University Of Tx Md Anderson Can Ctr

NIH Research Projects · FY 2025 · 2022-09

Mutant KRAS drives human cancers from several sites, including pancreatic ductal adenocarcinoma (PDAC) and low-grade serous ovarian cancer (LGSOC). Despite the prevalence of RAS mutations in different cancers, effective RAS-targeted treatment remains a challenge. KRAS monomers form homodimers and nanoclusters in the cell membrane to optimize signaling and to transform cells efficiently. Dimerization of KRAS is required for RAS-driven transformation and cancer growth. Agents that disrupt mutant RAS dimers and clusters can block oncogenic activity. Recent evidence indicates that inhibition of RAS signaling induces autophagy and enhances the response to anti-autophagic therapy. Despite four decades of effort, development of effective strategies for the treatment for mutant KRAS-driven cancers remains a work in progress. Our laboratory has discovered a novel endogenous physiological RAS inhibitor designated DIRAS3, a 26 KDa GTPase sharing 50-60% homology with classical RAS family members, but with a distinctive 34 amino acid N-terminal extension that reverses RAS function. Like RAS, DIRAS3 is prenylated at the C-terminal CAAX site, binds GTP with high affinity, exhibits weak GTPase activity, and requires membrane association for its biological function. DIRAS3 is downregulated in a number of cancers including PDAC and LGSOC, and re-expression of DIRAS3 blocks cancer cell proliferation, inhibits motility, and, importantly, induces autophagy by multiple mechanisms. Recently we have found that DIRAS3 and a DIRAS3-derived stapled peptide from its α5 domain interact directly with mutant KRAS, reducing KRAS dimerization and nanoclustering, and inhibiting KRAS signaling. Both intact DIRAS3 and DIRAS3-derived stapled peptide induce autophagy and potentiate the pro-apoptotic activity of autophagy inhibitors in PDAC and LGSOC cells. In this proposal, we will study the effect of DIRAS3 on KRAS-dependent cell growth, migration and effector signaling in MEF cells and genetically engineered mouse model with mutant KRAS, as well as in KRAS-driven PDAC and LGSOC, better defining the mechanism by which DIRAS3 inhibits KRAS (Aim 1). We will investigate the mechanisms by which DIRAS3 induces autophagy in KRAS-driven PDAC and LGSOC (Aim 2). Finally, we will test the ability of DIRAS3 or a DIRAS3-derived stapled peptide in combination with autophagy inhibitors (CQ/DC661) to enhance apoptosis and growth inhibition in PDAC and LGSOC (Aim3). These studies will not only lay the groundwork for exploring new therapeutic strategies targeting KRAS-mutant cancers, but also contribute to a fundamental understanding of the mechanisms by which DIRAS3, as a tumor suppressor, inhibits mutant KRAS activity and induces autophagy.

NIH Research Projects · FY 2024 · 2022-09

Current therapeutics for neuropathic pain are poorly efficacious. One likely reason is that current therapies target only a single biochemical mechanism or pathway while chronic pain is a multi-mechanism problem. Fumarates, such as monomethyl fumarate (MMF), are uniquely positioned to treat the multi-mechanism problem of pain due to their combined antioxidant and anti-inflammatory profile. However, the risk-benefit ratio of fumarates is unacceptable for treatment of chronic pain, given their poor gastrointestinal tolerability and indiscriminate immune interference. This research project proposes to build on the documented analgesic efficacy of fumarates by designing new MMF prodrugs that have improved tolerability through design of non-reactive prodrugs. To reduce systemic immune interference, prodrugs will further be designed to preferentially release MMF, the active metabolite, only where ROS/RNS are overproduced in the pain neuraxis. The overall objective is to develop a 1,2-dicarbonyl prodrug that preferentially releases MMF at sites of pathology to alleviate neuropathic pain, while reducing adverse effects caused by systemic distribution of fumarates. Preliminary data indicate that the antioxidant master regulator nuclear factor erythroid 2-related factor 2 (NRF2) is a major therapeutic target. Using validated NRF2 activators and standard of care therapeutics as comparators, the UG3 specific aims are: 1) Complete initial characterization of the lead 1,2-dicarbonyl MMF prodrug; 2) Optimize lead compound, and design and evaluate backup 1,2-dicarbonyl MMF prodrugs; and 3) Characterize safety, validate analgesic efficacy of lead and backup 1,2-dicarbonyl MMF prodrugs. The exploratory milestones will identify a lead and backup molecules. These compounds will be advanced through IND-enabling studies under the following UH3 phase specific aims: 4) Extensive PK/PD characterization of candidates; 5) demonstrate in vivo efficacy in established rodent models of neuropathic pain; 6) Dose range finding to establish maximal tolerated dose (MTD) and cardiovascular, respiratory and gastrointestinal safety and toxicity studies; and 7) establish a Lead Development Team to finalize an IND application. BPN resources will be engaged for CMC manufacture and scale up, and development of a phase 1 clinical trial plan. Successful completion of this project will have major impact by first-in-class target activated delivery of a non-opioid therapeutic agent (MMF) to localized regions of pain pathology. Site-specific release of MMF will reduce its adverse effects, eliminating the final hurdles to MMF- based treatment of neuropathic pain.

NIH Research Projects · FY 2025 · 2022-09

Overall Summary Approximately 50% of cancer patients are treated with radiation therapy (RT), but local recurrence can still occur even with the use of advanced RT techniques. This local recurrence, which commonly develops in 30-50% of cancer cases, is exacerbated by the acquisition of RT resistance. This RT resistance is especially true for patients with locally advanced thoracic cancers, such as lung and esophageal cancers. RT can lead to an iron- dependent cell death modality, called ferroptosis, but whether ferroptosis resistance occurs within tumors giving rise to acquired RT resistance is not known and is the central theme of the proposed Acquired Resistance to Therapy and Iron (ARTI) Center. The overarching goals of the ARTI Center are: 1) to bridge the basic science mechanisms of ferroptosis in acquired resistance with translational research in preclinical models and human patient samples; 2) to identify cohorts of patients who are at greatest risk to develop acquired RT resistance; and 3) to investigate the ability of novel therapeutic agents to re-sensitize lung and esophageal cancer cells to radiation by inducing ferroptosis. The ARTI Center comprises two basic/mechanistic projects (Project 1 and Project 2), one preclinical/translational project (Project 3), and one shared resource core (Molecular Imaging Core [MIC]). Project 1 will focus on elucidating whether ferroptosis evasion is a key driver in acquired RT resistance using radioresistant lung cancer and esophageal cancer cell lines and xenograft models that will be used in Project 2. Project 2 will test the hypothesis that hypoxia, a long-recognized driver of tumor radioresistance, suppresses ferroptosis induction during RT and contributes to RT-induced acquired resistance to ferroptosis. Furthermore, expression of hypoxia-related genes and other targets of acquired RT resistance will be analyzed by single-cell sequencing in Project 3. Project 3 investigates changes in immune cells in the tumor microenvironment of humanized tumor models derived from chemoradiation therapy-responsive or -non- responsive esophageal adenocarcinoma patients. These ferroptosis-mediated immunologic changes in the tumor microenvironment may serve as prognostic biomarkers for identifying tumors that may acquire RT resistance and predicting cancer patient outcomes, which could, in the future, be modulated by the ferroptosis- inducing agents tested in Projects 1 and 2. Projects 1, 2, and 3 will be supported by the MIC that utilizes bioluminescence imaging to monitor tumor growth, positron emission tomography (PET) tracers to monitor cystine transporter activity and to identify hypoxic regions within tumors, as well as novel, redox-tuned PET tracers for identifying activated innate immune cells. The ARTI Center will develop an Administrative Core for effective communication and collaboration between the ARTI Center Project and Core Leaders and Co-Leaders with National Cancer Institute (NCI) of Acquired Resistance to Therapy Network (ARTNet) program staff as well as other ARTNet centers to synergize ARTI Center-related activities.

NIH Research Projects · FY 2025 · 2022-09

Overall SUMMARY Head and neck squamous cell carcinoma (HNSCC) remains a leading cause of cancer deaths worldwide with ~500,000 cases/year. Cisplatin is the gold standard systemic agent for HNSCC. Cisplatin resistance, both intrinsic and acquired, has been described in preclinical models and is frequently encountered in clinical practice; when it occurs it is deadly. The overarching goal of H-CARR is to develop a robust biological understanding of the key drivers of cisplatin resistance in HNSCC and develop the means of detecting it early in development and overcoming it once it arises. We previously showed that: 1) cellular processing of cisplatin generated metabolic stress is a critical driver of sensitivity and/or resistance and 2) coordinated genomic (TP53 mutation) and transcriptomic (Nrf-2 activation) reprogramming is essential to organizing the metabolic response to cisplatin generated stress. H-CARR brings together our biological and metabolic models of cisplatin resistance and our translational capabilities to image tumor metabolism non-invasively and detect biological shifts using circulating tumor cells (CTCs), to provide a comprehensive window into acquisition of cisplatin resistance as outlined in the Projects listed below, supported by a robust administrative and analytical infrastructure organized into 3 Cores. Project 1 will use state of the art metabolomic studies to identify the critical metabolic dependencies of cisplatin resistant HNSCC, identify opportunities for effective metabolic inhibition and improve our understanding of the cross-talk between the acquisition of cisplatin resistance and modulation of the tumor immune microenvironment. Project 2 will explore the genomic and transcriptomic reprogramming required to sustain the metabolic shifts which accompany development of resistance and interrogate how Nrf-2 dependent and independent signaling drives resistance and enhanced distant metastasis through intrinsic cellular mechanisms and paracrine signaling between tumor cells and adrenergic neurons. Project 3 will test whether the metabolic reprogramming outlined in Project 1 is detectable via non-invasive imaging (hyperpolarized magnetic resonance imaging) and whether the biological shifts outlined in Project 2 due to clonal extinction and expansion can be detected using CTC analysis in patients undergoing cisplatin-based treatment. H-CARR has the potential to realize the full clinical utility of cisplatin by identifying acquisition of resistance early during treatment and developing the means to overcome this and associated phenotypes such as enhanced distant metastasis. Successful completion of the proposed experiments will generate the new clinical standard for precision oncology approaches to clinical utilization of cisplatin in HNSCC and related upper- aerodigestive tract cancers of the lung and esophagus and therefore have a major impact on cancer survival worldwide.

NIH Research Projects · FY 2025 · 2022-09

The vision for this program is to develop tumor forecasting methods to predict and optimize the response of glioblastoma multiforme to standard-of-care therapies—and do so on a tumor-specific basis. A fundamental challenge in the care of patients with brain tumors is the limitation of standard radiographic methods to accurately evaluate, let alone predict, patient response. We propose to address this shortcoming by developing predictive, biologically-based mathematical models that incorporate the hallmark characteristics of brain tumor growth (e.g., tumor induced angiogenesis, hypoxia, necrosis, proliferation, invasion, and resistance to therapy) that can be initialized using advanced, subject-specific imaging data. This project will address two critical gaps in the care of patients battling brain cancer. First, our imaging-based, mathematical framework accounts for subject-specific characteristics and treatment regimens on model predictions. Second, in most studies, the ground truth used for validation of the predictive model is whether the model can predict future regional contrast enhancement, despite the well-known limitations of this qualitative MRI feature. Thus, while prior human studies have demonstrated the potential of predictive modeling, its translation into a realistic radiologic tool is fundamentally hindered by lack of systematic, pre-clinical validation where critical tumor characteristics (e.g., tumor heterogeneity and whole brain tumor cell distribution) can be precisely known and rigorously controlled. To overcome these limitations, we aim to: 1) establish the accuracy of tumor-specific modeling to predict spatiotemporal progression and 2) establish the accuracy of tumor-specific modeling to predict therapeutic response. Experimentally, we will construct a family of mathematical models that employ quantitative MRI data to capture the fundamental biological features of glioblastoma. These data are longitudinally acquired in patient derived xenografts that are treatment naïve or undergoing radiotherapy and/or chemotherapy. The model family is then calibrated with these data and a novel model selection strategy is employed to choose the most parsimonious model for predicting the spatio-temporal evolution of each tumor which is then compared to MRI data collected at future time points. Model predictions of tumor progression will be validated via registration to 3D fluorescent images of cleared ex vivo tissue, a technique that enables visualization of whole brain tumor burden. We will provide the clinical and scientific community with a validated mathematical description of glioma progression that can reliably predict progression and therapy response across a range of relevant glioma signaling pathways and can be readily applied to the clinical setting.

NIH Research Projects · FY 2025 · 2022-09

Project Summary Early diagnosis of cancer can improve therapeutic effect and prolong patient survival. The increasingly sensitive and widely adopted early cancer screening technologies have led to significantly more detection of early lesions that may or may not progress to cancer. Elucidating the mechanisms that drive or restrain early cancer would allow differentiation of aggressive cancer versus indolent types, improving personalized treatment and avoiding over-diagnosis and over-treatment. Whether an early lesion progress to cancer or not is not solely decided by the molecular profile of the lesion but also is impacted by the surrounding microenvironment and mediated by other epidemiologic factors. Meanwhile, the progression of an early lesion to malignancy is a complex process that may take years to occur. The complexities of the problem highlight the unmet needs for researchers from basic to translational science to collaborate and coordinate in the research of the underlying mechanisms between early lesion and cancer development. In response to RFA-CA-21-055, we propose a Coordinating and Data Management Center (CDMC) for the Translational and Basic Science Research in Early Lesions (TBEL) Program. The CDMC interacts closely with other entities of the Program, including the Steering Committee, the Research Centers, biospecimen and image repository, pathology centers, sequencing facilities, Data and Safety Monitoring Board (DSMB), and NCI, and provides critical scientific, administrative, regulatory, managerial, logistic, and data-analytic support to the TBEL Program. Our proposed CDMC infrastructure and operating procedures have been time-tested in an ongoing NIH-funded Consortium for the Study of Chronic Pancreatitis, Diabetes and Pancreatic Cancer. Specifically, the proposed work includes the three aspects of required responsibilities: consortium coordination (Aim 1), statistical and computational support (Aim 2), and data management, study protocol development and implementation (Aim 3). Our team of experts include information technology specialists who have been supporting and developing innovative software tools for numerous basic and translation cancer studies, experienced research coordinators who have worked on both NIH- and industry-funded multicenter studies, and faculty statisticians and bioinformaticians who have led CDMC work for large NIH consortiums and are well- known experts in biostatistics and bioinformatics methodological research areas closely related to biomarker development, risk prediction, single cell analysis, image analysis, machine learning, and clinical trials.

NIH Research Projects · FY 2025 · 2022-09

Abstract Trillions of exosomes, a subset of extracellular vesicles, are naturally present in the blood and tissue, and all cells secrete them into the extracellular milieu. Exosomes are on average around 100 nm in diameter and are speculated to have an endosomal origin, with a membrane lipid bilayer of similar orientation as the plasma membrane of the cells they originate from, and with capacity to enter other cells to deliver their constituents. The cargo exosomes include proteins, lipids, mRNA, non-coding RNAs (ncRNAs), microRNAs (miRNAs), and genomic DNA; and their production and content can vary depending on the cellular source. Exosomes may represent a novel mean of intercellular communication, but their biogenesis, trafficking, organ tropism, and impact on normal physiology and pathology in the in vivo setting is largely unknown. Similarly, the function of exosomes in pancreatic cancer remains understudied. Our research program is currently designed to address key unanswered scientific questions related to the basic biology of exosomes, their functional role in pathogenesis and therapeutic intervention of pancreatic cancer, a disease with dismal prognosis, and on the rise in the United States. Employing newly engineered mice, we propose to address the role of exosomes in the complex interplay between cancer cells, and the cells of the tumor microenvironment. Additionally, employing our established platform for clinical grade exosomes production, we will investigate the potential of engineered exosomes to target driver oncogenes and modulate the pancreatic cancer immune microenvironment/tumor immunity to facilitate therapeutic response. The proposed focus areas are rooted in the expertise we have developed within our research program over the last decade, and our track record provides the rationale and guidance for a higher chance of feasibility of the proposed studies, with potential for translational application.

NIH Research Projects · FY 2025 · 2022-09

Project Summary: It is well established that defects in DNA damage response (DDR) pathways accelerate tumorigenesis. Significant efforts have been devoted to target defective DDR pathways to improve outcome for cancer patients. These efforts led to the FDA’s approval of PARP inhibitors for the treatment of cancers carrying BRCA1/2 mutations and also the approval of immunotherapy for cancers with mismatch repair deficiency. Moreover, many inhibitors targeting DNA repair and/or cell cycle checkpoints have also entered clinical trials. Thus, there is an urgent need to understand how to effectively use these existing and new therapies for cancer treatment. We now know that targeting DDR pathways and/or DDR defects not only affect intrinsic tumor proliferation, but also change tumor-microenvironment interactions. Thus, we are expanding our DDR studies from in vitro to in vivo settings. In this project, we will determine mechanistically how several essential DDR genes/proteins control cell proliferation and DNA damage repair. We plan to establish separation of function mutations to further elucidate the key roles of these DDR genes and pathways both in vitro and in vivo. Additionally, we will investigate DDR defects in cancer therapy in vivo. Our recent success with in vivo CRISPR screens provides us an opportunity to explore avenues to target DDR pathways and DDR defects for cancer treatment in vivo. We anticipate that knowledge gained from these studies will help us design better treatment strategies for cancer patients.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY/ABSTRACT Head and neck squamous cell carcinoma (HNSCC) remains a leading cause of cancer deaths worldwide. Genotoxic agents, including radiation therapy (RT) and cisplatin (CDDP), are treatments that damage cellular DNA. RT and CDDP are the current standard of care in multiple solid tumors, including HNSCC. CDDP is the most commonly used chemotherapeutic agent in HNSCC proving superior to novel targeted agents in recent large randomized trials. Despite this, high rates of treatment failure persist in patients who develop resistance following this toxic chemotherapy. Treatment failure is uniformly fatal. However, no robust predictors of acquired cisplatin resistance or tumor response exist. Given this critical unmet need, we have focused our efforts on the assessment of tumor response using minimally invasive quantitative imaging (hyperpolarized magnetic resonance imaging; HP-MRI) while patients are undergoing therapy. We showed that CDDP and other genotoxic agents trigger measurable fluctuations in tumor cell metabolism detectable through HP-MRI with [1-13C]-pyruvate in real time (confirmed by conventional biochemical assays). Genotoxic stress suppresses the apparent rate of pyruvate conversion into lactate (kPL) via lactate dehydrogenase (LDH) in a manner that correlates with anti- tumor effectiveness. We therefore hypothesize that changes in kPL provide unique insight into metabolic changes induced by cisplatin that can be used to optimize response to therapy in HNSCC. In Aim 1, we will characterize baseline HP-MRI parameters such as kPL across the spectrum of HNSCC subtypes and validate the relationship between CDDP and associated shifts in carbon flux. We will also identify metabolomic differences in HNSCC models that affect baseline values of metabolic imaging biomarkers and modulate apparent changes induced by cisplatin. In Aim 2, we will integrate the dose-response data from Aim 1 to develop a predictive model of response to CDDP based on metabolic imaging parameters. We will use a simple algorithm to adjust therapeutic dose based on HP-MRI data in animal models of HNSCC to maximize tumor growth delay, and test whether thresholds suggestive of strong response can be used to select the more effective treatment regimen when multiple regimens are tested in parallel. In Aim 3, we will conduct a first-in- human evaluation of changes in HP-MRI to detect shifts in carbon flux following CDDP in HNSCC patients. We will correlate changes in metabolic imaging parameters with the baseline metabolic phenotype of tumors as determined from metabolomic analysis and direct measurements of tumor LDH. Successful completion of this study will establish HP-MRI as a non-invasive imaging modality able to predict response to treatment, which will be a noteworthy first step towards a precision oncology approach that we have been seeking for nearly half a century. Thus, the proposed research is relevant to the part of the NIH’s mission that pertains to developing and applying fundamental knowledge that will help to reduce the burdens of human illness and addresses directly the recently published “Notice of Special Interest: Precision Imaging of Oral Lesions” (NOT-DE-21-010).

NIH Research Projects · FY 2025 · 2022-09

Abstract The lung cancer early detection CVC has two main goals: Specific Aim 1 is to develop a blood- based biomarker panel for personalized risk assessment, modeled for its cost effectiveness. To this effect, substantial validation work in phase 3 studies has been done using retrospective longitudinal cohorts to test the performance of a four-marker protein panel (4MP) as a means to determine lung cancer risk and need for CT screening. The goal moving forward is to test the 4MP alone and in combinations with other types of markers in the screening setting, using lung cancer screening cohorts available to the CVC. The resulting marker panel, in combination with subject characteristics, would identify subjects who are currently not eligible based on USPSTF criteria that would benefit from CT screening based on their risk, ultimately leading to a utility trial for which a concept has been presented at a recent EDRN scientific meeting. The utility trial concept also includes as an objective to test the value of biomarkers in informing subjects who are currently eligible but not decided to undergo CT screening, about their risk through a decision sharing process. Specific Aim 2 will test the use of biomarkers and AI for interpretation of CT images and to personalize the screening frequency and duration. Sub Aim 1 is intended to validate the macrovasculature surrounding a nodule (vessel number) previously developed as a biomarker, in an independent screening cohort. Sub Aim 2 is intended to develop a validated integrative computational model for improved early lung cancer detection that includes blood- based biomarkers, CT features such as emphysema, presence or absence of a nodule, small airways and subject characteristics for interpretation of CT images and to determine screening frequency. The model will be subjected to a cost effectiveness analysis compared to current lung cancer screening guidelines. The CVC represents a multi-institution, multi-investigator effort with expertise in cancer biomarkers and statistics; pulmonology and lung cancer; epidemiology; radiomics, bioinformatics and artificial intelligence; and clinical trial design, simulation modeling and cost-effectiveness analysis. The CVC brings in substantial accomplishments in biomarker discovery and validation related to lung cancer screening and in CT image analysis. In pursuit of its aims, the CVC has access to samples from a multitude of cohorts for validation studies.

NIH Research Projects · FY 2025 · 2022-09

Project Summary Radiation therapy delivered at ultra-high dose rates may be becoming a breakthrough treatment option for cancer patients. Targeting cancers with ultra-high radiation dose rates produces a FLASH effect, wherein control of tumor growth is maintained similarly to conventional (CONV) radiation dose rates, but normal tissue toxicity is significantly reduced. Although FLASH irradiation has been shown to evoke strong, reproducible responses across many different organ systems (e.g., brain, lungs, gastrointestinal [GI] tract, skin) across multiple species, some studies have shown that ultra-high-dose rate irradiation to have either no effect or detrimental effects on normal tissue. This discrepancy is not clear; however, it likely stems from inconsistency in the physical radiation beam and fractionation parameters. Furthermore, although previous studies have shown either no change in or improved tumor responses from FLASH irradiation as compared with CONV dose rate irradiation, no studies have looked beyond simple tumor growth delay when evaluating tumor responses. A more relevant analysis for preclinical tumor responses to radiotherapy is the Tumor Control (TCD50) assay, and to date, no comparisons between FLASH and CONV dose rate irradiation on the dose required to cure 50% of tumors (TCD50) have been performed. The lack of comparisons of radiation types, the lack of consistency between physical radiation beam parameters and fractionation, and the lack of accurate measurements of tumor control in previous FLASH irradiation studies provides impetus to conduct this rigorous, high throughput, multi-institutional study to provide confirmatory evidence of the reproducibility of FLASH effects. This proposed project will test the hypothesis that there is an optimal set of physical beam parameters that will maximize the FLASH effect, and that under the same dose parameters and the same physical dose, the FLASH effect dose response will be the same between different radiation types. In order to test the hypothesis, Aim 1 will focus on determining whether radiation type (e.g., electrons, photons, and photons) alters abdominal FLASH-mediated normal tissue-sparing effects, with the expectation of similar responses to the different radiation types. In order to optimize the physical beam and fractionation parameters to maximally reduce normal tissue toxicity, physical beam parameters (e.g., mean dose rate, dose per pulse, pulse duration, overall delivery time, priming dose, and oxygen tension) as well as fractionation will be systematically changed and tested (Aim 2). Aim 3 will focus on establishing the therapeutic effects of FLASH dose rate irradiation mediate similar control of syngeneic, heterotopic tumors of three different cancer cell lines using the more relevant TCD50 assay. The overarching goal of this project is to minimize side effects for all cancer patients receiving radiation therapy, which will inevitably improve quality of life. Preventing the post-treatment effects of radiation therapy in cancer patients so that individuals can live longer, and more fulfilling lives is in direct alignment with the mission of the National Cancer Institute.

NIH Research Projects · FY 2024 · 2022-09

PROJECT SUMMARY/ABSTRACT Research. Oral cavity and oropharyngeal (OC/OPC) cancers afflict more than 53,000 individuals in the United States annually. Despite advancements in oncologic therapies, the majority of patients will experience significant toxicity burden during and after therapy, including moderate-severe xerostomia, dysphagia, reduced mouth opening (i.e. trismus), periodontal disease, and osteoradionecrosis. Remote electronic symptom monitoring through standardized assessment tools for patient reported outcomes (ePROs) is an evidence-based best practice, particularly in the COVID-19 era, yet few clinical practices have demonstrated sustainability of implementation efforts. To date, acute and chronic orodental complications afflicting OC/OPC survivors are largely managed on empirical knowledge with wide inter-provider management variability based on provider experience and available clinical information which is often incomplete, incorrect, or nonexistent. Therefore, standardization of electronic data capture of PROs and objective measures of provider-assessed orodental toxicity severity remains an unmet public health need. Our central hypothesis is that synchronous optimization of machine-readable patient- and provider-generated data collection can be achieved through prioritization of effective implementation strategies for longitudinal oro-systemic ePRO data collection (Aim 1) and creation of novel dental standards for accurate orodental toxicity reporting in both electronic health and dental records (Aim 2). As a subcomponent to Aim 2, we will also design and pilot a novel radiation odontogram to enhance treatment communication between providers. Accurate risk predictions of high-morbidity high-prevalence post-therapy orodental sequelae using high-quality electronic data from Aims 1 and 2 will be incorporated into a statistically robust machine-learning based model (Aim 3). In summary, the PROHEALER proposal fosters innovative and novel informatics approaches for data-driven risk assessment and algorithmic prevention and management of treatment-related oral health diseases afflicting OC/OPC survivors. Career Development & Training. Dr. Moreno's overarching goal is to become an internationally recognized independent research investigator with domain expertise in advanced radiation therapy techniques, clinical informatics and rigorous toxicity modeling methodologies as they pertain to improving patient quality of life and promoting precision prevention and risk-based interventions for orodental complications. This proposal presents Dr. Moreno's 5-year mentored career development plan which includes mentorship from prominent Established NIH Investigators who have committed to overseeing the progress of the proposed projects and Dr. Moreno's overall professional development. The outlined training activities build upon Dr. Moreno's clinical expertise as a Head and Neck Cancer Radiation Oncologist and her prior work in EHR utility enhancement with the inclusion of a comprehensive didactic and project-based curriculum focused on domain knowledge expansion in dental informatics, implementation science, and advanced statistical methods in risk prediction modeling.

NIH Research Projects · FY 2025 · 2022-09

ABSTRACT While monoclonal antibodies inhibiting immune checkpoints (ICI) and CAR T-cell therapies have dramatically changed the therapeutic options for many cancer patients, up to 60% of patients will experience immune-related adverse events (irAE) depending on the tumor type and immunotherapy. Thus, many patients treated with immunotherapy will not see long term benefit and therefore only suffer potential side effects (and possibly hyper progression), and the importance of predicting and managing immunotherapy-related adverse events has already been identified as a critical gap in knowledge and clinical practice. Significantly, a common molecular node of integration between the various inflammatory mechanisms of irAE, particularly in the subacute setting, focuses on spurious activation of the innate immune system. Recently, we reported the synthesis and validation of 4-[18F]fluoro-1-naphthol ([18F]4FN), a novel redox-tuned radiopharmaceutical for the selective imaging of high energy oxygen and nitrogen radical species (RONS) by PET/CT. [18F]4FN provides a convenient reagent for rapid, quantitative whole-body imaging to identify and monitor inflammatory foci generated by NADPH oxidase-2 (NOX2) and myeloperoxidase (MPO) of the innate immune system and multi-organ inflammation, including immunotherapy-mediated irAE. Monitoring irAE by PET imaging is the long-term clinical imaging goal of our line of investigation. Near-term, we propose to investigate the role of activated innate immunity in vivo by combined molecular imaging and multiplexed analysis of tissues in mechanism-based pre-clinical murine models of irAE for which we can enhance our understanding of the activation dynamics of innate immunity and gain signals of efficacy.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY A significant fraction of lung adenocarcinomas (LUADs) in lifetime smokers harbor somatic mutations in the KRAS oncogene (KM-LUADs). Due to enhanced screening, KM-LUAD is increasingly being detected at earlier pathological stages, thus posing a growing public health burden that warrants improved early treatment. Despite this urgency, early changes that conceive KM-LUAD and that would thus likely comprise ideal targets for interception remain poorly characterized. Previously, our group and others have shown that tobacco exposure leads to a pervasive field of injury that is composed of molecular (e.g., KRAS mutations) and inflammatory changes in normal-appearing epithelium and in the lung, and that are prevalent in the LUADs themselves. We and others have also previously described molecular and immune changes, including a decrement in host immunity, that are associated with development of lung premalignant lesions (PMLs) and KM-LUAD. These earlier studies have shed light on events that are likely implicated in early lung tumor development. Yet, especially for a cancer like KM-LUAD that is causally related to smoking, the identity and properties of specific cell populations that trigger a field of injury as well as its progression to PML and KM- LUAD are not known. In our preliminary efforts, we performed single-cell RNA-sequencing of lung tissues from a human-relevant mouse model of tobacco-associated KM-LUAD. We found a population of Krt8+ alveolar cells (KACs) that was greatly increased early on in lungs exposed to tobacco carcinogen but not control saline and that were also associated with tumor cell onset. KACs displayed intriguing properties that allow us to surmise that they perhaps represent KM-LUAD progenitors: they amassed the same driver Kras mutations found in the resultant LUADs; they expressed transcriptomic programs and cell-cell interactions that are highly pertinent to KM-LUAD including augmented p53 as well as pro-inflammatory IL-1β and NF-κB signaling; and their expression profiles were highly enriched in human PMLs and LUADs. We also found that KACs were markedly increased in the human LUAD ecosystem relative to matched normal lung. Our preliminary findings motivate the hypothesis that oncogenesis of KACs in concert with pro-inflammatory signaling mediated by IL-1β/NF-κB underlie initiation and development of PML and KM-LUAD. To address our hypothesis we will 1) characterize at single-cell resolution evolution of KACs to PML and KM-LUAD, as well as determine the role of p53 signaling in this process; 2) discern the role of pro-inflammatory signaling in promoting evolution of KACs to PML and KM-LUAD; and 3) use multiple approaches including drug screening to determine whether targeting KACs will intercept PML and KM-LUAD development. At the conclusion of our studies, we will have unraveled novel paths in the phenotypic evolution of KM-LUAD as well as laid the foundation for development of new strategies that inhibit the inception of this dire malignancy.

NIH Research Projects · FY 2025 · 2022-08

ABSTRACT Lynch Syndrome (LS) is the most common cause of hereditary colorectal cancer (CRC) affecting >1 million Americans. LS is caused by germline mutations in one of four DNA mismatch repair (MMR) genes. Normal colorectal epithelial cells in LS patients become MMR deficient after a somatic ‘second’ hit generating the accu- mulation of hundreds of insertion-deletion mutations (indels) in microsatellite sequences. These indels generate frameshift peptides (FSP) that become neoantigens (neoAg) and stimulate the adaptive immune system. We have previously reported that adaptive immune genes are highly expressed in LS pre-cancers, and we have generated a detailed neoAg catalog with >1,000 FSP neoAg from a cohort of LS pre-cancers and early-stage CRCs using next-generation sequencing tools coupled with a state-of-the-art bioinformatics pipeline. In addition, we have published the results of a phase Ib chemoprevention clinical trial in LS patients using naproxen showing immune-activation of colorectal mucosa resident cells. Taken together, these results point strongly towards the development of a vaccine for ‘recurrent’ and ‘shared’ LS-associated tumor neoAg combined with naproxen for pan-cancer prevention in the LS population. However, the main knowledge gap remains to select the most opti- mal neoAg peptides and to establish the efficacy and safety of the vaccination in a reliable animal model that allows immediate human translation. Rhesus macaques are a promising non-human primate (NHP) model dis- playing the closest genomic resemblance to humans. Our research team has reported the first colony of spon- taneous LS in rhesus carrying a germline mutation in MLH1. In addition, we are partnering with industry collab- orators in AMAL Therapeutics that have developed an innovative vaccination platform called KISIMA, integrating several selected FSP in tandem with a cell-permeable peptide fostering cell penetration, and a toll-like receptor agonist that acts as a self-adjuvant. Our central hypothesis is that our state-of-the-art bioinformatics pipeline for neoAg prediction will lead to the identification of the most immunogenic, recurrent across tumors, and shared among LS-associated tumor types FSP neoAg to be integrated in the KISIMA self-adjuvant vaccine platform, which will render a strong immunogenicity in combination with naproxen. To explore this hypothesis, we propose three specific aims: 1. To validate in vitro the immunogenicity of the top 150 recurrent neoAg shared by LS non-colorectal tumors using ELISpot, ELISA, and cytokine assays using PBMCs and CD8+ T cells from healthy human donors; 2. To develop artificial antigen-presenting cells (aAPC) expressing human LS neoAg to validate the cytotoxicity of neoAg-enriched T cells; and 3. To assess the immunogenicity of a neoAg combination using the novel self-adjuvant vaccine platform KISIMA alone and in combination with naproxen in a co-clinical trial in LS rhesus. The proposal is highly innovative because is developing a novel self-adjuavnt vaccine platform in a unique spontaneous NHP model of LS. The proposed research will significantly impact the field because it is a stepping stone to develop a Phase I first-in-human clinical trial to test a novel CRC vaccine for LS patients.

NIH Research Projects · FY 2025 · 2022-08

Project Summary Chemotherapy is an extremely effective treatment for cancer; however, along with chemotherapy’s benefits there are many undesirable consequences including chemotherapy-related cognitive impairments (CRCI) and accelerated aging. Despite impacts on quality of life, these areas are relatively understudied and there are currently no FDA approved treatments available. A sophisticated animal model to address this problem will be extremely valuable, both to our understanding of the long-term consequences and to the development of potential treatments. Currently, CRCI and chemotherapy-related accelerated aging is studied in rodents, but a nonhuman primate model will significantly enhance the validity of studies addressing this still under-researched phenomenon. In this proposed study, we aim to establish a baboon model of CRCI and accelerated aging. We will further define healthy aging in a baboon sample, then investigate whether chemotherapy induces cognitive decline and accelerates brain aging in baboons as it does in humans and test an intervention which has been effective in mice. We will collect brain imaging, cognitive, and aging molecular biomarker data from baboons prior to treatment with cisplatin only or cisplatin followed by an HDAC6 inhibitor. We will then collect further brain, cognitive, and aging data post-treatment to examine changes related to the chemotherapy regimen and whether the HDAC6 inhibitor reverses cognitive declines and neurodegeneration. These data will be used to propose further studies as well as techniques to potentially mitigate these consequences of chemotherapy in humans.

NIH Research Projects · FY 2025 · 2022-08

Development of treatment resistance is a major impediment to effective cancer therapy. KRASG12C inhibitors (KRASi) have been recently approved by FDA and represents a promising new targeted therapy for cancer types with KRAS mutation, one of the most frequent genetic mutations observed in human cancers. However, as is the case with standard therapy, majority of tumors that initially respond to KRASi quickly develop resistant disease. Unfortunately, the underlying mechanisms of KRASi resistance are poorly understood. To tackle this challenging problem, this application will capitalize on the PI's track record and expertise in the area of KRAS signaling and biology and propose a comprehensive research program focusing on understanding the mechanisms of acquired resistance to KRAS targeted therapy. This application will identify new actionable therapeutic targets and approaches to overcome resistance, thus greatly improving the clinical outcome of patients with KRAS mutated cancers. To achieve this, the Yao laboratory has developed in vitro and in vivo model systems that will be employed to investigate hypotheses regarding mechanisms driving the development of KRASi resistance. The in vitro model system consists of KRAS dependent and independent cell cultures derived from genetically engineered mouse models of pancreatic cancer as well as human cancer cell lines of pancreatic and colon origin. The in vivo model system consists of patient-derived colon cancer models that developed resistance to KRASi, as well as genetically engineered mouse models of pancreatic cancer driven by KRASG12C. The major knowledge gaps to be addressed are that: i) what molecular events are activated to enable bypass of KRAS dependency in tumors treated with KRASi; ii) how these molecular events function to drive tumor survival and maintenance in response to KRASi; and iii) whether pharmaceutically targeting the resistant mechanism can achieve durable therapeutic responses in combination with KRASi. These studies will unveil a previously unknown mechanism by which tumor cells become resistant to KRAS targeted therapy and may define an innovative therapeutic option for KRASi-resistant patients. The proposed work comprises an essential step toward our long-term goal of developing effective therapy for patients with KRAS mutated cancer, in align with the mission of the National Cancer Institute (NCI) RAS initiative (“Kill RAS”).

NIH Research Projects · FY 2025 · 2022-08

PROJECT SUMMARY Medulloblastoma (MB) is the most common malignant primary brain tumor in children. MB is frequently induced by the alterations of cellular signaling pathways, such as sonic hedgehog and wingless pathways, which have been extensively characterized. Nevertheless, current treatment of MB causes severe life-long side effects and fails to cure many patients. Thus, there is an unmet need for a new mechanistic understanding that would be helpful for designing a mechanism-based approach for MB treatment. Epigenetic aberrations, which are heritable aberrations in gene expression or cellular phenotypes without accompanying changes in DNA sequences, are a major factor for tumorigenesis. Epigenetic modifiers often harbor DNA alterations, such as mutations and deletions, in human MB. However, the roles of epigenetic modifiers in MB development remain largely unknown. Histone lysine methylation, a type of histone posttranslational modification, is a hallmark of epigenetic and transcriptional regulation of gene expression and is reversibly modified by histone methyltransferases and demethylases. Of histone lysine methylation, methylations at histone H3 lysine 4 (H3K4) are key gene-activating epigenomic marks. For example, monomethyl H3K4 is a mark for enhancers, which activate genes by interacting with gene promoters. In addition, trimethyl H3K4 occupies as much as 75 % of all human gene-regulatory regions, and broad trimethyl H3K4 is a gene-activating signature that denotes tumor suppressor and cell identity genes. We have previously reported that the H3K4 methyltransferase KMT2D (also called MLL4, ALR, and MLL2; a transcriptional coactivator) is required for retinoic acid-induced neuronal differentiation of human neuron-lineage NT2/D1 stem cells. Notably, our other study showed that homozygous loss of Kmt2d in the mouse brain developed spontaneous MB in the cerebellum, a brain region that controls motor coordination and balance. Strikingly, our additional results showed that heterozygous loss (single-allelic) of Kmt2d highly promoted MB. Based on these compelling findings, our long-term goal is to define the oncogenic role of heterozygous loss of Kmt2d in MB pathogenesis. Our central hypothesis is that heterozygous loss of Kmt2d causes epigenomic alterations to downregulate tumor suppressor genes and thereby promotes MB. Here, we propose to study to 1) characterize the MB-promoting effect of heterozygous Kmt2d loss using genetically engineered mouse models; 2) define the molecular mechanism by which heterozygous Kmt2d loss promotes MB; and 3) determine how heterozygous Kmt2d loss causes epigenomic alterations. Because KMT2D is one of the most frequently mutated genes in MB and a majority of KMT2D mutations in MB are heterozygous and truncations, our proposed studies are significant and clinically relevant. In addition, our studies using genetically engineered mouse models will define an in vivo MB-promoting role for heterozygous Kmt2d loss. Furthermore, our results will uncover the previously unappreciated epigenetic mechanism underlying MB pathogenesis and provide valuable information for the rational design of a therapeutic approach for MB treatment.

NIH Research Projects · FY 2025 · 2022-08

Most patients with head and neck squamous cell carcinoma (HNSCC) present with locally advanced disease and are treated with a combination of surgery, radiotherapy, and chemotherapy; however, recurrences occur in about half. Despite recent success with immune checkpoint inhibitors, 82% of patients with recurrent or metastatic (RM) disease do not respond to immunotherapy and only a very small subset have complete responses. Therefore, there is an urgent need for new approaches to treat RM HNSCC, including approaches that can be combined with anti-PD-1 therapy to increase durable responses. Increased levels of activated STAT3 (pY-STAT3) or nuclear STAT3 have been reported in 37-75% of HNSCC tumor samples, which correlated with poor prognosis. STAT3 contributes to cancer cell proliferation and immune resistance in HNSCC by promoting the development and function of several immunosuppressive cells, most notably myeloid derived suppressor cells (MDSC), within the tumor microenvironment (TME). Importantly, resistance to immune checkpoint inhibitor (ICI) therapy in HNSCC patients is linked to increased numbers of MDSC; the addition of danvatirsen (STAT3 anti-sense oligonucleotide) enhanced ICI responses in HNSCC patients but was associated with episodes of severe thrombocytopenia and liver injury. Of note, targeting of STAT3 also may reduce immune-related severe adverse events (irSAE) of ICI therapy. The Tweardy group, working with a clinical-stage biotechnology company (Tvardi Therapeutics, Inc.), used computer-based docking and lead-compound optimization strategies to identify TTI-101, a potent, non-toxic and orally bioavailable inhibitor of STAT3 that in a Phase I study in patients with solid tumors at MD Anderson Cancer Center determined an RP2D and showed no TTI-101-attributable adverse events; TTI-101 administration reduced levels of pY-STAT3 within tumors and was beneficial in 9 of 18 evaluable patients, with two patients having partial responses of 40% and 62%. Thus, TTI-101 is the most promising smallmolecule STAT3 inhibitor currently in clinical development for cancer treatment. The main objective of this proposal is to determine if STAT3 targeting with TTI-101 is beneficial in treatment of RM HNSCC. The specific aims are: 1) to determine the effects of TTI-101 alone and in combination with anti-PD-1 therapy in syngeneic mouse models of HNSCC, and 2) to determine the effects of STAT3 inhibition on the TME and STAT3 gene targets in a window-of-opportunity (WOO) clinical trial of TTI-101 in HNSCC patients undergoing surgical resection. We will obtain tumor biopsies pre- and post-treatment in the WOO and Phase II trials and perform correlative studies in the two Aims that include determining the relationships between pharmacokinetics, pharmacodynamics, the TME, and clinical responses in patients and mice with HNSCC treated with TTI-101 alone or in combination with anti-PD-1 therapy. If successful, our research will lead to improved survival and decreased toxicity for patients with advanced HNSCC.

NIH Research Projects · FY 2025 · 2022-08

PROJECT SUMMARY Burkitt lymphoma (BL) and diffuse large B-cell lymphoma (DLBCL) are highly aggressive B-cell malignancies that are commonly treated with chemotherapy plus an anti-CD20 antibody, Rituximab. High-intensity chemotherapy is required in BL patients, which is associated with severe toxicity and treatment-related mortality in 10% of patients. Of the patients able to endure therapy, 36% will suffer disease progression and have a dismal outcome, with only a 1% 3-year progression-free-survival in patients that are primary-refractory. A similar fraction of DLBCL patients progress during/following first-line therapy and have a median overall survival of 6.3 months. Recent genomic studies have identified co-occurring genetic alterations that are highly-recurrent in BL and DLBCL tumors. However, detailed functional analyses have not been performed for the majority of these driver mutations, and hence there are currently no available targeted therapeutic strategies in either disease. Mutations of the SMARCA4 and ARID1A genes are together found in approximately 40% of BL tumors, and 12% of DLBCL tumors. These genes encode two components of a multi-subunit complex, the BAF (aka SWI/SNF) complex, which functions to activate gene expression by “unpacking” closed and silent states to become open and active genes. Mutations of SMARCA4 perturb its activity by affecting the catalytic domain, and mutations of ARID1A lead to loss of protein expression, together representing two alternative mechanisms for loss of function in the BAF complex. Although the function of the BAF complex has been recently described in other malignancies, its function during B-cell development, and therefore the consequence of its inactivation in B-cell lymphoma, remains to be explored. We have developed animal and cell line models of SMARCA4 and ARID1A inactivation and found that they regulate distinct processes in B-cell development. We will leverage these models and cutting-edge genomics approaches to understand both the molecular and immunological consequences of BAF complex deregulation in B-cell lymphoma. By contrasting and comparing the roles of two key components of the BAF complex, SMARCA4 and ARID1A, we hope to gain detailed insight into the role of discrete BAF complexes and their redundant and non-redundant roles. This work will uncover the biology of BL and DLBCL tumors carrying BAF complex mutations, which can lead to advances in precision medicine targeting and therapies for this disease, as well as for other cancers.

NIH Research Projects · FY 2025 · 2022-08

Abstract. Pancreatic ductal adenocarcinoma (pancreatic cancer, PDAC) is the classic example of a recalcitrant tumor that is extremely challenging to treat. Therapeutic strategies which can bypass the desmoplasia `fortress' and access hypoxic microenvironments without significantly affecting healthy cells would address the critical issues presented by PDAC physiology. Localized therapies are a critical component of PDAC treatment and there is strong interest in innovative ways to intensify radiation therapy (RT). A novel approach to enhancing the radiation dose delivered to tumors is to increase the radiation-interaction probability of the target tissues by delivering high atomic number (Z) nanoparticles (e.g., gold or hafnium oxide) to tumor cells. However, due to the exuberant desmoplasia characteristic of PDAC, the diffusion of even the smallest nanoparticles is limited by the dense stroma making delivery to cancer cells exceedingly challenging. Furthermore, recent evidence confirms that depleting the stroma may not be the answer since the stroma restrains and confines cancer cells to within the pancreas rather than promotes the growth of cancer cells. We contend that the ideal strategy is to penetrate the stroma without destroying it. Here we address this delivery barrier standing in the way of effective radiosensitization of PDAC tumors by employing a recently reported process of in situ gold biomineralization by mammalian cancer cells. This strategy will allow replacement of pre-synthetized radiosensitizing gold nanoparticles (GNPs) with ionic gold atoms thus achieving the smallest possible size of a therapeutic agent – a single ionic atom. Our hypothesis is that small gold ions (i) will uniformly distribute throughout the tumor as their diffusion is not likely to be impeded by the stroma, (ii) will be reduced to GNPs via the process of in situ biomineralization after specific uptake by cancer cells, and (iii) will radiosensitize the tumor while sparing adjacent normal tissue. This hypothesis is based on our preliminary data demonstrating a strong radiosensitization effect in PDAC cells by the intracellularly synthetized GNPs both in vitro and in vivo. In addition, normal pancreatic cells synthesized significantly fewer GNPs than cancer cells, consistent with recent reports showing higher efficiency of in situ gold biomineralization in cancer vs. non-cancer cells. Further, a long history of the clinical use of gold-salt based drugs in treatment of rheumatoid arthritis can provide a clear path towards clinical translation. In this project we have devised comprehensive mechanistic studies to evaluate and to optimize in situ biomineralization for efficient radiosensitization of PDAC. To this end we will carry out studies in three synergistic Aims: (1) to determine the mechanism of and to optimize conditions for intracellular synthesis of GNPs by PDAC cells; (2) to evaluate the determinants of radiosensitization efficacy in vitro and in vivo; and (3) to develop a predictive biological effect computational model of radiosensitization by in situ synthetized GNPs. These studies will provide the framework for continued development of a readily deployable radiosensitization strategy for pancreatic cancer that can be extended to other unresectable solid tumors.

NIH Research Projects · FY 2025 · 2022-08

PROJECT SUMMARY/ABSTRACT To maximize their growth and metastatic potential, solid tumors promote the formation of new nerve fibers in the tumor microenvironment (TME). In patients with oral, prostate, breast, gastric, pancreatic, and other cancers, high densities of nerve fibers in the TME are associated with poor clinical outcomes. We proved that oral cancer cells induce a unique heterogeneous composition of tumor-associated neurons (TANs) in the TME. The nervous system plays important roles in homeostasis and inflammatory responses in tissues. However, the regulation of immune cells by nerves remains largely unclear. Our long-term goal is to elucidate the reciprocal nerve-cancer signals that drive cancer progression to identify novel targets for therapy and for overcoming immunotherapy resistance. Our preliminary data show that neurons communicate with immune cells directly through the expression of immunomodulatory molecules and indirectly through paracrine, adrenergic-dependent cancer cell signaling. The overall hypothesis that we will test in the proposed project is that TANs induce a maladaptive immune response that supports tumor progression. These newly formed, reprogrammed TANs regulate the immune response through a multistep mechanism that includes the transformation of quiescent neurons into sprouting cells that can infiltrate and interact with other cell types, release adrenergic neuroactive molecules, and support the development of an immunosuppressive microenvironment. Each of these steps may promote tumor progression and therapy resistance. The proposed research is innovative because it will capitalize on new concepts in immunology and cancer biology using advanced model systems to yield insights into the mechanisms of tumor progression and identify new targets for cancer therapy based on neuro-immune crosstalk. This cross-disciplinary proposal will combine expertise from oncology, immunology, cell biology, neurobiology, cancer genetics, pathology, and biostatistics in two specific aims across the two labs (Amit and Calin). Aim 1: Determine the mechanisms by which neuron-dependent cancer cell signaling regulates cytotoxic T-cell function. We will use pharmacological and genetic approaches combined with advanced spatial imaging techniques (for both protein and RNA) in syngeneic mouse models to understand how reprogrammed neurons regulate cytotoxic T-cell antitumor activity. Deciphering how TANs exert both antitumor immune activation and suppression activity through adrenergic signaling and immune checkpoint expression respectively, will allow us to leverage safe, affordable and well established neuromodulatory approaches to overcome immunosuppression in cancer. Aim 2: Identify the extracellular vesicle-shuttled driver miRNAs of TAN reprogramming and their roles in oral cancer progression. Using human-derived sensory neurons and functional genomic approaches, we will investigate the miRNA-dependent functional plasticity of immunomodulatory genes in TANs. The completion of the proposed studies will pave the way for treatment strategies that target the neuronal mechanisms associated with immunosuppression and reverse resistance to immunotherapy. Therapeutic approaches targeting this critical component of tumor biology are anticipated to improve patients' survival, treatment responses, and quality of life.

NIH Research Projects · FY 2026 · 2022-08

Project Summary ARID1A, encoding a subunit of the SWI/SNF chromatin-remodeling complex, is the most frequently mutated epigenetic regulator across human cancers. Most notably, inactivating mutations in ARID1A occur in ~50% of ovarian clear cell carcinomas (OCCC) and ~30% of ovarian endometrioid carcinomas (OEC). There is an unmet need for effective treatment modalities for ARID1A-mutated ovarian cancers. For example, OCCC is generally refractory to standard agents used to treat epithelial ovarian cancer, and when diagnosed in advanced stages, OCCC carries the worst prognosis of all ovarian cancer subtypes. The overall goal of this proposal is to develop a novel therapeutic strategy for ARID1A-mutated ovarian cancers by combining a clinically applicable metabolic glutaminase inhibitor with an immune checkpoint blockade. We show that the ARID1A inactivation creates a dependence on the glutamine metabolism. We also show that ARID1A inactivation sensitizes ovarian cancer to anti-PD-L1 treatment. The objectives of this application are to investigate the mechanisms underlying the dependence on the glutamine metabolism created by ARID1A inactivation and to investigate a combination therapeutic strategy for ARID1A-mutated ovarian cancer. Our central hypothesis is that ARID1A-mutated ovarian cancer can be therapeutically eradicated by the combination of a clinically applicable glutaminase inhibitor such as CB-839 and an anti-PD-L1 immune checkpoint blockade. Two Specific Aims are proposed: Aim 1 is to investigate the mechanism underlying the dependence of ARID1A mutation on the glutamine metabolism; and Aim 2 will develop a novel therapeutic approach for ARID1A-mutated ovarian cancer by combining a clinically applicable glutaminase inhibitor and anti-PD-L1. The proposed studies are highly innovative because they challenge current research/clinical paradigms and utilize innovative methods to explore new intervention strategies for ARID1A-mutated ovarian cancers. The research proposed is of high impact because it will provide a scientific rationale for developing urgently needed novel therapeutic strategies by repurposing the clinically applicable glutaminase inhibitor CB- 839 and an FDA-approved immune checkpoint blockade for ARID1A-mutated ovarian cancer, a disease that currently has no effective therapy. Since ARID1A is the most frequently mutated epigenetic regulator across human cancers, the mechanistic insights gained from the current studies will have broad implications for many different types of cancers as well.

NIH Research Projects · FY 2025 · 2022-08

PROJECT SUMMARY Hepatocellular carcinoma (HCC) is increasing in prevalence, yet chemotherapy options remain very limited. Resistance to systemic therapy and recurrence at local and distant sites following systemic therapy are major problems in HCC. Innovative combination therapy is an appealing strategy for treating HCC. Cancer stem cells (CSC) are a small subset of self-renewing, pluripotent, slowly proliferating malignant cells that play key roles in tumor initiation, metastasis, recurrence, and multidrug resistance in various cancer types, including HCC. Targeting CSCs may offer a promising avenue for effective anti-HCC therapy. However, targeting CSC alone would be insufficient because new CSCs could be constantly derived from non-CSC tumor cells through activation of epithelial-to-mesenchymal transition (EMT). It is therefore critical to simultaneously eliminate CSCs and proliferating non-CSC tumor cells. To date, pharmacologic approaches toward this goal remain largely unsatisfactory. This proposal seeks to develop a polymeric micelle-based solution to drug resistance in HCC. The polymeric micellar delivery system is co-formulated with cyclopamine (CPA), a naturally occurring hedgehog (Hh) signaling inhibitor capable of eliminating CSCs, and paclitaxel (PTX), a cytotoxic chemotherapeutic agent that blocks the progression of mitosis and triggers apoptosis. In preliminary studies, we have demonstrated that polymeric micelles containing both CPA and PTX, termed M-CPA/PTX, significantly prolonged median survival of transgenic mice with spontaneous c-Myc-driven HCC in both preventive and therapeutic settings. Moreover, M-CPA/PTX was significantly more efficacious than PTX alone, CPA alone, or a physical mixture of CPA + PTX. M-CPA/PTX downregulated c-Myc, suppressed tumor spheroid formation, inhibited EMT, and decreased components of extracellular matrix in favor of vastly improved tumor disposition of both drugs. Taken together, our preliminary warrant further pre-IND development of the M-CPA/PTX technology. There are two primary goals for this project: to de-risk clinical translation of M-CPA/PTX and to obtain a robust IND package. To achieve our goals, we will pursue 3 specific aims: 1) to scale up synthesis of high-quality M-CPA/PTX and fully characterize the resulting products; 2) to determine the antitumor efficacy, biodistribution, pharmacokinetics (PK), toxicity, pharmacodynamics (PD), and PK/PD correlation of M-CPA/PTX in preclinical models of HCC; 3) to evaluate the mechanism by which M-CPA/PTX exerts its anti-HCC activity. Our ultimate goal is to translate M-CPA/PTX into the clinic as a safe and effective therapy that improves survival of patients with HCC. This project will have exceptional impact because it will pave the path for clinical translation of a potentially effective systemic therapy for HCC patients and provide insight into the role of CSCs and tumor stroma in resistance to anti-HCC therapy.

NIH Research Projects · FY 2025 · 2022-08

A well-prepared health workforce is essential to conducting cancer research that serves all individuals affected by or at risk for cancer regardless of their background as they seek comprehensive cancer care, including prevention, screening, treatment, and survivorship support. Research on Americans has revealed significant differences in cancer outcomes across groups. Individuals with multiple risk-related characteristics often face greater exposure to cancer risk factors, are more susceptible to certain cancer types, and tend to experience poorer outcomes. These individuals have a variety of backgrounds and experiences, resulting in complex and heterogeneous care needs. As a result, they are often understudied and challenged in accessing care. While research focused on these populations continues to grow, gaps persist in our knowledge of cancer risk and cancer treatment experiences. These critical gaps must be addressed to develop evidence-based interventions that optimize oncology care across the entire cancer care continuum—from prevention to survivorship. As no nationally available resource routinely trains and prepares the oncology and biomedical workforce for such cancer research, our short-term goal is to promote and annually offer a foundational curriculum that prepares and orients early-career and junior investigators for conducting cancer research and fosters fruitful collaborations. The curriculum has been designed by a multidisciplinary, multi-institutional project team and expert advisory committee that bring national perspective and experience with understudied and access-challenged populations and in-depth knowledge across key areas, including cancer research, clinical care, curriculum development, education, and health care advocacy. Specific Aims: (1) Offer the course annually to 30 early-career investigators recruited nationally from a broad audience of professionals interested in cancer research across the cancer continuum and relevant to understudied and access-challenged groups; (2) Facilitate their professional development and research success by providing orientation and post-workshop seminars to complement the course. These sessions will focus on new topics in under-researched areas of cancer, as well as provide participants with exposure to potential cancer research mentors and career development guidance; and (3) Maximize the efficacy of the course by evaluating and updating curricular enhancements and teaching innovations for multimodal dissemination. Core modules include clinical cancer research; behavioral sciences and interventions; epidemiology and population-level research; community based participatory approaches for cancer research; and data collection and using digital tools and platforms. This course design will be adaptable for in-person modality, blended learning, as well as online-only formats in response to the research and education restrictions related to the COVID-19 pandemic. The long-term goal of the course is to increase the number of professionals able to lead and advocate for research to reduce cancer disparities and to advance health for all Americans at risk for and suffering from cancer.