Weill Medical Coll Of Cornell Univ

NIH Research Projects · FY 2025 · 2024-06

Project Summary For breast cancer, skeletal metastases are a major source of the pain, disability and mortality associated with disease progression. As with other solid tumors metastasizing to bone, breast cancer displays a striking tropism for vertebral bone, with approximately 2-5 vertebral metastases for every one long bone metastasis, implying that vertebral-specific factors are key drivers of metastasis. Passive blood flow cannot explain the increased metastatic tropism for vertebral versus long bones, indicating that unrecognized biologic factors are likely to drive the high rates of vertebral metastasis observed clinically. We have here identified a new skeletal stem cell responsible for generating the osteoblasts that mineralize and remodel the vertebrae (vertebral skeletal stem cells, vSSCs) and provide evidence that this cell is a major determinant of vertebral-specific pathology, including the high rates of vertebral metastases observed clinically. Even after normalizing all anatomic factors through the use of in vivo bone organoids, vSSC-derived bone tissue recruited tumor cells more efficiently than comparable stem cells from long bones. Targeting loss- of-function to vSSCs using a mouse cre line developed for this project selectively reduces vertebral metastasis rates. In both organoid systems and native vertebrae, vSSCs drive high rates of initial vertebral seeding with tumor cells, indicating that vSSC-driven tropism is a major contributing factor behind the high vertebral metastatic rates observed in breast cancer. Building upon this observation, we have identified candidate trophic factors expressed by vSSCs and have generated evidence implicating specific vSSC-derived factors in vertebral metastatic tropism and outgrowth. Here, we will perform key studies needed to advance the rigor, underlying mechanism, clinical relevance and translational therapeutic impact of this discovery. First, (Aim 1), we will establish the metastatic functions of this vSSC in the native vertebral environment, including investigating how vSSCs may organize the entire vertebral metastatic niche and which vSSC-derived cell types contribute to metastasis. Next (Aim 2), we have identified a candidate vSSC-derived mediator driving metastatic tropism and will here determine how secretion of this factor is regulated and how it signals to tumor cells to impact metastasis. Lastly (Aim 3), we will develop the therapeutic impact of this discovery by determining if the human counterparts of vSSCs display a conserved metastatic function in xenograft systems and by conducting proof-of-concept therapeutic studies of blocking candidate vSSC-derived metastatic mediators. Altogether, this study will establish that a new stem cell responsible for forming the vertebrae drives vertebral metastases, thereby offering a new model for the site specificity of skeletal metastases, an explanation for the high rates of vertebral metastases observed clinically and new therapeutic opportunities.

NIH Research Projects · FY 2025 · 2024-06

Modified Project Summary/Abstract Section: Research: Emerging studies have characterized intricate neuro-immune interactions that influence immunity and tissue homeostasis at various barrier surfaces. Thus, synergistic targeting of neuro-immune pathways may be a novel therapeutic approach for treating various inflammatory diseases, including inflammatory bowel diseases (IBD). My recent study demonstrated that a cholinergic neuropeptide, neuromedin U (NMU), and intestinal tissue-resident group 2 innate lymphoid cells (ILC2s) cooperatively regulate the pathogenesis of IBD. However, the etiology of IBD is highly complex, and patients with extraintestinal inflammation, such as chronic inflammatory skin diseases, also present gastrointestinal complications and are at increased risk for IBD. Based on my new preliminary data combined with my recent work, this proposal will investigate the role of NMU and ILC2s in regulating such skin-gut inflammatory circuits. Completion of proposed studies will allow for the rational design and development of novel therapeutics and preventative medicine for IBD and other inflammatory diseases. Career Goals: My overarching career goal is to become an independent investigator at an academic institution to study novel approaches to harness neuro-immune circuitries to treat inflammatory diseases. Furthermore, I believe that training future scientists is also an essential aspect of being an independent investigator. Thus, I will strive to become an inspirational mentor for my future trainees. Career Development Plan: To successfully transition to independence, I must develop various scientific, professional, and personal skills, including broadening my expertise in murine models of skin inflammation and molecular and cellular techniques to modulate neuro-immune interactions in the skin and the intestine. I must also cultivate collaborations and skills in writing, communicating, mentoring, and laboratory management. In addition to the support provided by mentors and advisory committee members, I will participate in workshops and programs led by distinguished scientists at Weill Cornell Medicine (WCM) and the New York Academy of Sciences to aid in developing skills in specialized experimental techniques and laboratory management. Career Development Environment: I will conduct the K99 phase of the proposed research in the Artis lab (WCM), which is located at the heart of the Tri-Institutional campus, comprising the WCM, The Rockefeller, and Memorial Sloan Kettering Cancer Center. The laboratory has access to all the state-of-art equipment and core facilities required to complete the experiments proposed in this proposal, including transcriptomic sequencing, optical and high-throughput imaging, and the animal facility. Thus, the WCM campus is an ideal environment for my K99 mentored research training phase.

NIH Research Projects · FY 2026 · 2024-06

PROJECT SUMMARY/ABSTRACT: Microglial-mediated neuroinflammation has long been recognized as a pathological hallmark of the progression of Alzheimer's disease (AD). Although many anti-inflammatory drug candidates have undergone clinical trials as potential AD therapeutics, most have failed. Triggering receptor expressed on myeloid cells 2 (TREM2) is a microglia-specific receptor that mediates intracellular cascades to modulate the production of inflammatory cytokines and the phagocytosis of amyloid  (A) plaques. The interaction of TREM2 with galectin-3 (Gal-3) stimulates proinflammatory activation of microglia and represents a promising target for developing AD therapeutics that can alleviate neuroinflammation. In addition, stabilizing TREM2/A interaction has been reported to enhance TREM2-mediated A phagocytosis in vivo. However, targeting TREM2 is currently restricted to antibodies (Abs), and there are no small molecules in existence that target TREM2. To fill this gap, we have developed an innovative platform, Small Molecules from Antibody Pharmacophores (SMAPs), that can identify small molecule ligands for immune cell receptors with high binding affinity and selectivity. Our SMAPs platform is based on utilizing cocrystal structures of immune cell receptors with Abs in building pharmacophore maps from clusters of key interacting residues of Abs with immune cell receptors to identify small molecules that modulate the function of immune cell receptors. We propose to establish a new microglial modulating strategy to treat AD based on therapeutic targeting of TREM2 with small molecules identified from the SMAPs platform. We identified a focused chemical library using our pharmacophore-based virtual screening approach (SMAPs) based on interactions derived from a cocrystal structure of TREM2 and anti-TREM2 Ab single-chain variable fragment (scFv) (PDB ID: 6Y6C). In comparison to Abs, small molecules can readily cross the blood-brain barrier (BBB) and are amenable to pharmacokinetic optimization, which may enable avoiding immune-related adverse events associated with Abs. Building on our successful work in establishing screening platforms for TREM2-targeted small molecules, we hypothesize that small molecules can bind a novel druggable binding site in TREM2, consequently enabling therapeutic modulation of TREM2 interaction with both Gal-3 and A. We will test our hypothesis and attain our objective via the following specific aims: (1) screening the focused chemical using a panel of cell-free and cell-based assays, followed by hit-to- lead optimization, and (2) evaluation of the optimized leads in an AD mouse model following an assessment of their pharmacokinetic (PK) profiles. This research will lay the groundwork for the therapeutic modulation of TREM2 function using small molecules to develop new AD therapeutics.

NIH Research Projects · FY 2025 · 2024-06

Project Summary Tuberculosis (TB) is a leading killer among infectious diseases, and global progress against this pandemic has been set back by COVID-19. Major difficulties in combating TB are the need for months of multidrug therapy to ensure a high probability of relapse-free cure, and its ability to survive in the air as infectious aerosols and create a large pool of infected people. The requirement for prolonged drug treatment is likely due to the minority of bacteria that are able to survive antibiotics for a long duration, even in the absence of genetically-encoded resistance. Such bacteria are termed persisters. Similarly, transmission events rely on the subset of bacteria that are able to survive aerosolization stress until it reaches a new host. While recent strides have been made in understanding these subpopulations of cells, a way to interrogate their transcriptional states at a single-cell resolution in a high-throughput, unbiased manner does not exist. This limits understanding of the biology of these cells over time, and restricts identification of vulnerabilities that could be exploited to shorten TB treatment duration or reduce transmission. The aim of this proposal is to apply a method of single-cell RNA sequencing (sc-RNAseq) that allows for simultaneous comparison of the transcriptomes of thousands of cells of Mycobacterium tuberculosis (Mtb), the causative agent of TB. Preliminary data demonstrate that the protocol, which relies on in situ reverse transcription and barcoding with combinatorial indexing, is feasible in Mtb. In this application, the first aim is to test the limitations and characteristics of the technique in pre-defined mixtures of Mtb containing different plasmids. Over the first year of the grant, we will evaluate (1) the number of genes detected per cell, (2) the detection threshold of the smallest predefined subpopulation, and (3) the concordance of the aggregation of the single-cell transcriptome to bulk transcriptomes. With these parameters defined, the sc-RNAseq will be applied to biologically relevant in vitro conditions in the second year, including Mtb exposed to the front-line TB drug rifampin (RIF) and Mtb undergoing desiccation stress, a model of aerosolization. Here, the clusters of subpopulation transcriptomes found by the method will be compared with orthogonal, observable, and predictable phenotypes including cultivability and RIF resistance over time. These two aims will validate baseline characteristics of the assay, lay the foundation for leveraging this method against more complex biological samples in the future, and begin defining the fundamental biology of Mtb subpopulation behavior in response to stress.

NIH Research Projects · FY 2025 · 2024-05

PROJECT SUMMARY Kidney transplantation improves survival in patients with end-stage kidney disease. Immunosuppression, however, leads to infectious complications which cause significant morbidity and mortality in this fragile population. Infections are mostly diagnosed at the time of clinical presentation and clinical factors do not adequately anticipate infections. Predicting infectious complications is thus an important goal in kidney transplantation to prevent significant morbidity and mortality. The long term goal of this study is to develop comprehensive microbial biomarkers in the stool, blood, and the urine for diagnosing and predicting infections in kidney transplant recipients. Our objectives are: 1) Develop a fecal SCFA assay that predicts the risk for infections 2) Develop blood and urine cfDNA assays that can comprehensively diagnose infections as well as predict the risk for infections. These objectives are inspired by observations from several recent pilot studies: 1) We have found that the fecal abundance of short- chain fatty acid (SCFA) producing bacteria is associated with decreased future development of bacterial and viral infections 2) We have demonstrated that cell-free DNA (cfDNA) profiling in the blood and in the urine can simultaneously detect changes in the microbiome and virome over time and detect infections in transplant recipients. Importantly, with respect to cfDNA profiling, we have developed a novel technique called SIFT-seq to overcome the challenge of environmental contamination in low biomass specimens. By biochemically tagging the biological specimen prior to downstream DNA isolation, we can bioinformatically remove DNA introduced during sample preparation and accurately identify the microbiome and virome in low biomass specimens. In this study, we propose to recruit 300 kidney transplant recipients at the time of transplantation for serial fecal, urine, and blood specimen collections during the first year after transplantation. We will profile the gut microbiome using metagenomic sequencing to identify taxa at the species level and using metabolomic SCFA profiling. In addition, we will profile the blood and urine specimens using our novel technique, SIFT-seq, to identify the blood and urine microbiome and virome with high sensitivity and specificity. In Aim 1, we will determine the fecal bacterial and SCFA profiles that predict the risk of infections in kidney transplant recipients. In Aim 2, we will determine the blood and urine cfDNA profiles that are diagnostic and predictive of infections in kidney transplant recipients. Significance. Establishing microbial biomarkers as predictive of infections will allow for identifying kidney transplant recipients at high risk for infections and will allow for future clinical trials involving preemptive changes in immunosuppression, preemptive antibiotic/antiviral therapies, and/or potential gut microbiota-based therapies to prevent infections in these high risk individuals.

NIH Research Projects · FY 2026 · 2024-05

PROJECT SUMMARY/ABSTRACT: Glioblastoma (GBM), the most common malignant primary brain tumor, is the most aggressive diffuse glioma of the astrocytic lineage. Despite recent advances in multimodality therapy for GBM incorporating surgery, radiotherapy, chemotherapy, and supportive care, the overall prognosis remains poor, and long-term survival is rare. Immunotherapy has the potential to harness the immune system to kill brain tumor cells; however, the outcome of ongoing clinical trials with immunotherapies for GBM has been unsatisfactory. The highly immunosuppressive nature of GBM represents a key resistance mechanism to immunotherapy as glioma cells escape effective antitumor immunity by programming the tumor microenvironment (TME). The inability of immune-based therapies to cross the blood-brain barrier (BBB) represents another roadblock to the success of immunotherapy in the treatment of GBM. Therefore, there is an unmet need for the development of immunomodulators of the GBM TME that can cross the BBB. Chitinase-3 like-protein-1 (CHI3L1) is a key mediator of an immunosuppressive GBM microenvironment by reprogramming tumor-associated macrophages (TAMs). The binding of CHI3L1 to galectin-3 (Gal-3) selectively promotes TAM migration and infiltration with a protumor M2-like, but not an antitumor M1-like, phenotype in vitro and in vivo. Silencing CHI3L1 in syngeneic GBM mouse models results in increased tumor-infiltrating lymphocytes (TILs), tumor size reduction, and improved animal survival, which is reversed by the overexpression of CHI3L1. However, there are no drug candidates in existence that can block the immunosuppressive function of CHI3L1 in GBM. We aim to establish a new immunotherapeutic strategy for GBM based on targeting CHI3L1/Gal-3 interaction with small molecules, which may synergize with immune checkpoint inhibitors to effectively promote tumor regression for GBM patients. We hypothesize that small molecule-based blockade of CHI3L1/Gal-3 will reverse immune suppression and attenuate tumor progression in preclinical models of GBM. Our hybrid expertise in the development of small molecule immunomodulators, hit-to-lead optimization, translational drug discovery research, and immunopharmacology uniquely positions us to achieve this goal. We propose to perform optimization studies of a small molecule inhibitor of CHI3L1/Gal-3 interaction to develop optimized leads with single-digit nanomolar potency in cell-based assays (Aim 1). Subsequently, we will perform in vitro and in vivo pharmacokinetic (PK) evaluation of the optimized leads in order to select the top three lead compounds for further assessment of their therapeutic potential using two mouse models of GBM (Aim 2). The proximal expected outcome of this work is the introduction of clinically translatable small molecule inhibitors of CHI3L1/Gal-3 interaction that can be further implemented in combination therapy approaches to overcome the immunosuppressive nature of GBM.

NIH Research Projects · FY 2025 · 2024-05

Abstract. Alpha 1-antitrypsin (AAT) deficiency, an autosomal recessive disorder, manifests in lung as early onset panacinar emphysema. AAT functions to inhibit neutrophil elastase (NE) and other neutrophil serine proteases. AAT is produced predominantly in liver and diffuses into the lung from the circulation. Low AAT levels in AAT deficiency are associated with an imbalance between AAT and neutrophil-released proteases allowing slow destruction of the lung parenchyma. AAT deficiency is caused by mutation in the SERPINA1 gene; M alleles are normal alleles, Z (E342K) homozygotes account for >95% cases. Z AAT polymerizes in hepatocytes, limiting secretion, resulting in plasma levels 10-15% of normal. AAT inhibits serine proteases through its active site centered at methionine (M) 351 and 358. The active site methionines are modified by oxidants including cigarette smoke, air pollutants and endogenous oxidants from activated inflammatory cells, reducing AAT function. AAT deficiency therapy is currently treated with weekly infusions of AAT purified from human plasma; the infused AAT normalizes lung AAT levels, protecting alveoli from destruction. While AAT augmentation therapy reduces the rate of lung destruction, it is susceptible to oxidation, requiring excess “reserve” AAT to protect the lung. The focus of this proposal is to translate to humans gene therapy for AAT deficiency that circumvents both weekly requirements for protein therapy and the susceptibility of the therapeutic AAT to oxidation inactivation. We demonstrated that replacing M351 with valine (V) and M358 with leucine (L) on a normal M1 alanine (A)213 background provides anti-protease protection despite oxidant stress. One-time intravenous (IV) administration to mice of AAV8hAAT(AVL), a serotype 8 adeno-associated virus vector coding for the oxidation resistant variant hAAT(AVL) maintains high, dose-dependent anti-protease activity in serum and lung under oxidant stress compared with normal AAT. A toxicology study over 6 months in C57Bl/6 mice demonstrated that IV administration of AAV8hAAT(AVL) is safe. Based on the preclinical efficacy and safety data, we propose a phase 1 safety/dose ranging study, with IV administration of AAV8hAAT(AVL) at each of 3 doses to n=5 AAT Z homozygotes at each dose. The highest dose [2x1013 genome copies (gc)/kg] is ~ ½ log lower than the highest dose in the toxicity study. Aim 1, R61. Prepare and submit an Investigational New Drug package and gain approval from the FDA and other regulatory groups to initiate a Phase 1 clinical trial. Aim 2, R61. Optimize AAV8hAAT(AVL) production. Aim 3, R33. Manufacture clinical grade AAV8hAAT(AVL) for the Phase 1 safety/dose-ranging clinical trial. Aim 4, R33. Carry out a Phase 1 safety/dose-ranging clinical trial to determine the highest tolerable dose and preliminary biologic efficacy of AAV8hAAT(AVL) therapy for AAT deficiency.

NIH Research Projects · FY 2026 · 2024-05

PROJECT SUMMARY/ABSTRACT Tauopathies are a growing group of neurodegenerative disorders, including Alzheimer’s disease and mul- tiple related dementias, and are characterized by intracellular deposition of tau aggregates. Absence of tau mutations in the vast majority of tauopathy patients, and all of Alzheimer’s disease cases, highlights a critical need to understand why the tau protein fails to retain its native conformation, and forms pathogenic aggre- gates. The objective of the proposed research is to identify a common molecular mechanism underlying tau proteostasis, and to determine in vivo how this mechanism modifies tau aggregation and neurodegeneration: Chaperones are key mediators of proteostasis, and the chaperone protein Hsc70 is known to bind tau directly. However, the fate of Hsc70-bound clients is determined by co-chaperones, assembled into chaperone com- plexes. We have identified two chaperone-complexes that affect tau in diametrically opposite directions: CSPα/Hsc70/SGT/Hsp90 containing “foldase” complex and Hsj1/Hsc70/CHIP containing “degradase” complex. This study will test the hypothesis that tau protein levels, aggregation and neurodegeneration are regulated by these two chaperone machines/complexes: the foldase complex stabilizes tau and the degradase complex tar- gets tau for degradation, and tau pathology can be manipulated by these activities. We will test this hypothesis in vitro, in primary neurons, and in mouse models of tauopathies in vivo. Completion of these studies is ex- pected to identify two new molecular processes which affect tau aggregation, as well as to reveal how these proteostatic activities affect tau pathology in vivo. The proposed study is innovative, because two new func- tions of the foldase and degradase chaperone complexes on their new client tau are proposed, and because of the use of a comprehensive experimental approach, including: a) a new primary neuron tau aggregation model, b) viral approaches for CRISPR/Cas9-knockout of co-chaperones in primary neurons, c) stereotactic injections to virally regulate the co-chaperone levels in vivo, d) using patient brain samples to establish rele- vance of our findings to tauopathy in Alzheimer’s disease. The proposed study is significant because a) the concept of multifaceted proteostasis, where tau aggregation is affected by the equilibrium between “degradase” and “foldase” mechanisms, can be readily translated to other proteins related to Alzheimer’s dis- ease and related dementias (e.g. TDP43 or α-synuclein), b) understanding how neurons prevent tau from ag- gregating will establish novel targets for treating tauopathies, and c) understanding cellular proteostasis mech- anisms of misfolding-prone proteins may reveal clues common to the sporadic versions of tauopathies, includ- ing Alzheimer’s disease, for which tau mutations are not the most robust or common risk factor. This grant thus addresses the Notice of Special Interest NOT-AG-21-037: Common Mechanisms and Interactions in Neuro- degenerative Diseases.

NIH Research Projects · FY 2026 · 2024-05

PROJECT SUMMARY/ABSTRACT My career goal is to be an independent investigator in clinical trials of transcranial magnetic stimulation (TMS) focusing on the relationship between depressive symptoms and the blood-brain barrier (BBB). The central hypothesis of this proposal is that BBB function is significantly abnormal in treatment-resistant depression (TRD) and is associated with depressive symptoms, with region specificity that underwrites core depressive symptom domains, and are clinically improvable using TMS. The specific aims are to: 1) test if BBB abnormalities occur in specific symptom domains of TRD; and 2) test if TMS-induced BBB functional improvements explain symptom responses in TRD patients. To investigate these aims, the hypotheses are: 1) the TRD group will have specific BBB dysfunctions compared to a group of age-, sex-, and metabolically-matched controls, and will be correlated to core depressive symptom domains; and 2) BBB function, in subsets of patients, will predict core depressive symptom domain outcomes to accelerated TMS (aTMS) treatment a priori. Underlying regional BBB patterns will be tested with using measurements obtained before and after a course of aTMS. We propose that aTMS improves BBB function in the target region (dorsolateral prefrontal cortex or connected frontostriatal/limbic areas) and that the same regions will not reveal BBB changes similarly in non-responders. More broadly, my goal is to uncover BBB functions implicating a biological mechanism linked to TRD and improved by aTMS to build new, testable, mechanistic insights for future clinical trial work. This Mentored Patient-Oriented Research Career Development Award (K23) builds upon a firm background in BBB cellular and molecular biology and clinical expertise in psychiatry to bridge these areas in highly translational ways. Weill Cornell Medicine/New York- Presbyterian Hospital is a premier institution, with top-ranked clinical and research programs on which to support and build my career. The primary mentor, Dr. Conor Liston, is a psychiatrist who has made major scientific contributions to psychiatric neuroimaging and TMS research, detailing mechanisms and that have propelled the field forward. The co-mentor, Dr. Danny J. J. Wang, is an expert in MRI physics/engineering widely prized in the field, with key advancements to methods of BBB measurements used in this study. Critical progress to our understanding of BBB functioning in TRD, symptom domains, and its ability to be improved by aTMS will be known by study conclusion, along with a potentially detectable subtype of TRD most amenable to these findings and interventions.

NIH Research Projects · FY 2026 · 2024-05

PROJECT SUMMARY & ABSTRACT Candidate. Dr. Puja Chebrolu is a physician-scientist who has spent the past five years conducting research on HIV and diabetes in India. She has conducted longitudinal research, developed strong scientific collaborations with Indian scientists, trained an Indian research team, and authored 10 publications. She has documented that HIV increases the risk of gestational diabetes (GDM). She hypothesizes that HIV will also increase the risk of prediabetes (preDM) and type 2 diabetes (T2DM) development postpartum. Career Development Plan. Dr. Chebrolu's immediate and long-term goals are the following: 1) Increase her understanding of HIV-associated diabetes epidemiology and pathogenesis. 2) Increase her skills in advanced biostatistics for longitudinal data 3) Obtain training in the conduct of clinical trials 4) Develop professional skills including leadership and mentorship Dr. Chebrolu will develop these skills through coursework and implementation of her longitudinal study on time course and predictors of progression to postpartum preDM and T2DM in women living with HIV and GDM. Environment. The proposed research and training will take place at Weill Cornell Medicine (USA) and at Byramjee Jeejeebhoy Government Medical College (India). Dr. Chebrolu's mentors have expertise that encompass her goals. They have worked with her and each other previously. Research. Over data two-thirds of women who develop GDM will progress to T2DM within 10 years. suggests that HIV doubles the risk of GDM Dr. Chebrolu's , and is associated with increased risk of progression to T2DM. The impact of HIV on postpartum progression to preDM and T2DM is not known. Preliminary data suggests that South Asian women progress to T2DM faster than obese populations due to low baseline pancreatic beta cell mass. HIV increases beta cell stress through multiple mechanisms and may further accelerate the rate of progression to T2DM. Understanding progression its the impact of HIV on the time course and pathophysiology of T2DM will help inform the timing and type of interventions to prevent or delay progression to T2DM and devastating sequelae. Aim 1: Estimate the time to progression to postpartum prediabetes and T2DM in 180 women with a history of GDM (60 women with HIV and 120 women without HIV). We will test for prediabetes and T2DM at 5 timepoints over 2 years postpartum. This aim will test the hypothesis that hazard of progression to prediabetes or T2DM will be significantly higher in women with GDM and HIV compared to women with GDM and without HIV. Aim 2: Determine the effect of HIV on longitudinal beta cell function in women with a history GDM. The primary hypothesis is that women with HIV and GDM will have a greater rate of decline in pancreatic beta cell function over 2 years postpartum compared to women with GDM and without HIV.

NIH Research Projects · FY 2026 · 2024-05

PROJECT SUMMARY Glioblastoma (GBM) is a highly aggressive and incurable brain tumor. The inter-patient and intra-tumoral heterogeneity of GBM, resulting from genetic alterations and epigenetic plasticity, poses a major challenge in its treatment. GBM IDH-wt tumors are composed by different proportions of transcriptionally defined cell states, which although resemble neurodevelopmental cell types, are highly plastic and interconvertible -rather than hierarchic- as we and others have shown. This suggests the presence of a core regulatory logic that enables toggling among different transcriptional states, endowing GBM tumors with increased phenotypic plasticity and fitness. Here, we aim to unravel core regulatory modules that are critical for GBM programs with a particular focus on enhancers, which together with transcription factors govern cell-state specific programs. Enhancer dysregulation by genetic variants and epigenetic mechanisms is increasingly appreciated as a key process in oncogenic transformation and drug resistance. However, dissecting and modulating enhancer function remains very challenging due to the large number of putative enhancers and the complex ways they control their target genes in the context of the three-dimensional (3D) genome. By constructing 3D enhancer-promoter interaction networks in four patient-derived glioma stem-like cells (GSCs) and normal neuronal stem cells -as controls- we have identified a subset of GSC-specific hyperconnected enhancers, which we coin "3D regulatory hubs”. 3D hubs harbor genes with robust and coordinated transcriptional levels that enrich for oncogenic pathways and are associated with worse patient outcomes. Importantly, epigenetic perturbation of a highly-recurrent enhancer hub in GSCs resulted in concordant donwregulation of multiple hub-connected genes, leading into significant shifts in the transcriptional states and altered clonogenic and proliferation capacities. Building on this foundational work, we propose that de novo 3D regulatory hubs (established by genetic or epigenetic mechanisms) lie in the core of GBM networks, where they connect and control multiple target genes, resulting in non- linear effects on the transcriptional program and oncogenic behavior. To address this hypothesis, our interdisciplinary team will combine advanced chromatin topology assays, computational modeling and network analysis with state-of-the-art epigenetic engineering and proteomics tools as well as powerful ex vivo and in vivo functional assays. Specifically, we will pursue the following aims: (i) characterizing the inter-patient and intra- tumoral heterogeneity of 3D regulatory networks and identifying conserved structures across patients and states, (ii) predicting and targeting candidate central hubs and interrogating the molecular and functional consequences and (iii) uncovering critical players of hub organization and unique vulnerabilities. The findings will provide insights into enhancer-based reprogramming of cancer fate, opening new avenues for therapeutic targeting of GBM and establishing a paradigm for identifying and prioritizing key enhancers and regulatory factors in oncogenic programs. 1

NIH Research Projects · FY 2026 · 2024-05

Abstract Neuromodulatory GPCRs are critical regulators of neuronal activity that function within complex networks of accessory proteins which shape their signaling across space and time. Proteins in the potassium channel tetramerization domain (KCTD) family are important accessory subunits of the GABAB receptor, serving to regulate the intensity and duration of signaling in response to the inhibitory neurotransmitter, GABA. However, a mechanistic understanding of KCTD regulation of GABAB receptors is lacking and whether KCTDs regulate other neuromodulatory GPCRs is unknown. In aim 1, we will investigate GABAB receptor/KCTD coupling at the molecular level, using a combination of structural studies by cryo-electron microscopy and biophysical analysis in vitro. In aim 2, we will probe the cellular mechanisms of KCTD regulation of GABAB receptor signaling with functional studies by electrophysiology and fluorescence microscopy and proximity labeling mass spectrometry both in cell lines and primary neurons. In aim 3, we will build on preliminary data identifying the M5 muscarinic acetylcholine receptor as a novel KCTD target by characterizing this novel signaling complex using in vitro biochemical and biophysical techniques and live cell functional assays. Collectively, the results of this study will serve to develop a detailed mechanistic understanding of KCTD modulation of GPCR signaling as well as laying a foundation for investigation of newly discovered regulatory proteins and signaling components. Proposed technology development efforts may also find broader utility in other areas of molecular neurobiology, further enhancing the potential impact of this work.

NIH Research Projects · FY 2026 · 2024-05

Among individuals who survive ≥5 years from a cancer diagnosis, cardiovascular disease (CVD) is the leading cause of death. Indeed, adults with cancer have more than twice the risk of poor CVD outcomes as adults without cancer. This difference is partially due to the incidence of cardiotoxic effects from cancer therapies, but we hypothesize that upstream factors (e.g., area-level indicators of social context and individual socio-economic position) are also responsible for CVD outcomes in cancer survivors. We have shown that having ≥1 adverse socio-economic indicators increases an individual’s risk of coronary heart disease, stroke, and heart failure outside of cancer. However, underlying mechanisms of how upstream factors may lead to worse CVD outcomes in cancer survivors are not well understood. Area-level factors such as high school completion rates, unemployment rates, and public health infrastructure have policy implications and understanding their downstream influences on CVD can point the way to solutions to improve CVD health for all Americans. The objective of this proposal is to determine the role of area-level social context and individual socio-economic position on CVD outcomes among a community-based cohort of 8,000 adults with cancer. We seek to leverage a novel link between data from the REGARDS study to cancer registries from the Virtual Pooled Registry Cancer Linkage System (VPR-CLS). REGARDS is a national, prospective, longitudinal cohort study that recruited 30,239 English-speaking individuals, at least 45 years of age from the 48 contiguous US states in 2003-7 and follows participants today. The VPR, coordinated by the North American Association of Cancer Registries, includes 45 registries covering 95% of the U.S. The novel combination of data sources will leverage area-level social context measures and individual-level measures of socio-economic status, cancer biologic factors, health behaviors, self-rated health, and expert adjudicated CVD outcomes to determine among cancer survivors: 1) associations between multi-level socio-economic indicators and CVD outcomes; 2) the role of cancer biologic, behavioral, and psychosocial factors in the relationship between multi-level socio-economic indicators and CVD; and 3) the role of cancer and non-cancer health service use in the relationship between multi-level socio-economic indicators and CVD. Our long-term goal is to develop strategies to improve CVD health among all cancer survivors. We have assembled a multi-disciplinary team of experts in CVD and cancer epidemiology, health services research, cardiology, oncology, and biostatistics. Together, we will generate evidence to inform and develop interventions to support CVD health during cancer survivorship.

NIH Research Projects · FY 2026 · 2024-05

Project Summary Early life is a critical period for the proper development of the immune system. As mammalian hosts have co- evolved with their symbiotic bacteria, signals from the commensal microbiota are crucial for the prevention of allergy, inflammatory disorders, and protection against enteric pathogens. Our lab has recently uncovered that commensal antigens are trafficked to the thymus by dendritic cells, where they are presented to developing thymocytes. Rather than inducing deletion or Treg development, this results in an expansion of naïve microbiota- specific T cell populations, which are functional and egress to populate the peripheral tissue. This process is limited to early life, suggesting that it is critical for the establishment of the commensal-reactive T cell repertoire. Analysis of thymic 16S sequencing from mice at weaning reveals a stark enrichment of select taxa, such as members of Escherichia. When considering the vast diversity of the intestinal microbiome, these data imply an antigenic filter through which only select microbes are trafficked to the thymus. This proposal seeks to understand how the unique environment of the early-life intestine promotes commensal ferrying to the thymus, and the microbial and host factors that curate specific taxa for sampling by sub-epithelial dendritic cells. My central hypothesis is that this process has evolved to prioritize the generation of naïve T cell clones capable of recognizing antigens associated with epithelial invasion. We posit that the signals promoting commensal antigen trafficking to the thymus are 1) adherence to the intestinal epithelium and 2) opsonization by maternal IgG. As recent publications have identified commensal-specific IgG as targeting microbes with the ability to translocate, both of these signals indicate the capacity for epithelial invasion – whether defined by the neonate or by maternal cues. Preliminary data indicate that non-adherent commensals are significantly underrepresented in the thymic tissue. I have also found increased expression of the IgG receptor FcRn in migratory thymic DCs in early life, indicating their enhanced ability to recognize and process IgG-opsonized commensals. In Aim 1, I will quantify the epithelial adherence of a model commensal E. coli throughout early development. I will genetically target adhesins in this commensal to determine how these factors impact epithelial adherence and thymic trafficking in vivo. I will utilize 16S sequencing to compare the mucosa-associated microbiome with that of the thymus. Aim 2 will focus on the role of maternal IgG opsonization, and will quantify thymic trafficking in B cell- and antibody- deficient mouse models throughout early life development. I will manipulate maternal IgG titers through transfer of commensal-specific IgG into IgG-deficient dams, and the impacts on thymic trafficking in offspring will be examined. The proposed experiments will reveal crucial elements of how the commensal-reactive T cell repertoire is curated in early life, and will have important insight into how the microbiome affects the development of the mucosal immune system.

NIH Research Projects · FY 2026 · 2024-04

PROJECT SUMMARY/ABSTRACT: Despite the availability of current biologics, such as anti-tumor necrosis factor (anti-TNF), anti-integrins, anti-interleukins, and small molecules such as tofacitinib, the rates of primary and secondary treatment failure remain high in inflammatory bowel disease (IBD). This highlights the unmet need for the identification of rational combinations of therapies for IBD with complementary mechanisms of action. Dual targeting of cluster of differentiation 28 (CD28) and inducible T cell costimulation (ICOS), closely related costimulatory molecules that play partially overlapping roles in the pathogenesis of IBD, has revealed remarkable success as a potential therapeutic strategy for IBD as well as other inflammatory diseases. However, dual CD28/ICOS blockade is currently restricted to protein-based therapeutics associated with unwanted immunogenicity and an increased risk of adverse events (AEs). In comparison to therapeutic proteins, small molecules will minimize the immunogenicity risk and enable better management of AEs based on their amenability for pharmacokinetic optimization. However, there are no small molecules in existence that target ICOS or CD28. To fill this gap, we have developed an innovative platform, Small Molecules from Antibody Pharmacophores (SMAPs), that can identify small molecule inhibitors of immune cell receptors with high binding affinity and selectivity. Our SMAPs platform is based on utilizing cocrystal structures of immune cell receptors with antibodies (Abs) in building pharmacophore maps from clusters of key interacting residues of Abs with immune cell receptors to identify small molecules that function as Ab-mimetics. Building on our successful work in drugging immune cell receptors with small molecule inhibitors, we propose to perform preclinical validation of small molecule-based dual CD28/ICOS inhibition as a therapeutic strategy for IBD based on small molecule CD28 and ICOS inhibitors identified from the SMAPs platform. We hypothesize that small molecule-based dual CD28/ICOS inhibition will result in improved therapeutic outcomes in preclinical models of IBD in comparison to single- targeted CD28 and ICOS small molecules. We will test our hypothesis and attain our objective via the following specific aims: (1) optimization of small molecule CD28 inhibitors, followed by validation using biophysical screening and cell-based assays, (2) assessment of the pharmacokinetic profiles of the optimized CD28 leads, followed by preclinical validation of small molecule-based dual CD28/ICOS blockade using clinical samples from IBD patients as well as a mouse T cell transfer model of chronic colitis. Collectively, the proposed investigations have the potential to identify clinically translatable CD28 inhibitors that can be used in combination therapy for IBD with small molecule ICOS inhibitors. Successful completion of this work will enable the initiation of Investigational New Drug (IND)-enabling studies for the optimized therapeutic leads.

NIH Research Projects · FY 2026 · 2024-04

Project Summary Approximately .5% of children are born missing a limb, and 2.1 million in the US alone are living with limb loss. Current management for these patients largely relies on the use of prostheses, however most prosthetic use is marked by significant patient dissatisfaction due to functional limitations. Only regeneration of an intact limb can offer optimal function and quality of life for these patients. While it is well known that select non- mammalian organisms have remarkable capabilities for complete limb regeneration, it is sometimes overlooked that similar capacity for regeneration, at least in latent or incomplete form, exists in mammals including humans and mice. Digit tip amputation in both humans and mice can lead to blastema formation and composite regeneration of the digit tip. Understanding the mechanism of these mammalian limb regenerative responses is a necessary step to inducing or extending these responses to promote limb regeneration. However, methods to achieve this goal have been elusive, in part because the specific stem cells mediating regenerative blastema formation and organizing the regenerative response were unknown. Recently, we have identified a new stem cell forming the skeleton that is specialized for skeletal healing responses, and we find that these stem cells are the cell of origin of the blastema forming after digit amputation, undergoing injury induced plasticity that then confers upon them the capability to not only mediate bone regeneration, but to more over orchestrate a composite tissue regeneration program. We further find that when this specific stem cell is harvested from blastema tissue and transplanted to a site of tibial amputation, they the capacity to mediate true limb regeneration at this new site, thereby allowing initiation of a true limb regeneration response to be initiated at any amputation site. Here, we will develop these observations into a new approach for limb regeneration, first (Aim 1) we will determine how this stem cell acts in the blastema forming after digit tip amputation orchestrate multi-tissue regeneration and how these cells are transcriptionally reprogrammed in the blastema to acquire the capacity to orchestrate regenerative responses. Next (Aim 2), we will further develop the method of blastema stem cell implantation into amputation sites to initiate regeneration, evaluating whether the true multi-tissue regeneration occurs at the transplantation site, and whether new joint tissue can be patterned in the transplanted cells, thereby allowing for true limb reconstruction, including joints. Ultimately, this proposal will develop key initial elements of a new approach for the regeneration of amputated limbs, by transducing a specific population of autologous skeletal stem cells to induce a blastema-like state, and then implanting them into the amputation site to initiate regeneration of the missing limb.

NIH Research Projects · FY 2025 · 2024-04

Project Summary BRCA1 and BRCA2 are tumor suppressor genes frequently inactivated in cancer, particularly in ovarian and breast cancers. Despite recent advances in therapy, many patients with BRCA1/2-deficient cancers eventually die of the disease and, therefore, new treatment options are needed. The goal of this proposal is to develop a novel targeted therapy against BRCA1/2-deficient cancers that combines high doses of tetrahydrofolate (THF) and vitamin C (VC) to generate metabolic genotoxins in cancer cells. The rationale of the proposed strategy stems from the recent discovery that high doses of THF, the active form of folate, oxidize generating the potent crosslinker formaldehyde (FA), which kills cells deficient in DNA crosslink repair such as BRCA1/2-mutant cells. In addition, preliminary data show that vitamin C (VC) promotes the selective killing of BRCA1/2-deficient cells by THF. VC and THF synergize at generating reactive oxygen species (ROS), suggesting formaldehyde and ROS generation might drive the cytotoxic effects in BRCA-mutant cells. In contrast, VC reduces THF toxicity in normal tissues in mice, which allows the administration of higher doses of THF. Furthermore, treatment with THF and VC in combination prolongs the survival of mice with BRCA1-mutant tumors. Based on these findings, this proposal aims to: (1) Understand the mechanism of selectivity of the vitamin combination against BRCA1/2- deficient cells, by using isogenic cell lines, metabolic tools, DNA damage assays and molecular and cell biology techniques. (2) Characterize the interaction between THF and VC in mice, by employing metabolomics, ultrasensitive mass-spectrometry analysis of DNA adducts, immunohistochemistry and infrared microscopy. (3) Assess the efficacy of THF/VC therapy against BRCA-mutant tumors in mice, by using orthotopic syngeneic models of ovarian and breast cancer and measuring the impact of THF/VC in tumor growth and overall survival. By using state-of-the-art tools and cancer models, this project could provide the foundations of a novel therapeutic strategy for the treatment of BRCA1/2-mutant cancers. In addition, it could support an unconventional approach of harnessing metabolism for cancer therapy that manipulates metabolism to generate metabolic genotoxins in cancer cells.

NIH Research Projects · FY 2026 · 2024-04

Project Summary Low-value care (LVC), defined as care where harms or costs outweigh the benefits, is common and costly. For patients, LVC can result in physical harm from the service itself, unnecessary follow-up tests/procedures, and needless out-of-pocket costs. Patients with coronary artery disease (CAD) are at high risk of receiving LVC: studies estimate that up to 50% of tests and 15% of procedures performed on patients with CAD may be LVC. To identify and reduce LVC, accurate measurement is critical. Despite widespread adoption of administrative claims-based measures to assess LVC, most measures have not been tested against the “gold standard” of chart review (i.e., they do not have good criterion validity). Using inaccurate LVC measures undermines quality reporting and leads to inaccuracies in understanding the frequency, costs, and trends in use of LVC services; a critical knowledge gap in understanding which LVC services should be prioritized for reduction. Bundled payment models (BPMs), which pay organizations a fixed price for a 90-day episode of care such as elective percutaneous coronary intervention (PCI), could reduce LVC but could also have unintended consequences. Because BPMs provide a fixed payment to providers independent of patient risk, BPM providers might perform more PCIs on low-risk patients for whom medical therapy alone is the guideline- recommended treatment (i.e., they may perform more LVC PCIs). BPM providers might also perform fewer appropriate PCIs for high-risk patients, or those perceived to be high-risk such as racial/ethnic minorities. To assess the impacts of the elective PCI BPM, we will use data from the National Cardiovascular Data Registry, which contains a reliable measure to classify PCIs as LVC or appropriate using clinical chart review data. We have assembled an experienced team of cardiologists, economists, and outcomes researchers. In prior and pilot work we have validated 3 claims-based LVC measures, but also found that 1 measure was not valid. In Aim 1, we will use chart review as a “gold-standard” to assess the validity of commonly-used claims-based LVC measures relevant for CAD patients. In Aim 2, we will use validated LVC measures to assess LVC frequency, costs, and trends among a national sample of Medicare beneficiaries with CAD, and provide policymakers with a priority list of LVC services to target for reduction. In Aim 3, we will conduct a difference-in- differences analysis to assess the impact of elective PCI BPMs on a) LVC PCIs and healthcare disparities for appropriate PCIs using detailed clinical data from the NCDR, and b) other validated LVC services using linked Medicare claims. For patients with CAD, this research will 1) establish valid LVC measures which are needed for quality improvement and accountability programs, 2) guide efforts to reduce LVC services, and 3) evaluate the effect (and potential unintended consequences) of a policy intended to reduce LVC.

NIH Research Projects · FY 2026 · 2024-04

Project Summary/Abstract α-Actinin-4 (ACTN4) is a force-sensitive actin-binding protein that interacts with F-actin via a catch bond, which is defined as a protein-protein interaction that strengthens under tensile force. The K255E ACTN4 variant, which causes focal segmental glomerulosclerosis (FSGS), exhibits ultrahigh F-actin binding affinity and is rendered force-insensitive because it cannot catch bond with F-actin. Although actin-binding proteins have well- established mechanosensing functions, force-dependent conformational changes in actin-binding proteins have never been visualized. Our central hypothesis is that K255E ACTN4 binds F-actin in a constitutively strong state that structurally resembles the force-stabilized state of wild-type ACTN4. The long-term goal of this research is to elucidate structural mechanisms underlying deficient actin mechanosensing in hereditary podocytopathies. This project’s immediate objective is to determine the structural basis for actin mechanosensing by ACTN4 via in vitro force reconstitution assays, total internal reflection fluorescence microscopy, and cryo-electron microscopy (cryo-EM). In Specific Aim 1, a structural mechanism for deficient actin mechanosensing in FSGS caused by ACTN4 mutations will be revealed via a cryo-EM structure of the force-insensitive mutant K255E ACTN4 bound to F-actin. In Specific Aim 2, a structural mechanism for force-stabilized actin-binding by ACTN4 will be revealed via cryo-EM structures of wild-type ACTN4 bound to F-actin with and without mechanical force. Overall, this project will: (1) advance our insights into the structural basis for actin mechanosensing by ACTN4 and lack thereof in familial FSGS, (2) provide the first structure of a catch bonding protein under force, and (3) pioneer methods for introducing physiological mechanical forces in a manner amenable to cryo-EM. Dr. Gregory M. Alushin, PhD, an expert on cryo-EM studies of actin mechanobiology who practices active mentorship, and Dr. A. James Hudspeth, MD, PhD, an expert on molecular motors and mechanotransduction who has a proven track record of training successful physician-scientists, are co-sponsoring this proposal. The research will be conducted at The Rockefeller University within the deeply supportive Weill Cornell/Rockefeller/Sloan Kettering Tri-Institutional MD-PhD Program. This fellowship constitutes an important career milestone for dual-degree students seeking to become independent investigators.

NIH Research Projects · FY 2026 · 2024-04

Project Summary/Abstract The objective of this proposal is to utilize quantitative susceptibility mapping (QSM) to establish a clinically feasible, accurate, and robust treatment biomarker for chronic inflammation in multiple sclerosis (MS). Chronic active lesions (CALs) characterized by persistent smoldering inflammation significantly contribute to progressive cognitive and ambulatory decline in MS. Within a subset of CALs, a distinct subgroup has been identified as paramagnetic rim lesions (PRLs), where iron-laden pro-inflammatory immune cells are localized at the edge of the lesion. Leveraging the quantitative capabilities of QSM, we have made a significant observation regarding the inflammatory trajectory of PRLs. These lesions exhibit an initial increase in susceptibility, followed by a subsequent decrease, signifying the transition from a chronic active to a chronic inactive MS lesion. Notably, our findings have also indicated that QSM can detect the longitudinal susceptibility changes in PRLs resulting from the administration of current disease modifying therapies. Building on our promising preliminary data, our primary objectives are as follows: 1) develop a high-resolution, clinically feasible approach to accurately detect PRLs and identify their iron source; 2) provide in vivo validation for precise detection and quantification of inflammatory changes within individual PRLs; 3) identify existing disease- modifying therapies that target innate immune burden within lesions and 4) establish the positive impact of reducing inflammation in PRLs on clinical disability. To achieve these objectives, we have developed the Learned Acquisition and Reconstruction Optimization (LARO) approach, enabling rapid high-resolution QSM for PRL detection. We have also implemented a QSM post-processing method to eliminate the confounding effect of myelin, thereby enhancing rim iron detection sensitivity and quantification accuracy. In Aim 1, we hypothesize that these advancements in QSM technology will enhance PRL detection. Similarly, in Aim 2, we anticipate that our new approach will enable accurate quantification of the longitudinal inflammatory trajectory in PRLs, which we will validate through longitudinal in vivo monitoring of inflammation using TSPO PET. By building upon our preliminary work, in Aim 3, we will identify and evaluate current therapies that can effectively benefit patients with PRLs lesions, thereby advancing QSM as a treatment biomarker to target innate immune activity within chronic lesions. The overall impact of this proposal is to provide an innovative treatment target for chronic innate immune activity in MS, resulting in a paradigm shift in the care of patients with MS.

NIH Research Projects · FY 2026 · 2024-04

Prostate cancer (PCa) shows a striking degree of clinical variability, with most aging men harboring indolent, low risk PCa that will not threaten their health during their natural lifetime. Definitive treatment for these indolent, low risk cancers with surgery or radiation therapy (RT) risks unnecessary cost and treatment-related toxicity. Because of this, active surveillance (AS), a management strategy that avoids or defers treatment, has emerged as a standard of care for low risk PCa. AS consists of monitoring low risk, clinically insignificant PCa patients, with treatment with curative intent ONLY with progression to clinically significant PCa (csPCa). Determining when a patient progresses on AS consists a variable mix of serial prostate specific antigen (PSA) measurements, imaging, and prostate biopsies . However, no consensus exists as to the ideal set of tests and monitoring frequencies during AS. Prostate biopsy itself has measurable morbidity and is associated with infection, urinary retention, pain and worsening of urinary symptoms. There is therefore a need for optimization of AS protocols and to improve the detection of csPCa for men on AS. Prostate imaging with magnetic resonance imaging (MRI) has become an integral tool in the diagnosis of PCa and monitoring of men on AS as it improves the detection of clinically significant PCa compared to prostate biopsy without MRI targeting, however accuracy is limited. A negative MRI still misses approximately 20% of csPCa and a positive MRI will result in a negative biopsy 50% of the time. Better imaging is required to improve the detection of csPCa. Prostate Specific Membrane Antigen Positron Emission Tomography-Computed Tomography (PSMA-PET CT) is the most sensitive technique available for detection of metastatic prostate cancer. The accuracy of PSMA-PET CT in localized disease is limited; however, it has been shown that PSMA-PET CT adds diagnostic value to prostate MRI for detection of csPCa, specifically improving negative predictive value. We hypothesize that PSMA-PET CT in AS patients will improve diagnostic accuracy of prostate MRI such that a negative study will obviate the need for surveillance prostate biopsy. The Evaluation of Prostate Specific Membrane Antigen Positron Emission Tomography-Computed Tomography in Active Surveillance for Prostate CancEr (ESCAPE) trial is a single arm, prospective multi-institutional clinical trial assessing the negative predictive value of PSMA PET CT in detecting clinically significant prostate cancer. We hypothesize that PSMA-PET CT can rule out clinically significant prostate in AS patients due to its high negative predictive value.

NIH Research Projects · FY 2026 · 2024-04

PROJECT SUMMARY Given the recent SARS CoV2 pandemic, and monkeypox outbreak, there is an urgent unmet need to understand immunity in our barrier tissues (e.g. skin, lung, and gut) where viruses are encountered. Tissue specific memory is needed for long-lived protective immunity, including immunization strategies that target infections and cancers of the tissue. T resident memory cells are long-lived memory populations generated by infections, cancers, and vaccines. Tissue resident memory T cells maintain long-term protective immunity to re-encountered pathogens (which include CoV2 and influenza in lung, and herpes, monkeypox, and smallpox in skin). Tissue resident memory T cells also survey against primary cancers and metastases. However, in pathogenic contexts tissue resident memory cells drive autoimmune memory recall. This proposal tests intervenable regulatory mechanisms when skin-specific T resident memory cells are formed, maintained, and governed, and a regulatory axis with local tissue Dendritic Cells. Our goal is foundational: to understand the basic principles by which barrier immunity, and T cell receptor repertoire is generated and shaped in the tissues, like skin. We apply our findings to important and relevant in vivo mouse models for infection and test the consequences for tissue inflammation, autoimmunity, and protective memory recall. Establishing a mechanistic groundwork and robust preclinical modeling is needed to later test interventions. This work is also likely to offer basic insight into the foundational mechanisms by which specific immune-modulatory drugs drive tissue-specific toxicities in the skin and other peripheral organs of patients, known as immune-related adverse events (irAEs).

NIH Research Projects · FY 2026 · 2024-03

Project Summary/Abstract The goals of this K24 application are for Dr. Jennifer Downs to train physician-scientists in the conduct of patient-oriented research on schistosomiasis and women's reproductive health in Tanzania and Lebanon, and to conduct a new study on human papillomavirus (HPV) persistence in women with schistosome infections. The candidate has spent the past 16 years conducting research in Tanzania on: (1) the female genital tract and gut mucosal effects of schistosome infections and how these affect host immune response to viral infections and vaccines, and (2) implementation science to improve women's reproductive health in resource limited settings. She has also established longstanding partnerships with a leading research organization (MITU) and medical school (Weill Bugando School of Medicine) in Tanzania. These partnerships, and her ongoing research studies, provide an outstanding research environment to study schistosomiasis and women's reproductive health while providing mentorship to the next generation of patient-oriented investigators. The training that Dr. Downs offers leverages ongoing funded research in Tanzania in Aims 2-4 (2 NIH R01s and foundation awards) and a newly funded women's reproductive health research project among Syrian refugees in Lebanon (Aim 5). United States (US) trainees from Weill Cornell will be supported by NIH research training grants and spend >50% of their time at their international sites in Tanzania or Lebanon. The projects include: 1. HPV persistence and cervicovaginal dysbiosis in women with schistosome infections 2. Genital mucosal, immune, and viral effects of female genital schistosomiasis 3. Effects of Schistosoma mansoni on the gut mucosal response to oral poliovirus vaccine and gut immune cell populations 4. Implementation science to promote women's reproductive health through religious institutions 5. Implementation science to develop a mobile reproductive health intervention for Syrian refugee women Research training will focus on a diverse pool of clinical trainees ranging from medical students to junior faculty. Dr. Downs has a strong track record of mentoring patient-oriented researchers. She founded and leads the Weill Cornell Women in Global Health Research Initiative. Her trainees will participate in this longitudinal mentorship initiative and work in US-international partnerships on research projects in Tanzania (Aims 1-4) and Lebanon (Aim 5). These 5 projects offer robust clinical, translational, and implementation science training. The K24 award will enable Dr. Downs to decrease her administrative responsibilities and commit 50% effort to research and mentorship. She will seek formal training in mentorship and strengthen her skills in HPV research, working towards her long-term goal of building a world-class research and training program to improve care for schistosomiasis and women's reproductive health among the world's poorest populations.

NIH Research Projects · FY 2026 · 2024-03

PROJECT SUMMARY / ABSTRACT Aberrant development of the prefrontal cortex (PFC), including the axonal connections it receives, are hallmarks of neurodevelopmental and neuropsychiatric disorders (NDDs). Foundational studies have shown that axons that project to the PFC, particularly from thalamic nuclei, influence PFC development and function. Notably, these axons are often disrupted in NDDs. We hypothesize that aberrant thalamocortical connectivity seen in these disorders results from abnormal axon pruning, which in turn leads to deficits in PFC maturation and function. To test this hypothesis, I will use a novel mouse model to inhibit axonal pruning in this circuit and determine its role in PFC development. I will use a combination of neural tracers, genomic, and molecular analysis to quantify axon pruning within the thalamocortical circuit to determine whether its disruption in our mouse models affects critical aspects of PFC development including lamination, neuronal number, interneuron migration, and electrical activity. This proposal, undertaken with the intellectual support of the Tri-Institutional research community and guidance of a mentorship team, will address a significant gap in understanding of how axon refinement regulates normal cortical development and can lead to cortical dysfunction seen in NDDs.

NIH Research Projects · FY 2026 · 2024-03

Project Summary Pancreatic beta cells (β-cells) produce and secrete insulin in response to acute elevations in blood glucose. Diminished β-cell insulin production and secretion deregulate whole-body glucose homeostasis and are hallmarks of Type 2 Diabetes (T2D). Endoplasmic Reticulum (ER) stress and Unfolded Protein Response (UPR) activation are required steps in β-cell dysfunction. Activating Transcription Factor 6 alpha (ATF6α) is a UPR-effector protein that helps increase cellular tolerance to stress by inducing expression of genes involved in ER function. Based on extensive literature documenting the beneficial role of ATF6α, we hypothesized in vivo β-cell ATF6α activation would protect mice against diabetes. Using destabilized domain technology provided by Luke Wiseman’s lab at the Scripps Institute, the Alonso lab generated knock-in mice (C57BL/6) expressing the N-terminal fragment of human ATF6α fused to a destabilized variant of E. coli Dihydrofolate Reductase (DHFR-ATF6α). Upon constitutive expression, the destabilized DHFR domain directs DHFR-ATF6α towards proteasomal degradation until exposure to the pharmacologic chaperone, Trimethoprim (TMP), which stabilizes DHFR and turns on DHFR-ATF6α transcriptional activity. These mice allow cell-type-specific (via Cre recombinase) and temporal (via TMP) activation of ATF6α in β-cells. Surprisingly, contrary to our hypothesis in vivo β-cell DHFR-ATF6α activation caused marked glucose intolerance during an intraperitoneal (i.p.) glucose challenge. There was no loss of insulin responsiveness and β-cell mass remained intact, suggesting β-cell DHFR-ATF6α activation causes insulin insufficiency. Indeed, our preliminary data indicate loss of glucose stimulated insulin secretion in vivo and a reduced islet insulin content. Intriguingly, loss of insulin secretion preceded reduced islet insulin content, suggesting β-cell DHFR-ATF6α activation impacts these processes by distinct molecular mechanisms. In Aim1, I propose to examine the impact of in vivo β-cell DHFR-ATF6α activation on insulin production; specifically, proinsulin production, proinsulin processing, and insulin granule maturation using transmission electron microscopy. In Aim 2, I propose to examine the impact of in vivo β-cell DHFR-ATF6α activation on the insulin secretion pathway; specifically, glucose uptake, glucose metabolism, ATP production, cAMP signaling, ATP-sensitive potassium channel closure, calcium induced insulin release, and cortical actin remodeling, using the gold- standard islet perifusion assay. These aims will contribute valuable knowledge for developing therapies that help preserve and/or rescue β-cell function in T2D. ATF6α is a key component of the chronic stress response that is thought to contribute to β-cell dysfunction. Understanding the impact of continuous, inappropriate β- cell ATF6α signaling may be useful for uncovering novel molecular mechanisms that explain one part of the damaging effect of chronic stress that precedes diminished β-cell insulin production and secretion in T2D.