University Of Tx Md Anderson Can Ctr

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Melanoma is the fifth most common cancer in the United States. While immunotherapy has improved treatment options, patients who do not respond to immune checkpoint inhibitors (ICI) continue to have poor prognosis. Developing a strategy to identify patients with high likelihood of response to treatment intensification with a novel combination of neoadjuvant ICI and radiotherapy (RT) could significantly improve outcomes. In this proposal, we plan to create a novel, multimodality platform to predict for responders to ICI+RT by leveraging a pilot, phase 2 clinical trial evaluating pathologic response and long-term outcomes after neoadjuvant ICI+RT, in mucosal melanoma, a subtype of melanoma enriched for immunotherapy refractory patients. As part of the “Preoperative Radiotherapy and Immunotherapy in Sinonasal and Anorectal Melanoma (PRISAM)” study, sinonasal and anorectal melanoma patients with non-metastatic disease receive 1-2 cycles of neoadjuvant ICI before starting neoadjuvant RT (with tissue-, blood-, and advanced imaging-based biomarkers collected before each change in treatment modality). This sequential therapy approach with serial biomarker collection provides a unique opportunity to prospectively evaluate how each treatment modality changes each patient’s tumor microenvironment and systemic response. In Aim 1 we focus on longitudinally evaluating tumor tissue using multiplex immunofluorescence and RNA expression analysis. We also plan to evaluate the systemic response to therapy by profiling peripheral mononuclear blood cells and T-cell receptor sequences as well as quantifying patient-specific circulating tumor DNA. In Aim 2, we plan to longitudinally evaluate multiparametric MRI sequences to characterize dynamic changes in cellularity and microvascular function using DWI/ADC and DCE- MRI. Our overall objective is to determine if baseline features as well as early ICI-induced changes measured by the various biomarker assays independently and/or together predict for pathologic response as well as overall outcomes. Ultimately, this proposal serves as a pilot study to evaluate the potential of our multimodality biomarker platform to predict for response to neoadjuvant ICI+RT. We anticipate the results will serve as a first step in the design of the next generation of neoadjuvant immunotherapy based clinical trials, whereby an optimized, longitudinal biomarker platform can help select the therapeutic pathway most likely to yield treatment response.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Type 1 conventional dendritic cell (cDC1) vaccination is a novel pre-clinical cell therapy that elicits systemic immunity in several murine tumor models, including melanoma, and augments response to immune checkpoint blockade (ICB). cDC1s possess the ability to cross-present tumor antigen to CD8+ T cells, migrate from tumors to tumor-draining lymph nodes, and enhance naïve CD8+ T cell priming; however, mechanisms by which cDC1s elicit effective anti-tumor CD8+ T cell responses in the context of vaccination remain unclear. My project will explore the potential of cDC1 vaccination alone and in combination with αCTLA4 ICB to generate diverse and effective CD8+ T cell responses. Specifically, I will study the ability of cDC1s to induce antigen spread, which is defined as induction and expansion of T cell clones that were not a direct target of or present in the initial targeted therapy. Importantly, antigen spread has been observed in patients responsive to immunotherapy and may present an opportunity to counteract tumor antigen loss that can occur during tumor evolution. It is thought to occur after tumor antigens, released from dying tumor cells, are phagocytosed by antigen presenting cells (APCs) and cross-presented to naïve T cells to stimulate their activation. In addition, CTLA4 blockade can induce antigen spread by promoting T cell costimulation and lowering the activation threshold for naïve T cells during priming. The specialized functions of cDC1s create an ideal setting to study antigen spread. My project investigates the ability of a target-antigen-loaded cDC1 vaccine alone and with αCTLA4 to control target antigen- expressing (antigen-positive) and target antigen non-expressing (antigen-negative) tumors using B16F10 and YUMM1.7 murine melanoma lines expressing ovalbumin (B16F10) and mLama4 and mItgb1 (YUMM1.7) as target antigens. I hypothesize that cDC1 vaccination elicits antigen spread, and that combination treatment with cDC1 vaccination and αCTLA4 further potentiates tumor control through increasing CD8+ T cell diversity. In AIM 1, I will test the hypothesis that the target antigen-loaded cDC1 vaccine will control target antigen-positive and target antigen-negative tumors, and that cDC1s enhance αCTLA4 efficacy against target antigen-negative tumors by enhancing the quantity and quality of the CD8+ T cell response. In AIM 2, I will test the hypothesis that cDC1 vaccination with αCTLA4 will result in the most diverse CD8+ effector T cell repertoire, and that cDC1 vaccination elicits a CD8+ T cell transcriptional state that promotes tumor control. The Watowich and Yee laboratories at MD Anderson provide the ideal environment to conduct my proposed studies, as I will learn cutting-edge immunotherapy techniques, strengthen my fundamental immunology knowledge, and acquire pertinent bioinformatics analysis skills under the direct guidance of my mentors. This training environment will afford me the chance to learn from world leaders in tumor immunology and grant access to state-of-the-art facilities and resources that are integral to my further development of my career as a physician-scientist.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Recent paradigm-shifting discoveries demonstrated that neurons are critical drivers of malignancies (cancer), leading to clinical trials to test neuromodulatory drugs in cancer patients. Many types of cancer are associated with severe pain which indicates sensory neuron hyperactivity, raising the intriguing possibility that cancer pain- associated sensory neuron activity accelerates cancer progression. Our long-term objective is to understand the role of neuronal activity in cancer pathophysiology and identify therapeutic targets to improve cancer patient survival and quality of life. Leveraging preclinical models of malignant peripheral nerve sheath tumor (MPNST), where the neoplasm develops within peripheral nerves, our preliminary data reveal that MPNST cells induce sensory neuron hyperactivity and pain, and that sensory neuron activity accelerates tumor growth. Based on these findings, our central hypothesis is that a feedforward mechanism exists between cancer pain-associated sensory neuron hyperactivity and MPNST growth. We will address the hypothesis by uncovering how MPNST cells activate sensory neurons to promote cancer pain via fibroblasts (Aim 1) and determining the mechanisms underlying pain-associated sensory neuron activity-mediated tumor progression in a neuronal subtype-dependent manner (Aim 2). To achieve these aims, we have assembled a multi-disciplinary team consisting of cancer neuroscientists (Yuan Pan, PhD and Xiaofan Guo, MD-PhD), pain researchers (Peter Grace, PhD and Andrew Shepherd, PhD), an MPNST physician-scientist (Angela Hirbe, MD-PhD), a pathologist (John Chrisinger, MD), a bioinformatician (Rajasekaran Mahalingam, PhD), biostatisticians (Chongliang Luo, PhD and Roland Bassett, PhD), and a data management and sharing expert (Robert Allaway, PhD). Together, we will leverage genetically engineered mice, mouse and human MPNST cells, human MPNST tumor specimens, and neuroscience approaches to test the hypothesis. Successful completion of these studies will uncover the mechanisms under which cancer pain-associated neuronal activity interacts with MPNST cells to drive malignancy. Findings from this study will open a new avenue for predicting prognosis and improving the survivorship and quality of life of patients with these deadly tumors.

NIH Research Projects · FY 2025 · 2025-09

The concept of synthetic lethality holds great promise in cancer targeting, as a critical principle for conceptualizing how to effectively target tumors with combinatorial therapies. However, the vastness of the pairwise gene search space demands integration of advanced computational and experimental approaches. The complete map of genetic interactions in yeast has yielded insight into the functional organization of the eukaryotic cell and extending this approach into mammalian cells promises to both elucidate the “wiring diagram” of a normal cell and reveal how that network is rewired in cancers and in genetic diseases more broadly. CRISPR technologies have made genetic screens in mammalian cells tractable but, to date, even CRISPR-mediated genetic interaction assays do not scale effectively due to the inherent combinatorial nature of a gene-vs-gene search space. Moreover, the necessity of measuring interactions in different cell types renders the task at least two orders of magnitude larger in human cells than in yeast. To realize the potential of synthetic lethal interactions in cancer, the field requires major improvements in both assay efficiency and in predictive models that narrow the search space. We have developed a CRISPR/Cas12a based multiplexing system called IN4MER that accurately quantitates genetic interactions with five-fold fewer reagents than the current state of the art. In parallel, we reverse engineer the yeast functional interaction network to predict regions of the search space enriched for genetic interactions and apply these findings to human data to guide experimental design. We hypothesize that, by integrating our computational and experimental approaches, will identify biologically relevant functional modules that are enriched for genetic interactions. With our highly efficient IN4MER platform, we can assay all pairwise combinations of hundreds of target genes at a scale comparable to a standard genome-wide monogenic knockout screen. By doing so across dozens of relevant cell lines, we can understand how genetic interaction networks vary across lineage and mutation state, identifying candidate synthetic lethalities for tumor-specific therapeutic exploitation. Further, screening data will feed back into the predictive model, iteratively increasing the efficiency and discovery potential of this approach. This proposal offers the potential for a major advance in understanding the basic biology implications of genetic interactions and the translational opportunities for exploiting context-specific synthetic lethalities in cancer and beyond.

NIH Research Projects · FY 2025 · 2025-09

Cervical cancer is a chronic condition for both cancer survivors who experience long–term toxicity from radiation therapy (RT) and those who experience relapse and require treatment for the remaining years of their lives. Standard treatment includes chemotherapy, external and internal RT. The Food and Drug Administration (FDA), recently approved use of the immune checkpoint inhibitor pembrolizumab for patients with International Federation of Gynecology and Obstetrics (FIGO) 2014 Stage III-IVA cervical cancer. There are currently no means to perform risk stratification to personalize this complex and often toxic treatment regimen. We propose that widely implementable response-based biomarkers (e.g., imaging techniques and blood biomarkers) hold great promise for risk stratifying cervical cancer patients in the immediate future. Almost all cervical cancers are associated with prior infection with human papillomavirus (HPV), and despite an uptick of worldwide vaccination rates for HPV, cervical cancer remains the fourth most common malignancy amongst women. Women in low-resource settings lack advanced imaging techniques, with access only to computed tomography (CT) imaging, and current efforts to develop predictive and prognostic biomarkers for cervical cancer involve state-of the- art next generation sequencing and other “omics” platforms that are not easily implementable in low-resource areas with the highest cervical cancer incidence. There are no well-established predictors of treatment response to inform RT dose decisions to balance the risks of toxicity to disease control. Thus, there is a need to develop evidence-based, globally applicable, early-treatment biomarkers so that treatment regimens and RT doses can be optimized. The proposed project hypothesizes that a globally accessible and pragmatic predictive signature (GAPPS) for risk stratification utilizing serially sampled circulating biomarkers along with imaging response and clinical FIGO stage will accurately predict recurrence after CRT and response to immune checkpoint blockade with the ultimate goal of ensuring that patients requiring intensified treatments receive them, while those effectively treated with standard RT are spared from unnecessary, chronic debilitating toxicity and cost. To test this hypothesis, Aim 1 will focus on identifying a GAPPS using pre-treatment and early response circulating blood-based and CT-based biomarkers. Next, in Aim 2, the utility of a GAPPS will be validated in prospectively predicting two-year OS in a multi-site cohort of patients treated with CRT including pembrolizumab when indicated and available. Successful completion of the project will have an immediate impact on women with cervical cancer in different resource settings. Risk stratification tailored to individual patients will render treatments more effective and affordable with compounding benefits

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT / PROJECT SUMMARY Allogeneic hematopoietic cell transplant (alloHCT) is a curative therapy for many life-threatening hematologic malignancies but its full therapeutic potential is undermined by complications such as chronic graft-versus-host disease (GVHD). Chronic GVHD is the greatest cause of non- relapse mortality and develops in 25-50% of long-term survivors of alloHCT. Novel approaches are needed to diagnose and treat chronic GVHD after alloHCT. 4-[18F]fluoro-1-naphthol ([18F]4FN) is a novel radiotracer that is specific to high energy reactive oxygen and nitrogen species produced during the respiratory burst in the active innate immune system. The proposed pilot study will apply [18F]4FN PET/CT to 16 patients with joint and other manifestations of chronic GVHD to obtain preliminary estimates of the anatomic and temporal correlations between [18F]4FN PET/CT signal intensity / location and chronic GVHD manifestations. Although PET, CT, and MRI have previously been used to image chronic GVHD, our approach differs by the use of [18F]4FN which reflects the functional activity of the innate immunity as indicated by respiratory burst intensity in contrast to glucose uptake as in [18F]fluoro-2-deoxyglucose (FDG) PET or anatomical changes as in CT and MRI. Patients will receive one to three [18F]4FN PET/CT scans and six to twelve Chronic GVHD Assessments. Data from these endpoints will be used to calculate preliminary estimates of the utility of [18F]4FN PET/CT as a predictive and prognostic / pharmacodynamic biomarker. We will test the following hypotheses with our specific aims: 1) [18F]4FN PET/CT images correlate anatomically with chronic GVHD manifestations and selected [18F]4FN PET/CT images will precede chronic GVHD manifestation changes. 2) [18F]4FN PET/CT images will provide pharmacodynamic metrics for treatment of chronic GVHD. Information gained from this pilot study will form the scientific basis for a subsequent larger phase II imaging trial to evaluate [18F]4FN PET/CT as a quantitative, predictive, and prognostic / pharmacodynamic biomarker for chronic GVHD. [18F]4FN PET/CT could be applied to other inflammatory conditions such as acute GVHD and cytokine release syndrome / immune effector cell associated neurotoxicity syndrome after CAR T cell infusion, and rheumatologic diseases. This multiple principal investigator project combines the unique, complementary expertise of Drs. Chen (alloHCT, chronic GVHD) and Piwnica-Worms (molecular imaging, radiopharmaceuticals). In 2022-2023, MD Anderson Cancer Center performed 416 adult alloHCTs of which 13.5% developed chronic GVHD within 1 year.

NIH Research Projects · FY 2025 · 2025-09

Individuals living with cancer and HIV face unique challenges due to the confluence of dual stigma at individual, provider, community, and/or structural levels. Addressing stigma at multiple levels is critical to improving their cancer outcomes. We propose to use a randomized controlled trial to test the effects of a multilevel, culturally adapted storytelling intervention to address stigma at both patient and provider levels. The proposed project will recruit patients (i.e., Zambian women living with cancer and HIV, WLC+H) and healthcare workers at the Cancer Diseases Hospital (CDH). In the patient-level intervention, patients (N=240) will be randomly assigned to one of four conditions in a 2 (storytelling videos vs neutral videos) by 2 (storytelling disclosure vs neutral disclosure) factorial design. The storytelling videos are inspirational videos featuring actresses portraying Zambian WLC+H and sharing their stories and the storytelling disclosure involves privately speaking or writing about their experiences as WLC+H. Participants in all four conditions will attend four weekly 20-minute sessions, involving video watching and disclosure. Patient outcomes will be assessed at baseline, immediately after the intervention, and at 1-, 3-, 6- and 12-month follow-ups. We hypothesize that the intervention groups will experience improved quality of life and reduced depressive and anxiety symptoms through reduced internalized stigma and improved coping self-efficacy. In the provider-level intervention, we will explore the impacts of the intervention on workers and patients. Healthcare workers (N=100) will be randomly assigned to either an intervention group (N=50) watching storytelling videos or a waitlist control group (N=50) watching neutral videos. Worker outcomes will be self-reported at pre- and post-intervention. An anonymous patient survey will be administered to WLC+H to compare outcomes between those treated before (control cohort, N=50) and after (intervention cohort, N=50) the intervention's implementation. We hypothesize that the intervention will reduce social and enacted stigma and increase compassion among healthcare workers. It will also reduce perceived stigma and increase care satisfaction among patients. Should the interventions prove effective, an immediate outcome will be evidence-based, culturally responsive, and widely accessible interventions that support individuals experiencing dual stigmas of HIV and cancer beyond Zambia and across the globe, ultimately improving healthcare for all populations.

NIH Research Projects · FY 2025 · 2025-09

HIV persistence during ART is primarily due to the establishment of long-lived viral reservoirs in resting immune cells and mucosal and lymphoid tissues. Recently, we showed that blocking TGFβ in a non-human primate model (NHP) of mucosal SIV infection leads to enhanced T cell differentiation toward effectors, which in turn leads to SIV latency reversal. TGFβ, the master regulator of tissue immunity, is increased in HIV-1/SIV infection and its levels remain elevated with ART. We found an increased TGFβ signature at ART initiation (ARTi) including an increase in quiescent/resting memory T cells which parallel a decrease in effectors with waning immune responses and decreasing viral loads. Herein, we propose to use TGFβ blockade during ART initiation to reverse the resting/quiescent status of T cells and support viral transcription while ART blocks new infection. We hypothesize that this will decrease latency establishment and reduce the levels of the already established reservoir especially when TGFβ blockade is used in combination with a broadly neutralizing antibody (bNAb). As proof-of-principle we will test this combination in a non-human model of SHIV-AD8 infection with 4 groups of monkeys: 1) TGFβ blockade with galunisertib, 2) HIV-bNAbs, 3) Galunisertib+bNAbs, 4) Isotype control. Moreover, in order to investigate whether galunisertib’ s ability to stimulate the immune system and enhance the vaccinal effect of the bNAb, we will label HIV-bNAbs with Cy5 and follow their dynamics in vivo. PET/CT-guided collection of tissue areas of viral reactivation will allow us to study the impact of bNAbs in tissues and determine whether galunisertib increases their ADCC activity. Overall, we expect to be able to reduce viral reservoir and stimulate immune responses so that at the end of ~1 year of ART, viral rebound upon ART interruption will be delayed or controlled.

NIH Research Projects · FY 2025 · 2025-09

Project Summary While most cases of lung cancer are related to tobacco use, with effective tobacco control, lung cancer in those with light or no smoking history is becoming a greater public health concern. It is estimated that lung cancer in those who have never smoked (LCINS) would be the fifth-leading cause of cancer death worldwide and the seventh-leading cause in the United States. No widely accepted early detection strategy exists for those who are not high risk for lung cancer based on a tobacco use history. Efforts to broaden low-dose CT based lung cancer screening to those at lower risk have been likely met with increasing rates of overdiagnosis. Our team has an extensive track record of developing and validating blood-based biomarkers for cancer risk, including a 4-protein biomarker panel (4MP) that has been extensively validated to improve prediction of lung cancer risk over clinical risk factors alone. Here we demonstrate that the 4MP and a newly validated 4-metabolite biomarker panel (4MetP) have similar performance in LCITNS. Our translational objective of this proposal is to test the predictive performance of the 4MP and 4MetP, individually and in combination with clinical risk models, for 1- year risk prediction of lung cancer among those no smoking history. We hypothesize that a composite panel of the 4MP, 4MetP, with addition of clinical risk characteristics, can identify those with no smoking history and at a high enough risk that they may benefit from lung cancer screening. Samples from those with no smoking history from several MD Anderson cohorts and the Prostate, Lung, Colorectal, and Ovarian (PLCO) trial will be utilized to build a composite panel consisting of blood-based biomarkers and clinical characteristics to identify those at risk for lung cancer. Aim 1: Evaluate the predictive performance of the 4MP and 4MetP for 1-year risk of lung cancer among never-smoking individuals. We will build on preliminary data and evaluate the performance of the 4MP and 4MetP in cohorts of LCINS and matched controls from MD Anderson. We will measure the 4MP and 4MetP in 12-month prediagnostic LCINS samples from the PLCO (n=74) as well as ten times the number of unmatched controls to externally validate the biomarker panels. We will assess the performance of these panels and any composite biomarker panel to predict 1-year risk of developing lung cancer as well as risk of lung cancer mortality. Aim 2: Assess whether an algorithm that consider repeat biomarker testing improves sensitivity and lead-time detection of lung cancer among never-smoking participants in the PLCO cohort. We will obtain all available longitudinally collected samples from the cases and controls specified in Aim 1. We will evaluate whether an individualized algorithm accounting for biomarker trends can improve upon a single threshold measurement to improve diagnostic performance.

NIH Research Projects · FY 2025 · 2025-09

Project Summary Background: The interaction between T cell receptors (TCRs) and antigens presented by MHC proteins is crucial for initiating antigen-specific T cell responses in various biomedical contexts. Understanding these interactions can provide insights into adaptive immunity, antigen discovery, therapeutic TCRs, and the impact of T cells on immune checkpoint inhibitor (ICI) and other immunotherapy treatments. However, traditional experimental methods for studying TCR-antigen interactions are inefficient, costly, and labor-intensive. Solution: We have developed a deep foundation model, pMTnet-omni, which accurately predicts TCR-antigen interactions for both human and mouse, and for class I and II pMHCs. This tool overcomes the caveats of traditional experimental approaches. We will innovate upon this foundation model to (a) discover TCRs for therapeutic use, (b) perform antigen discovery from spatial-TCR-sequencing data, and (c) develop TCR-based biomarkers for predicting immune-related adverse events (irAEs) for patients on ICI treatments. Team: The multidisciplinary team includes bioinformaticians, immunologists, and clinical researchers, ensuring comprehensive development and application of the proposed aims. We have developed AI-based techniques in predicting TCR-antigen interactions and have done pioneering works in multi-instance learning, spatially resolved transcriptomics, T cell engagers, biomarker development, which laid the foundation for this proposal. Aim 1: We will use AI (pMTnet-omni) to discover and optimize TCRs for T cell engagers, against a KRAS neoantigen. AI, coupled with limited experimental validation, will reduce the cost and time spent on engineering TCRs and will result in better TCRs. We will experimentally validate these TCRs’ specificity and efficacy. Aim 2: We will apply multi-instance learning and pMTnet-omni to spatial-TCR-sequencing data to identify novel antigens, based on spatial location, gene expression and TCR sequences. The discovered antigens can serve as druggable targets by therapeutic TCRs, or be incorporated into tumor vaccines. Aim 3: We will develop a biomarker model using pMTnet-omni, TCR-sequencing data generated from patient peripheral blood, and autoantigens of healthy organs to predict irAEs in ICI-treated patients. This biomarker approach is highly generalizable as we can develop this biomarker for any antigen of interest in any disease. Significance: (1) The project aims to create platform technologies and resources, based on the deep foundation model, for broad research and translational uses. (2) We will prove that AI can accelerate the discovery of therapeutic TCRs, which will ultimately benefit patients suffering from various diseases. (3) Significant resources have been spent on inventing spatially resolved transcriptomics (including spatial TCR- seq). We will showcase the significant value that can be derived from such expensive data. (4) Our biomarker method will provide an interpretable and non-invasive approach to extract clinically actionable information from patient TCR repertoire. (5) Our research outcome will be shared publicly through webservers and databases.

NIH Research Projects · FY 2025 · 2025-09

Overall Summary This P01 program project is focused on bladder cancer, a common human malignancy with significant morbidity and mortality in the United States and across the world. The goal is to develop new and more effective therapies for both early and advanced disease. This goal will be accomplished with molecular characterization of mucosal field effects and their evolution to aggressive and therapy-resistant bladder cancer. Whole-organ mapping studies will be complemented by molecular characterization of two mechanisms based on downregulation of lysophosphatidic acid receptor 6 (LPAR6) and calcium-binding protein 39-like (CAB39L). These studies will be supplemented by in-depth investigations of the role of luminal and basal phenotypic switch in the urothelium and its role in therapeutic responses of bladder cancer cells. This P01 project has three well-defined goals: 1) development of molecular profiles of bladder cancer evolution from field effects to clinically aggressive disease on a whole-organ scale and investigating LPAR6 and CAB39L role, which contribute to bladder carcinogenesis by dysregulating the urothelial differentiation program; 2) characterization of the role of luminal differentiation program and its role in tumor formation; and 3) characterization of the role of luminal to basal plasticity in response to interferon therapy. These goals will be accomplished in three projects. Project 1: Understanding the Evolution of Bladder Cancer from Field Effects; Project 2: The Role of Nuclear Receptors Signaling in Bladder Cancer; and Project 3: Identifying Molecular Vulnerabilities to Improve Interferon Gene Therapy in Bladder Cancer. These projects will be supported by the Administrative, Pathology and Biostatistics and Bioinformatics Cores. Collaborative studies by Projects 1 and 2 as well as by Projects 1 and 3 will focus on the role of luminal to basal plasticity and dysregulated urothelial differentiation in therapeutic responses of bladder cancer. Projects 1 and 2 will focus on the role of luminal dysregulation controlled by nuclear receptors such as Pparg in the evolution of bladder cancer from mucosal field effects. Projects 1 and 3 will investigate the expression pattern of IFN target genes in the evolution of bladder cancer from mucosal field effects as well as the role of luminal to basal plasticity in therapeutic response. The work is proposed by a multi-institutional team of scientists who have worked on bladder cancer for decades and have documented collaborations focused on bladder cancer. The studies proposed in this P01 program project are highly promising options for early detection, prevention, and treatment of bladder cancer.

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT The recalcitrance of high-grade serous ovarian cancer (HGSOC) to available therapies is largely enabled by the disease’s ability to fast disseminate into the peritoneal cavity and its intrinsic molecular heterogeneity. A first-line platinum-taxane regimen, in combination with PARP inhibitors (PARPi) as maintenance therapy has improved patients’ survival and revolutionized HGSOC management, especially for those carrying mutations in BRCA1/2 genes. Recently, the sensitivity to PARPi has been associated not only with BRCA1/2 mutations, but with a broader homologous recombination deficiency (HRD) signature, which accounts for nearly 50% of HGSOCs. Surprisingly, PARPi sensitivity has also been observed in homologous recombination-proficient (HRP) tumors. Despite successful, the emergence of cross-resistance to platinum/PARPi treatment strongly hindered the long- term clinical benefit, posing a significant clinical challenge. The genetic and epigenetic instability of HGSOCs strongly contributes to the development of cross-resistant clones, which undergo lineage expansion, leading to treatment failure and disease relapse. Furthermore, the mechanisms driving the selection of Platinum/PARPi cross-resistant clones, and their evolution remain poorly understood. A critical question is whether recurrent disease is driven by pre-existing resistant clonal populations or by de novo (acquired) molecular scars selected during the treatment. Addressing this question is crucial for understanding HGSOC progression and developing more effective therapies. To investigate these issues, we have developed a novel somatic mosaic genetically-engineered mouse model (smGEMM) of HGSOC combined with a molecular barcoding technology. This approach allows for the high-throughput isolation and detailed characterization of specific cell lineages, including therapy cross-resistant clones, enabling us to track clonal dynamics and to identify molecular drivers of resistance. To assess this hypothesis, we will pursue specific aims that 1) generate longitudinal lineage-traced models of HRP and HRD HGSOCs using smGEMM models, 2) functionally characterize the clonal dynamics of platinum/PARPi resistance and 3) perform cross-species validation of molecular dependencies driving platinum/PARPi resistance. Our findings will be significant because they will 1) uncover the transcriptomic and genomic mechanisms behind platinum/PARPi cross-resistance in HRP and HRD tumors, 2) reconstruct the evolutionary pathways of aggressive HGSOCs, 3) identify the cross-species molecular signatures of platinum/PARPi resistance. Importantly, this research has the potential to extend beyond HGSOC, as Platinum/PARPi combinaiton is used in other HRD cancers, such as breast and pancreatic cancer. Furthermore, our CRT and sm-GEMM platforms are adaptable to other gynecological cancers, hence benefiting the broader scientific community. Ultimately, this study will address urgent clinical needs by identifying molecular signatures of platinum/PARPi resistance and discovering biomarkers for patient stratification and therapeutic targeting.

NIH Research Projects · FY 2025 · 2025-09

PROJECT SUMMARY Research objectives: Influenza- and respiratory syncytial virus-associated pulmonary aspergillosis (IAPA/RSV- APA) are deadly secondary mold infections, especially in immunocompromised patients. Promising preliminary data suggest that restoration and enhancement of epithelial resilience and attenuation of post-viral immune pa- ralysis with locally delivered (inhaled) immunomodulators may provide an innovative therapeutic avenue to im- prove the often-detrimental outcomes of post-viral mold pneumonias. This approach will be studied using nu- anced sequential infection models in mice with underlying immunosuppressive conditions, i.e., remission-induc- tion chemotherapy for acute myeloid leukemia or high-dose glucocorticosteroids. Aim 1 will provide a temporally and spatially resolved characterization of host-associated, global post-viral, and virus-specific (RSV- or influ- enza-specific) impairment of pulmonary and systemic anti-Aspergillus immune defense. Aim 2 will characterize protective immunity against IAPA and RSV-APA after nebulized immunotherapy (TLR2/6/9 agonists or GM-CSF) given at different timepoints, with or without systemic antiviral or antifungal therapy. Detailed immunologic studies and dual-RNA sequencing will unveil key components of protective immune augmentation, fungal adaptation to an altered host environment after multimodal therapy, and host biomarkers associated with favorable therapeutic outcomes. Collectively, these studies will provide comprehensive preclinical data to facilitate future clinical trans- lation of nebulized immunomodulators to protect high-risk patients from deadly secondary mold pneumonias. Candidate and career development plan: Dr. Wurster’s postgraduate training in molecular oncology combined with his postdoctoral and junior faculty experience in experimental mycology and translational immunology pro- vides him with an ideal and unique background to study fungal pathogenesis, host defense, and immunomodu- latory therapy in the context of underlying malignancy, virus-induced immune dysfunction, and opportunistic mold infection. Supported by an outstanding team of mentors and advisors with internationally recognized expertise in fungal pathogenesis and preclinical infection models (Dr. Kontoyiannis), pulmonary immunobiology and im- munotherapy (Dr. Evans), viral immunology (Dr. Chemaly), spatial transcriptomics (Dr. Wang), and dual host/fun- gal transcriptomics (Dr. Bruno), this data science-focused K01 project will provide an ideal setting for thorough training in state-of-the-art transcriptomics techniques and advanced immuno-informatics approaches for multidi- mensional data integration. These training objectives, along with dedicated leadership training, will substantially enhance Dr. Wurster’s competitiveness as an emerging independent investigator and future thought leader in experimental and translational mycology. Moreover, the strategic alignment of this K01 project with Dr. Wurster’s career goal of establishing a host-targeted precision medicine program for fungal and polymicrobial infections in immunocompromised patients, and the institutional focus on personalized care programs, will provide an optimal framework for him to succeed as a future independent group leader at MD Anderson Cancer Center.

NIH Research Projects · FY 2025 · 2025-08

Across lymphoma subtypes, African American (AA) and Hispanic patients experience marked differences in age of onset, disease presentation, treatment, and survival. Biologic and patient-level factors contribute to inferior outcomes for specific patients but remain poorly understood. These groups are understudied in clinicopathologic cohorts examining lymphoma pathogenesis and therapeutic targets, in lymphoma clinical trials and are also more likely to receive care at under-resourced community practices, where lack of onsite hematopathology can lead to delayed or inaccurate lymphoma diagnosis. We have developed unparalleled resources to examine and address the multilevel factors influencing lymphoma outcomes. We established the multicenter Lymphoma Epidemiology of Outcomes Cohort Study, prospectively enrolling >13,000 lymphoma patients with banked tumor tissue, germline DNA, and linked pathology, demographic, treatment, and survival data on all patients. We performed the first large-scale genomic analyses of the most common lymphoma subtype, diffuse large B-cell lymphoma (DLBCL), from patients with African ancestry and identified that in addition to patient-level factors such as inferior access to care, AA patients with DLBCL have specific tumor mutations such as SETD2 and microenvironment phenotypes that may impact clinical outcomes. By developing murine and human SETD2 mutation models, we found these lymphomas feature genomic instability, a senescence-associated secretory phenotype, and an inflammatory microenvironment that can be effectively therapeutically targeted using SETD2 inhibitors available for use in humans in a trial. We hypothesize that these and other novel patient subsets can be readily diagnosed in the community from simple H&E slides, through deconvolution of complex tissue architecture. We contend that these data are clinically actionable through evidence-based community interventions, so that precision therapy for patients with lymphoma can be successfully implemented in clinical trials and in routine care, guided by their relevant predictive biomarkers. The overarching goal of this SPORE is to overcome gaps in knowledge, survival, and access to cancer care through a comprehensive bench-to-bedside-to-community approach that interrogates the relationships between ancestry, lymphoma biology, and patient outcomes; evaluates strategies to improve clinical trial participation and survival rates; and develops population-specific models, accessible diagnostic tools, targeted therapies, and active engagement with community oncology practices to improve care and outcomes for all patients.

NIH Research Projects · FY 2025 · 2025-08

Accelerated understanding of nervous system functions and cancer has led to cancer neuroscience evolving into a new frontier of discovery in cancer biology and therapy. The 3rd Cancer Neuroscience: Crossing Disciplines to Revolutionize Cancer Treatment biennial symposium. It will build on the remarkable success of our previous meetings, which attracted over 2,400 participants from 70 countries and resulted in high-impact publications in Advanced Biology (2022) and Cancer Cell (2024). Survey results of past symposia have indicated overwhelming satisfaction with the meeting and have consistently identified a critical, unmet need for the field: increased understanding of neuro-immune interactions in cancer. The 2026 meeting will address this by making neuroimmuno-oncology its centerpiece at a time when both immunotherapy and neuromodulation are demonstrating unprecedented clinical potential. This focus reflects the symposium's commitment to remaining responsive to participant feedback while driving forward cutting-edge research at the nexus of these disciplines. Building on our track record of fostering cross-disciplinary dialogue, we will create a platform that brings together oncologists, neuroscientists, immunologists, cancer biologists, and a wide array of other disciplines. The program will feature keynote sessions on neural regulation of anti-tumor immunity, in both central and peripheral nervous systems, pioneering research on neuro-immune crosstalk in the tumor microenvironment, and translation of findings to clinical applications that address healthcare disparities for marginalized populations. Our overall goal is to accelerate advances in cancer neuroscience by facilitating dialogue between established leaders while nurturing a new generation of scientists. We aim to 1) advance understanding of neuroimmune interactions in cancer through cross-disciplinary integration; 2) support early-career investigators through mentorship and career development; and 3) Accelerate scientific discovery through strategic cross-institutional collaboration. This meeting comes at a critical time when understanding neural regulation of cancer immunity could transform treatment. The Cancer Neuroscience Symposium will catalyze discoveries at the intersection of neurobiology, immunology, and cancer biology. By breaking down the barriers that have separated these fields, we will ultimately improve outcomes for all patients.

NIH Research Projects · FY 2025 · 2025-08

The overarching goal of The University of Texas MD Anderson Cancer Center (MD Anderson) Sarcoma SPORE is to reduce the morbidity and mortality from sarcomas through innovative translational research focused on immunologic and targeted treatments. To do this, we first address two critical needs in this arena. First, sarcomas are a complex assembly of collectively rare entities that, no different from the more prevalent carcinomas, require a multidisciplinary team-based approach in both clinical and pre-clinical research. Secondly, sarcomas have the added dimension of spanning the pediatric, adolescent-young adult, and adult populations. Thus, any translational research effort must be assembled to address this patient distribution. We bring together a multidisciplinary team of clinicians, physician scientists, statistical and computational experts, and basic and translational scientists who have substantial track records in the research and treatment of sarcomas across the age spectrum. Specifically, we propose the following: Project 2 will develop an effective systemic adoptive cellular therapy platform for chordoma patients by pursuing a clinical trial of adoptive T cells targeting a well validated immunogenic epitope of Brachyury, a well-established tumorigenic chordoma protein. Project 3 will evaluate, optimize, and refine novel antibodies, which have previously been used for cell therapies and which target proteins specifically for osteosarcoma, as antibody drug conjugates, linking them to appropriate payloads for treatment in the context of a phase 2 clinical trial. Common themes emerge from these projects, including exploration of the immune system and tumor microenvironment as targets for therapeutic activity. These projects place an emphasis on osteosarcoma, the most common tumor of bone and one where we have made virtually no treatment progress in 30+ years. All projects, working in concert with a Pathology and Tissue core of sarcoma experts, incorporate state-of-the-art molecular profiling of either patient biopsies or model organism samples. The data analyses and identification of risk stratification factors will be explored with a Biostatistics and Bioinformatics Core of experienced statisticians, bioinformaticians, and computational biologists. The team is drawn from multiple departments and divisions at MD Anderson spanning pediatrics and adult sarcoma practices and will be bolstered by an active patient advocate group, a team of expert external advisors with immense experience in sarcoma research and care and a core team of administrative expertise.

NIH Research Projects · FY 2025 · 2025-08

ABSTRACT: Overall Head and neck cancer afflicts nearly 65,000 persons annually in the United States, with an explosive growth, in what has now reached epidemic levels, human papilloma virus associated (HPV+) squamous carcinoma. Advances in therapy and changes in etiology have resulted in long-term survival for the majority of head and neck cancer patients; however, nearly all patients report therapy-related side effects from locoregional therapy (i.e., surgery or radiotherapy) due to normal tissue injury, such as xerostomia, osteoradionecrosis and deglutitive muscle damage, with a majority reporting one or more moderate-severe symptoms that impair quality of life. For highly curable HPV+ cancers, competing risk of therapy associated aspiration morbidity may match or exceed cancer-specific mortality, for example. Consequently, innovative strategies to identify, monitor, and prevent therapy-related normal tissue injury are a significant unmet need in public health. Because classic staging and therapy selection have focused necessarily on tumor characteristics, strategies for normal tissue injury reduction via surgical or radiotherapy modification have largely not been patient specific. Similarly, assessment and diagnosis of normal tissue damage has been symptomatic-driven and qualitative, as a reactive post-therapy measure. Using advanced imaging and computational image analysis, how best to execute image-guided interventions to reduce normal tissue injury and quantitative monitoring of spatial patient- specific normal tissue injury, remains a knowledge gap to be answered in the current proposal. Our central hypothesis is that through computationally assisted analysis of advanced imaging, we can better predict, detect, prevent, and manage normal tissue injury, and simultaneously reduce normal tissue injury from image-guided surgery/radiotherapy. This will be investigated by leveraging novel functional and anatomic imaging of spatially distinct normal tissue injury, using state-of-the art computational approaches to improve patient risk stratification and extending post-therapy mitigation of adverse late effects through image-guided surgical and chemoprevention efforts.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY Generating antitumor immune responses requires the phagocytosis of tumor cells and subsequent cross-presentation of tumor-derived antigens by antigen-presenting cells. However, these processes are impeded by phagocytosis checkpoints and inefficient cytosolic transport of antigenic peptides from phagolysosomes. CD47, an integrin family member, is highly expressed in multiple human cancer cells and plays an important role in helping cancer cells evade phagocytosis by professional antigen-presenting cells (APCs). Even in the cases where tumor cells are phagocytosed, escape of cancer cell-derived proteins and peptides from phagolysosomes into the cytosol where they can be further processed by proteasomes and loaded onto MHC molecules for cross-presentation to T cells is limited. Therefore, strategies to promote tumor cell phagocytosis and subsequent phagolysosomal escape of tumor cell-derived cellular components in antigen-presenting cells, are critical for activating innate immune functionalities of APCs and their ability to prime effector T cell responses. Here, we propose to engineer a novel antibody-drug/toxin conjugate (ATC) inspired by the phagolysosomal escape mechanisms of intracellular infectious pathogens including Listeria monocytogenes. We linked the anti-CD47 antibody with the L. monocytogenes toxin LLO via a cleavable linker (CD47-LLO). The antibody binds to CD47-expressing tumor cells, promoting their phagocytosis by phagocytes. Once exposed to the reducing conditions within the phagolysosomes, the linker between CD47-LLO is cleaved, and the released LLO is activated in the acidic environment to form pores on the phagosome membrane. This creates a channel for digested tumor and protein fragments to leak into the cytosol to promote the cross-presentation of tumor antigens. We hypothesize that CD47-LLO, as a novel cancer immunotherapeutic, promotes innate and adaptive immune cell interactions to produce potent and durable antitumor responses against localized and metastatic cancers. To test this central hypothesis, we will first in Aim 1 determine the mechanisms of action of CD47-LLO, including the impact of the ATC treatment on antigen presentation in APCs and the diversity of T cell clones. In Aim 2, we will study the impact of CD47- LLO treatment on immune cell populations and their interactions with tumor cells and one another within the tumor microenvironment. Finally, in Aim 3, we will test a humanized version of CD47-LLO against patient tumors established in mice reconstituted with a functional human immune system, as a single agent or in combination with checkpoint blockade. If successful, our proposed research would pave the way for developing a first-in-class, non-cytotoxic ATC that promotes immune activation to generate potent antitumor responses.

NIH Research Projects · FY 2025 · 2025-08

PROJECT SUMMARY/ABSTRACT B cells are the master regulators of humoral immune responses, and many autoimmune diseases arise due to the breakdown of self-tolerance in B cells. Understanding the signaling and genetic control of B cell tolerance will lead to the development of new tools for the early detection and treatment of B cell mediated autoimmunity. There is increased evidence on the potential contribution of the E3 ubiquitin ligase gene related to anergy in lymphocyte (GRAIL) in B cell mediated autoimmune diseases; however, to date, GRAIL expression in B cells and its role in B cell mediated immunity and inflammation remains undefined. Our data provides the first evidence of GRAIL expression in both mouse and human B cells, with higher expression particularly in anergic B cells, suggesting that GRAIL may contribute to establishment of B cell tolerance. In fact, GRAIL deficient B cells were hyper-responsive in terms of proliferation upon antigenic stimulation. In addition, aged GRAIL B cell conditional knockout (BcKO) mice developed lupus-like symptoms characterized by high titers of anti-double stranded DNA in the sera, accumulation of T follicular helper and B cells in the lymphoid tissues and spontaneous germinal center formation. Moreover, GRAIL BcKO mice are more susceptible to autoimmune diseases such as lupus and rheumatoid arthritis (RA). Importantly, we detected significantly reduced GRAIL expression in B cells from patients with RA, lupus, and immune checkpoint inhibitor (ICI)-induced arthritis compared to healthy donors, further indicating that GRAIL down-regulation could serve as a marker for onset of B cell mediated autoimmunity. Based on these findings, we hypothesize that regulation of B cell activation and tolerance by GRAIL may be an important checkpoint in censoring and elimination of autoreactive B cells; thus, GRAIL function is crucial to control the onset and development of B-cell mediated autoimmunity. Here in Aim 1, we propose to determine the molecular mechanisms responsible for regulation and function of GRAIL in B cell tolerance by utilizing conditional gene knockdown approaches and in vivo B cell tolerance models. We will identify the exact target(s) of GRAIL, which will determine its function in B cell activation and tolerance. In Aim 2, we will determine the role of GRAIL in antibody-dependent and - independent functions of B cells. The physiological significance of this finding will be assessed in a novel ICI- arthritis model. By utilizing B cell specific GRAIL targeting approach, we will assess whether GRAIL functions in B cells to control ICI-arthritis at the onset and/or progressive stages. In addition, cellular, global transcriptomic, and genome-wide analysis of biospecimens from cancer patients with ICI-arthritis will help to unmask B cell specific role of GRAIL in immune-related adverse event pathogenesis. The proposed research will provide new significant insight into mechanisms underlying B cell tolerance that will lead to development of pharmacological approaches in controlling B cell mediated autoimmunity.

NIH Research Projects · FY 2025 · 2025-08

Abstract Tumor cell of origin is a major determinant of cancer evolution. While the cell of origin of several tumor types has been successfully identified, glioblastoma, a malignant primary brain tumor, remains deprived of such biological information. Although rigorous studies have identified potential candidates among central nervous system cytotypes, a clear consensus of the early tumorigenic steps in glioblastoma has yet to be achieved. Unveiling glioblastoma cell of origin will shed a light on its evolutionary trajectories and, ultimately, better explain disease heterogeneity and poor clinical outcomes. To investigate cell of origin of glioblastoma, I generated multiple technological tools leveraging i) CRISPR/Cas9 genetic engineering, ii) human induced pluripotent stem cells derived cerebral organoids, iii) adeno-associated- and lentiviral gene delivery, iv) genome-tagging dynamic-barcoding. This work laid the foundation of the generation of human pre-clinical model of spontaneous gliomagenesis, thus allowing to trace and characterize early steps of human gliomagenesis. The overarching hypothesis of this research plan is that cell-specific molecular programs are permissive to malignant transformation under the selective pressure of known genetic drivers. To test this hypothesis, the Aim 1 of this work will be focused on characterizing histopathological and transcriptomic alterations following key driver events to provide detailed annotations of early glioblastoma tumorigenesis. To complement Aim 1 and improve our understanding of glioblastoma tumorigenesis, Aim 2 will be focused on developing a reliable platform that will permit to lineage trace glioblastoma cell of origin thanks to a sophisticated CRISPR/Cas9 barcoding technology. Single cell RNA sequencing of these models will generate a computational framework able to identify normal cells susceptible to gliomagenesis and transcriptional programs that lead malignant transformation. Comparative analysis with publicly available single cell RNA sequencing dataset of patient derived glioblastoma, will further advance our knowledge regarding tumor progression. This study will define a comprehensive panoramic of glioblastoma tumorigenesis and clarify its cell of origin, opening new avenues for future clinical therapeutic strategies.

NIH Research Projects · FY 2025 · 2025-08

Project Summary Radiation-induced lymphopenia (RIL) is a common adverse effect of radiation therapy. Recent studies have shown that the development of severe (grades 3 or 4) RIL has a significant detrimental association with overall survival. RIL is also associated with reduced efficacy of anti-PD1 immunotherapy, which is concerning because of the considerable potential of combining radiation therapy with immunotherapy in order to take advantage of the immunostimulatory effects of radiation. Therefore, it is important to better understand the mechanisms of RIL in order to reduce patient risk. Clinical studies by our team have demonstrated that proton therapy, because of its compact dose distributions, leads to significant reduction in severe RIL and to improved outcomes. The long- term goal of this research is to be able to predict which patients are more likely to develop severe RIL and produce optimized proton treatment plans that will reduce this risk. The objective of this study is to understand how proton linear energy transfer (LET), in addition to dose, contributes to the death of lymphocytes and their subtypes. The compact dose distribution from proton therapy should theoretically reduce the incidence of RIL, but higher LET outside of the treatment site may diminish this advantage. This may explain why some studies have shown reduced incidence of RIL when the patient is treated with protons while other studies have not shown this association. The role of LET in lymphocyte death and RIL has been largely ignored. Currently, there are no data, to our knowledge, on the sensitivity of lymphocytes to LET. Further, none of the current lymphopenia risk models incorporate LET and it is not used as a parameter for plan evaluation at most proton centers. In Aim 1, I will characterize the sensitivity of lymphocytes and their subtypes to radiation dose and LET using a specialized irradiation device to expose lymphocytes simultaneously to a range of doses and LETs. In Aim 2, I will incorporate LET into the RIL risk prediction model that has been developed by our lab and test if this improves the model’s predictive power. In Aim 3, I will develop the methodology to utilize LET-guided treatment optimization, considering differential radiosensitivity of lymphocyte subtypes on LET, and assess the effectiveness of this approach for RIL risk reduction. With well regarded expertise in proton therapy and a strong medical physicist training record, the Mohan lab is an ideal environment for me to complete the proposed research. Throughout my graduate training, I will work collaboratively with several renowned scientists in multiple fields including medical physics, immunology, radiobiology, and bioinformatics from MD Anderson and our collaborating institutions. I will have access to world-class facilities including a proton center to conduct my research. Additionally, I will submit four first-authored publications, present my work regularly, and attend national and international conferences. I will also take additional coursework on immunology and bioinformatics and learn about the clinical roles of medical physicists through specialized clinical training and shadowing.

NIH Research Projects · FY 2025 · 2025-08

Project Summary Prostate cancer (PCa) is the second most common cancer-related mortality cause in American males. Although therapies targeting androgen receptors are generally effective in treating advanced PCa, no curative treatments currently exist for castration-resistant prostate cancer (CRPC). Recent research reveals CRPC’s vulnerabilities to oxidative stress due to its unique metabolic reprogramming. This elevated susceptibility creates an environment favorable for a form of cell death called ferroptosis. Ferroptosis is brought about by the intracellular accumulation of toxic lipid hydroperoxides. Hence, ferroptosis induction represents a promising novel therapeutic approach for drug-resistant cancers. Despite increased sensitivity to ferroptosis, CRPC manages to avert this cellular fate. In this proposal, we have identified a key enzyme involved in lipid metabolism, adipose triglyceride lipase (ATGL), that engenders metastatic castration-resistant prostate cancer cells with the ability to resist ferroptosis. Our preliminary data demonstrates that ATGL facilitates prostate cancer growth, migration, and invasion. In CRPC cell lines, ATGL deletion sensitizes prostate cancer cells to ferroptosis, likely through regulating the GPX4-mediated ferroptotic control pathway. However, the exact mechanism linking ferroptosis and ATGL in CRPC is unknown. Moreover, while ferroptosis inducers (FINs) are hopeful candidates for anticancer agents under development, their use for treatment and how to enhance their efficacy remains underexplored in CRPC. Our central hypothesis is that ATGL promotes the resistance of CRPC to ferroptosis inducers in an oxidizable lipid-dependent manner. Therefore, Aim 1, the F99 phase, is designed to (1) identify the underlying mechanisms by which ATGL modulates the regulatory pathways that lead to ferroptosis, and (2) evaluate the efficacy of a combined therapy employing FINs, ATGL inhibitors, and anti-androgens as well as investigate the underlying mechanisms behind the observed synergy between anti-androgens and ferroptosis inducers in the context of ATGL ablation. Dr. Daniel Frigo, Ph.D., in the Department of Cancer Systems Imaging at MD Anderson Cancer Center is the primary sponsor of this F99 phase. Aim 2, the K00 phase, is to demonstrate the role of ATGL’s transacylation activity and its product, fatty acid esters of hydroxy fatty acid (FAHFA) in prostate cancer. Considering the K00 proposal focuses on metabolism and cancer progression, it will be conducted at a laboratory that excels in these fields. The fulfillment of these aims holds promise to deepen our knowledge of cancer biology and guide the development of innovative complementary therapeutic approaches that enhance the effectiveness of anti- androgens, thus improving the clinical outcomes of patients suffering from advanced prostate cancer. The proposal also contains a training plan that details further scientific, technical, and professional training. The plan is formulated to ensure the successful completion of the project and a smooth transition to a future career as an independent investigator.

NIH Research Projects · FY 2025 · 2025-08

ABSTRACT Lymphoid malignancies are common forms of cancer and leading causes of death that currently have no approach for prevention nor interception. Early data support clonal hematopoiesis in the form of mosaic chromosomal alterations (mCAs) as a promising premalignant and prognostic biomarker. mCAs are somatic megabase-scale alterations that can be detected with high sensitivity from array-based genotyping of blood-derived DNA. Prior studies have found mCAs confer a 10-fold increased risk of developing a lymphoid malignancy and 2-fold increased risk of mortality. However, prior studies have not been powered to precisely investigate mCAs in major lymphoid subtypes. Deeper characterization by subtype is needed. The overall objective of this application is to determine the role of mCAs in risk and prognosis of lymphoid malignancies. Our central hypothesis is that mCA carriers have distinctive etiologies and inferior outcomes. The rationale for this project is that mCA events can be readily detected in peripheral blood and could allow for targeted screening and earlier detection of at-risk individuals, promoting future prevention. We will utilize our existing studies, the most diverse prospective cohorts of lymphoma survivors in the world, to evaluate mCAs across lymphoid malignancies. Aim 1 will evaluate the association between mCAs and risk of lymphoid malignancy subtypes. Our hypothesis is that certain mCA loci are associated with risk of specific lymphoid subtypes. We will first evaluate the effect of mCAs across subtypes in 34,519 prevalent cases compared to 63,103 controls. Then we will validate these results in 5,029 incident cases and >500,000 controls. Aim 2 will determine the prognostic role of mCAs in lymphoid malignancies. Our hypothesis is that patients with an mCA have worse outcomes than those without an mCA. We will examine outcomes in 10,353 patients from the MER and LEO cohorts, accounting for therapy, known prognostic factors, mCA loci, and ancestry. Aim 3 will elucidate the blood cell lineage of mCAs in patients with a lymphoid malignancy. We will compare mCA events from peripheral blood with chromosomal alterations in matched tumor tissue in 2,164 patients. Then we will generate data to explore if mCAs are restricted to lymphoid cells in lymphoid malignancy patients. Completing these aims will provide a deeper understanding of the relationship between mCAs and lymphoid malignancy subtypes. Together, these studies will pave the way for future modeling of lymphoid malignancy risk and prognosis. We anticipate that the results of this project will generate critical preliminary data for future research on mCAs, specifically regarding (a) mechanisms underlying the risk of lymphoid malignancies, (b) therapeutic and prognostic implications, and (c) broader outcomes, such as susceptibility to infections and cardiovascular disease. The resulting preliminary data and additional mentored training from this award will propel me towards my long-term goal of establishing an independent research program to improve prevention, interception, and outcomes of blood cancers.

NIH Research Projects · FY 2025 · 2025-08

Project Summary Tumor infiltrating B cells (TIL-B) express B cell receptors (BCR) and antibodies (Abs) that can have a role in antitumor function. What tumor antigens they target can give insight into anti-tumor immune function and potentially leveraged for therapeutic strategies. Public tumor RNA-seq databases such as the TCGA, represent a wide and deep data source that include TIL-B BCRs. Our analysis of these datasets reveals the surprising finding of BCRs with the same V, J and complementarity-determining region 3 (CDR3) shared across patients. The occurrence of this phenomena by random chance is extremely rare (as low as one in 10 trillion) and therefore suggests a convergent response shared in patient tumor types. Although BCR repertoire has been analyzed in small groups of patients, a wide BCR analysis within and across tumor types has not been done. Analyzing convergent BCRs could inform what common antigen targets the immune system is focusing on. The bulk of tumor sequencing data exists as NGS RNA-seq. A challenge for reconstructing a BCR is matching the immunoglobulin heavy and light chains. To accomplish this, we have developed a comprehensive algorithm based on our Frequency Inference Model (FIM) to predict the paired heavy and light chains from sequencing datasets. We propose to refine our sequence analysis by leveraging machine learning algorithms to inform our ability to clone, produce and characterize Abs shared across patient tumor samples. This proposal will characterize antibodies reconstructed from RNA-seq datasets and test their tumor and antigen recognition characteristics. Abs will be tested for anti-tumor potential and antigen identification will be done on a small cohort with the following aims. Aim 1: Analyze immunoglobulins from different cancer types in TCGA. Convergent immunoglobulins will be identified, and matching chains identified by leveraging the deep learning algorithms. Aim 2: Produce shared Abs using the MDACC Recombinant Antibody Production Platform based on the computationally derived immunoglobulins. The reactivity to cell lines and tumor samples will be analyzed. Aim 3: Elucidate targets of TIL-Bs and their cognate BCRs and evaluate the anti-tumor function the antibodies. A pipeline to identify antigen epitopes will be used. The anti-tumor function of the antibodies will be assessed. With the completion of these aims we will have a proof-of-concept that our pipeline can recapitulate shared BCRs and assess tumor targeting potential. This platform would enable identifying shared patient antigen response that could be potentially leveraged to develop therapeutic targeting strategies.

NIH Research Projects · FY 2025 · 2025-08

Project Summary/Abstract Radiation-induced heart disease affects 15-30% of cancer patients who undergo radiation therapy (RT) to the chest and encompasses a broad range of cardiovascular disease (CVD) with potentially profound prognostic implications. Despite its impact, the exact causes of RICVD remain uncertain, and limited preventive treatments are available. Addressing this gap is an urgent requirement. Ionizing radiation (IR) has been shown to promote a persistent senescence-associated secretory phenotype (SASP) in myeloid cells, which plays a key role in the development of CVD. IR-induced SASP and accumulating senescent myeloid cells in tissues can trigger prolonged inflammation, but the mechanisms by which IR induces SASP remain unclear. Our long-term goals are to determine the fundamental mechanisms contributing to RICVD, identify patients at high risk for RICVD, and establish novel therapeutic approaches. It has become evident that glutamate metabolism plays a key role in promoting the survival of senescent cells and SASP induction. We conducted a metabolite profiling and found that both glutamate (Glu) and amino sugar metabolisms were upregulated in human monocyte-derived macrophages from patients with RICVD compared to those from non-RICVD patients. Glutamine-fructose-6-phosphate transaminase 2 (GFPT2) is a crucial enzyme that upregulates both glutamate and amino sugar (a precursor of glycosaminoglycan (GAGs)); our preliminary data showed that GFPT2 is acetylated at K17 and K254, which stabilizes GFPT2 expression after IR, and that this acetylation is regulated by NAD+-dependent sirtuin 6 ( SIRT6) deactivation. We hypothesize that IR-induced telomeric DNA damage initiates a positive feedback loop in which PARP activation leads to TOP2 degradation via TOP2- PARylation. The sustained TOP2 reduction stabilizes GFPT2 by deactivating SIRT6 and upregulating acetylation, promoting Glu/amino sugar/GAGs, SASP, and thus RICVD. We will test our hypothesis by pursuing the following three specific aims: In Aim 1, we will Investigate the role of the PARP-TOP2β positive feedback loop in provoking Glu and GAGs and subsequent SASP after IR in vitro. In Aim 2, we will determine how SIRT6 deactivation-mediated GFPT2 acetylation/stabilization and subsequent Glu/GAGs contribute to the persistent induction of SASP following IR in vitro . In Aim 3, we will determine the role of PARP1-TOP2β and Glu/GAGs in IR-induced atherosclerosis and atrial fibrillation (AF) in an in vivo setting. By elucidating the role of the PARP-TOP2β axis and NAD+-dependent GFPT2 acetylation/stabilization and its subsequent impact on Glu metabolism and SASP induction, this study a/GAGsims to enhance our understanding of the mechanisms underlying the persistent induction of SASP after radiation in myeloid cells. Additionally, the investigation of GFPT2 activation and Glu/amino sugar/GAG’s regulatory role may provide insights into therapeutic strategies for mitigating RICVD.