molecular biology with a focus on cell fate regulation. The goals of this award are to refine and develop skills that will be necessary for a successful career as an independent investigator including expertise in epigenomics, mammalian gene editing, and advanced bioinformatic analysis. The mentor for this award is Dr. Edward Morrisey, an internationally renowned expert in lung regeneration and repair with an outstanding and expansive training record. Furthermore, an advisory committee of complementary and diverse scientists has been assembled to provide breadth and depth to the training plan. The applicant will benefit from the unparalleled mentoring, resources and scientific community at the University of Pennsylvania and the unreserved support of her institution. The aims of this proposal are focused on expanding our knowledge of an understudied region of the human lung, the respiratory bronchioles. Respiratory bronchioles are absent in mouse and hence their role in lung injury, regeneration and repair is essentially unknown and often over-looked. Recent data demonstrate that respiratory bronchioles are a site of injury in COPD, highlighting the need to understand the molecular regulation and function of cell populations within this niche. This project will examine the regulation of a novel cell type recently identified in the human respiratory bronchioles, RASCs. RASCs can serve as progenitors for AT2 cells, and are transcriptionally altered in COPD, suggesting that their function could be altered in, and contribute to, the pathogenesis of this highly prevalent disease. This highlights the scientific need to investigate this cell type and its response to injury. Successful completion of these proposed studies will address the central hypothesis that the cell fate and progenitor function of RASCs is regulated through a combination of Notch signaling and the transcription factor SOX4, and that this progenitor function is altered in COPD. This will be accomplished through multiple techniques including human embryonic stem cell modeling, next-generation sequencing, and advanced epigenetic analysis of primary human lung tissue. These studies will provide significant insight into a novel lung progenitor cell and have a high potential for therapeutic impact in COPD and lung regeneration more broadly.

NIH Research Projects · FY 2025 · 2022-04

PROJECT SUMMARY The goal of this proposal is to study and control nephron induction and assembly towards the formation of replacement renal tissue. Kidney organoids re-create an astonishing cellular diversity comparable to the early fetal kidney. However, structural connectivity of urine-producing nephrons and their drainage network formed by ureteric epithelium (UE) is required to avoid rapid pathology, yet has not been achieved. Accordingly, there is an urgent need to achieve connectivity between nephrons and ureteric epithelium before kidney organoids can achieve their potential in regenerative medicine. Our long-term goal is to construct ‘higher-order’ synthetic kidney tissues using human autologous stem cell lineages and assembly technologies that mimic the outcomes of morphogenesis. Our overall objectives at this stage are firstly to gain spatial control over nephron formation by determining how the mechanical microenvironment contributes to their induction sites and maturation. Its second objective is to direct nephron fusion with UE at many spatial sites through a controlled invasive process. Achieving these objectives will mark a transformative advance towards creating replacement kidney tissue. Our central hypothesis is that mechanical compaction of mesenchymal cells during kidney morphogenesis permits nephron induction, and subsequently that tight spatiotemporal control over WNT signaling events is necessary for their efficient fusion with UE. We plan to achieve the objectives through two specific aims. Firstly, we will determine the mechanical basis of nephrogenesis and use it to specify nephron positions. We will study mechanical compaction of the nephrogenic mesenchyme, assess biophysical properties of early nephron cells, and optimize nephrogenesis at specific locations using micropatterning technology. Secondly, we will program WNT-induced fusion of nephrons with ureteric epithelium. We will optimize fusion in nephron-ureteric epithelial co-cultures using optogenetic control over WNT signaling, and then trigger nephron assembly with UE spheroids after transferring them from micropatterned surfaces. The proposed research is innovative because we create fundamental knowledge while creating tissues that are biomaterial- free, human-derived (compatible with patient-derived autologous cell strategies), and therefore open to future development for transplantation. The proposed research is significant because higher-order assembly of human kidney tissue will create a step-change in renal replacement technology beyond dialysis, transplant, and “abiotic” filtration. We expect these efforts to have significant positive impact in the areas of fundamental biological discovery, drug target screening, and regenerative medicine.

NIH Research Projects · FY 2026 · 2022-04

Project Summary Gain-of-function Notch mutations are among the most frequent mutations in small B cell lymphomas, including chronic lymphocytic leukemia (CLL) and mantle cell lymphoma (MCL). Harboring Notch mutation is also a predictor of poor outcomes in small B cell lymphomas, which despite recent advances remain incurable with chemotherapy. Together, these observations provide a compelling rationale for focusing on Notch inhibitors as potential therapeutic options in small B cell lymphomas. Unfortunately, progress in treating patients with Notch inhibitors has been stymied partly due to a limited understanding of underlying mechanisms of sensitivity and resistance to Notch inhibitors. Here, we represent a concerted effort to overcome these limitations and fill major gaps in current knowledge by testing our central hypothesis that Notch-driven activation and positioning of enhancers synergize Notch and several crucial signaling pathways to co-activate pro-growth and survival genes in small B cell lymphomas. This premise is based on our preliminary data showing an important consequence of Notch inhibition is decommissioning and repositioning of putative enhancer elements associated with key genes in primary small B cell lymphoma samples, including several components of B cell receptor (BCR) and cytokine signaling pathways. We will test this hypothesis by using cutting-edge functional genomics, chromatin conformation capture, genome engineering, and single-cell resolution genomics and optical assays. We will systematically determine: i) the Notch-dependent enhancer activation and nuclear positioning, and their genomic requirements that together mediate crosstalk between Notch, BCR and cytokine signaling pathways; ii) the rewiring of Notch-driven epigenetic program that enables drug-resistant cells to bypass effects of Notch inhibitors and maintain the expression of critical Notch targets. Together, these complementary studies will greatly expand understanding of epigenetic mechanisms underpinning sensitivity and resistance to Notch inhibitors and provide a clearer rationale for targeting Notch in small B cell lymphoma.

NIH Research Projects · FY 2026 · 2022-04

ABSTRACT Knee osteoarthritis (OA) is a painful and debilitating joint disease that can result in joint pain, loss of joint function, and deleterious effects on the quality of daily life. Despite recent advances in drug development, there is no disease-modifying drug available to delay OA progression or reverse the disease. Over 600,000 knee replacements are performed each year in the US. Many preclinical and clinical studies have revealed that various inflammatory mediators have been implicated in OA pathogenesis. Despite the strong association between inflammation and OA, a major challenge is how to resolve the inflammatory state since common anti-inflammatory drugs have demonstrated limited utility to slow or reverse OA progression. Moreover, rapid joint clearance and poor penetration into avascular cartilage tissues further limit the clinical application of many promising OA drugs. Therefore, there remains a critical need to identify new therapeutic targets and to develop effective drug delivery systems for OA patients. Secreted phospholipase A2 (sPLA2) enzyme specifically hydrolyzes the sn-2 ester bond of phospholipids, releasing free fatty acids and lysophospholipids. These products are well-known upstream inflammation mediators in many chronic inflammatory diseases. However, few studies have explored the role of sPLA2 in OA and the cause-effect relationship between sPLA2 upregulation and OA progression. Our recent studies found that sPLA2 level is drastically increased in full-thickness articular cartilage from both human patients and animal models of OA. To explore the potential of targeting sPLA2 pathway (i.e. by sPLA2 inhibitor, sPLA2i) for OA treatment and overcome the challenges of small sPLA2i delivery within joints (i.e. rapid clearance and poor cartilage penetration), we engineered sPLA2i (thioetheramide-PC)-loaded phospholipid micellar nanoparticles (thioetheramide-NPs) and provided the first evidence that thioetheramide-NPs, but not free sPLA2i, can effectively reduce joint inflammation, alleviate join pain and prevent OA progression in a mouse OA model. While these data are promising, thioetheramide-PC is not a clinically-approved drug and has a rather poor IC50 (~2 μM). The overall goal of this proposal is to use a clinically tested and more potent sPLA,i varespladib (~210-fold lower IC50 than thioetheramide-PC) to construct sPLA2-responsive varespladib-NPs and test their efficacy in clinically relevant animal models. We believe the proposed work will result in a clinically translatable nanotechnology that could alter the standard of care for knee OA. The specific aims for the proposal are 1) synthesize and optimize sPLA2-responsive varespladib-NPs; 2) evaluate the efficacy of varespladib-NPs in injury-induced mouse OA models; and 3) evaluate the efficacy of varespladib-NPs in the guinea pig model of spontaneous OA.

NIH Research Projects · FY 2025 · 2022-04

Dynamic Longitudinal Functional Models with Applications to the CRIC Study The goals of this project are to develop novel dynamic longitudinal functional models and to apply them to the electrocardiographic (ECG) data that are measured repeatedly over time in the Chronic Renal Insufficiency Cohort (CRIC) study of individuals with chronic kidney disease (CKD). We will also develop real-time risk prediction algorithms to identify individuals at high-risk of cardiovascular diseases (CVD). The CRIC study is an ongoing study of individuals with chronic kidney diseases (CKD), funded by the National Institute of Diabetes, Digestive, and Kidney Diseases (NIDDK) since 2001. The CRIC study has recorded standard twelve-lead electrocardiograms (ECG) annually in all participants recruited from seven clinical centers. Our primary objective is to evaluate whether longitudinal ECG patterns are precursors to CVD and thus can be used to identity high-risk individuals. We propose new statistical methods to extract novel features from the raw ECG tracing both at baseline and in terms of longitudinal changes that are predictive of complications from CVD such as hospitalizations for heart failure (HF), myocardial infarction (MI), stroke, atrial fibrillation (AFib), and cardiovascular death. The information will be incorporated in the proposed real-time, computationally efficient risk prediction algorithms. We will also validate our discovery using an external cohort of CKD patients collected from the University of Pennsylvania Health System (UPHS).The proposed methods are not restricted to ECG data analysis and have a wide range of applications. We will develop user- friendly software packages for the new statistical models and risk prediction algorithms and share the validation data to promote their use in both statistical and clinical communities.

NIH Research Projects · FY 2025 · 2022-04

Abstract Applying CAR T cell therapy to solid tumors such as ovarian cancer (OvCa) is widely considered a major opportunity but also a major challenge and thus the focus of this proposal. Here, we seek to develop new clinical strategies for OvCa and other solid tumors using CAR T cells specific for folate receptor-alpha (FRα), as the target of next generation CAR T cell therapy. FRα is a surface protein that is expressed in 80-90% of OvCa cases and associated with poor prognosis. FRα is known to be a safe, “druggable” therapeutic target in trials of antibody drug conjugates and bispecific antibody armed T cells in platinum-resistant OvCa patients, with clinical response rates of 26% and 27%, respectively. These agents are short-lived in patients and thus responses are non-enduring; CAR T cells however have the capacity for persistence and maintained activity in vivo. In multiple preclinical models, human FRα CAR T cells exhibit potent anti-tumor efficacy against human solid tumor xenografts that express FRα. Here, we propose to test the central hypothesis that lentivirus engineered FRα-specific CAR T cells can achieve clinically meaningful tumor responses in patients with recurrent OvCa without untoward toxicity. We propose to (1) determine the feasibility, safety and tumor response following intraperitoneal injection of autologous FRα lentivirus CAR T cells in patients with confirmed FRα-overexpressing recurrent OvCa in a phase I dose escalation trial (NCT03585764), (2) determine FRα CAR persistence, immunological potency and mechanism of FRα-specific CAR T cell activity in treated patients to understand the immune reaction and other modulations in the OvCa microenvironment following CAR T cell injection, and (3) determine the scope, breadth, and duration of induced systemic immune responses, as a foundation for anticipated future combinatorial therapies. In this line, a secondary hypothesis is that schedule-dependent preconditioning of the tumor microenvironment (TME) is a requirement for effective therapy. In OvCa and other cancers, tumor associated macrophage (TAM) accumulation is associated with poor outcome and resistance to immunotherapy, suggesting that TAM depletion or disruption may improve patient outcome. We have now developed novel CAR T cell technology that mediates deep and highly selective depletion of immunosuppressive M2-like TAMs. In preclinical studies, we find that anti-TAM CAR T cells augment endogenous CD8+ T cell antitumor responses, spares M1-like macrophages, re-educates the TME, and inhibits tumor progression in vivo in three independent mouse tumor models, suggesting that synergy with FRα CAR T cell therapy may be achieved, particularly when applied as a preparative preconditioning regimen. We are positioned to test this novel hypothesis by evaluating and optimizing this novel TAM depletion regimen, and other established approaches, in the context of FRα CAR T cells therapy in various preclinical tumor models, allowing us to capitalize on the discovery of a novel effective combination as a bridge to next generation clinical trials for patients with OvCa and solid tumors bearing TAM accumulation.

NIH Research Projects · FY 2026 · 2022-03

Project Summary Prevention of HIV-associated neurocognitive impairment (HIV-NCI) remains elusive, despite the efficacy of cART in suppressing viral replication within the CNS. Although cART initiated immediately after HIV infection (INSIGHT START study) profoundly reduced disease progression, there was no neurocognitive advantage over delayed cART. Acute brain injury occurs within weeks of infection (HIV, SIV), before cART suppression is typically achieved. Spontaneous limited recovery may occur thereafter, suggesting a therapeutic window for rapid-acting neuroprotective treatments. These have not yet been tested. Our overall objective is to determine the ability of a rapidly-assimilated neuroprotective drug (dimethyl fumarate/DMF, FDA-approved), in combination with cART, to reduce injury and promote recovery in acute SIV infection in rhesus macaques. SIV/HIV injury is linked to oxidative stress and inflammation, which DMF can target through enhancing Nrf2- driven antioxidant enzyme expression and associated antioxidative/anti-inflammatory pathways. In our human brain autopsy studies, HIV-NCI associated with reduced expression of heme oxygenase-1 (HO-1), an antioxidant enzyme with two isoforms (HO-1 and -2), and with increased neuroinflammation. Moreover, HIV infection without HIV-NCI associated with increased HO-1 levels, consistent with a neuroprotective role for HO. In a separate cohort of persons living with HIV (PWH), we showed that an HO-1 promoter variation ((GT)n dinucleotide repeat)) that enhances HO-1 expression, associates with lower neuroinflammation and lower HIV- NCI risk. In acute HIV infection (in vitro) we showed that DMF induces HO-1 and other Nrf2 antioxidant enzymes in infected macrophages, and reduces TNF and glutamate release, thus linking enhanced enzyme expression with neuroprotection. In acute SIV infection in rhesus macaques, we defined a potential therapeutic window for DMF enhancement of antioxidant responses. We identified unique patterns of acute synaptic injury linked to low antioxidant enzyme levels, and changes in expression. Brainstem injury associated with higher neuroinflammation, lower enzyme levels, and progressive loss of HO-2. Recovery associated with stable HO-2 and increasing HO-1 levels. In our pilot macaque treatment study, DMF induced brain antioxidant enzymes, including HO-1, reduced oxidation of DNA and proteins, and produced a less-oxidized brain redox state. These findings support testing DMF as an adjunct to early cART. We hypothesize that DMF therapy concurrently with cART in acute SIV infection will reduce oxidative stress and acute neuronal injury while enhancing neuronal recovery throughout the brain. We will determine effects of concurrent DMF/cART on: (Aim 1) regional brain, oxidative injury, inflammation, neuronal integrity, signaling and recovery, and association with plasma markers of injury, oxidative stress and microbial translocation; (Aim 2) brain localization of immune cell infiltration, cell activation and oxidative injury in immune, endothelial, glial, and neuronal subtypes; and (Aim 3) infiltration of SIV-infected immune cells in brain and lymphatic tissue, in acute SIV infection of rhesus macaques.

NIH Research Projects · FY 2025 · 2022-03

PROJECT SUMMARY Hypertension (HTN) prevalence increases with aging and is a leading risk factor for several chronic illnesses including Alzheimer's disease and related dementias (ADRD), cardiovascular disease (CVD), and several cancers, as well as mortality. Angiotensin II receptor blockers (ARBs) and angiotensin-converting enzyme inhibitors (ACEIs) are two of the most commonly prescribed anti-HTN classes, used by ~40 million US adults. ARBs and ACEIs and have distinctive beneficial downstream effects on physiologic abnormalities in HTN, including vasoconstriction, inflammation, fibrosis, and oxidative stress, which in turn may result in different long-term risks of ADRD and multimorbidity associated with aging. However, current HTN guidelines recommend prescribing ARBs and ACEIs interchangeably due to presumed equivalent benefit and safety. Our goal is to optimize initial anti-HTN medication prescribing by clarifying the optimal first choice RAS-blocker between ARBs vs. ACEIs. Because ~23 million US adults are currently taking an ACEI and physiologic evidence supports differences in downstream effects of these medications, even if ARBs are only 15% more effective, the long-term population health impact of switching first-line RAS-blockade from ACEI to ARB would be enormous. We will leverage data from the Veterans Health Administration (VHA) and Kaiser Permanente Southern California (KP SoCal) to evaluate the effects of ARBs vs. ACEIs on the risk of ADRD, multimorbidity, frailty, and health-adjusted life expectancy (HALE; the amount of time one can expect to live accounting for one's cumulative morbidity burden). The VHA and KP SoCal are ideal data sources to perform this research because they include comprehensive healthcare information for >10 million patients, collect detailed information on medication use and health outcomes, and have high patient retention with >10 years of follow- up. The specific aims are to determine long-term comparative effects, including duration of use, of ARB- vs. ACEI-based anti-HTN medication regimens on (Aim 1) the incidence of ADRD, CVD (stroke, myocardial infarction, coronary revascularization, or heart failure), and cancers, separately and (Aim 2) the patient- centered outcome of frailty and the population-centered outcome of HALE. We will use an active comparator, new-user design accounting for medication adherence, as well as natural language processing to ascertain ADRD more accurately in the electronic health record over using administrative codes alone. Our team is well- suited to perform the study given considerable prior experience analyzing VHA and KP data, including pharmacoepidemiologic analyses of anti-HTN medication use; assessment of ADRD, CVD incidence, cancer incidence, and multimorbidity; and application of causal inference methods. Our project could support a paradigm shift of first-choice RAS-blockade. Current projections indicate that ADRD will affect >115 million people by 2050 and cancer incidence will be 27 million per year by 2040. The potential public health benefit of addressing these knowledge gaps and, thereby, improving the quality and length of life is enormous.

NIH Research Projects · FY 2026 · 2022-03

PROJECT SUMMARY Clostridioides difficile infection (CDI) is one of the most common causes of healthcare-associated infectious diarrhea and results in significant morbidity and mortality. CDI occurs when the native gut microbiome is disrupted, most often following antimicrobial therapy, and the consequent dysbiosis results in a decrease in microbial diversity, changes in abundance of certain bacterial taxa, and loss of colonization resistance against C. difficile. Restoration of a “functionally intact” gut microbiome is critical to clearing C. difficile, and inadequate restoration can lead to recurrent CDI. The recovery of the gut microbiome from dysbiosis is poorly understood, and factors associated with having and re-gaining a microbiome capable of providing colonization resistance against C. difficile are not well known. While animal reservoirs can serve as potential sources of pathogenic bacteria, studies by the candidate and other investigators found that pet ownership protects against colonization and re-infection with C. difficile. Moreover, microbiota are shared between pets and their owners, and the microbiomes of pets contain bacterial taxa that provide colonization resistance against C. difficile. Based on these data, the proposed research will 1) test the hypothesis that the observed protective effects of pet ownership are due to sharing of microbiota that provide colonization resistance against C. difficile between pets and owners; 2) determine whether pet contact mitigates antimicrobial-associated disruption of the gut microbiome and enhances its recovery; and 3) assess whether pet contact decreases the likelihood of colonization and infection with C. difficile following antimicrobial therapy. This will be accomplished though longitudinal sampling of the gut microbiome within the patient/pet unit among patients older than 60 years (i.e., at greatest risk of CDI) receiving prophylactic antimicrobials for non-enteric indications (dental implants). The study will further define epidemiologic and pathophysiologic characteristics of CDI that could enhance therapeutic options for this disease. The underlying premise that animals are a source of protective microbiota rather than a reservoir of C. difficile represents a paradigm shift in CDI epidemiology that may identify animal contact as a novel microbiome-based form of therapy. The proposed study will form a foundation for an independent career in patient-oriented research dedicated to understanding and mitigating antimicrobial-associated dysbiosis and CDI. The candidate will acquire experience in directing a large, observational microbiome study and essential training in the advanced statistical and bioinformatics methods necessary to analyze the interaction between patient-level factors and microbial ecology. The research proposal is paired with a career development plan that makes use of the extensive resources of the University of Pennsylvania and that capitalizes on a superlative mentoring committee with broad, complementary expertise in infectious diseases epidemiology, deep sequencing methods, bioinformatics, and microbiology.

NIH Research Projects · FY 2026 · 2022-03

Project Summary/Abstract Binge eating disorder is the most common eating disorder afflicting up to 10% of US adults. It is characterized by recurring episodes of eating large sums with a sense of loss of control. Preclinical studies have implicated dysregulation of the nucleus accumbens (NAc) in loss of control eating. To provide a deeper understanding as to how the NAc underlies LOC eating in humans, we leverage data from an early feasibility trial entitled Responsive Neurostimulation for Loss of Control Eating (NCT03868670). This UH3 study aims to examine the safety, feasibility, and preliminary efficacy of responsive neurostimulation of the NAc for LOC eating. Two subjects are currently enrolled and successfully underwent bilateral surgical implantation of the responsive neurostimulation system (NeuroPace, Inc.) into the NAc. Both subjects are undergoing responsive DBS guided by NAc delta (2-4Hz) oscillations (i.e. “delta-responsive DBS”) and have exhibited less LOC (primary endpoint). A supplement to enroll the 4 additional subjects is under FDA review. Here, we intend to characterize the role of NAc delta signaling in human LOC eating behaviors. Preliminary data suggests that ventral NAc delta power and bilateral connectivity elevate during anticipatory and LOC-like cravings states, specific to palatable foods. Futhermore, delta power and ventral bilateral NAc connectivity are reduced following delta-responsive DBS, and that this effect is sensitive to stimulation parameter optimization. Our central hypothesis suggests the NAc as a key region in LOC eating, and that aberrant delta signaling underlies LOC eating. As such, we predict that delta-responsive DBS acts on LOC behaviors through a rescue of the NAc’s aberrant delta signaling. Our aims are to determine how NAc delta power and connectivity can differentiate LOC eating from normal eating, and identify neural underpinnings of behavioral response to NAc delta-responsive DBS. As LOC is common to all binges and is a transdiagnostic feature of many mental health disorders, this work will provide invaluable insight relevant to much of Psychiatry.

NIH Research Projects · FY 2026 · 2022-03

Breaks in the structure of DNA are a persistent stress on the integrity of the genome, and they pose a substantial risk of chromosomal rearrangement and genetic mutation that can challenge the well-being of an organism and promote the development of cancer. There are several cellular mechanisms that monitor the state of the genome and rapidly initiate repair mechanisms in response to DNA damage so that a healthy genome is passed on to the next generation. Poly(ADP-ribose) Polymerase-1, or PARP-1, is a primary responder to breaks in the structure of DNA. PARP-1 has a unique catalytic activity that synthesizes polymers of ADP- ribose as a posttranslational modification on target proteins, primarily on PARP-1 itself (automodification). Upon binding to DNA breaks, PARP-1 activity is “turned on” to modulate DNA damage repair pathways and thereby promote cell survival. In contrast, excessive DNA damage leads to an elevated level of PARP-1 activity that results in cell death. Regulation of PARP-1 activity is therefore a critical factor in determining the fate of a cell. Importantly, inhibitors of PARP-1 (PARPi) have recently emerged as promising therapeutic agents for the treatment of cancer and inflammation. Despite a growing interest in PARPi and the discovery of expanded roles for PARP-1 activity in DNA repair, transcriptional regulation, and apoptotic signaling, there are still limited insights into the mechanism of PARP-1 catalytic activity and regulation. The objective of this research program is to fill major gaps in our knowledge of how PARP-1 is activated, modulated by a critical accessory protein (HPF1), and subsequently silenced in the process of detecting DNA damage in healthy cells and how it can be best inhibited by small molecules in current efforts to target PARP-1 in cancer and inflammation. Hydrogen-deuterium exchange coupled with mass spectrometry (HXMS) and crystallography are the major structural tools that we will apply to understand the impact of PARPi on PARP-1 dynamics and how PARP-1 in the DNA damage response is initially activated and then subsequently silenced through automodification. The structural and protein dynamics experiments will be combined with biochemical analysis of PARP-1 catalysis and DNA binding, and cell-based analysis of PARP-1 function based on our structural and biochemical work. In addition, medicinal chemistry will be employed to engineer allosteric PARP-1 “trapping” into PARPi compounds in order to increase the efficacy of targeting this enzyme in the cancer clinic. Moreover, an emerging PARPi-based imaging approach using established tumor assays with breast cancer patient-derived xenografts will determine the degree to which the PARPi compounds that we generate engage/kill cancer cells and impact survival of mice carrying tumor xenografts. The proposed studies of PARP-1 activity and regulation will advance current models of PARP-1 biological functions and generate new small molecule tools that will advance the understanding of PARP-1 biology and potentially represent new medicines for the cancer clinic.

NIH Research Projects · FY 2026 · 2022-03

PROJECT SUMMARY Oral squamous cell carcinoma (OSCC) is the most common type of head & neck cancer and the 10th most frequent human malignancy worldwide. Over the last several decades, the overall survival rate of OSCC patients has stagnated between 40~55% despite some progress in diagnosis and therapy. The invasive growth or progression of OSCC relies on the aggressiveness of cancer cells and their unique microenvironment, whereby cancer stem cells (CSCs) and infiltrated tumor associated macrophages (TAMs) play pivotal roles. Previous studies have demonstrated that polyamines (PA) are commonly elevated in tumor microenvironment (TME) and have long been proven to be necessary for transformation and progression of various types of cancers. eIF5A2, an isoform of a highly conserved translational factor, is overexpressed in many types of cancer. Remarkably, spermidine-mediated eIF5A hypusination (eIF5Ahpu) that is implemented by two highly specialized enzymes, deoxyhypusine synthase (DHS/DHPS) and deoxyhypusine hydroxylase (DOHH), appears to be essential to most, if not all, of eIF5A’s biological functions, including its important role in regulating cancer cell proliferation, epithelial-mesenchymal transition (EMT), and CSC properties as well as immune cell functions, thus rationally emerging as a potential target for both therapy and prevention of cancer. Our analysis of TCGA dataset indicated an overall upregulation in the mRNA expression of eIF5A2 and several key enzymes involved in PA metabolism in HNSCC, which was confirmed by Western blot and IHC studies. Our studies showed that blocking DHPS/eIF5Ahpu remarkably inhibited proliferation and CSC properties of OSCC cells, which correlated a downregulation of TWIST1-BMI1 expression and NOTCH1/HES1 signaling. Meanwhile, we found that blocking DHPS/eIF5Ahpu robustly inhibited OSCC-induced polarization of M2-like TAMs and reversed the immunosuppressive effects conferred by OSCC-induced TAMs on T cell activation in vitro. More Importantly, we found that blocking DHPS/eIF5Ahpu dramatically retarded tumor growth and infiltration/polarization of M2-like TAM in an orthotopic syngeneic mouse tongue SCC model. Based on these compelling preliminary studies, we hypothesize that eIF5Ahpu might play a critical role in OSCC growth and progression due to its dual functions in regulating proliferation/CSC properties of OSCC cells and OSCC-induced M2-like TAM polarization. To test our hypothesis, we propose three specific aims: 1) Elucidate mechanism by which eIF5Ahpu regulates proliferation and CSC properties in OSCC; 2) Determine whether eIF5Ahpu plays a critical role in OSCC-induced polarization of M2-like TAMs; 3) Target eIF5Ahpu to suppress tumor growth and immunosuppressive TAMs in OSCC in vivo. New findings from this application might not only shed light on elucidating the function of eIF5Ahpu activation in development and progression of OSCC, but also hold promises for identifying novel therapeutic targets for treatment and prevention of OSCC.

NIH Research Projects · FY 2026 · 2022-03

Dr. Willis is a neurologist, neuroepidemiologist, and health services researcher who has established a nationally- and internationally- recognized, independently funded research program committed to patient-oriented research focused on drug effects, care process and outcome differences, and health service use in older adults with neurological disease. She has a successful record in mentoring trainees who approach POR from multiple educational disciplines and clinical specialties. Proposal aims: (1) to conduct analytical epidemiology research examining the impact of prescribing practices, drug exposures, drug-drug and drug-disease interactions on clinical, patient-reported and health care utilization outcomes in older adults, particularly those with Parkinson Disease + Dementia; Dementia with Lewy Bodies (which together comprise Lewy Body Dementia, an Alzheimer’s Disease Related Dementia/ADRD), Alzheimer’s disease (AD), Mild cognitive impairment and chronic neurological disease; (2) to conduct health services research in older adults with neurological disease, particularly those with the above ADRDs and AD, to gain actionable insights into systematic barriers to receiving high quality, appropriate care and achievement of best health outcomes; (3a) to develop new studies at the intersection of aging, neurology, and pharmacoepidemiology focused on the biological, clinical, and qualitative effects of deprescribing and safety-guided CNS drug prescribing on cognitive function and cognitive decline in individuals with the above ADRDs, AD, and MCI; (3b) to develop new studies at the intersection of aging, disparities, and health services research, focused on describing and decomposing the relative contributions of individual, neighborhood, and social factors on disparities in guideline adherent treatment initiation, escalation, and monitoring; treatment adherence and outcomes in older adults with neurological disease, including ADRDs and AD; (4) to use Dr. Willis’s research program to train and mentor new investigators in research methods that will improve health care outcomes for older adults with ADRDs, AD and chronic neurological disorders through neuroprotective prescribing, more effective care delivery. A K24 award will provide Dr. Willis with the protected time needed to develop new research focused on neuroprotective prescribing and health outcomes generate knowledge on the drivers and effects of medical care and drug therapies for neurological disorders. A K24 will also allow Dr. Willis to increase her mentoring expertise and develop a formal mentoring program for engaging young investigators drawn from neurology, epidemiology, geriatrics, psychiatry, pharmacology, informatics, and biostatistics. Dr. Willis’s mentoring program will (1) provide mentees with direct exposure to primary data collection and secondary data analysis (2) create an interdisciplinary environment where trainees from a range of specialties will learn directly from each other (3) provide mentees with individualized research and career mentoring from her. The outstanding institutional environment and the K24 Advisory Team Dr. Willis has assembled to monitor and guide her progress at the University of Pennsylvania support the success of this proposal.

NIH Research Projects · FY 2026 · 2022-03

PROJECT SUMMARY This application proposes to translate activatable fluorophores targeted to cytosolic phospholipase A2α (cPLA2α) for intraoperative surgical detection in lung cancer. Lung cancer is the leading cancer-related cause of death and the third most diagnosed cancer in the United States. Non-small cell lung carcinomas (NSCLC), represent 90% of lung cancers, and surgical resection continues to be the most effective approach to cure patients with low stage premetastatic disease. Surgeons typically use visual inspection and finger palpation to define solid tumor margins. However, this approach is often insufficient to detect residual disease, leading to recurrence in up to 40% of patients and significantly reduced 5-year survival. Improving intraoperative detection of tumor margins is imperative, because small foci of residual disease at or close to the resection margins are the most common cause of local recurrence. The most important prognostic indicator following cancer surgery is complete resection, which prolongs patient survival and improves post- surgical quality of life. Contrast enhancement of tumors using near-infrared (NIR) imaging with targeted fluorophores can identify small lesions that are not detectable by visual observation or palpation. NIR imaging offers high resolution and sensitivity and can be performed in real-time during surgery. In this application, we propose to translate DDAO-arachidonate (DDAO-A), an activatable probe designed to specifically target cPLA2α, a critical signaling enzyme upregulated early in tumorigenesis and overexpressed in more than 47% of NSCLC. Preliminary Studies indicate that DDAO-A is selectively activated by murine and human NSCLC tumors, and can detect small foci in both mouse models and excised human lung tumor tissue. DDAO-A will be synthesized under GLP conditions and dispersed in clinically approved liposomal formulations for delivery. Selected mouse studies will be performed to confirm probe activation, tumor targeting and the ability to detect tumor margins. Systemic toxicity will be evaluated in mice and in a small cohort of canines. We will utilize an established platform for clinical translation of NIR fluorophores, employing a canine clinical trial for intraoperative surgery. We will recruit a patient cohort of 30 companion animals with spontaneous NSCLC presenting to our Veterinary School. DDAO-A will be used for detection of primary tumors and identification of tumor margins during intraoperative fluorescence-guided surgery (FGS) in canine patients. We will investigate both systemic injection and topical application for detection of residual tumor. These data will be compared to control surgical cohorts using the non-targeted indocyanine green (ICG) or without NIR fluorescence guidance. If successful, this study will improve surgical outcomes by improving localization of small tumor deposits in the lung, lymph nodes and margins. The completion of these studies will form the basis for an investigational new drug (IND) application in support of a clinical trial to assess DDAO-A-guided surgery in human lung cancer patients.

NIH Research Projects · FY 2026 · 2022-03

PROJECT SUMMARY Visual hallucinations affect approximately 20% of Alzheimer disease and 50% of all Parkinson disease patients. Hallucinations are a leading source of patient and caregiver distress and are an independent risk factor for injury, nursing home placement, and mortality. Because treatment options for hallucinations are limited and have significant adverse effect risks, the prevention of hallucinations would have a transformative public health impact for older adults with neurodegenerative disease. Visual impairment is a risk factor for hallucinations, and since up to half of all vision loss in the U.S. is preventable or treatable, the prevention and treatment of ophthalmic disease could prevent or reduce the severity of hallucinations in older adults. However, studies of specific age-related eye diseases and hallucination outcomes are lacking, limiting these improvements in healthcare. This proposal requests support for a mentored career development award for Dr. Ali Hamedani, a neurology-trained neuro-ophthalmologist and epidemiologist at the University of Pennsylvania. The overarching goal of this project is to understand how visual pathway structure, function, diseases, and treatments contribute to hallucinations in older adults. In Aim 1, Dr. Hamedani will analyze longitudinal data from two Medicare-linked national health surveys (the National Health and Aging Trends Study and Health and Retirement Study) to determine whether age-related macular degeneration, primary open-angle glaucoma, and cataract surgery are associated with the incidence of hallucinations in a nationally representative sample of 5,200 high-risk older adults using advanced survival analysis with marginal structural models to account for time-dependent confounding. In Aim 2, Dr. Hamedani will recruit a prospective cohort of Parkinson disease patients who are beginning a medication for hallucinations to determine whether low-contrast acuity and retinal ganglion cell thickness are associated with hallucination severity and treatment response. In executing these aims, Dr. Hamedani will be obtain additional training in ophthalmic epidemiology, retinal imaging, and biostatistics under the mentorship of experts in optical coherence tomography and ophthalmic clinical investigation (Joel Schuman, MD) and neurodegenerative disease epidemiology and health services research (Allison Willis, MD MSCI). The results of this project will provide fundamental knowledge about the visual system’s role in causing hallucinations and pave the way for future studies to test visual impairment and ophthalmic disease as a prevention target for hallucinations and other neurocognitive outcomes in older adults. Through the research training and mentorship experience gained during this career development award, Dr. Hamedani will establish himself as an independent investigator in the applied epidemiology and outcomes research of ophthalmology in aging and neurodegenerative disease.

NIH Research Projects · FY 2026 · 2022-03

PROJECT SUMMARY/ABSTRACT The cornea is the most highly innervated structure in the body, supplied by the ophthalmic branch of the trigeminal nerve. As part of the peripheral nervous system, corneal nerves respond to pain, temperature, mechanical and chemical stimuli. They also secrete various trophic and growth factors, which are essential to the health and function of the cornea. However, corneal nerves are highly susceptible to injury through various mechanisms that include trauma, infections, metabolic imbalances, and therapeutic interventions such as refractive surgeries. Once injured, they fail to reestablish their baseline density or morphology, contributing to corneal dysfunction. Currently, there are no targeted treatments specific for corneal nerve regeneration. The long-term goal of this proposal is to develop therapies for corneal nerve regeneration. The objective is to determine key molecular mechanisms involved in corneal nerve regeneration to help inform new experimental and therapeutic interventions. The central hypothesis is the N-Methyl-D-aspartate receptors (NMDAR), a type of glutamate receptor, help restore corneal nerve density and morphology, and therefore, corneal function. The rationale underlying this proposal is that NMDARs have been shown to enhance nerve regeneration in other analogous peripheral nervous systems. However, their role in corneal nerve regeneration remains unknown. Additional justification for investigating the role of NMDARs in corneal nerve regeneration is based on other published findings: 1) NMDARs are expressed throughout the nervous system, including the trigeminal nerves; 2) they have been shown to regulate neuronal maintenance and plasticity; 3) they regulate Schwann cell activity, which are supporting cells essential to nerve regeneration; and 4) NMDARs cooperate with other signaling molecules that have been shown to regulate corneal nerve regeneration such as LDL-receptor-related protein-1 and Ephrin type-B receptor 2. Therefore, we propose three aims to support our hypothesis. AIM 1 will determine the role of NMDAR in corneal nerve maintenance and regeneration by conditionally deleting NMDAR in sensory nerves and Schwann cells independently. AIM 2 will determine the effect of modulating NMDAR levels on corneal nerve regeneration. AIM 3 will determine key downstream effectors, including the EphB2-Sox2 axis, with spatial transcriptomics, correlated with protein levels and morphologic changes during corneal nerve regeneration. We will pursue these aims using innovative genetic mouse models, intravital imaging, and spatial genomics. The proposed aims are significant because they will define new molecular pathways that will inform the development of future therapies. The immediate expected outcome of this work is rigorous interrogation of key pathways in corneal nerve regeneration in vivo and contribution to our fundamental understanding of peripheral nerve regeneration. The results will have an important direct positive impact because they will interrogate new experimental approaches and inform the development of targeted therapies for corneal nerve regeneration.

NIH Research Projects · FY 2025 · 2022-03

Project summary Post-acute care (PAC) is increasingly common and costly. One in five Medicare beneficiaries receives care after hospitalization in a skilled nursing facility at a cost of more than $28 billion annually. Unfortunately, more than 1 in 4 Medicare beneficiaries are readmitted to the hospital within 30 days, and these readmissions are associated with increased mortality. The Skilled Nursing Facility Value-Based Purchasing program (SNF VBP) ties Medicare reimbursements to SNF to their 30-day all-cause hospital readmission rates. Determining the effect of SNF VBP on patient outcomes is crucial for patients, health systems, and policymakers, and will inform the development and implementation of similar programs in other post-acute care settings. Evaluating potential unintended consequences of this policy is especially important because SNFs face significant financial pressure, and SNFs that care for large proportions of patients who are especially vulnerable to adverse outcomes (e.g. frail, cognitively impaired, poor, or racial and ethnic minority populations) are most likely to be penalized under the program, potentially leading to increased disparities. The COVID-19 pandemic may have magnified the effects of SNF VBP, acting as a second “stress” on SNFs already stressed by SNF VBP. There is an urgent need to determine the effect of SNF VBP on patient outcomes and on disparities, especially given the magnifying effect of COVID-19. Our long-term goal is to drive the delivery of high-value care for all older adults leaving the hospital. SNF VBP is among the first pay-for-performance programs in post- acute care settings. In order to improve the development and success of these policies, it is crucial to understand how their design and implementation influences outcomes. Our central hypothesis, based on preliminary data, is that SNF VBP achieves its intended effects at SNFs that were already high-performing, but has unintended and negative effects at low-performing SNFs. Our specific aims are to: 1) Determine the impact of SNF VBP on intended outcomes prior to the COVID-19 pandemic; 2) Determine the effect of SNF VBP on disparities in outcomes in vulnerable populations; 3) Determine how financial penalties from SNF VBP impacted COVID-19 readiness and outcomes; and 4) Assess key aspects of organizational context among SNFs that improved performance in SNF VBP and explore how this impacted their response to COVID-19. Accomplishing these aims will improve the design of future VBP initiatives, and lead to higher-value care for the growing number of vulnerable older adults receiving SNF care.

NIH Research Projects · FY 2026 · 2022-02

Traumatic Brain Injury (TBI) is associated with accelerated neurodegeneration and is a recognized cause of late-life dementia, a type of Alzheimer’s Disease Related Dementia (ADRD). Although the driving pathological mechanisms are incompletely understood, preclinical evidence increasingly points to microvascular injury as a key component of TBI neuropathology, indicating that TBI-related Neurodegeneration (TreND) shares important features including a vascular contribution to cognitive impairment and dementia (VCID) with other ADRDs. While neuroimaging investigations of TBI to date have primarily focused on structural damage to neuronal cell bodies and axons, there remains an unmet need to characterize the evolution of microvascular dysfunction after TBI in humans and establish the role of TBI-related VCID. An improved understanding of how microvascular injury contributes to long-term neurodegeneration holds potential to improve patient outcome prediction and help overcome persistent barriers to the development of effective therapies to ameliorate this disease process. Thus, there is a great need to develop reliable imaging biomarkers of TBI-related VCID to better inform prognosis, classify injury endophenotypes, monitor long-term recovery, and identify treatment targets. The central hypothesis of this proposal is that chronic microvascular dysfunction as a consequence of TBI represents a type of VCID and links the initial brain injury to subsequent slowly progressive neuronal loss occurring in the ensuing months to years, ultimately contributing to brain atrophy and long-term cognitive decline. To test this hypothesis, we propose a multi-timepoint neuroimaging study to determine the natural history of microvascular dysfunction over the first 3 years post-TBI, in which we will establishing the pattern of longitudinal change in MRI-based measures of microvascular function including cerebral blood flow, cerebrovascular reactivity, blood-brain-barrier dysfunction, and extracellular free water in relation to brain atrophy. We expect that TBI patients will exhibit brain atrophy and microvascular dysfunction in excess of that associated with normal aging, which will be determined by comparison to a longitudinally examined healthy control group. We additionally hypothesize that microvascular imaging measures assessed in the early postinjury period will predict the magnitude of subsequent brain atrophy and cognitive decline during the first 3 years after TBI. Ultimately, this project will yield novel insights into the role of VCID in TBI pathophysiology, a potentially treatable yet understudied endophenotype of TBI.

NIH Research Projects · FY 2026 · 2022-02

Project Summary Although targeted therapy and immune checkpoint inhibitors have made a major impact on survival for some patients with advanced cancer, the majority of patients do not respond to standard of care treatments. Abundant evidence indicates autophagy is induced by chemotherapy and targeted therapy, and also limits the efficacy of immunotherapy. Clinical trials testing autophagy inhibitor combinations show encouraging preliminary results with increased response rates when compared to standard of care approaches. New autophagy inhibitors are entering clinical trials. Preclinical studies and the available clinical data indicate that tumors can overcome autophagy modulating therapies producing resistance. There is a critical unmet need to understand mechanisms of resistance to autophagy-modulating therapy. Using melanoma as a model we have discovered that extensive lipid raft induction is induced by autophagy modulating therapy. This is especially pronounced with lysosomal autophagy inhibition, which induces the expression of key proteins (LDLR, SR-B1, and UGCG) in the cholesterol and sphingolipid salvage pathways (CSSP). At least one of these enzymes, UGCG, can be targeted with an FDA approved therapy eliglustat and preliminary results indicate combined autophagy inhibition and UGCG inhibition produces synergistic antitumor activity in vivo. This proposal will test the hypothesis that the increased expression of CSSP and subsequent lipid raft formation induced by autophagy-modulating therapy promotes cell survival, and may be a key druggable vulnerability that can be targeted to improve therapeutic outcomes in cancer. To test this hypothesis, we will leverage the longstanding collaboration between Dr. Amaravadi (oncologist, autophagy expert) and Dr. Speicher (systems biology expert). We also recruited Dr. Meenhard Herlyn, a melanoma expert who has developed a humanized mouse model and bank of patient-derived xenografts, as well as Dr. Phyllis Ginotty, a biostatistician who has worked closely with this team for years. In Aim 1 we will define the mechanism by which autophagy modulation regulates the cholesterol and sphingolipid scavenging pathways (CSSP). We will determine the effects of chemical or genetic manipulation inhibition of key CSSP genes in lipid-depleted and precisely reconstituted media on tumor cell survival. In Aim 2 we will determine the role of UGCG as a driver of resistance across melanoma therapy combinations in in vivo models. We will utilize a panel of patient-derived xenograft (PDX) models generated from BRAF mutant and NRAS mutant melanoma patients to determine if targeting UGCG results in decreased lipid raft assembly, that overcomes resistance to clinically relevant therapies. Impact: These studies will determine how two key resistance mechanisms to cancer therapies, autophagy and altered lipid metabolism, intersect. Our results should uncover new therapeutic vulnerabilities in melanoma as well as other cancers and should identify new therapeutic combinations incorporating CSSP inhibitors to be tested in future clinical trials, which could significantly improve outcomes for cancer patients.

NIH Research Projects · FY 2026 · 2022-02

PROJECT SUMMARY Radiation therapy (RT) is used in the curative setting for many cancers including sarcomas and lung and pancreatic cancer. Despite significant improvements over the past few decades, there is still much room for improvement as patients still develop RT-induced injuries or second malignant neoplasms. FLASH radiotherapy, which delivers a large dose of radiation at an ultra-high dose rate could potentially reduce toxicity. Our overall hypothesis is that Proton/Carbon Particle FLASH RT is superior to Standard Particle RT in protecting normal tissues while the two modalities will be equipotent in controlling malignant growth. Project 1, which focuses on pancreatic cancer, will define the dosimetric and biophysical parameters that will maximally spare normal intestine tissues using FLASH proton therapy (F-PRT) without compromising antitumor effects. It will delineate mechanistic aspects of differential response of normal intestinal tissues, by focusing on the relative sparing of the stem/progenitor cell population. Project 1 will also employ p53+/- transgenic mouse models to dissect the genetic determinants of differential GI toxicity of Standard proton therapy (S-PRT) vs F-PRT. Project 2 will explore the ability of F-PRT to ameliorate adverse events (inflammation, fibrosis, lymphedema, changes to bone structure, radiation-induced cancers) that pose barriers to the treatment of sarcomas with RT. We will also carry out a phase 1/2 trial that will treat canine patients with osteosarcomas definitively with F-PRT. Project 3 will compare the efficacy of FLASH-RT given with carbon ion radiotherapy (C- RT) vs. standard dose rate and compare it to electron F-RT. Studies will focus on the mitigation of normal tissue injury in NSCLC with an emphasis on the impact of normal tissue and intratumoral hypoxia to response following C-RT. Lastly, Project 4 will develop and validate the use of pencil beam scanning (PBS) technology for particle F-RT. It will analyze spatiotemporal variations and SOBP (spread out Bragg peak) vs. shoot through PBS and develop dose delivery algorithms for modeling biological effects for PBS-based FLASH proton therapy. These tools will be incorporated in the experimental plans of project 1-3. These Projects are supported by 4 Cores including an Administrative Core (Core A), Physics-Dosimetry Core (Core B), which will offer infrastructure services to harmonize dosimetry between various sites and a Comparative Pathology core (Core C) for tissue preparation for histopathological evaluation. Statistical services will be provided by Core D. Collectively, this highly integrated effort led by recognized leaders in Radiation and Tumor Biology, aims to define the biological, dosimetric and biophysical parameters and molecular mechanisms under which FLASH RT is most effective in tumors and tissues we deem the most likely to be first tested in clinical trials. It is our belief that only by acquiring this knowledge will this exciting and novel modality be ushered into the clinic in a safe and effective manner to improve therapeutic outcome and quality of life of cancer patients.

NIH Research Projects · FY 2026 · 2022-02

SUMMARY Messenger RNA (mRNA) vaccines represent a powerful vaccine approach for the induction of protective immune responses against severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2). Since this vaccine platform was approved for human use in 2020 for the first time, little is known about the durability and mechanisms of induction of the immune responses elicited by SARS-CoV-2 mRNA vaccines. In mice, the generation of SARS- CoV-2 neutralizing antibodies (nAbs) driven by mRNA vaccines is associated with the formation of robust germinal centers (GCs). GCs are sophisticated processes during which antigen-specific B cells give rise to high- affinity Ab-secreting cells and memory B cells. The GC reaction is tightly regulated by T follicular helper (Tfh) cells, which are also efficiently generated during SARS-CoV-2 mRNA vaccination. In this grant proposal, we seek to address the following 3 fundamental questions related to SARS-CoV-2 mRNA vaccines: 1) How durable are the GC-derived B cell responses to SARS-CoV-2 mRNA vaccines in mice and humans?; 2) What types of antigen presenting cells (APCs) promote Tfh cell differentiation in SARS-CoV-2 mRNA vaccination? And how do these APCs sense these mRNA vaccines?; and 3) How do SARS-CoV-2 mRNA vaccines induce GC B cell responses? Overall, the studies that we propose here will allow us to determine the longevity of GC-derived B cell responses and to shed light on the mechanisms by which SARS-CoV-2 mRNA vaccines ensure a powerful elicitation of Tfh and GC B cells. The knowledge acquired here will be important to inform future boosting strategies for SARS-CoV-2 mRNA vaccination, as well as the rational design of next generation vaccines for difficult-to-neutralize pathogens.

NIH Research Projects · FY 2026 · 2022-02

Project Summary/Abstract Tendon injuries are challenging clinical problems due to slow, incomplete healing with fibrovascular scar formation, which reduces tendon function and causes chronic complications such as pain and tendon ruptures. The limited understanding of the regulatory mechanisms underlying fibrovascular scar formation is a significant gap in knowledge, hindering the development of effective treatment modalities for tendon diseases. Fibrovascular scar tissue is characterized by a disorganized extracellular matrix (ECM) with high cellularity and neovascularization. Therefore, there is a critical need to understand the key factors regulating tendon cells and ECM maturation during tendon healing to develop regenerative medicine. Recent studies have demonstrated that mTORC1 (mechanistic target of rapamycin complex 1) signaling is a critical regulator for postnatal tendon maturation and is associated with pathogenic tendon conditions such as fibrotic adhesion of flexor tendon, fatty infiltration after rotator cuff tears, and human tendinopathy. However, the function of mTORC1 in fibrovascular scar formation and its downstream molecular mechanisms are not known. Stat3 (signal transducer and activator of transcription 3) is involved in fibrotic scar formation in multiple tissues and is known as a downstream target of mTORC1 signaling in cancer cells. However, there is no reported evidence showing that Stat3 mediates the function of mTORC1 in fibrovascular scar formation in tendons. Our overall objective is to define the function of the mTORC1/Stat3 signaling cascade in fibrovascular scar formation and evaluate the beneficial effects of mTORC1/Stat3 modulation on regenerative tendon healing. The central hypothesis of the proposed research is that (i) injury-induced mTORC1 signaling governs fibrovascular scar formation during tendon healing, and (ii) Stat3 mediates mTORC1 function in fibrovascular scar formation via regulation of ECM organization. We will test the hypothesis using innovative multidisciplinary approaches, including mouse genetics, a surgical injury model, advanced molecular/imaging analyses, and a biomechanical test. The goal of Aim1 is to determine the function of mTORC1 in fibrovascular scar formation during tendon healing. The goal of Aim2 is to define Stat3 as a mediator of mTORC1 function in fibrovascular scar formation in tendons. The success of the proposed research will significantly advance the mechanistic understanding of fibrovascular scar formation during tendon healing and provide a new platform to develop translational and clinical researches targeting the mTORC1/Stat3 signaling cascade for the treatment of debilitating tendon diseases.

NIH Research Projects · FY 2026 · 2022-02

In vivo Mapping of Muscle Specific Metabolism ABSTRACT Phosphorus-31 magnetic resonance spectroscopy (31P-MRS) has long been the method of choice to study muscle bioenergetics in humans. 31P-MRS measures relative amounts of phosphocreatine (PCr) and adenosine triphosphate (ATP), and can be used to estimate muscle creatine kinase (CK) kinetics. During exercise, PCr, a high-energy “reserve” source of ATP, is depleted to meet transient energy demands. The rate of PCr re-synthesis after exercise is commonly used as a measure of skeletal muscle oxidative phosphorylation (OXPHOS) capacity. Studies using 31P-MRS and multiple other modalities have suggested that abnormal creatine metabolism and deficient OXPHOS may contribute to the pathophysiology of aging. In addition, it is well established that muscle groups vary with respect to these metabolic properties, and also in their response to aging. Despite its strengths, 31P-MRS has low anatomic resolution, and does not readily provide muscle group specific estimates of creatine metabolism. The currently available option for measuring muscle group specific metabolism is invasive biopsy. Thus, there is a clear unmet need for high-resolution, non-invasive strategies to assess muscle metabolism simultaneously across heterogeneous muscle groups. Our group recently introduced a new magnetic resonance imaging (MRI) technique known as the Cr-amine chemical exchange saturation transfer (CrCEST), which measures free Cr formed from PCr utilization. CrCEST provides over three orders of magnitude higher sensitivity compared to 31P-MRS and can also be used to investigate CK kinetics and muscle bioenergetics. In this proposal, we further develop and optimize the CrCEST for use in humans, by improving time resolution, characterizing reproducibility, and assessing the effects of pH. As a critical part of interrogating the optimized CrCEST technique, we will test the effects of age, sex, race and physical activity on high-resolution CrCEST in healthy adults. We expect to demonstrate muscle group specific differences in creatine metabolism and OXPHOS capacity using non-invasive techniques that were not feasible until now. Successful accomplishment of this project will i) yield quantitative imaging biomarkers of muscle creatine metabolism, lactate metabolism and OXPHOS capacity that provide anatomic specificity without the invasiveness of biopsy-based approaches; ii) provide reference data to support future studies using CrCEST signal as a non-invasive index of muscle quality in aging and other conditions, including but not limited to diabetes, muscular dystrophy, peripheral arterial disease, and genetic mitochondrial disorders. We anticipate that CrCEST (iii) will also serve as a non-invasive muscle group specific monitoring tool to evaluate response to potential therapies targeting abnormal muscle metabolism in aging, in neuromuscular disorders, and myriad other conditions. Thus, CrCEST based technologies have the potential to fill a number of important unmet needs and are expected to exert sustained impact on the field.

NIH Research Projects · FY 2026 · 2022-02

Title: Structure and Function of TRPV channels The long-term objective of this research program is to understand the molecular mechanisms of gating and cellular function of the two physiologically important members of the TRPV subfamily: transient receptor potential vanilloid 2 (TRPV2) and transient receptor potential vanilloid 5 (TRPV5). These channels belong to the same subfamily of TRP channels, nevertheless they exhibit striking differences in their tissue distribution, physiological function, cellular localization, ion permeation and selectivity, mechanisms of channel gating and pharmacology. TRPV2 is broadly expressed Ca2+-permeable non-selective cation channel, which plays a vital role in neuronal development, immunity, cardiovascular physiology, and cancer. TRPV5 is a highly selective Ca2+ channel, that is only expressed on the apical membrane of kidney distal convoluted tubule epithelial cells and plays a critical role in Ca2+ homeostasis in the human body. This proposal is built on the major advances achieved by my group in understanding TRPV2 and TRPV5 channels structures, molecular details of gating and cellular function in the last 10 years. The overall goal of this proposal is to further understand how TRPV2 and TRPV5 channels are gated by endogenous and exogenous modulators at the atomic level; and to elucidate TRPV2 channel precise molecular function in neuronal development and immunity. To answer these questions, we will employ cutting- edge multidisciplinary approaches available to our laboratory, including membrane protein biochemistry and biophysics, single-particle cryo-EM, peroxidase-catalyzed proximity labeling and mass spectrometry, cryoAPEX for electron microscopy imaging, flow cytometry, confocal fluorescence microscopy, and functional characterization by electrophysiological and cell biological methods. This information would be critical in our understanding of TRPV channels physiological and pathophysiological roles.