Weill Medical Coll Of Cornell Univ

NIH Research Projects · FY 2026 · 2025-06

PROJECT SUMMARY/ABSTRACT Prostate cancer (PCa) is a clinically and molecular heterogenous disease, with biologically distinct subtypes driven by characteristic genomic alterations in both early, untreated disease and treatment resistant castration- resistant prostate cancer (CRPC). However, whether early alterations affect fidelity to prostate lineage, shape the specific resistance patterns that emerge with treatment, and the underlying mechanisms, remain unclear. We hypothesize that specific molecular features of early, untreated PCa establish distinct pathways to progression and therapeutic resistance through transcriptional control and fidelity to luminal prostate lineage. Preliminary data generated by our multi-institutional, multidisciplinary, collaborative group suggest specifically that the subclass of PCa defined by recurrent mutations in SPOP maintain strong fidelity to prostate lineage, and are therefore preferentially reliant on androgen receptor (AR) signaling and possibly resistant to conversion to AR-indifferent subtypes of CRPC. The overall objective of this proposal is to define the propensity of prostate cancers harboring SPOP mutations to progress to specific subtypes of treatment resistant CRPC, and to define the mechanisms that shape these resistance patterns. Using novel models and human prostate cancer samples, our preliminary data demonstrate that SPOP mutation reprograms AR function, altering chromatin accessibility and transcription driven by AR and making these cancers highly reliant on AR activity. In contrast, N-Myc induction combined with RB1-loss is a strong driver of AR-indifferent CRPC that functions to rewire and deactivate the AR transcriptional program. Our central hypothesis is that opposing effects on rewiring of the AR-directed epigenomic and transcriptional programs mediate the downstream impact on biology and therapeutic sensitivity shown with SPOP mutation and drivers of AR-indifferent disease. This project will elucidate the molecular details underlying these phenomena through the following Aims: 1) defining the propensity of SPOP mutant PCa to progress to AR-indifferent disease in response to specific driver alterations, 2) mechanistically, determining the contribution of FOXA1 and TRIM24 in lineage fidelity and plasticity downstream of SPOP, and 3) establishing if drivers of luminal lineage fidelity can prevent or reverse resistance to AR-targeting therapies. To accomplish this, we will leverage unique, biologically and clinically relevant model systems, innovative approaches to epigenomic and transcriptomic discovery, and data from human prostate cancer samples. This project will define the critical transcriptional processes in specific subtypes of prostate cancer and the broader applicability to treatment resistance, and provide the foundation for precision clinical trials.

NIH Research Projects · FY 2025 · 2025-06

Project Summary Myopathy is one of the most common manifestations of mitochondrial diseases. Although genetic and bioenergetic causes of Oxidative Phosphorylation (OxPhos) impairment are well established, there is a limited understanding of the metabolic drivers of muscle degeneration. This knowledge gap contributes to the lack of effective treatments for these disorders. Our recently published studies indicate that OxPhos defective muscle initiates an integrated systemic, multiorgan metabolic response coordinated by endocrine signals, which contributes to the pathogenesis of mitochondrial myopathy. In the COX10 KO mouse model of mitochondrial myopathy, increasing plasma levels of the myokine GDF15 activate central and peripheral neurocircuits through GFRAL signaling, which over time reduce caloric intake, induce mobilization of lipid from adipose tissue, and promote energy-consuming futile cycles in muscle. This chronic GDF15-driven fat and muscle wasting result in a cachectic phenotype which aggravates the myopathy. Therefore, we hypothesize that inhibiting the GDF15-GFRAL signaling with established anti-GDF15 and anti- GFRAL antibodies will increase caloric intake, decrease lipid mobilization and attenuate energy expenditure, thus improving body weight and muscle function. In aim 1 of this application, we will target GDF15-GFRAL signaling to prevent cachexia in pre-symptomatic COX10 KO mice. In aim 2 we will target GDF15-GFRAL signaling to treat cachexia in symptomatic COX10 KO mice. This study will establish if anti-GDF15-GFRAL Ab therapy improves mitochondrial myopathy and assess whether the Ab treatment can reverse symptoms of cachexia, setting the stage for testing anti-GDF15-GFRAL Ab therapy in human mitochondrial myopathies.

NIH Research Projects · FY 2026 · 2025-06

The accumulation and propagation of tau aggregates are key drivers of cognitive decline in Alzheimer’s disease (AD) and frontotemporal dementia (FTD). However, our knowledge of underlying mechanisms is not yet refined enough to meet the urgent clinical need for treatment. There is an urgent need to understand the mechanisms underlying propagation of tau aggregates. Ratios of 3R/4R are altered by FTD mutations, suggesting differential roles of 3R and 4R in tau pathogenesis. We recently showed that Tau-seeded 4R-P301S human neurons develop highly robust and progressively Tau aggregation , recapitulating many aspects of Tau pathology in AD. We also uncovered the critical roles of retromer in limiting and UFMylation cascade in promoting tau propagation. However, the cellular mechanisms underlying Tau propagation in AD are mostly unknown though likely affected by amyloid β (Aβ). Our overall objective is to discover AD-relevant cellular machinery underlying the propagation of misfolded tau aggregates in human neurons using unbiased proteogenomic approaches. To this end, we propose to combine ascorbic acid peroxidase (APEX), affinity purification mass spectrometry (AP-MS), and transcriptomics with our newly generated knockin hiPSC lines that express 3R/4R and 4R-P301S Tau. We propose three Specific Aims. Aim 1 will determine Tau interactomes influenced by amyloid and Tau seeding in Tau isoforms, using advanced proteomic techniques and deep learning analyses to prioritize genetic candidates. This will be validated in induced neurons and AD brains. Aim 2 will focus on the role of VPS29 in tau aggregation, examining its impact on proteomic and transcriptomic profiles, and integrate this data with human AD datasets to identify key molecular drivers. Aim 3 will explore how UFMylation regulates tau aggregation, mapping its effects on proteomic profiles and interactions with the retromer pathway, aiming to identify and test potential chemical interventions in AD human neuronal models. The proposed work will be led by three teams with complementary expertise in close collaboration. Gan lab will create proposed human iPSC neuronal and directly-programmed iN models, conduct transcriptomic profiling, and execute various cell treatments alongside interactome purification and validation experiments. Yu lab will perform AP-IP, APEX2-MS, and proteomics. Cheng lab will integrate genetic and proteomic data to prioritize genetics-driven candidate genes and pathways influencing Tau pathology using network topology-based deep learning analyses and Mendelian randomization techniques. Successful completion of the proposed study will provide unprecedented insight into AD-related tau pathology and pave the way for novel therapeutic strategies.

NIH Research Projects · FY 2026 · 2025-05

PROJECT SUMMARY/ABSTRACT HIV infection remains an unmet global health and socioeconomic burden lacking a safe and scalable cure or a vaccine for prevention. While in the last decades the development of highly effective antiretroviral therapy (ART) has revolutionized infection outcome drastically decreasing transmission and AIDS-related mortality/morbidity among infected individuals, treatment remains a lifelong burden. This is due to the inability of ART and the immune system to clear rare intact, transcriptionally competent and persistent proviruses already integrated into the genome of infected cells – the HIV reservoir. Viral latency is a crucial barrier towards eliminating the HIV reservoir, enabling viral immune escape through transcriptional silence during ART. However, recent lines of evidence have demonstrated that low level HIV production persists over years of ART and correlates with HIV- specific cytotoxic T-lymphocyte (CTL) responses that shape the HIV reservoir. This suggests that rare cells with transcriptionally active proviruses are actively surveilled by HIV-specific CTL responses but are unable to be eliminated. Profiling of the HIV reservoir has also demonstrated that persistent HIV-harboring cells become preferentially clonal and express molecules that can inhibit CTL cytolytic interactions with infected target cells (BCL-2, SERPINB9 and PVR), further implicating CTL resistance as a mechanism of infected cell persistence. Our preliminary work using an in vitro infection model designed to evaluate CTL resistance in infected cells has demonstrated that a rare subset of HIV-expressing cells is preferentially resistant to CTL elimination. Multiomic analysis of these rare cells has confirmed the presence of previously suggested features of CTL resistance, and further revealed a vastly shared transcriptional and surface protein signature denoting dampened levels of metabolic flux and oxidative stress. Here, we aim to demonstrate the presence of a metabolic profile skewed towards quiescence in CTL resistant, HIV expressing infected cells. We hypothesize that lower levels of metabolic flux puts CTL resistant infected cells at a lower threshold of oxidative stress than the general population of infected cells, making them harder targets to push over the edge of irreversible and lethal damage. The experiments of this proposal will evaluate the therapeutic value of pharmacologically inducing oxidative stress in HIV-infected cells to improve CTL mediated elimination of infected cells and HIV reservoirs.

NIH Research Projects · FY 2026 · 2025-05

PROJECT SUMMARY/ABSTRACT Isolated congenital asplenia (ICA) is characterized by the absence of a spleen at birth without any detectable associated developmental abnormalities (OMIM #271400). It is a life-threatening disorder, as affected patients are prone to invasive bacterial disease, especially prior to the diagnosis of ICA. We previously discovered germline heterozygous variants in the coding region and 5'UTR of RPSA (encoding ribosomal protein SA) as the first genetic etiology of ICA, and demonstrated that RPSA haploinsufficiency underlies autosomal dominant ICA. Follow-up analysis of the splenic relatives of ICA patients in our cohort revealed that some coding RPSA variants and all 5'UTR RPSA variants display incomplete penetrance for ICA. We now hypothesize that sufficiently high levels of wild-type (WT) RPSA expression may compensate for deleterious RPSA variants, and that variation in WT RPSA expression between individuals underlies the observed incomplete penetrance. Furthermore, we hypothesize that variants in the 5'UTR of RPSA impair, but do not ablate, the translation of RPSA mRNA, thereby lowering the threshold of WT RPSA expression required to compensate and explaining why the observed 5'UTR variants all lead to ICA with incomplete penetrance. To test our hypothesis, we will first assess the impact of the 5'UTR variants on RPSA translation using ribosome-profiling, polysome analyses, and mRNA secondary structure probing and mutagenesis experiments. We will then search for sequence variation between the splenic and asplenic members of our cohort at genetic positions that modulate the expression of WT RPSA. Finally, we will develop assays to precisely quantify the allele specific expression of RPSA mRNA and then compare WT RPSA mRNA expression between splenic and asplenic carriers of deleterious RPSA variants. We have already sequenced the whole genome of 69 RPSA heterozygotes in our cohort. Our preliminary data are exciting, as we have delineated three haplotypes at the WT RPSA locus that seem to tightly correlate with the presence or absence of a spleen in individuals heterozygous for a variant RPSA allele. An understanding of why some RPSA genotypes lead to ICA with incomplete penetrance will prove useful in our understanding of a life-threatening immunodeficiency, and will provide a roadmap for the study of incomplete penetrance in other genetic diseases driven by haploinsufficiency. I am an MD/PhD student at the Weill Cornell/Memorial Sloan Kettering/Rockefeller Tri-Institutional Program, performing the proposed research in the laboratory of Dr. Jean-Laurent Casanova at The Rockefeller University. My long-term goal is to become a physician scientist who balances patient care with running an independent research program at an academic institution. The plan outlined in this proposal, along with the support and mentorship provided by Dr. Casanova, Dr. Boisson, my thesis research committee, and the Tri-Institutional administrative faculty will help me achieve my career goals.

NIH Research Projects · FY 2025 · 2025-05

Mycobacterium tuberculosis (Mtb), the causative agent of tuberculosis (TB), continues to be a severe global health problem. TB is multifaceted, but a central clinical and microbiologic feature of the disease is the ability of Mtb to resist complete elimination, both by the host immune system and by chemotherapeutic agents with potent growth inhibitory activity. This persistence in the face of immunologic and antibiotic pressure underlies several important facets of TB disease, including 1) the existence of latent tuberculosis infection (LTBI) and, in the setting of immunologic failure of LTBI control, its role in the genesis of active TB and 2) the prolonged course of TB antibiotic therapy, which requires 6 months of multidrug therapy to achieve reliable clinical cure. Rather than producing complete bacterial eradication in all treated subjects, cure following TB chemotherapy is now understood to be an antibiotic induced paucibacillary state in which prevention of relapse depends in part on poorly understood host factors. The host and bacterial determinants that mediate these two interrelated types of persistence are only partially understood, a knowledge gap the Tri-I-TBRU aims to fill. We propose a set of 3 intersecting projects and 3 cores all focused on different facets of the problem of paucibacillary TB, both post treatment and LTBI. The projects will use samples and clinical data from TB cohorts at our clinical site at GHESKIO in Port Au Prince Haiti, to examine the immunologic, microbiomic, transcriptomic, pharmacokinetic, and genetic factors that influence or predict the transition points between paucibacillary states of TB disease and active transmissible infection. These human studies will be compared and contrasted with a new mouse model of paucibacillary infection that will allow us to test mechanistic hypotheses about the host and bacterial determinants of paucibacillary disease. This work with be conducted by a team of highly collaborative investigators who have who have worked well together for several years.

NIH Research Projects · FY 2026 · 2025-04

Abstract Despite the significant reduction in HIV-1 vertical transmission with the implementation of antiretroviral therapy (ART) to pregnant and breastfeeding parents, in 2022, 130,000 new pediatric HIV-1 infections occurred, disproportionately impacting infants who constitute 10% of new infections but just 2% of the global population. More than half of these infections resulted from breastmilk transmission. Thus, there is a critical need for new strategies that could complement ART-based interventions for prevention of breastmilk HIV-1 transmission. One of the most promising strategies under development is passive immunization with broadly neutralizing antibodies (bnAbs). The safety and pharmacokinetics of bnAbs in infants were recently evaluated in the IMPAACT P1112 trial, one of only a handful of trials of its kind. IMPAACT P1112 was an open-label, dose-escalating, phase 1, multicenter study of the safety and pharmacokinetics of the CD4 binding site (CD4bs) bnAbs VRC01, VRC01LS, and VRC07-523LS in HIV-1-exposed infants. The bnAbs were well tolerated and high concentrations were achieved in serum, but the ability of the serum from the passively immunized infants to neutralize viruses found in breastmilk has not been evaluated. Importantly, two efficacy trials evaluating the VRC01 bnAb to prevent HIV- 1 in adults established the Predicted serum neutralization 80% inhibitory dilution titer (PT80) as an excellent predictor of the level of prevention efficacy conferred by that bnAb. Moreover, this biomarker that combines the concentration of the bnAb with its in vitro potency was highly correlated with the serum neutralization in the adult trials. The overall goal of this study is to determine if HIV-1 exposed infants passively immunized with CD4-bs bnAbs in the IMPAACT P1112 trial attained high serum neutralization titers against HIV-1 viruses transmitted through breastmilk and if the level of infant serum neutralization can be predicted by a biomarker. Identifying a biomarker that predicts serum neutralization will guide the down selection of bnAb regimens and their evaluation in pediatric clinical trials. Because HIV-1 exposed infants have preexisting maternal antibodies, whether the tight correlation observed between the PT80 and the measured 80% neutralization titer (ID80) seen in adults will hold in HIV-1 exposed infants, or if another biomarker will better predict serum neutralization, remains unknown. We hypothesize that the PT80 predicts the serum neutralization in HIV-1 exposed infants passively immunized with a bnAb, but that this correlation is strong only after maternal antibodies have waned. Leveraging existing samples from P1112 participants, we will test our hypothesis by 1) Defining the serum antiviral functional profile of HIV-1 exposed infants passively administered a bnAb; and 2) Evaluating the PT80 as a predictor of serum neutralization in bnAb passively immunized HIV-1 exposed infants. Our proposal to thoroughly characterize the antiviral responses elicited in P1112 participants and develop methodology for predicting the neutralization function of bnAb regimens will fill critical gaps for identifying the most promising bnAb regimens to advance in the clinical pipeline and designing clinical trials for their evaluation.

NIH Research Projects · FY 2026 · 2025-04

PROJECT SUMMARY Genomic instability creates heterogeneity in cancer cells, fueling evolution and treatment resistance. However, the costs of genomic damage can lead to cell death or immune detection. In lung cancer, somatic mutagenesis driven by the APOBEC3 (A3) family of cytosine deaminases is enriched in 75% of nonsmoking patients. A3 mutagenesis is associated with driver mutations, resistance to targeted therapies, and poor prognosis. Yet, the precise A3 family members (A3A/B/C/D/F/G/H) responsible for lung cancer mutagenesis are still unknown. Thus, the etiology of many somatic mutations in lung cancer remains poorly understood, a critical barrier to improved therapeutics and prognostics. To dissect the mechanisms of A3 mutagenesis in cancer, my lab developed a novel pipeline that measures endogenous A3 mutagenesis in human cancer cell lines using whole genome sequencing (WGS) and in vitro propagation. I aim to study A3 mutagenesis in lung cancer. Based on preliminary data, I hypothesize that multiple A3s are critical in lung cancer mutagenesis (Aim 1). To identify the A3 mutators in LUAD, I established A3 KO cell lines in multiple LUAD cell lines. I will use them in our de novo A3 mutagenesis detection protocol to determine the major mutators (Aim 1.1). Additionally, I will test proposed surrogate measures of active A3 mutagenesis using my WGS data set as a “gold standard” to evaluate A3 mutagenesis (Aim 1.2). With this toolkit, I will test the prevalence of A3 mutagenesis in a broad range of LUAD cell lines (Aim 1.3). A3 enzymes play a key protective role in healthy tissues by regulating genotoxic retroelements, but this role has been unexplored in cancer biology. Endogenous retroelements can activate during cancer and drive genome instability. They are active in lung cancers, including LUAD, and particularly in Squamous cell lung carcinoma (LUSC). Exogenous expression of A3s can inhibit genome-destabilizing retroelements, including LINE-1 and Alu. However, a link between A3 activity and retroelement regulation in cancer has been completely overlooked. I hypothesize that A3s limit genome instability and protect cancer cells by inhibiting retrotransposition (Aim 2) . To explore retroelement regulation by A3 in lung cancer, I will use LINE-1 and Alu Retrotransposition Assays to determine if endogenous A3s can repress retroelements (aim 2.1); Next, I will compare the levels of retrotransposition breakpoints in the A3 KO and wildtype LUAD WGS datasets I have, to directly test endogenous A3s as regulators of retroelements (aim 2.2). Finally, I will investigate the impact of A3-dependent regulation of retrotransposition by exploring immune signaling and DNA damage driven by LINE-1 and Alu in the absence of A3s in the NSLC cell lines (aim 2.3). This aim will expand the repertoire of A3s beyond somatic mutagenesis to include protecting genomic integrity. The data collected will dissect paralogs driving A3 mutagenesis, evaluate accurate surrogate methods for assessing A3 mutagenesis, and explore A3 activity outside of cancer mutagenesis. This will pave the way for research on specific targets and tools for treatment and prognosis.

NIH Research Projects · FY 2026 · 2025-04

Abstract: Pituitary neuroendocrine tumors (pitNET), also known as pituitary adenomas, represent 15% of all intracranial tumors diagnosed in the United States. Postoperative MRI can underestimate residual disease extent due to postsurgical changes such as disruption of tissue planes, autologous bone and fat grafts, and free mucosal flaps. Patients with hormone-producing pitNET who are not cured with surgery may be treated medically or with radiation therapy (RT), most commonly stereotactic radiosurgery (SRS). Suppressive therapy requires lifelong administration of often-expensive medications that may have substantial off-target side effects. RT will stop the growth of residual pitNET, but only achieves long-term biochemical remission in approximately 50% of patients at 10 years. While stereotactic radiosurgery (SRS) can be more effective, MRI-based SRS that does not treat all of the residual pitNET will fail to control tumor growth and hypersecretion. PitNET express somatostatin receptor 2 (SSTR2), however the expected range of SSTR2 density and factors influencing SSTR2 expression in pitNET are not well understood. SSTR2 can be targeted with [68Ga] DOTATATE, a clinically approved PET radiotracer. Our team has pioneered a dynamic [68Ga]-DOTATATE brain PET/MRI acquisition protocol, allowing differentiation of SSTR2-positive brain and skull base neoplasms, most notably meningioma, from post-surgical and post-RT change, using SUV analysis and Patlak modeling. Given our extensive experience in clinical translation of DOTATATE PET/MRI in SSTR2-positive brain tumors, we are uniquely positioned to translate this approach in patients with pitNET. DOTATATE PET/MRI has the potential to improve diagnostic accuracy, increase our understanding of pitNET biology, and improve SRS targeting, thereby increasing local control and biochemical remission rates in pitNET. Our preliminary findings suggest that DOTATATE PET/MRI-based extent delineation, particularly of the extrasellar component, may improve radiosurgical targeting and treatment success in pitNET. However, present knowledge regarding DOTATATE PET utility in pitNET is limited. We will characterize the range of DOTATATE PET SUV in the context of clinical, demographic, and endocrine variables. We will compare PET- and MRI-guided RT planning, and evaluate the potential of PET/MRI to local control and endocrine remission rates relative to MRI-based literature benchmarks and relative to a separate MRI-based prospective institutional observational control cohort. We will determine distribution of DOTATATE PET SUV in subtotally resected pitNET, examine association with demographic variables and hormonal lineage, and compare PET/MRI- to MRI-based tumor extent delineation. We will further determine short- and intermediate-term clinical outcomes of DOTATATE PET/MRI guided SRS for residual/ recurrent pitNET. Our proposed study has the potential of improving clinical outcomes in patients with pitNET by increasing diagnostic accuracy and improving extent delineation of extrasellar disease, which will optimize radiation targeting thereby achieving greater local control and higher endocrine remission rates.

NIH Research Projects · FY 2026 · 2025-04

Project Summary Olfaction is the primary sensory modality that many animals use to navigate. However, navigating through the chemical world is inherently challenging due to the complexity of odor plumes, which can vary in both composition and concentration. Tracking a plume to its source therefore requires the integration of complex sensory cues with continuous spatial decision-making, posing a distinct challenge to the flexibility of the nervous system. Dopamine has been extensively studied as a neuromodulator that confers flexibility to nervous systems, yet the anatomical heterogeneity and genetic inaccessibility of mammalian dopaminergic systems have precluded a detailed understanding of dopamine’s role at the molecular and circuit level. In comparison, the simple neural architecture and genetic tractability of Drosophila melanogaster provide an exceptional model to study the role of dopaminergic modulation during flexible, naturalistic behaviors like olfactory navigation, as flies robustly track odor plumes using neural circuits that are both genetically accessible and comprehensively mapped at synaptic resolution. In preliminary data, I demonstrated that dopamine neurons in the Drosophila mushroom body, an associative brain center essential for olfactory learning and memory, are continuously engaged during olfactory navigation and can rapidly and bidirectionally bias a fly’s navigational strategy. I also showed that Kenyon cells, the post-synaptic targets of dopaminergic modulation whose activity is required for associative plasticity in the mushroom body, are also required for sustained olfactory pursuit. Using in vivo two-photon imaging, I showed that the strength of signaling from mushroom body output neurons can be rapidly modulated as flies navigate within a virtual olfactory landscape. These results support the hypothesis that the same dopaminergic plasticity mechanisms underlying associative learning are also continuously engaged to regulate and dynamically shape ongoing olfactory navigation. In Aim 1, I will leverage in vivo two-photon imaging and optogenetic perturbations of neural activity to define both correlational and causal relationships between the activity of dopamine neurons and their downstream mushroom body output neurons. In Aim 2, I will use a targeted genetic knockdown strategy in combination with optogenetics to identify the dopamine receptor signaling pathways mediating dopaminergic modulation during olfactory navigation. Together, these studies will provide an updated framework for understanding how dopamine dynamics can flexibly guide naturalistic behaviors like navigation, in circuits with fully characterized cell types and synaptic connections.

NIH Research Projects · FY 2026 · 2025-04

PROJECT SUMMARY Glucose homeostasis is critical during embryogenesis; fetal hyper- or hypoglycemia can cause neurodevelopmental disorders, including impaired neurulation, altered brain growth, epilepsy, Autism, and Attention Deficit Disorder. GLUT1-Deficiency Syndrome (GLUT1-DS) causes a variety of neurological disorders, including infantile seizures, developmental delay, postnatal microcephaly, and intellectual disability. To better understand the role of Glut1 during cortical development and the contribution of neural progenitor cell (NPC)- specific GLUT1 loss to GLUT1-DS, we aim to determine how NPCs regulate glucose uptake during neurogenesis and how glucose metabolism influences NPC self-renewal and cell fate decisions. Our preliminary data make us hypothesize that 1) Glut1 expression in cortical NPCs (termed radial glia; RG) regulates glucose uptake and metabolism dynamics throughout neurogenesis to direct cell fate decisions. 2) During human development, early changes in RG proliferation due to a reduction in GLUT1 expression impact neurogenesis and contribute to the etiology of GLUT1-DS. To test these hypotheses, we will pursue two parallel aims in mouse and human models of cortical development. Aim 1 will explore the function of Glut1 in RG during mouse corticogenesis. We will conditionally remove Glut1 from RG and determine the impact on proliferation, cell cycle dynamics and differentiation using a combination of approaches including immunohistochemistry, single cell transcriptomics, and fluorescent in situ hybridization. We will assess the metabolomic changes associated with loss of Glut1 through metabolic flux and proteomic assays. Aim 2 will use cerebral organoids generated from human pluripotent stem cells harboring GLUT1 mutations and examine the impact on cortical differentiation and neuronal function. Using a cerebral organoid model will enable us to explore the neural contribution to GLUT1- DS and potential evolutionary differences between progenitor metabolism in mouse and humans. Experiments will combine cutting edge technologies to determine the link between RG metabolism and cell fate decisions using in vivo and in vitro metabolomic assays along with single-cell transcriptomic and imaging methods. Pursuing these aims will open new avenues of inquiry into the metabolic regulation of RG fate and differentiation in health and disease. Our long-term objective is to understand the combinatorial actions of environmental and genetic factors during cortical development and identify potential therapeutic pathways.

NIH Research Projects · FY 2026 · 2025-03

PROJECT ABSTRACT RNA oxidation is a hallmark of neurodegeneration, and may contribute to the altered gene expression seen in neurodegenerative disease. The major oxidative modification is 8-hydroxyguanosine (8-OHG), which impairs translation and other aspects of RNA function. Reactive oxygen species (ROS), generated by mitochondria through cellular respiration, are the drivers of RNA oxidation. Although ROS is generally thought to nonspecifically modify nucleic acids, microarray analysis of 8-OHG-containing mRNA from Alzheimer's patient samples found that only certain mRNAs underwent oxidation. However, it is currently unclear how ROS can selectively modify specific mRNA. To understand the basis for selective RNA oxidation, I performed an 8-OHG mapping study using antibodies that bind 8-OHG. I found that these antibodies bind to regions which appear to correspond to G-quadruplexes. In vitro studies previously showed that G-quadruplexes can bind heme, converting heme into a highly efficient ROS generator. Based on these prior studies, I asked if heme can bind to cellular mRNA. I developed Heme-seq, a method for mapping heme-binding sites in mRNA, and found that heme-binding sequences are found throughout the transcriptome and correspond to G-quadruplex regions. Additionally, I developed 8-OHG-seq, a new single-nucleotide 8-OHG mapping method, which shows 8-OHG enrichment in G-rich regions consistent with G-quadruplexes. My 8-OHG mapping data supports the idea that 8-OHG is a selective modification. Furthermore, my data suggest that heme binds to G-quadruplex sequences in cellular mRNA, leading to localized ROS generation and RNA oxidation. In order to test this hypothesis, the aims of this project are: Aim 1: To determine the role of heme-binding events on 8-OHG formation in mRNA. I will use small molecules that displace heme from G-quadruplexes to determine if heme must directly contact G-quadruplexes in RNA to catalyze 8-OHG formation. I found that cell stress leads to increased heme binding. I will determine if this increased binding occurs at G-quadruplexes and is associated with formation of new 8- OHG sites. These experiments will address whether heme-binding to G-quadruplexes is the mechanism of specificity for RNA oxidation. Aim 2: To determine the role of cell stress on the folding of heme-binding sites. Cell stress markedly increases the folding of G-quadruplexes in cells. I will map the location of endogenous heme-binding sites induced by serum deprivation and determine if these sequences are G-quadruplexes that fold in response to stress. These aims will provide insights into the role of heme-binding to RNA as a mechanism in RNA oxidation. These studies will also allow me to achieve training goals related to acquiring knowledge and skills in RNA biology, molecular mechanisms related to neurodegeneration, and developing bioinformatic skills, which will help me develop the skills to become an independent investigator.

NIH Research Projects · FY 2026 · 2025-03

PROJECT SUMMARY/ABSTRACT Despite recent progress, most patients with solid tumors still do not respond to immunotherapies. Responders often experience immune-related adverse events (irAEs), which can be life-threatening and require treatment discontinuation. Current immunotherapies focus on enhancing the fitness of anti-tumor effector T cells, but do not simultaneously engage innate immune responses. Neutrophils infiltrate tumors and can exhibit either anti- or pro-tumorigenic properties, depending on the context of tissue inflammation. While chronic inflammation favors pro-tumorigenic neutrophils, we and others have demonstrated that acute inflammation induced by T cell therapies promotes anti-tumorigenic neutrophils that help eradicate the tumor. However, it remains unclear how neutrophils become tumoricidal following immunotherapy. In melanoma, neutrophils also infiltrate healthy tissues of immunotherapy-responders with irAEs, suggesting potential pathogenic properties. Nonetheless, the direct contribution of neutrophils to development of irAEs has not been addressed. Understanding these mechanisms is crucial to develop new combination treatments that activate synergies between the innate and adaptive immune systems to potentiate anti-tumor responses while preventing immunological toxicities. To investigate the precise role of neutrophils in anti-tumor immunity and irAEs, we propose: Aim 1. Characterize gene pathways and tumor-derived factors defining functional subsets of anti-tumorigenic neutrophils. We will identify key molecular targets underlying neutrophil tumoricidal activity by integrating scRNA- seq and proteomics across models of anti- and pro-tumorigenic neutrophils. We will evaluate whether these targets can promote neutrophil-mediated tumor control in preclinical melanoma and breast cancer models. Aim 2. Identify clinically relevant mechanisms of tumor cell destruction employed by neutrophils. We will evaluate the contribution of known neutrophil killing pathways, including ROS and NETosis, to immunotherapies using genetic models and inhibitors targeting key mediators of these pathways. We will assess if neutrophils selectively kill tumor cells. Furthermore, we will evaluate the clinical relevance of gene signatures, molecular cues, and killing mechanisms in biospecimens from patients with solid tumors undergoing immunotherapies. Aim 3. Uncover the role of neutrophils in development of irAEs. Preliminary data suggest that neutrophil depletion alleviates skin immunological toxicities associated with immunotherapy. We will use this model along with scRNA-seq studies to identify actionable targets that uncouple neutrophil tumoricidal activity from pathogenic behavior and validate findings in patient samples. We will also evaluate the contribution of VISTA and NETosis to development of irAEs. We expect to find immunotherapeutic interventions based on innate immune profiles and reliable biomarkers for cancer prognosis. Our long-term goal is to develop approaches targeting neutrophils that enhance anti-tumor immunity while preventing associated toxicities to advance the precision and safety of immunotherapies.

NIH Research Projects · FY 2026 · 2025-03

PROJECT SUMMARY/ABSTRACT The successes of immune checkpoint inhibitors (ICI) have demonstrated the power of T cells to reject tumors, but responses are limited to a subset of patients. Recent data show that cancer cell-intrinsic cGAS/STING signaling is a prerequisite for effective immunotherapy, suggesting that treatments such as radiation therapy (RT) that activate this pathway should synergize with ICI. Consistently, we have previously shown that RT- induced cancer cell-intrinsic cGAS/STING signaling is required for the activation of systemic anti-tumor T cell responses in combination with ICI. Despite the substantial preclinical data that RT increases responses to ICI, clinical evidence has been inconsistent with a mixture of positive, negative and inconclusive results in clinical studies, challenging initial assumptions that RT could be broadly beneficial in unselected patient populations. These results highlight the need to identify the determinants of RT’s ability to convert a tumor into a hub for in situ immunization. Efforts have largely focused on investigating the barriers at the level of the tumor microenvironment and on improving RT use (e.g., dose and delivery schedule) to maximize its pro-immunogenic effects. The proposed studies represent a shift in focus from the search for actionable targets in the immune compartment to the role of the most frequently mutated gene in human tumors, p53, as a central regulator of the pro-immunogenic effects of RT. Recent data demonstrate that p53 modulates the IFN-I response induced by cytosolic DNA via cGAS/STING, with opposite effects of wild type p53 (p53wt), which increases the response by promoting TREX1 degradation, and mutant p53, which decreases it by hindering the interaction of TBK1 with STING and IRF3. The central hypothesis of this application is that the p53 mutational status of a tumor is a key determinant of the ability of RT to induce cancer cell-intrinsic IFN-I pathway activation and in situ immunization. To test this hypothesis, we will use several p53-isogenic mouse and human cancer cell lines and primary patient-derived tumor organoids (PDOs) that carry the most common mutations in p53 gene. Firstly, we will determine if RT-induced IFN-I is increased by p53wt and decreased by different p53 mutants in human and mouse cancer cells in vitro. Secondly, the effect of p53 status on RT-induced immune activation will be determined in syngeneic immunocompetent mouse models. Thirdly, we will evaluate RT-induced IFN-I pathway activation in a cohort of 38 PDOs (derived from breast, colorectal, and lung cancers) with different p53 mutations and 38 tumor type-matched p53 wild type control PDOs.

NIH Research Projects · FY 2026 · 2025-03

PROJECT SUMMARY/ABSTRACT Cancer cells must initiate a telomere maintenance mechanism to continuously proliferate. While the majority of tumors (85%) reactivate telomerase, a subset (15%) employs homologous recombination in a process termed alternative lengthening of telomeres (ALT). ALT primarily manifests in tumors originating from mesenchymal tissues and is characterized by long and heterogeneous telomeres, the presence of extrachromosomal telomeric repeats, localization of telomeres in nuclear PML bodies, and telomere sister chromatid exchanges. Chromatin dynamics play a crucial role in ALT; loss of chromatin remodeler ATRX is almost always observed in ALT cells, and exogenous expression of ATRX suppress ALT. However, ATRX is a large multifunctional protein, and the mechanism by which it suppresses ALT remains elusive. Furthermore, H3K9 methylation at telomeres has been implicated in promoting ALT phenotypes, and loss of H3K9 methylation represses ALT phenotypes. How H3K9 methylation regulates ALT is also unknown. I hypothesize that ATRX loss promotes ALT induction by disrupting its role in histone variant H3.3 deposition at telomeres. Further, I hypothesize that loss of H3.3 disturbs the H3K9 methylation balance at the telomeres, leading to increased H3K9 methylation, R-loop stabilization, replication stress, and double-stranded breaks driving ALT homologous recombination. The goal of this proposal is to investigate the chromatin landscape at ALT telomeres and elucidate its impact on ALT through two distinct but related objectives. In Aim 1, I will focus on the role of ATRX in ALT suppression. I will employ a base editing screen to create mutations spanning the coding sequence of ATRX. Using this screen, I will identify ATRX domains and functions that suppress ALT. In Aim 2, I will study the effect of H3K9 methylation on ALT by manipulating H3K9 methylation specifically at the telomeres. I will then assess ALT phenotypes and explore the mechanism behind H3K9 methylation’s impact on ALT with a focus on R-loop stabilization. This work will provide novel insight into how the chromatin state governs ALT and offer potential avenues to target ALT for cancer therapeutics. I am an MD/PhD student at the Weill Cornell/Memorial Sloan Kettering/Rockefeller Tri-Institutional Program, performing the proposed research in the laboratory of Dr. Agnel Sfeir at Memorial Sloan Kettering Cancer Center (MSKCC). My long-term goal is to become a physician scientist who balances patient care with running an independent research program at an academic institution. The plan outlined in this proposal, along with the support and mentorship provided by Dr. Sfeir, my thesis research committee, and the Tri-Institutional administrative faculty will help me achieve my career goals.

NIH Research Projects · FY 2026 · 2025-03

PROJECT SUMMARY / ABSTRACT The androgen receptor (AR) is the central determinant of prostate tissue identity and differentiation, controlling normal, growth-suppressive gene expression associated with differentiation of prostate cells. It is also a key driver of prostate tumorigenesis, becoming “hijacked” to drive oncogenic transcription. However, the key factors that mediate this switch, and whether the growth suppressive program can be reactivated to inhibit prostate cancer, remain unknown. Critically, AR also remains the key therapeutic target for prostate cancer, even in advanced, treatment resistant disease, where genomic alterations such as massive amplification, overexpression, mutations, and splice variants of AR drive continued reliance on androgen signaling. Therefore, understanding how AR is reprogrammed from its normal growth suppressive role to an oncogenic factor has major impact on our understanding of context-specific regulation of lineage-specific transcription factors and responses to prostate cancer therapies targeting AR. Our work has shown, in both published and unpublished preliminary data, that specific factors mediate the balance of AR activity between normal and oncogenic transcriptional programs. In unpublished preliminary data, we use a novel system to modulate canonical androgen receptor response element (ARE) motifs to show that the normal AR transcriptional program is governed by AREs, and reactivation of this normal program is growth suppressive in prostate cancer cells. We hypothesize that the growth-suppressive AR transcriptional program can be reactivated in advanced prostate cancer, and specific genomic and epigenetic modulators will mediate the transition between normal and oncogenic AR activity. We will test this hypothesis with three Specific Aims: 1) Determine the context that allows engagement of growth suppressive AR transcriptional programs in prostate cancer, 2) Establish the translational implications of oncogenic and growth suppressive AR programs in human prostate cancer patients, and 3) Define the key cofactors and modifiers that mediate the switch between normal and oncogenic AR programs. With a combination of novel epigenomic modulation strategies, functional genomics, genetically engineered mouse models, patient derived model systems, and human patient data, we will establish the potential of reactivating the AR growth suppressive program in vitro and in vivo, examine the implications for patients, and define novel AR cofactors and therapeutic targets. Together, these studies will evaluate an innovative, potentially transformative treatment strategy for lethal prostate cancer, expose new biology and new therapeutic targets, and define a new paradigm for cancers dependent on lineage-specific, multifunctional transcription factors.

NIH Research Projects · FY 2026 · 2025-03

PROJECT SUMMARY Most proteins have more than one function and participate in a wide range of biological processes. This compli- cates our ability to infer protein function from sequence. This problem, otherwise known as genotype-to-pheno- type mapping, represents one of biology’s most intractable challenges. One way by which proteins diversify their functionality is through the evolution of modular domains, each of which can engage in distinct functions. Satu- ration mutagenesis, which systematically alters each amino acid in a given protein, is a powerful technique to map diverse functions to individual domains by generating separation-of-function mutants. However, the utility of this approach remains severely limited by excessive labor, prohibitive costs, and, most importantly, low-di- mensional readouts. The vast majority of saturation mutagenesis libraries have thus far been screened in vitro using biochemical or simple cellular assays. There are currently no approaches to probe the complex cellular and in vivo functions of saturation mutagenesis libraries at scale. This inspired our development of SatSeq – a modular breakthrough technology that couples the power of single cell sequencing with saturation mutagenesis and DNA barcoding – to map the structure-function landscape of multimodal proteins in complex and dynamic biological systems both in vivo and in vitro. As a proof of principle, we will develop SatSeq to interrogate the opposing functions of an immune-related protein called Stimulator of Interferon Genes (STING) in cancer pro- gression. STING is a master regulator of innate immunity, and it has been proposed to activate anti-tumor im- mune responses early in disease progression. Yet, recent evidence suggests that its chronic activation in cancer cells can drive metastasis. We hypothesize that this duality is achieved through distinct functional elements. Coupling genome-wide, single cell transcriptional profiling with in vivo interrogation of saturation mutagenesis barcodes, we will test the modular and time-dependent functions of STING during cancer progression. We will then utilize SatSeq as a target validation platform for STING modulators in clinical development, given its ability to readily identify resistance mutations that prevent drug-target binding without impacting target protein activity (i.e. gatekeeper mutations). The identification of such mutations is the gold standard for validating on-target drug effects, which will be applied here to a set of small molecule activators and inhibitors of STING. This is particularly salient since the first STING agonist to be used in clinical trials (DMXAA) was ineffective due to its inability to bind its target. In sum, we expect SatSeq will have significant impact at the fundamental and translational levels. First, it will expand our understanding of STING’s diverse functions during cancer progression – revealing the context in STING which activation vs. inhibition is therapeutically beneficial and creating new avenues for patient selection. Second, as a high-throughput platform to validate on-target drug activity, SatSeq will determine whether emerging STING modulators act through on-target effects and suggest mechanisms of drug action.

NIH Research Projects · FY 2025 · 2025-02

Project Summary/Abstract (limited 30 lines of text): In humans, cellular diversity and phenotypic variation stem not only from differential gene expression but also from the expression of different gene isoforms across distinct cellular contexts. Across various cancer types, including both liquid and solid tumors, mutations and expression imbalances in splicing factors contribute significantly to disease progression and heterogeneity. The lack of comprehensive understanding of splicing aberrations and their impact on individual cell phenotypes in cancers like myelodysplastic syndromes (MDS) and high-grade serous ovarian cancer (HGSOC) presents a critical gap in knowledge. The proposed research aims to explore the role of aberrant splicing in cancer cell heterogeneity and its potential as targets for novel immunotherapeutic strategies. Aim 1 will utilize GoT-Splice to study mutations in splicing factors such as SRSF2, U2AF1, and ZRSR2 in MDS, enhancing the characterization of retained introns and identify tumor-specific isoforms with translational potential. Aim 2 will explore the cell-to-cell variability of isoform usage in HGSOC, leveraging single-cell long-read sequencing data to elucidate the relationship between splicing regulation, genomic instability, and immune interactions within the tumor microenvironment. Finally, Aim 3 seeks to identify splicing-derived neoantigens for immunotherapy, utilizing multi-omic characterization of single-cell splicing variation to develop models that distinguish between tumor-specific and normal isoforms predicting their immunological potential. The proposal emphasizes the importance of integrating advanced computational frameworks with multimodal single-cell approaches to comprehensively characterize splicing aberrations and their functional implications in cancer. It also highlights the potential of splicing-derived neoantigens as promising targets for precision immunotherapies, while addressing challenges related to neoantigen off-target effects and HLA variability. Overall, this proposal seeks to provide a comprehensive understanding of isoform regulation in cancer, offering insights into potential targets for precision medicine approaches in cancer immunotherapy.

NIH Research Projects · FY 2026 · 2025-02

1 Project Summary 2 3 Autologous fat grafting is the transfer of fat from one anatomical region to another and is commonly used in 4 breast reconstruction after mastectomy. Accurate prediction of graft retention remains an enduring clinical 5 problem, as resorption is reported to range between 10% to 50% in the literature. While there are several devices 6 available for fat grafting, no randomized, controlled trials have been completed to compare outcomes between 7 them, including volume retention and factors that would affect volume retention. Given the lack of evidence- 8 based guidelines, the decision of how best to process graftable fat remains up to physician preference. Moreover, 9 much work fails to assess the microscopic alterations to the grafted tissue resulting from each processing 10 technique, the study of which might reveal important predictive factors of complications or graft take. In response, 11 we are seeking to compare outcomes between three fat grafting devices which process the lipoaspirate in different 12 ways before it is injected into the graft site. We will utilize fat samples collected from patients undergoing fat grafting 13 who have been enrolled in an ongoing randomized, controlled trial to receive fat processed by techniques that 14 actively wash and filter the fat, known as “active closed wash and filtration” (ACWF), passively filter the fat 15 through a collecting system with minimal processing, known as a “passive low pressure closed system” (LPCS), 16 or actively wash and filter the fat with the addition of surfactant, known as “active wash and filtration plus 17 surfactant” (AWFPS). Originally, the ACWF and LPCS devices were compared to standard decantation; however 18 interim analysis showed superior volume retention with the ACWF and LPCS devices. Standard decantation has been 19 replaced with AWFPS to identify the optimal fat grafting device to use for patients. We will take the lipoaspirate that has 20 been harvested and processed via ACWF, LPCS, or AWPS, and inject a predefined quantity into the dorsa of 21 immunocompromised mice. We will assess the volume and projection of the grafted material in the mice at 1, 2, 22 and 3 months, as the literature states that reabsorption happens within the first 3 months and then the graft 23 becomes stable. We will also characterize the cellular environment around and within each graft after 3 months, 24 including any immune or inflammatory response as well as the viability and potential hormonal activity of the 25 grafted fat. Further, we will compare the retention rates and complication profiles observed in our mouse models 26 with those of their patient counterparts in the companion trial to provide further confirmation of any preliminary 27 conclusions drawn. Lastly, we will complete this analysis with a small subset of humanized mice as well in order 28 to compare outcomes and the ability to measure volume retention in immunocompromised versus humanized 29 mice. Measurement of volume retention of human lipoaspirate has not yet been documented in humanized mice, 30 and including a pilot study would allow us to assess immunogenicity effects on volume retention of lipoaspirate 31 and whether future studies of this nature could be completed in humanized mice. Immunogenicity effects will be 32 assessed by measuring angiogenesis in the grafted fat after 3 months and comparing between mouse species.

NIH Research Projects · FY 2026 · 2025-02

PROJECT SUMMARY/ABSTRACT Humans face many types of environmental genotoxic agents, such as various pollutants and radiation, that can cause deleterious mutations and diseases. To survive and adapt to genotoxic conditions, cells rely on many highly conserved and complex molecular strategies. One strategy to withstand genotoxic stress is the use of error-prone repair mechanisms to increase mutation rates, which can potentiate new functions for better adaptation to toxic environments. Another mechanism is the DNA damage checkpoint (DDC) system, which controls the arrest and resumption of the cell cycle in accordance with the burden of genomic lesions. While the frameworks of both strategies are largely established, important mechanistic details are lacking. To address these gaps in knowledge, I will capitalize on the budding yeast model organism’s highly conserved DDC and repair systems. Aim 1 will build on the recent observation that the desumoylase Ulp1 is sumoylated upon genotoxin treatment and that mutating its two sumoylation sites results in a phenotype indicative of increased homologous recombination (HR). I thus hypothesize that Ulp1 sumoylation suppresses HR to favor error-prone repair. I will test this hypothesis and define the broader SUMO-based control of DNA repair pathway usage in response to genotoxins. Aim 2 will address the recent findings that RPA is downregulated after genotoxin exposure. I hypothesize that RPA downregulation can induce DDC recovery. To test this, I will elucidate how RPA degradation is regulated and how this impacts DDC recovery. Defects in either of the DNA repair pathway choices and DDC responses can lead to genome instability syndromes, characterized by increased sensitivity to environmental genotoxins, developmental abnormalities, and increased tumorigenesis. This work will lay the foundation for elucidating cellular adaptation strategies and can inform disease prevention and treatment strategies. It will also provide important training for me to reach my long-term career goal of becoming a principle investigator studying how human cells response to environmental genotoxins.

NIH Research Projects · FY 2026 · 2025-01

Project Summary/Abstract Atrial fibrillation (AF) is the most common sustained arrhythmia with approximately one quarter of all adults developing AF by the age of 80. AF can be symptomatic with people experiencing palpitations, racing heart, syncope, congestive heart failure, and cardioembolic stroke. Epidemiologically, aging is associated with AF and increasing obesity confers a stepwise higher lifetime risk of AF. Atrial myopathy and AF are thus expected to become more rampant with the obesity and type 2 diabetes epidemics and the aging population. The molecular mechanism(s) by which obesity leads to atrial fibrillation is poorly understood. We have developed a new mouse model of spontaneous atrial fibrillation triggered by obesity. We found that nicotinamide adenine dinucleotide (NAD+) and the Atf6 ER stress pathway are perturbed in obesity-induced atrial fibrillation. Importantly, correcting NAD+ levels in the heart ameliorates atrial fibrillation. In this proposal, we seek to follow up on these studies and assess the mechanisms by which NAD+ pathways protect against atrial fibrillation. We will pursue the following specific aims: 1. Dissect the role of the NAD+ pathway in obesity- induced AF. 2. Assess the role of the Atf6 ER stress pathway in obesity-induced AF. The overall goal of these studies is to gain greater insight into the molecular pathogenesis of metabolic diseases and AF that may ultimately lead to new treatments to prevent or ameliorate AF.

NIH Research Projects · FY 2026 · 2025-01

Our key objective is to develop renal quantitative susceptibility mapping (QSM) for reliable detection and quantification of hemorrhagic cysts as a prognostic biomarker of kidney function decline in autosomal dominant polycystic kidney disease (ADPKD). ADPKD is the most common hereditary kidney disease affecting over 500,000 people in the US, and characterized by formation of cysts causing compression of nephrons and progressive decline of glomerular filtration rate (GFR). The critical task in management of ADPKD is the identification of patients at higher risk of rapid kidney function decline. The only available drug to slow ADPKD progression is vasopressin V2 receptor antagonist tolvaptan. Because disease progression is highly variable and because tolvaptan has long-term adverse aquaretic and hepatotoxic effects, it is important to identify patients who are most likely to benefit from the drug. The only FDA-approved biomarker for this risk stratification is height- adjusted total kidney volume (htTKV). But htTKV is only a global measure and doesn't inform on pathogenic factors such as cyst types, distribution, fibrosis and inflammation. Patient risk stratification by renal function decline remains a critical and unanswered need. We and others have demonstrated that the hemorrhagic cystsare strongly associated with rapid progression of chronic kidney disease. Hemorrhage in ADPKD is the product of the vascular endothelial growth factor (VEGF) expression by the cystic epithelium leading to formation of highly permeable vasculature within the cyst wall. Hemorrhage and increased vessel permeability facilitate cyst volume increase. The inflammatory responses triggered by hemorrhage contribute to the pathogenesis of tubulointerstitial fibrosis. Identification and quantification of hemorrhagic activity in ADPKD provides an early window into ongoing cystogenesis and tissue remodeling before their consequences can be detected in conventional clinical imaging. At present, hemorrhagic cysts are detected as hyperintense on T1-weighted (T1w) and hypointense on T2w images in conventional MRI. However, conventional MRI intensity characteristics are unspecific and unable to differentiate between hemorrhagic and proteinaceous cyst. Breathhold MRI acquisitions cause low slice resolution and misregistration between T1w and T2w images, complicating visualization and classification of cysts. Non-contrast CT and ultrasound have low specificity for identifying hemorrhage The lack of reliable modality for the detection of hemorrhagic cysts is an unmet gap in clinical imaging. We have preliminary cross-sectional data demonstrating association between QSM and decreased eGFR in ADPKD patients, but a validation of these findings in a robustly designed longitudinal study is needed. Our central hypothesis is that detection of hemorrhage in ADPKD by QSM will allow precise identification of patients at high risk of eGFR decline benefiting most from therapeutic intervention. Our research plan has 3 specific aims: 1. Develop robust and accurate renal QSM; 2. Validate renal QSM using biochemical measurements and histology in kidney explants; 3. Validate renal QSM for eGFR decline risk stratification in ADPKD patients.

NIH Research Projects · FY 2025 · 2025-01

PROJECT SUMMARY Inflammatory bowel diseases (IBD), which include ulcerative colitis and Crohn's disease, are estimated to affect 3 million individuals in the United States, and the number of people living with IBD continues to rise. Currently available treatment options are ineffective for some patients, costly and pose serious long-term complications, such as opportunistic infections, autoimmunity, and cancer. Thus, there is an urgent need to improve our understanding of the modulators of intestinal inflammation and repair in order to identify novel therapeutic targets to treat and prevent IBD. Emerging studies from multiple groups have demonstrated that the peripheral and enteric nervous systems regulate intestinal immunity to infection and inflammation via complex neuro-immune circuits, including neuropeptides neuromedin U (NMU) and calcitonin gene-related peptide (CGRP) that directly regulate inflammation and restore tissue homeostasis in the gastrointestinal mucosa. However, little is known whether and how these immunomodulatory neuropeptides play a pivotal role in the pathogenesis of IBD. In new preliminary studies, we identified that adrenomedullin 2 (ADM2), an understudied neuropeptide hormone, was highly upregulated in inflamed intestines of both humans and mice. While ADM2 utilizes ADM1 receptor (ADM1R) and ADM2 receptor (ADM2R) for signal transduction, mice deficient in ADM2R exhibited significantly increased susceptibility to a murine model of intestinal damage and inflammation. Further, therapeutic administration of exogenous rADM2 peptide substantially ameliorated the disease while concomitantly inducing a population expansion of a tissue-protective group 2 innate lymphoid cells (ILC2s) in the colon. Based on our new preliminary data, we hypothesize that the ADM2-ADM2R pathway drives non-redundant ILC2 responses to limit inflammation and promote tissue protection in the intestine. We propose to generate a detailed understanding of how the ADM2-ADM2R axis mediates tissue protection in both murine models of intestinal inflammation and human IBD. In Aim 1, we will test the hypothesis that during intestinal damage and tissue protection enteric neurons modulate their expression of ADM2. Further we will utilize chemogenetic manipulation of ADM2-expressing neurons to test how this impacts intestinal inflammation. In Aim 2, we will employ a novel mouse model that allows ILC2-specific deletion of ADM2R to directly test if ILC2-intrinsic ADM2R signaling is required for ADM2 mediated tissue protection and test if AREG is the specific molecular mediator that confers intestinal tissue protection. In addition to uncovering fundamental and novel neuropeptide biology of ADM2 and its roles in the pathogenesis of IBD, these studies could provide further insights to support the development of novel therapeutics to target this pathway.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY White adipose tissue (WAT) is a complex organ with functional roles beyond storing energy and insulating organs. Recent studies have demonstrated the critical role of WAT in supporting systemic health through secretion of biologically active substances (adipokines). WAT dysfunction can arise from tissue expansion through increased adipocyte size or number density. While the systemic expansion of WAT is typically caused by increased energy intake, localized diseases like cancerous tumors or coronary artery disease can affect the microstructure of nearby adipocytes. Systemic WAT dysfunction correlates with levels of certain adipokines measurable in blood serum, but local microstructural changes of adipocytes associated with local disease do not necessarily correlate with blood serum adipokine levels, requiring direct evaluation of WAT microstructure to quantify changes through biopsy or tissue resection. No methods exist to directly characterize the microstructural properties of adipocytes in WAT non- invasively and in vivo. Given the importance of WAT to human health and the relationship between WAT microstructural changes and dysfunction, there is clearly a need for methods to rapidly and non-invasively quantify adipocyte size, number density, and spatial organization in vivo for detecting and monitoring both systemic and local disease. We propose to develop quantitative ultrasound (QUS) imaging methods for this purpose. We hypothesize that 1) QUS parameters correlate with adipocyte size, number density, and spatial organization in WAT and 2) QUS can non-invasively detect local microstructural changes in adipocytes not measurable through blood serum biomarkers which are typically associated with systemic WAT dysfunction. Aim 1 will test the first hypothesis by comparing QUS parameters computed from WAT regions in breast tissues with adipocyte size and number density measurements computed from digital image analysis of H&E-stained tissue sections of the same regions. The second hypothesis will be tested in aim 2 by comparing QUS parameters from WAT regions near cancerous breast lesions – where microstructural properties of adipocytes are known to change locally – and several millimeters away from lesions (not affected by cancerous tumors) to measurements of adipokine levels in blood serum. Aim 1 validates QUS as a method to characterize adipocyte microstructure in vivo whereas Aim 2 demonstrates the efficacy of QUS parameters to detect local, not systemic, microstructural changes. The expected outcome of this work is the validation of QUS as a non-invasive method to characterize the microstructural properties of adipocytes in WAT. If successful, QUS could become a meaningful tool for routine clinical care and for supporting other research efforts attempting to uncover the relationship between alterations in WAT microstructure and disease.

NIH Research Projects · FY 2026 · 2025-01

PROJECT SUMMARY/ ABSTRACT Heart failure with preserved ejection fraction (HFpEF) affects more than 3 million people in the United States (U.S.) and contributes to substantial impairments in health-related quality of life (QOL). While there are proven benefits of ß-blockers in other phenotypes of HF (e.g., HF with reduced ejection fraction [HFrEF]), there is limited evidence of benefit and some evidence suggesting harm in the setting of HFpEF. Despite this, >80% of adults with HFpEF receive ß-blockers often without a clear guideline-based indication. While ß-blockers were originally postulated to improve cardiac output in HFpEF by slowing down the heart rate and improving left ventricular filling, there is emerging evidence that ß-blockers may cause more harm than good by exacerbating chronotropic incompetence, worsening cardiac output, and/or reducing exercise tolerance. Relatedly, ß- blockers may worsen several domains of health-related physical and mental QOL. Taken together, these findings suggest that ß-blockers are a compelling target for deprescribing. Yet, ß-blockers are not commonly deprescribed, even when physicians recognize there is no evident clinical indication. A critical barrier to deprescribing in HFpEF is a lack of rigorous evidence regarding safety and efficacy, supporting the urgent need for our proposal. Our central hypothesis is that deprescribing ß-blockers will lead to net benefit in older adults with HFpEF. This hypothesis is supported by our preliminary data showing that withdrawal of ß-blockers is safe and improves both physical and mental health. To establish evidence for the net benefit of deprescribing ß-blockers in HFpEF, we will conduct a double-blinded, placebo-controlled, pragmatic randomized clinical trial in 320 older adults with HFpEF. Our “Determining Evidence in a Placebo-Controlled, Randomized Experiment Studying Continuation vs. Removal of Inessential Beta-Blockers in Elders with Heart Failure with Preserved Ejection Fraction (DEPRESCRIBE-HFpEF) study” will be conducted at Kaiser Permanente Northern California with pragmatic design elements. We will use an innovative hierarchical endpoint (analyzed using the win-ratio) for net benefit that reflects clinical priorities, incorporating both safety and efficacy. Our team is led by two cardiologists with expertise in HFpEF—one has studied deprescribing ß- blockers for the last 5 years (supported by NIA K76), and one has significant experience designing and conducting pragmatic RCTs within a learning health system. Our team also includes senior scientific leaders with expertise in aging research, HFpEF, deprescribing, QOL, pragmatic RCTs, and biostatistics. The expected outcome of this proposal is foundational evidence that will impact clinical practice guidelines for the management of older adults with HFpEF. This research directly addresses NIA’s Strategic Directions Research Goal C-3 to “develop interventions for treating…or mitigating the impact of age-related disease and conditions.”