Sloan-Kettering Inst Can Research

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY/ABSTRACT T helper (Th) cell differentiation into specialized lineages is critical for effective immune responses against pathogens, but dysregulated Th differentiation can contribute to autoimmune diseases. Emerging evidence suggests that metabolism can influence Th cell development and function, but which metabolites and metabolic pathways are involved remains poorly understood. Somatic mutations in the isocitrate dehydrogenase (IDH) enzymes contribute to the pathogenesis of acute myeloid leukemia (AML) and other malignancies via production of the ‘oncometabolite’ D-2-hydroxyglutarate (D-2HG). D-2HG blocks differentiation of malignant cells by inhibiting alpha-ketoglutarate (⍺KG)-dependent enzymes that regulate chromatin structure and gene expression. Recent reports suggest that D-2HG can influence the balance of Th17 and Treg differentiation, but the findings are contradictory and inconclusive. Whether CD4+ cells lacking IDH mutations produce physiologically relevant levels of D-2HG remains unclear. This raises the question as to whether analogous metabolic pathways might influence physiologic Th cell differentiation. Intriguingly, 2HG is a chiral molecule that can exist in either the D- or L- enantiomer. Although cancer-associated IDH mutants exclusively produce D-2HG, our lab and others have shown that virtually all normal and malignant cells produce L-2HG in response to hypoxia. Hypoxia-induced L- 2HG potently inhibits ⍺KG-dependent enzymes, including chromatin modifiers and regulators of hypoxia- inducible factor (HIF) stability. Nonetheless, physiologic sources and functions of L-2HG remain poorly understood. I discovered that activated Th cells produce L-2HG and that L-2HG levels are enhanced by hypoxia and hydrogen sulfide (H2S), microenvironmental factors that are prominent within the intestine. Furthermore, I found that genetic manipulations to elevate endogenous L-2HG in Th cells can promote T helper 17 (Th17) and impair regulatory T cell (Treg) differentiation. Thus, I hypothesize that CD4+ helper T cells produce L-2HG in response to specific microenvironmental cues to fine-tune the balance of Th17 and Treg cells and enhance inflammatory responses. I will rigorously test this hypothesis in two Specific Aims. Aim 1 will dissect the molecular mechanisms by which L-2HG regulates the balance of Th17 and Treg cells in vitro. In this aim, I will use novel genetically engineered mouse models (GEMM) that allow for tissue-specific manipulation of endogenous L-2HG levels to determine the effects of L-2HG on metabolism, chromatin structure, and gene expression. Aim 2 will elucidate how both T cell-intrinsic and microenvironmental L-2HG regulate CD4+ T cell- mediated inflammatory immune responses in vivo. Here, I will use novel GEMM to elevate and deplete L-2HG in Th cells or intestinal epithelial cells in models of intestinal bacterial infection and autoimmune colitis. The proposed studies will provide important insights in the metabolic control of Th cell differentiation and potentially expose novel therapeutic strategies to boost defense against pathogens or treat autoimmune diseases.

NIH Research Projects · FY 2024 · 2024-08

Project Summary Skin cancers are the most common cancers in the U.S and often require biopsy for histopathology confirmation. However, ~70% of biopsies yield benign findings, representing an unnecessary healthcare burden. Biopsies are also invasive and associated with complications such as bleeding, infections, and scars, and histopathologic analysis is time-consuming, delaying treatment. Confocal microscopy (CM) images skin at cellular resolution. Two CM techniques are used to evaluate skin lesions: reflectance CM (RCM), which is performed noninvasively on intact skin, and ex vivo CM (EVCM), which allows rapid imaging of fresh ex vivo tissues without time-consuming tissue processing. RCM has better sensitivity (90–94%) and specificity (82–85%) for skin cancer diagnoses compared with dermoscopy and reduces benign biopsies by ~75%. Likewise, EVCM can detect residual carcinoma in surgical margins with high sensitivity of 96.6%% and specificity of 89.2%. These images are challenging for novices to read, posing a major barrier to their clinical adoption. This is compounded by a lack of experts and teaching resources in this field. To overcome this barrier, we have created a CME-accredited Annual Workshop on Confocal Microscopy for Cutaneous Diagnostics at Memorial Sloan Kettering Cancer Center (MSK). While the workshop focuses on CM, we will also familiarize attendees with advances in CM, including multimodal techniques such as full-field optical coherence tomography (FF- OCT), line-field confocal (LC-OCT), and RCM-OCT. The 2024 two-day workshop will be presented in person and virtually. We have gathered a world-renowned faculty with expertise in this field and anticipate global participation based on our previous 6 workshops. The workshop will be interactive, including lectures, hands- on image acquisition, case discussions, and competitive quizzes. R13 funds will be used to provide scholarships to attendees from underrepresented groups, women, minorities, and individuals with disabilities to attend the workshop and undertake an observership at MSK. We will further promote participation by these groups through our marketing and outreach efforts and by ensuring physical and virtual accessibility. The workshop’s learning objectives are to: (a) familiarize with underlying optical principles and associated terminology, (b) recognize features of neoplastic and inflammatory skin lesions, (c) develop skills in acquiring images, (d) understand to integrate CM in clinical and surgical workflows, and (e) gain familiarity and applications of novel multi-modal devices. The overall goal of the workshop is to encourage clinicians, especially those from underrepresented backgrounds, to integrate CM and multimodal devices into clinics.

NIH Research Projects · FY 2025 · 2024-08

Summary Antitumor immunosurveillance by cytotoxic lymphocytes is generally conceived as a biochemical process in which tumor specific markers are recognized by activating receptors on the lymphocyte surface. We have found, however, that cytotoxic T cells and natural killer cells also respond to the mechanical properties of cancer cells, preferentially destroying targets that are physically stiffer. This mechanical form of immunosurveillance, which we call mechanosurveillance, appears to be particularly relevant during metastasis, when cancer cells remodel their cytoskeleton to invade new organs. In this proposal, we will investigate the interplay between mechanosurveillance and the physical properties of the metastatic microenvironment. Cellular mechanics are modulated continuously by cell-extrinsic biophysical signals. A particularly important manifestation of this crosstalk, called mechanoreciprocity, induces cells in stiffer environments become stiffer themselves, and those in softer locales to become softer. Whether environmentally-induced stiffening might sensitize cancer cells to mechanosurveillance in vivo, however, has not been explored. This an interesting question because metastatic microenvironments vary widely in their physical properties, ranging from very rigid (e.g. bone) to very soft (e.g. lung). Enhanced mechanosurveillance in rigid microenvironments would establish a regime in which cytotoxic lymphocytes control the spectrum of metastatic site preference by disproportionately suppressing outgrowth in organs like the bone. Using a mouse model of metastasis, we have found that cancer cells colonizing the bone are significantly stiffer than cancer cells colonizing the lung, and that the in vivo expansion of bone metastasis is exquisitely sensitive to cytotoxic lymphocytes. Building on these preliminary observations, we propose that microenvironmental stiffness dictates the efficacy of mechanosurveillance and that this relationship shapes both metastatic site preference and the power of anti-tumor immunotherapy. We will investigate this hypothesis in three Specific Aims. Aim 1 will examine how distinct metastatic microenvironments affect cancer cell biomechanics and immune vulnerability in mice and humans. Aim 2 will determine if environmental stiffness can, as an independent variable, control the efficiency of mechanosurveillance. Finally, Aim 3 will apply state-of-the- art rigidity dependent cell sorting technology to identify novel mechanoregulators of metastasis in vivo. Our proposed studies are organized around the conceptually innovative idea that crosstalk between environmental mechanics and cellular cytotoxicity determines where metastases grow. In addition, we will employ highly innovative technologies, including suspended microchannel resonator (SMR) devices that rapidly measure cell deformability and sort cells based on stiffness. The successful completion of our Specific Aims could identify biomarkers for guiding antitumor immunotherapy and aid development of novel strategies for treating metastatic growth in specific target organs. As such, this work is highly relevant to the NIH mission in that it will contribute to the advancement of knowledge that could improve human health.

NIH Research Projects · FY 2025 · 2024-08

PROJECT SUMMARY Further, more than one-quarter of cancer patients in Mexico suffer from a depression disorder and about half have significant mood disorder symptoms. Also, research shows is a prevalence of 67-82% clinical depressive symptoms in patients with advanced cancer. Patients with advanced cancer have specific needs; among the most pressing are to receive help in dealing with their mortality fears, uncertainty about the future, and spiritual concerns. However, there are no targeted psychosocial or psychotherapeutic interventions available for Latinos diagnosed with advanced cancer. Meaning-Centered psychotherapy (MCP) is a novel psychotherapeutic intervention designed to help patients with advanced cancer sustain or enhance a sense of meaning, peace and purpose as they approach the end-of-life. Meaning Centered Psychotherapy has been culturally adapted and found feasible in a pilot study with heterogenous Spanish-speaking Latinos with advanced cancer, but not yet tested with Mexican cancer patients. This study is a collaboration of investigators from two major cancer centers, Memorial Sloan Kettering Cancer Center (New York, where MCP f or Latinos, MCP-L, was adapted) and the National Mexican Cancer Institute (INCan, Spanish Acronym), the major public cancer center in Mexico and Central American region serving approximately 1000 new annual patients. Implementation of this project in INCan is feasible given the resources (psycho-oncology service with 3 full time psychologists) and institutional support for psycho-oncology research. Informed by implementation science framework, the objective of this study is to conduct a type 1 hybrid (testing effects of an intervention while gathering information on implementation) randomized clinical trial. The first aim examines the impact of Meaning Centered Psychotherapy for Mexicans on improving depression and anxiety (primary outcomes) and the secondary outcomes: distress, hopelessness, symptom burden and quality of life, compared to the control condition (cognitive behavioral therapy) at one and three months after treatment. We will also examine the mediation effect of spiritual and existential (meaning) well-being and moderation effect of socioeconomic hardship and literacy. Guided by a dissemination and implementation model and using a mixed-methods approach, the second aim examines the factors influencing the intervention implementation at the external, organizational, and patient levels, and potential implementation strategies. The third aim is to increase and support clinical and research capacity by providing specialized training to the INCan team and cancer MH providers from Latin American. This study is significant in that it targets the emotional impact of a diagnosis of advanced cancer in Mexican patients, a prevalent yet understudied need in this population. This study is innovative because it will be the first evidence-based psychotherapy intervention adapted for a low- and middle-income country to treat patients with advanced cancer.

NIH Research Projects · FY 2025 · 2024-07

This proposal outlines a three-year training plan in cellular immunology and mechanobiology, offering the applicant diverse expertise in these fields. The sponsor's laboratory excels in modeling immune responses in relation to biophysical features of target cells, and the institutional strengths in immunology, mechanobiology, and cell biology create an exceptional training environment conducive to success. Metastasis occurs in a variety of organs with varying mechanical states. In recent years, the biophysical properties of these microenvironments have emerged as an influencer of metastatic site preference (MSP) via mutual biomechanical crosstalk where cancer cells mimic the stiffness of their environments’, otherwise known as mechanoreciprocity. Our preliminary data has shown that cytotoxic lymphocytes, comprising of cytotoxic T cells and natural killer cells, selectively destroy stiffer cancer cells during metastatic dissemination, a process that we call mechanosurveillance. Our study combines biophysical measurements with in vivo models, demonstrating that microenvironmental stiffness influences immune vulnerability, impacting the organ distribution of metastatic outgrowth. In-depth analyses of metastatic cells from different organ sites, utilizing atomic force microscopy and single-cell RNA sequencing, indicate that microenvironmental stiffness shapes immune discrimination. Specifically, bone metastases exhibit increased stiffness, with the gene Spp1 playing a role in maintaining this stiffness. Further studies using CRISPR-based knockout systems show that loss of Spp1 softens cancer cells. This suggests mechanosurveillance targets stiff organ metastases, forming a biophysical basis for metastatic site preference. Building on these preliminary observations we propose that microenvironmental stiffness dictates the efficacy of mechanosurveillance and that this relationship shapes both metastatic site preference and the power of anti-tumor immunotherapy. We will investigate this hypothesis in two specific aims. Aim 1 employs in vivo mouse models to examine how distinct microenvironments affect cancer cell mechanics, transcriptomics, and immune vulnerability. Specific genes, including Spp1, will be manipulated to explore their role in mechanosurveillance. Aim 2 utilizes synthetic cancer cell niches to investigate if environmental stiffness independently controls mechanosurveillance efficiency. The successful completion of these aims could identify biomarkers guiding antitumor immunotherapy and inform novel strategies for treating metastatic growth, advancing knowledge to enhance human health.

NIH Research Projects · FY 2025 · 2024-07

PROJECT ABSTRACT Small cell lung cancer (SCLC) and neuroendocrine prostate cancer (NEPC) are aggressive, high-grade neuroendocrine neoplasms (HG-NENs) that often metastasize and do not respond well to treatment. Lung cancer is the leading cause of cancer mortality worldwide and SCLC represents ~15% of lung cancers. Prostate cancer is the most common solid tumor and the second-leading cause of cancer-related death among men in the US. More than 20% of males with advanced prostate cancer eventually develop the highly aggressive NEPC form. Both SCLC and NEPC are characterized by similar genetic (RB1 and TP53 loss) and phenotypic features, as well as substantial intra-tumoral heterogeneity, which is implicated in therapeutic resistance. Identifying and developing novel treatment targets and strategies for SCLC and NEPC is a critical unmet need. Delta-like ligand 3 (DLL3) is an antigen expressed on the cell surface of HG-NENs, whereas in normal tissues expression is restricted to intracellular compartments, predominantly confined to the Golgi apparatus. DLL3 expression correlates with neuroendocrine marker expression, RB1 loss, and aggressive clinical features. Based on the over-expression of DLL3 in HG-NENs, we propose to utilize a DLL3-targeted radiolabeled monoclonal antibody (mAb) for non-invasive diagnosis using PET imaging (89Zr-DFO-SC16.56 Ab) as well as for image-based therapy (177Lu-DTPA-SC16.56). Our central hypothesis is that the overexpression of DLL3 on HG-NEN lesions can be exploited for diagnostic and therapeutic purposes. To examine this hypothesis, our phase I study will evaluate patients affected by HG-NENs with 89Zr-DFO-SC16.56 mAb PET/CT. Those who exhibit sufficient target expression in at least 80% of the growing lesions will be treated with 177Lu-DTPA- SC16.56. A key issue in such aggressive malignancies is the need to objectively determine treatment efficacy by quantifying the radiation dose delivered to the target and assessing tumor radiosensitivity, which engenders susceptibility to treatment. We will use molecular genomic tools to identify DLL3 transcripts and compare these with known molecular signatures in blood and tissue that can define neuroendocrine and prostate cancer tumors and correlate with response. The proposed study will investigate the safety and mechanistic basis of the efficacy of radiotheranostics specifically targeting DLL3, a novel target for lethal malignancies for which currently limited therapeutic tools exist. Our extensive experience in the development of DLL3-targeting PET agents and validation in SCLC and NEPC models, along with our preliminary clinical PET data in SCLC and NEPC patients, as well as our expertise in the ancillary use of molecular genomic tumor markers position us to successfully complete the aims of this study.

NIH Research Projects · FY 2025 · 2024-07

Metastatic lung cancer kills >160,000 people in the US annually, and 10%-50% of patients with surgically resected lung cancer will develop distant metastasis. There remains an unmet need to understand the tumor genomics that drive metastases. SMARCA4 is the most frequently mutated member of the SWI/SNF complex in lung adenocarcinoma (LUAD), and its alterations are associated with development of metastatic disease. Whereas SMARCA4 canonically acts as a chromatin remodeler, mechanistic investigations from our lab reveal a noncanonical ROR2/p-GSK3βS9 pathway involved in loss-of-SMARCA4–mediated transcriptional regulation and metastasis in LUAD. Specifically, knockdown of SMARCA4 in LUAD cells increases invasion and metastases. Increased SMARCA2 upregulates ROR2 and inhibition of ROR2 abrogates invasion and metastasis of SMARCA4-deficient LUAD. Moreover, ROR2 increases the abundance of transcription factor C/EBPa by inactivation of nuclear GSK3b. Using a GSK3β inhibitor, we rescued invasion of SMARCA4KD/ROR2KD cells. Our overarching goals are 1) to elucidate the mechanisms through which SMARCA4-deficient LUAD regulates transcription and promotes metastases and 2) to determine the role of ROR2/p-GSK3βS9 pathway in metastasis of SMARCA4-deficient LUAD. Two specific aims will test our hypotheses. Aim 1: Investigate the mechanisms through which deficiency of SMARCA4 promotes metastasis of LUAD. We will perform RNA-seq, ChIP-seq, ATAC-seq, and selected reaction monitoring–mass spectrometry using LUAD SMARCA4dTAG cells to measure gene transcription and chromatin accessibility and to quantify transcription factors in real time. An in vivo CRISPR/sgRNA screen will be performed to identify functional downstream transcription factors of ROR2/p-GSK3βS9 pathway. Next, we will use mass spectrometry and immunoFISH to decode the mechanism(s) through which SMARCA2 upregulates ROR2 in SMARCA4-deficient LUAD and to explore alternative target(s) to suppress metastases. Aim 2: Evaluate the therapeutic efficacy of targeting ROR2 to suppress metastasis in SMARCA4-deficient LUAD. We will assess the clinical relevance of ROR2/p-GSK3βS9 pathway in SMARCA4-altered LUAD by leveraging our clinically annotated biorepository of >2000 human LUAD specimens. We will then use our LUAD patient-derived organoid model to examine the requirement for p-GSK3βS9 for metastases of SMARCA4-altered LUAD and to test whether pharmacologic inhibition of ROR2 using an antibody-drug conjugate currently in clinical trials decreases metastases in SMARCA4-deficient LUAD. Finally, using a published KrasLSL-G12D/Trp53fl/fl/Smarca4fl/fl genetically engineered mouse model, we will investigate the cell-specific dependency of ROR2/p-GSK3βS9 pathway. Impact: Our work will provide mechanistic and translational evidence that will highlight deficiency of SMARCA4 as an important metastatic driver in LUAD through ROR2/p-GSK3βS9 pathway and will provide an important preclinical justification for future clinical trials targeting ROR2 in SMARCA4-deficient LUAD to decrease metastasis.

NIH Research Projects · FY 2024 · 2024-07

Abstract The overall goal of this line of research is to examine the psychosocial, ethical, and clinical implications of incidental medical findings of clonal hematopoiesis (CH) in healthy volunteer hematopoietic stem cell (HSC) donors. The proposed project focuses on feasibility. CH refers to the presence of mutations acquired during a person's lifetime in the hematopoietic stem cells. CH in healthy individuals is not considered a disease state, although it is associated with increased risk for cardiovascular diseases, increased risk for hematologic malignancies, and lower survival rates. Currently, most people become aware of CH as an incidental finding during workup for another disease. Many clinical questions about CH remain, including whether (1) different CH mutations are associated with different long-term health outcomes, (2) there is mutation frequency threshold associated with increased risks, and (3) there are any modifiable/lifestyle factors that alter the risks associated with CH. This proposal presents a unique opportunity to assess the psychosocial, clinical, and ethical implications of identifying incidental CH findings among healthy hematopoietic stem cell (HSC) donors at the National Marrow Donor Program (NMDP), which manages the largest worldwide registry of individuals who have volunteered to donate to unrelated patients in need of HSC transplants. After stem cell transplant, CH that originated from the donor may be identified in recipients. The NMDP recently formalized a process for notifying donors with potential CH. This investigation will evaluate the feasibility of tracking the psychosocial impact and clinical markers among this otherwise healthy group of donors. Specifically, we will (1) qualitatively examine the psychosocial impact of receiving information about incidental medical findings in a group of recent HSC donors by conducting a series of focus groups (N=30-40; Aim 1), (2) demonstrate the feasibility of conducting longitudinal structured health-related quality-of-life (HRQoL) telephone interviews with HSC donors diagnosed with CH (N=30; Aim 2), and (3) demonstrate the feasibility of prospectively tracking clinical markers (in blood, stool, and calcium scores) in HSC donors diagnosed with CH (N=30; Aim 3). This investigation is significant because it will lay the groundwork for a larger investigation of incidental findings of CH and will have implications for how such findings are communicated to individuals receiving this information through standard health screenings. The investigation is innovative because it (1) will be the first to investigate CH in a healthy donor population, (2) presents a unique, low-resource, low-burden opportunity to identify individuals with CH, (3) focuses on potential psychosocial issues in the delivery of incidental findings to healthy donors, and (4) demonstrates our ability to collect longitudinal clinical marker information from a young, healthy cohort. Findings from this study will lead directly to a larger investigation of incidental findings in this healthy donor group, which will have implications not only for HSC donors, but for a broad range of healthy and ill individuals in whom incidental medical findings are discovered.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY/ABSTRACT Chronic pain is a common symptom that affects 30-40% of cancer survivors, diminishing their quality of life, impairing physical functions, and increasing health care costs. Breast cancer survivors constitute the largest group of survivors with more than 4.1 million in the US. Nearly one in two breast cancer survivors who take a class of medication known as aromatase inhibitors are affected by a chronic painful condition, aromatase inhibitor-associated arthralgia (AIA). AIA is associated with reduced physical activity, increased risk of falls, and reduced AI adherence, leading to increased overall mortality. With insufficient relief from conventional treatments and a rapidly growing population of breast cancer survivors, there is an urgent need to develop novel, effective, and scalable pain management options. Mindfulness-Oriented Recovery Enhancement (MORE) is an innovative mindfulness-based intervention (MBI) rooted in affective neuroscience that integrates training in mindfulness, reappraisal, and savoring skills to specifically target chronic pain and related symptoms (e.g. psychological distress). Although the efficacy of MORE for pain in non-cancer populations has been established, the potential for this mindfulness intervention to address pain and comorbid symptoms in breast cancer survivors has yet to be confirmed. Therefore, a rigorous, adequately-powered randomized controlled trial is needed to conclusively determine whether MBIs such as MORE can alleviate AIA. To address this critical gap in research and clinical care, we have convened a multidisciplinary team to conduct the Enhanced Pain Coping in Cancer (EPIC) trial with the following specific aims: 1) to evaluate the specific efficacy of MORE for managing AIA among breast cancer survivors, 2) to evaluate the specific effects of MORE on comorbid symptoms, quality of life, and adherence to AIs, and 3) to elucidate the cognitive-affective mechanisms of MORE for pain management among breast cancer survivors. The MORE intervention that we will test aligns with a downward spiral of chronic pain model and neuroscience and is based on discoveries from preliminary studies. For this multisite, randomized controlled trial we will randomize 200 breast cancer survivors free of oncologic disease with AIA to one of two eight-week treatments: 1) MORE or 2) supportive group psychotherapy (SG). We will assess the primary outcome (pain-related functional interference) and secondary outcomes at baseline, week 8 (end of treatment), week 12 and week 24 (primary end point) using validated patient-reported outcomes. EPIC will address major methodological limitations of existing MBI trials including pain not being a primary outcome, no eligibility requirement for having pain, and the MBIs under investigation having been developed to target stress management rather than pain management. EPIC will provide timely clinical evidence for MORE, a neuroscience-informed, mindfulness-based intervention for pain and co-morbid symptoms, during cancer survivorship. Further, virtual delivery will ensure MORE is highly scalable with broad reach to improve pain management for millions of breast cancer survivors across the US.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY/ABSTRACT Immune-based treatments for acute myeloid leukemia (AML), an aggressive hematologic malignancy that remains fatal for over half of the 20,000 patients diagnosed in the United States annually, remain an unmet clinical need. The impetus to use T cell-based immunotherapy approaches for AML treatment is underscored by the curative potential of allogeneic hematopoietic cell transplantation for AML, arising from immune- mediated graft-versus-leukemia activity of donor T cells, and by emerging correlative data between the immune landscape and AML treatment efficacy. Understanding mechanisms of ineffective anti-leukemic T cell activity in the AML bone marrow (BM), the site where AML emerges, is fundamental for advancing immunotherapeutic approaches for AML. Our preliminary data highlight the immunosuppressive nature of T cells within the AML BM, the dynamic evolution of T cell clones with regard to states of T cell activation and exhaustion, and the immunomodulatory potential of malignant AML cells on T cells. We hypothesize that both malignant and non-malignant cells in the AML BM promote impaired T cell immunity, leading to distinct T cell compositions at different disease states. This career development program will address two specific aims: (1) determine T cell intrinsic features that affect the relationship between T cell phenotype, repertoire, and disease status in the AML BM, and (2) understand mechanistically how malignant (AML blasts) and non-malignant cells in the AML BM shape features of the T cell compartment. The candidate has developed this proposal as an extension of her ongoing T cell studies in hematologic malignancies and clinical experience in AML. During the award period, she will complete her research with close counsel from her primary mentor, Omar Abdel-Wahab, MD, an accomplished scientist in leukemia biology and genomics who is the Chair of the MSK Molecular Pharmacology Program. Additional input will be offered by her co-mentor, Marcel van den Brink, MD, PhD, an international expert in T cell biology in hematologic malignancies, and by her Advisory Committee. Drs. Abdel-Wahab and van den Brink are both mentors with outstanding records of navigating physician–scientists toward research independence. Under their guidance, the candidate will develop skills that are critical for her future laboratory program, including learning to generate and analyze syngeneic mouse models of AML, to build upon her knowledge of functional studies with primary AML samples, and to advance her computational and bioinformatics abilities in sequencing-based immunologic techniques. Completion of this proposal will provide the candidate with the training and mentorship required to cultivate and refine her expertise in T cell immunology and leukemia research and to establish her academic career as an independent laboratory-based clinical investigator dedicated to advancing immunologic treatments for AML.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY Lung cancer is the leading cause of cancer death among the African American population. Although many actionable biomarkers and targeted treatments are available to significantly prolong lung cancer survival, patients of non-European ancestry are less likely to undergo next-generation sequencing testing and received targeted therapies than their white counterparts. Structural and environmental factors likely contribute to this disparity; however, there is evidence suggesting that somatic genomic differences also contribute – several lung cancer drivers and targetable biomarkers have been found to have different alteration frequencies between populations. However, African American patients are severely underrepresented in research studies and clinical trials, and therefore, the genomic landscape of lung cancers with African ancestry remains poorly understood. Our preliminary analysis based on real-world, observational data of clinical tumor sequencing suggest that the prevalence of targetable alterations in KRAS and ROS1 fusion are different in patients of African ancestry, independent to smoking status. Interestingly, ancestry may modify the effect of smoking exposure on the lung cancer genome – TP53 mutations were significantly enriched in smokers but not in never smokers of African ancestry. Moreover, there is a significant enrichment of high tumor mutation burden in light smokers compared to their white counterparts, in line with previous studies showing higher risk of lung cancer in smokers of African ancestry, suggesting different genomic mechanism as the resultant influence of ancestry-smoking interaction. In this project, we will take three parallel approaches to comprehensively characterize lung cancer genomes in patients of African ancestry. In the first aim, we will identify and validate more genomic differences between patients with African and European ancestry, with a focus on drug targetable and clinically relevant biomarkers. We will also locus-specific germline ancestry to identify the genetic and non-genetic contributors to somatic differences. In the second aim, we will focus on investigating the interaction between African ancestry and smoking exposure on somatic alterations and develop a computational pipeline leveraging off-target reads from panel sequencing to study genome-wide germline influence. In the third aim, we will develop a deep learning model to allow us further to understand the generalizability and ancestry-specificity of immunotherapy outcome prediction. Finally, we will move beyond panel sequencing to identify new markers for patients of African ancestry, using whole-genome sequencing coupled with RNA-sequencing. The proposed study will broaden our knowledge about the complex relationship between genetic ancestry, environmental exposure and ancestry-environment interaction contributing to genomic differences. The findings will reveal germline factor associated with somatic phenotype, improve immunotherapy outcome prediction, and lead to future study and discovery of new treatment options for African American patients.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY/ABSTRACT RESEARCH: Natural Killer (NK) cells are crucial for the immediate defense against virus infections. To exert their function, NK cells need to receive signals through activating receptors and cytokines. Cytokines mediate their function via STAT transcription factors. We could previously show that STAT1, STAT2, STAT4 and STAT5 are all required for NK cell expansion in murine cytomegalovirus (MCMV) infection. Studying the role of STAT3 in NK cells, I found that STAT3 plays a context-dependent role. In regular-dose infection, STAT3 augmented proliferation of NK cells. However, in high-dose infection STAT3 appeared to impair NK cell expansion. This suggests that our model can be used to study the context-dependent role of STAT3, which has broad relevance for the fields of immunology and cancer biology. In this proposal, I seek to answer 1) how are different upstream signals altering the role of STAT3 for NK cell function and 2) how are identified downstream targets of STAT3 contributing to the context-specific role of STAT3. CANDIDATE: I am a postdoctoral fellow in the lab of Dr. Joseph Sun at Memorial Sloan Kettering Cancer Center (MSKCC). Prior to this position, I studied the differentiation of CD8+ T cells and NK cells. My interest is the mechanistic study of immune cell differentiation and function during viral infection. I joined the Sun lab as one of the leading groups studying the molecular mechanisms underlying NK cell function. Through the work outlined in this proposal I will 1) acquire technical expertise available in my current lab with a focus on the study of transcription factors, 2) develop a network of scientists that enable the collaborative type of research I seek to participate in and 3) develop skills necessary for career development such as mentoring, data presentation and grantsmanship skills. I have assembled an excellent Advisory Committee that is committed and capable to guide me on my path to independence: Dr. Rudensky for technical support, Dr. Leslie for mentorship in computational biology and Dr. Ivashkiv as an expert in cytokine and STAT biology. With the support of my mentor and Advisory Committee I will be able to acquire the necessary knowledge and expertise during the K99 phase to launch my own independent career. ENVIRONMENT: The Immunology Program at MSKCC is part of the Tri-Institutional network composed of MSKCC, Rockefeller University and Weill Cornell University. The highly collaborative culture promotes scientific exchange and has provided me with crucial technological support. Furthermore, I could begin to form a network with multiple groups at our campus. Regular presentations by renowned senior scientists enable me to expand the scope of my scientific questions. Weekly lab meetings and frequent personal meetings with my mentor Joseph Sun, who has successfully guided multiple former trainees in launching their own academic career, make my environment ideal for starting my path to independence.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY A prolonged hospital stay after cancer surgery is a common but potentially avoidable problem that negatively impacts quality of care. Goal-directed pathways for discharge are widely used in surgical oncology to avoid unnecessary extended lengths of stay, but discharge milestones are maintained by the clinical team without patient input. The resultant discharge pathways are rigid, with little or no personalization, and the necessary steps in the process are often unclear to patients. These gaps in communication and mismatch in expectations between clinical care team members, patients, and caregivers contribute to excessive lengths of stay. Integration of patient-reported outcomes (PROs) into clinical care is one strategy shown to improve communication and align expectations. There may be great opportunity to use transition-of-care–focused PROs items in the hospital setting to align patient and clinical team members during discharge planning to prevent prolonged hospital stays. Hypothesis: A discharge-focused electronic PROs tool for surgical oncology patients will decrease the length of stay after elective surgery. PROs including readiness for discharge and activation will also be measured. Jennifer Cracchiolo, MD, has developed an electronic PROs tool, “Goals to Discharge” (G2D), for oncology patients focused on discharge planning to augment currently established discharge pathways. While the G2D tool has been successfully employed during hospitalization and patients have responded, there have been challenges to implementing this technology, including helping patients navigate the digital tool; devising strategies to increase response rates and ensure inclusivity; optimizing the content; and developing PROs-prompted clinical decision support for clinicians. In the proposed project, Dr. Cracchiolo will refine the library of actionable PROs items that measure domains relevant to inpatients and their readiness for hospital discharge. She will include patients from surgical oncology (colorectal, thoracic, and gynecologic surgery) at Memorial Sloan Kettering Cancer Center, with the goal of actively engaging patients during the discharge process. Aim 1: Refine content for the G2D tool with PROs-based clinical decision support. Aim 2: Systematically plan for broad clinical adoption of G2D using an implementation mapping approach. Aim 3: Pilot test the effectiveness of the G2D tool compared with usual care. Results will lead to better quality of care, better informed patients, and better outcomes. This research, along with the career development plan for training in advanced qualitative methods, implementation science, and clinical trial study design by her multidisciplinary mentors, Deborah Schrag, MD, MPH; Jamie Ostroff, PhD; Andrew Vickers, PhD; and Thomas Atkinson, PhD, will provide Dr. Cracchiolo with the infrastructure needed to grow in her career and facilitate her progress toward independence as a researcher in clinical integration of PROs systems in surgical oncology care.

NIH Research Projects · FY 2025 · 2024-06

PROJECT SUMMARY/ABSTRACT The overarching goal for this project is to understand the events governing the earliest stages of mammalian embryo development. Specifically, we will investigate how the pluripotent epiblast which generates the embryo- proper, and its sister lineage the extra-embryonic (or primitive) endoderm, arise from a common progenitor population, the inner cell mass. Furthermore, we will determine how the cells of these two nascent lineages differentiate and organize themselves, as they sort into two adjacent tissue layers. We will use the mouse as an experimentally tractable animal model system to investigate a universal and critical stage of mammalian development. We will take an integrative approach across scales (from gleaning molecular details to tissue-level organization) by applying cutting-edge methods, including in toto light microscopic imaging, the analysis of gene expression and protein localization at the level of single cells across a population of cells, as well as performing perturbations, pharmacologic and optogenetic manipulations, within the spatiotemporal context of developing embryos. Our experiments will be coupled to computational analyses of data, and mathematical modeling. These contextual time and space resolved studies will shed insight into how a population of progenitors gives rise to two lineages each possessing a distinct identity and stereotypical tissue organization, and the mechanisms that ensure the robustness reproducibility and scalability of this process. An in-depth mechanistic understanding of critical events taking place in vivo in the mouse model provides the foundational knowledge for: extending our understanding to other mammalian species, the stem cell populations than can be derived from early mammalian embryos which are increasingly being used to generate embryoid (also referred to as synthetic embryo) models, and the differentiation of cells with distinct identities having therapeutic potential. Moreover, using a simple and robust in vivo paradigm of self-organization for decoding the dynamics of cell-cell communication and growth factor signaling will provide insights into how developmental mechanisms are hijacked during disease progression, and can be targeted for therapeutic intervention.

NIH Research Projects · FY 2025 · 2024-06

Abstract: eDyNAmiC (extrachromosomal DNA in Cancer) Human genes are arranged on 23 pairs of chromosomes, but in cancer, tumour-promoting genes can free themselves from chromosomes and relocate to circular, extrachromosomal pieces of DNA (ecDNA). These ecDNA do not follow the normal “rules” of chromosomal inheritance, enabling tumours to achieve far higher levels of cancer-causing oncogenes than would otherwise be possible, and licensing cancers with a way to evolve and change their genomes to evade treatments at rates that would be unthinkable for human cells. The altered circular architecture of ecDNAs also changes the way that the cancer-causing genes are regulated and expressed, further contributing to aggressive tumour growth. These unique features make ecDNA-containing cancers especially aggressive and difficult to treat. Cancer patients whose tumours harbour ecDNA have markedly shorter survival. Despite being first seen over fifty years ago, the critical importance of ecDNA has only recently come to light, and the scale of the problem is substantial. ecDNAs are present in nearly half of all human cancer types and potentially up-to a third of all cancer patients. The collective current understanding of how ecDNA form, how they function, how they move around the cell, how they evolve to resist treatment, how they impact the immune system, and how they can be effectively targeted are lacking. We bring together an internationally recognized, pioneering interdisciplinary team of cancer biologists, geneticists, computer scientists, evolutionary biologists, mathematicians, clinicians, and patient advocates to boldly create novel insights and resources and to provide transformative solutions to one of Cancer’s Grand Challenges. A core team of experienced and productive ecDNA investigators will work with new investigators in the ecDNA and cancer fields to bring completely new perspectives and approaches to this daunting challenge. By bridging cutting-edge and diverse approaches and insights from cancer genomics, yeast genetics, epigenomics, artificial genome synthesis, longitudinal patient tracking, combinatorial and machine learning algorithms, mathematical modelling, immunobiology, and innovative chemistry we will develop a new understanding of the role of ecDNA in cancer, and we will find new ways to drug the undruggable. This bold programme, which consists of 7 work packages and a committed international infrastructure, generates new and unusual collaborations that would simply be impossible under any other type of funding mechanism. Our programme endeavours to foster bold innovative solutions to one of the hardest problems in cancer and to one of the greatest challenges facing cancer patients.

NIH Research Projects · FY 2025 · 2024-06

PROJECT SUMMARY Quantitatively predicting drug-resistant mutations to improve precision oncology My work builds towards a mechanistically informed approach to model and predict drug-resistant kinase mutations that will enhance patient treatment regimens. Protein kinases are important signaling enzymes often dysregulated in cancer; their pharmacological value as drug targets exemplified by the clinical use of over 75 FDA-approved inhibitors. Unfortunately, multiple clinically observed kinase mutant resist inhibitors and drastically reduce patient survival rates. Precision oncology approaches, matching specific tumor profiles to optimally therapies, have proven useful thanks to tumor sequencing and mutation profiling. However, it remains challenging to identify drug-resistant mutants prior to treatment and develop regimens to circumvent them. A lack of mechanistic information describing clinically observed kinase mutants makes it difficult to predict whether a mutation will resist canonically used kinase inhibitors. Kinase mutations may decrease drug-binding affinity, increase kinase activity, tune inhibitor sensitivity profiles, or any combination of these mechanisms. Structure-based methods promise to help predict the impact of kinase mutations. I hypothesize that a kinase inhibitor's utility against drug-resistant mutants is expressed using physical, quantitative properties like structural state populations and binding affinities. My work quantitatively assesses the impact of clinical kinase mutations on inhibitor resistance, sensitivity, and susceptibility. Specifically, I will develop models that predict whether clinical kinase mutations perturb inhibitor-binding, increase kinase activity by stabilizing active configurations, or sensitize kinases to alternative inhibitors. In this proposal, I draw upon clinical mutation databases to study mutation-inhibitor pairs of c-Met kinase, the target in Non-Small-Cell lung cancers (NSCLCs), building upon previous studies of resistance mutations in Abl kinase. As a mentee (K99), I will use binding free-energy calculations to predict how clinical mutations reduce c-Met inhibitor affinity (Aim 1). As I transition to independence (K99/R00), I will use molecular simulations to biophysically evaluate whether clinical mutations increase kinase activity by shifting kinase populations to catalytically active conformations (Aim 2). Upon independence (R00), I will study whether clinical mutations sensitize kinases to rarely used alternative inhibitors (Aim 3). These computationally intensive calculations can often take years to collect sufficient data on a normal computer. Instead, I will run calculations on the planetary-scale Folding@home distributed computing platform in collaboration with high-throughput biophysical experiments that measure kinase activity and inhibitor binding affinity. Overall, my proposal, and future lab, will build towards a precision-oncology platform that helps clinicians plan treatment regimens.

NIH Research Projects · FY 2025 · 2024-06

PROJECT SUMMARY/ABSTRACT CANDIDATE: I am a postdoctoral research associate in the laboratory of Dr. Yu Chen in the Human Oncology and Pathogenesis Program (HOPP) at Memorial Sloan Kettering Cancer Center. My PhD training at Boston University School of Medicine under the supervision of Dr. Sam Thiagalingam allowed me to develop cellular and molecular biological skills to functionally characterize candidate genes involved in cancer progression and investigate therapeutic vulnerabilities. My current research focuses on utilizing patient-derived organoid models to define epigenetic subtypes of castration resistant prostate cancer (CRPC) and determining the dependency on transcription factors to mediate growth and lineage identity. My proposed research and mentoring plan will provide me with a strong foundation towards becoming an independent investigator in academia. My long-term career goal is to advance personalized cancer medicine, with specific interests in disease modeling, functional characterization of drivers of lineage plasticity, and evaluation of therapeutics to target oncogenic dependency. To achieve this goal, I have developed a career plan that will ensure my success to becoming an independent investigator by: 1) bolstering my scientific knowledge and technical expertise, 2) assembling an advisory committee to oversee my training progress, 3) improving my communication skills, 4) expanding my professional network, and 4) preparing me for leading and mentoring future trainees. RESEARCH: Prostate cancer depends on androgen receptor (AR) signaling for growth and survival. The advent of therapies targeting AR signaling has driven the disease towards AR-independence. Using a functional genomics approach, we have identified a new epigenetic subtype of CRPC called stem cell-like (SCL), which is driven by FOSL1 and lacks therapeutic options. Genetic perturbation of FOSL1 and its cooperating factors YAP/TAZ leads to impaired cell growth and loss of SCL lineage, suggesting that CRPC- SCL cells are dependent on FOSL1 for survival. Building on these discoveries, in this proposal, I aim to: 1) Define the role of FOSL1 in mediating lineage plasticity and resistance to enzalutamide in prostate cancer and 2) Evaluate therapeutics to target the YAP/TAZ/TEAD/FOSL1 pathway in the stem cell-like subtype of prostate cancer. ENVIRONMENT: The Yu Chen laboratory is a part of the HOPP under the leadership of Dr. Charles Sawyers at Memorial Sloan Kettering Cancer Center, a top-tier institute in cancer research. The primary mentor is Dr. Yu Chen, a pioneer in developing prostate cancer organoids from patients and an expert in the study of epigenetics and aberrantly activated transcriptional programs. Furthermore, my advisory committee and collaborator will provide additional support for the proposed research and career development plans.

NIH Research Projects · FY 2026 · 2024-06

SUMMARY. Neurodegenerative diseases, as age-dependent disorders, represent a rising hazard to human health in America, given the rapidly growing number of seniors in its population. For this reason, the development of novel therapies to treat neurodegenerative diseases has become a national priority. These disorders are often characterized by the incorrect folding of certain proteins, accumulation of cytotoxic macromolecular structures, and changes in intracellular stress response capacity - our proposal aims to identify a means to resolve these conditions. Numerous labs over the last decade have elucidated mechanisms underlying selective autophagy - the process by which damaged organelles, protein aggregates, or misfolded proteins are targeted to the autophagosome for degradation in the lysosome. Our understanding of autophagy is that it controls a large fraction of regulated protein homeostasis in human cells, particularly post-mitotic cells (i.e., a cell that does not divide) like neurons. While autophagy is broadly associated with the turnover of proteins or organelles, several autophagy-related genes have been specifically genetically linked with aging and neurodegenerative cellular signaling pathways. Despite these advances, our understanding of how neuronal cells use and rely on specific forms of autophagy via specialized proteins called receptors is incomplete for several reasons. First, most studies have focused on a small number of well-studied receptors, and systematic approaches aimed at uncovering the global contribution of known and other possible receptors are lacking, leaving major gaps in our understanding of the many roles these proteins may play. Second, the majority of studies have focused on processes in cancer cell lines in culture, and we know very little about the cell type-relevant roles of selective autophagy receptors in neuronal homeostasis. Third, although we know that selective autophagy receptors are regulated dynamically, we still have a relatively rudimentary understanding of the signaling pathways that control receptor engagement in physiological or stress-induced states. Here, we propose a series of experiments that seek to address limitations in our understanding of the neuronal selective autophagy pathways, their regulation, and their function. In the first series of experiments (AIM 1), we will build on preliminary data to elucidate the role of an endoplasmic reticulum membrane-bound receptor we identified to be regulated upon nutrient stress in neurons, and reveal the molecular mechanisms involved in its regulation and the nature of the substrate turned over. A second series of experiments (AIM 2) will be focused on investigating the cellular specificity across neuroimmune cell subtypes, response to pathological cues modeled in iPSCs, and functional and physiological relevance using knock-out mouse models. Together, these studies will quantitatively, mechanistically, and functionally address gaps in our understanding of the neuronal relevant functions of this selective autophagy pathway and the mechanisms by which substrates are selected for lysosomal recycling.

NIH Research Projects · FY 2025 · 2024-05

PROJECT SUMMARY/ABSTRACT CANDIDATE: I am a Postdoctoral Research Associate at Memorial Sloan Kettering Cancer Center (MSKCC). During my Ph.D. studies, I dedicated myself to developing biology-based mathematical models of bacterial metabolism. My current research extends my interest from single organisms to microbial communities, with a particular focus on the human intestinal microbiome. Since joining MSKCC, I have compiled a large longitudinal microbiome dataset from hospitalized patients who have undergone allogeneic hematopoietic cell transplantation (allo-HCT). I have also acquired bioinformatic skills and data-driven modeling techniques to profile microbiome compositions and quantify their associations with clinical outcomes. Built upon this dataset, my proposed research aligns well with my long-term career goal of establishing an independent laboratory to elucidate mechanistic links between the intestinal microbiota and infectious diseases. To prepare for my transition to independence and developing a competitive computational research program, I have developed a focused career plan to enhance my computational skills in metagenomic/metabolomic data analyses, community metabolic modeling, and development of neural network models. In parallel, I will improve my soft skills, including presentation, networking, grantsmanship, mentorship, leadership, and teaching. RESEARCH: Immunocompromised patients undergoing intensive antimicrobial therapy are at high risk for developing invasive fungal bloodstream infections (BSIs). Between 2016 and 2020, C. parapsilosis was responsible for the most breakthrough BSI cases among allo-HCT recipients at MSKCC. Typically, C. parapsilosis BSI occurs subsequent to its intestinal expansion. This proposal will leverage my mathematical modeling expertise and the vast microbiome dataset of our allo-HCT cohort to elucidate the ecological mechanisms underlying intestinal expansion of C. parapsilosis. My central hypothesis is that altered intestinal metabolic environment enables C. parapsilosis expansion. Specific Aim 1 will identify bacterial secreted metabolites that inhibit C. parasilosis. In Specific Aim 2, I will investigate the impacts of genomic variations across different C. parapsilosis isolates on their ability to utilize nutrients and grow in the human intestine. The Specific Aim 3 will involve building a machine-learning-powered computational framework for the risk assessment of C. parapsilosis expansion and the rational design of antifungal therapy to reduce the risk. ENVIRONMENT: I will complete the K99 phase of this grant in the Computational & Systems Biology Program at MSKCC, a state-of-the-art cancer research institute. My primary mentor, Dr. Joao Xavier, has a proven track record in mathematical modeling of bacterial microbiomes, while my co-mentor, Dr. Tobias Hohl, is an expert in fungal mycobiomes and infectious disease. The two labs will jointly provide a rich and complementary training and research environment that integrates computational, experimental, and clinical resources.

NIH Research Projects · FY 2026 · 2024-05

Abstract The mevalonate pathway enzymes produce precursors of sterols and isoprenoids, essential metabolites for cell signaling and membrane biogenesis. As such, cells employ complex transcriptional, translational, and post- translational processes to regulate mevalonate production. Specifically, the ubiquitin proteasome system (UPS) tightly controls the mevalonate enzymes on the ER membrane, converging on the ER-associated degradation pathway. Importantly, increased levels of distinct sterol metabolites in the ER membrane cause proteolysis of specific enzymes, suggesting that the rate-liming enzyme in the mevalonate pathway can change based on the stimuli. While studies have focused on the feedback regulation of the mevalonate pathway enzymes upon sterol stimulation, it is unclear whether other environmental changes, such as cell growth signaling or nutrient availability, can also tightly control the mevalonate synthesis through post-translational mechanisms. Here, we report that HMG-CoA Synthase1 (HMGCS1), the first committed enzyme in the mevalonate pathway, is the primary responder of the master regulator of cell growth, mTORC1, among ~20 mevalonate/cholesterol enzymes. Our quantitative degradomics data indicate that HMGCS1 is stable for several days in cells with active mTORC1 while substantially degraded within 5 hours of mTOR inhibition. We have determined the E3 ligase that is solely responsible for the degradation of HMGCS1. Reversing the stabilization of HMGCS1, by a targeted proteolysis approach, significantly inhibited the anchorage-dependent and independent cell proliferation in HMGCS1 copy-number-dependent manner. Altogether, our study suggests that HMGCS1 is an underappreciated, first gatekeeper of the mevalonate pathway flux and that the mTORC1-UPS-HMGCS1 axis dynamically controls the initiation of mevalonate production to meet the cellular metabolic demand. This pathway is distinct from the ER-associated degradation of HMGCR and Squalene monooxygenase (SQLE), which respond to cellular sterol levels.

NIH Research Projects · FY 2025 · 2024-05

Project Summary Pancreatic cancer is the fourth-most lethal cancer across both sexes, with five-year overall survival of all stages at 12%. In patients diagnosed with pancreatic cancer, first-line chemotherapy with gemcitabine and nab- paclitaxel with or without FOLFIRINOX often requires second-line chemotherapy combinations, with median survival ranging from 6 to 26 months. New therapeutic combinations are needed. One promising approach for overcoming this impasse centers on senescence. Cells can undergo senescence in response to replication, oncogene induction, or targeted drug therapy, including most chemotherapeutic regimens used for pancreatic cancer. However, while senescent cells have been implicated in tumorigenesis via pro-inflammatory factors seen in circulation as part of the senescence-associated secretory phenotype (SASP), their direct tumoral activity over time and distribution elsewhere is unknown. Moreover, current senescence imaging agents are small molecules measuring lysosome activity. An antibody-based approach, in contrast, offers greater targeting specificity and biological links to surface antigens. Noninvasive PET tools for senescence antigens will improve our ability to identify senescence with the option to change the isotope for targeted alpha therapy in the tumor. The members of the Scott Lowe Lab have pioneered senescence induction in pancreatic cancer with the combination of trametinib (T) and palbociclib (P), leading to the release of cytokines as SASP remodels the tumor microenvironment and beyond. Using immunoPET to study combination TP therapy in their models, we have found that shed antigens such as VEGF and IL-6 are decreased in the tumor environment while membrane- bound antigens like uPAR are elevated. We hypothesize that senescence induced by chemotherapy is temporal and immunoPET can be used noninvasively to quantify these dynamics during therapy. In collaboration with the Scott Lowe and Christine Iacobuzio-Donahue Labs at Memorial Sloan Kettering Cancer Center, we will use noninvasive immunoPET imaging to (1) quantify previously identified senescence markers during senescence- inducing therapy in human and murine pancreatic cancer models, (2) independently discover more senescence- specific markers, and (3) use targeted immunoPET agents for endoradiotherapy and improved senolytic delivery. In collaboration with Patricia Ribeiro Pereira at Washington University at St. Louis, we will also identify pharmacologic methods to prevent antigen shed, while advanced PET reconstruction with Joaquin Lopez- Herraiz at Complutense University Madrid will enable dual radiotracer immunoPET imaging of immune populations during therapy. Our efforts to quantify senescence in vivo with the proposed markers and enhance targeted alpha therapy with senescence will be guided by Dr. Lisa Bodei for clinical relevance and Andrea Schietinger for immunological insight. This work will unlock multiple directions in the precision theranostics of senescence, with great potential for career development and R01 research encompassing numerous cancers.

NIH Research Projects · FY 2026 · 2024-04

PROJECT SUMMARY The Fungal Colonization Resistance (FunCoRe) Program Project “Understanding the molecular mechanisms regulating fungal colonization and disease in the mammalian intestinal niche” focuses on developing an integrated and predictive model of the molecular, cellular, and metabolic determinants of Candida gastrointestinal (GI) colonization and its relationship to life-threatening invasive candidiasis. FunCoRE consists of 3 projects and 3 cores and combines in-depth studies of hematopoietic cell transplant patients with defined Candida colonization phenotypes and curated clinical data with experiments in reductionist experimental models of Candida GI colonization. The patient cohort serves as a source for a large Candida strain collection and for fecal metabolite abundance measurements that are harnessed in projects and cores. Project 1 emphasizes the role of morphology, virulence factors, and strain-specific genomic features on C. albicans and C. parapsilosis GI colonization. Project 2 characterizes immune signaling pathways and cell types (Paneth cells, innate lymphoid cells) that mediate Candida colonization resistance. Project 3 focuses on the bacterial microbiota and their biosynthetic capabilities, with a focus on short- and medium-chain fatty acids, to determine direct and indirect mechanisms of colonization resistance. A Gnotobiotic Core enables deep mechanistic dissection of the contribution of individual molecular, cellular, immune pathways, and commensal bacteria to colonization phenotypes. A Mathematical Modeling Core ties all 3 projects together and will generate predictive models of Candida GI colonization that incorporate specific fungal attributes, the bacterial microbiota and metabolites, and host immune parameters. Modeling outputs will be iteratively tested and refined in collaboration with the experimental projects to explore the overall hypothesis that the synergy of experimental and modeling approaches will reveal fungal, immune-mediated, and metabolic determinants of Candida GI colonization and colonization resistance which can inform and be harnessed for therapeutic strategies to limit invasive candidiasis. Aim 1 will investigate the Candida functional capacities that promote GI colonization; Aim 2 will interrogate host cell networks and colonization-relevant metabolites at the host-Candida interface; Aim 3 will develop an integrative model of Candida colonization resistance that identifies ecologic, immune, and metabolic targets to restore and enhance Candida colonization resistance. Successful completion of these aims will be supported by a strong Administrative Core management plan to facilitate communication and collaboration between projects and cores. The breadth and complementarity of expertise, multidisciplinary approaches, track record of collaboration, and commitment to establish a mentored undergraduate internship program represent signature features of FunCoRe. Insights will inform benchtop-to-bed strategies to formulate novel interventions that can shape the fungal communities in the GI niche to prevent invasive candidiasis and improve patient outcomes.

NIH Research Projects · FY 2026 · 2024-04

SUMMARY The International Skin Imaging Collaboration (ISIC)'s overarching goal is to leverage technology to reduce skin cancer mortality and morbidity. The ISIC Archive, created and led by Memorial Sloan Kettering Cancer Center since 2016, is the leading publicly available skin imaging repository. With >240 000 images, >6000 registered users, and 17-21 million image downloads per month, the ISIC Archive has made major contributions to dermatology teaching and the development of artificial intelligence (AI) for skin cancer diagnosis. ISIC has also been central to the development and adoption of standards for skin imaging to promote quality and interoperability. ISIC has hosted 5 machine learning Grand Challenges and hosts continuous Live Challenges using ISIC Archive images. ISIC Challenges have attracted 6818 participants and 25 326 submissions. ISIC has directly published 21 scientific papers, while another 260 peer reviewed papers and 3060 technical reports have relied on ISIC data. Here, we propose to develop the ISIC Archive into an even greater resource for research through major enhancements that focus on making the ISIC Archive fully compliant with FAIR principles (findability, accessibility, interoperability, and reusability), while expanding the Archive with additional support for broadly used data formats (eg, Digital Communications Standards in Medicine [DICOM]) and skin imaging modalities (clinical imaging, total body photography [TBP]). We propose to leverage ISIC's leadership in the DICOM community to improve software support for established skin imaging standards, while publishing new standards for TBP, and to improve quality and interoperability for these imaging modalities. We have organized key milestones for this proposal under 4 specific aims: (1) to bring the ISIC Archive into rigorous compliance with FAIR principles; (2) to overcome the challenges of ingesting and sharing clinical images and representations of TBP while maintaining patient privacy and addressing sensitivity; (3) to strengthen the interoperability of ISIC data through the use of DICOM; and (4) to implement approaches to facilitate scaling, disseminating, and sustaining the ISIC data repository. The resulting ISIC Archive will be a more scientifically robust, impactful, and sustainable resource. ISIC will continue to promote use of the Archive and engage the dermatology and computer science communities through Challenges, workshops, and publications. By promoting and supporting research to effectively leverage skin imaging and AI to diagnose skin cancer earlier and more accurately, we will save lives and reduce the morbidity associated with unnecessary biopsies and the treatment of advanced disease.

NIH Research Projects · FY 2026 · 2024-04

Only one in ten pancreatic cancer patients is alive 5 years after their diagnosis. Unfortunately, this outlook has not improved significantly in decades. This is in stark contrast to most other cancers, which have benefited tremendously from the recent advances in targeted therapies and immunotherapies. Therefore chemotherapies remain as the only option for pancreatic cancer patients, but these therapies are not particularly effective, and they produce significant side effects. Given that pancreatic cancer patients have received little benefit from conventional cancer therapies, there is a need for deeper understanding of the biology of pancreatic cancer. Here, we propose to investigate how low levels of oxygen (hypoxia) promote emergence of aggressive behavior of pancreatic cancer cells. Cancer cells in the same pancreas tumor can show substantial diversity at the molecular level and in how they respond to therapies. The inability to effectively target all cancer cell subsets within a tumor contributes to treatment failure. Pancreas tumors have regions of hypoxia, which are caused by variable access of cancer cells to blood vessels that deliver oxygen to tissues. Our initial results indicate that metabolic adaptations in cancer cells that reside in these hypoxic regions reprogram the cancer cells, so that they become resistant to chemotherapy. In other words, what does not kill the cancer cells makes them stronger. We propose to investigate other possible outcomes of this reprogramming process, including the ability of the cancer cells that experienced hypoxia to contribute to tumor growth as well as their ability to spread into distant parts of the body (metastasis) or resist therapy. The second part of the proposed study will focus on identifying the molecular changes that are induced in the cancer cells by hypoxia. In the third part, we will determine which of these molecular changes enable cancer cells to survive hypoxia and acquire more aggressive behavior. The project will identify fundamental drivers of cell state diversity in pancreatic cancer, which may point to a new generation of targeted therapies that will control the cellular composition of tumors to improve treatment response. We have devised a two-prong research strategy focusing on both the role of the hypoxic pancreatic cancer cells within the tumor and the molecular programs unlocked by hypoxia that promote tumor progression. If successful, this study will (i) fundamentally advance our understanding of the molecular and biological mechanisms underpinning intra-tumoral heterogeneity, tumor progression, and drug resistance; (ii) provide innovative experimental approaches for investigating phenotypic diversity; and (iii) elucidate novel strategies for targeting tumor heterogeneity, malignant progression, and drug resistance in pancreas cancer.

NIH Research Projects · FY 2025 · 2024-04

PROJECT SUMMARY/ABSTRACT: Radiation therapy has a definitive role in curing high risk, very high risk, and even oligometastatic or low- volume metastatic disease as phase III trials in these settings have demonstrated overall survival benefits. However, radiation therapy also involves incidental radiation dose to rectum, small bowel and urethra, which cause radiation dose-limiting toxicity. Locoregional control, particularly within pelvic lymph node regions where currently possible radiation doses are non-curative, can be a limiting factor in overall prostate curability in these settings. In this proposal, we seek to create state-of-the-art PSMA-targeting therapeutics, taking advantage of this relatively prostate-specific surface antigen. We will create two molecules consisting of the PSMA ligand linked to a lysosomal cleavable linker which is turn linked to radiation sensitizing small molecule inhibitors of the DNA damaging response. One molecule will involve a potent inhibitor of ATM and the second will involve an inhibitor of DNA-PK, two targets integral to radiation efficacy. In SA1 we will test these novel molecules with in vitro assessments of inhibition of the ATM and DNA-PK kinases in PSMA expressing prostate cancer cell lines compared to PSMA non-expressing cells. In SA2, we will test these molecules for in vivo efficacy in prostate cancer xenografts and determine target engagement and preliminary normal tissue toxicity. The overarching goal addressed by the proposed research project is to develop treatments that could improve survival for men with lethal prostate cancer by improving locoregional control, which does impact overall survival in these men. In addition, if these molecules successfully radiosensitize, it is also possible they will allow for external beam radiation dose de-escalation and enable less normal tissue toxicity in order to improve quality of life outcomes. Finally, in downstream work the molecules could be combined with Radium-223 to systemically radiosensitize a therapy proven to extend survival. In this pilot grant, we will determine the initial feasibility of this approach, laying the groundwork for further therapeutic development.