Fred Hutchinson Cancer Center

NIH Research Projects · FY 2025 · 2022-09

In biomedical research, the need for engaged and well-trained researchers in statistics and data science has never been greater. Yet, the requisite academic pipeline for training and mentoring such a workforce remains inadequate. The driving objective of SeattleStatSummer (SeattleStatSummer for Biomedical Data Science Research Training) is to simultaneously tackle these obstacles. To address the pipeline problem, we will develop and implement a new summer mentored research program for undergraduates. The program will offer an engaging and supportive mentored research experience that increases their awareness of and interest in pursuing careers in statistics and data science research and career development, community building, and social/emotional support activities. Our goal is that students will not only gain skills and confidence in statistics and data science, but they will feel engaged in the statistics and research communities. The program will also provide coaching to faculty mentors to optimize their chances of success and confidence in mentoring students and will engage faculty in activities that build community. The specific objectives of SeattleStatSummer are Educate through intensive training in R programming and statistics via an introductory didactic program followed by ongoing reinforcement and support; Engage: via carefully crafted mentored statistics projects; Enable: through career development workshops and a role-model program; Endure: foster ongoing connection and support students in pursuing research careers in the field via activities to build community and sense of belonging and career support post program. In addition, we propose to offer a program for faculty mentors that will improve mentoring skills, build confidence in supervising students, and increase awareness of the importance of fostering a sense engagement among students. The Fred Hutchinson Cancer Center and Kaiser Permanente Washington Health Research Institute offer world-renowned research environments and scientific faculty and prioritize training the next generation of biomedical researchers. The Puget Sound area is home to a large student population, which will be targeted for recruitment via a well-honed recruitment plan bolstered by longstanding academic partnerships.

NIH Research Projects · FY 2026 · 2022-09

Almost one in ten young adults report current e-cigarette use, putting them at risk of developing nicotine addiction and long-term health effects of exposure to inhaled toxicants. Despite the need for effective treatments to help these young users quit, very few treatments targeting any type of tobacco use among young adults have been evaluated, particularly for young adults who vape and have unique treatment needs. To address these needs, we propose to develop and evaluate an avatar-led, digital Acceptance and Commitment Therapy (ACT) program called ACT on Vaping for young adult e-cigarette users at all stages of readiness to quit. This program builds upon an intervention framework employed successfully in previous pilot work, with high user satisfaction and very promising rates of biochemically confirmed tobacco abstinence. In the UG3-phase of this study, we will conduct a pilot randomized controlled trial to preliminarily evaluate acceptability and preliminary efficacy of the intervention relative to a brief advice and education control condition. Go/no-go criteria for the UG3 phase include: (a) Completed app development. (b) Completed FDA Q-submission. (c) Completed pilot trial (n=60) showing satisfaction with ACT on Vaping averaging at least 3.5 out of 5; and (d) relative to control, evidence of better outcomes on at least 1 of 3 efficacy endpoints: change in readiness to quit (mean difference in Contemplation Ladder change scores ≥ 1), 24-hour quit attempts (≥ 5% difference), and cotinine-confirmed 30-day point prevalence abstinence from all nicotine/tobacco products at 3 months (≥ 5% difference). If these benchmarks are met, we will proceed to the UH3 phase—a fully-powered, randomized controlled trial (n=1178) of the two interventions, with a primary efficacy outcome of cotinine-confirmed 30-day point prevalence abstinence at 6 months post-randomization. We will also evaluate moderators and mediators of treatment effects. This project is significant: (1) it focuses on a growing population of young adults who vape, (2) e-cigarette users are at risk for developing nicotine dependence and are exposed to toxicants that negatively impact health; (3) standard care tobacco interventions don’t address the unique treatment needs of young adults who vape, (4) ACT has demonstrated better efficacy than standard care approaches for tobacco treatment, and, (5) there is high consumer demand for vaping cessation programs among young adults, as evidenced by 241,000 young adult enrollments to Truth Initiative’s This is Quitting text messaging program between January 2019 and November 2021. It is also innovative: (1) it is the first ACT-based program of any kind for young adult vaping, (2) a digital cessation program using avatars, interactive games, and other multimedia experiences as engagement strategies is substantially different than existing treatments, (3) there are no evidence-based smartphone apps targeting young adult vaping, despite a strong preference for this modality; and, (4) there are no digital programs of any type with demonstrated efficacy for young adult e-cigarette users at varying levels of readiness to quit.

NIH Research Projects · FY 2025 · 2022-09

PROJECT SUMMARY/ABSTRACT Esophageal adenocarcinoma (EAC) is one of the most lethal cancers, with a 5-year survival rate less than 20%. Incidence of EAC has risen sharply in the U.S. and other Western countries over the past four decades, largely due to rising prevalence of two risk factors – gastroesophageal reflux disease and obesity. EAC develops from Barrett’s esophagus (BE), a cancer precursor defined by a specialized columnar metaplasia of the distal esophagus. Although BE follows an indolent course in most patients, 5-10% eventually progress to cancer, and a sizable fraction of BE remain undetected in the population. A critical unmet need is to identify individuals with high-risk BE who are most likely to develop EAC and thus benefit from screening and endoscopic surveillance. Conversely, identifying the majority who are unlikely to progress will reduce risks and costs associated with unnecessarily frequent surveillance. Biomarker-assisted risk stratification, however, continues to be hindered by our limited understanding of the molecular pathways underlying early steps in the development of EAC. In recent years, genome-wide association studies (GWAS) have identified ~20 novel genetic susceptibility loci, yet most heritability remains unexplained, and only one SNP is linked specifically to BE→EAC progression. The epigenome, an important interface between the environment and the genome, has not been studied for its potential mediating roles in relation to genotypes, strong environmental risk factors and progression to EAC. To address these gaps, overcome sample size limitations for a rare cancer, and invigorate existing research efforts, newer molecular and statistical approaches are needed to systematically integrate multi-omics data with established disease exposures. In this project we will conduct the most comprehensive multi-omics study of BE, the key cancer precursor, profiling the transcriptome and methylome of 500 biopsies from the NCI-funded Barrett’s and Esophageal Adenocarcinoma Consortium and Roswell Park Comprehensive Cancer Center, and perform integrative analyses leveraging genotypes and environmental risk factors already available through the largest GWAS meta-analysis for BE/EAC (n≈27,000). The goal is to identify new genetic risk loci through eQTL mapping and transcriptome-wide association study (Aim 1), new epigenetic loci mediating and predicting the risk of BE progression to EAC (Aim 2), and develop risk prediction models integrating genome-wide polygenic risk score and environmental exposures (Aim 3). The unifying theme of the three aims is development and implementation of innovative analytical strategies, leveraging transcriptome and methylome data. Ultimately, genetics, epigenetics, and environmental exposures will be incorporated to identify high-risk populations for tailored screening and surveillance, and prevent cancer development.

NIH Research Projects · FY 2025 · 2022-09

Leukocytes are essential immune components protecting the body against foreign invaders. Adhesion, migration, extravasation, and cell-cell communication are mediated though the bidirectional signaling of β2 integrins, which are integral membrane proteins found on the leukocyte surface. Due to their complex and multifunctional roles, dysregulation of β2 integrins is linked to autoimmune, cardiac and pulmonary pathologies as well as infectious diseases and several cancers. Although integrins are prime therapeutic targets, drug development has been hindered due to unanticipated side effects that arise from large gaps in our understanding of the mechanisms that drive specificity and integrin activation. I will decipher the molecular basis for β2 integrin activation, ligand recognition, and bidirectional signaling using an integrative approach. I'll build on my expertise in cryoEM method development to capture high- resolution conformational snapshots of isolated β2 integrin and ligand-bound complexes to reveal the dynamic structural rearrangements associated with signal transduction and identify key residues mediating ligand specificity. Using cell-surface expressed integrins, I'll assess the functional consequences of mutating these residues on binding of ligands and conformation- specific antibodies and on adhesion, phagocytosis, and cell motility. To gain broad insight into integrin allostery in a near-native context, I'll develop a membrane mimetic system using next- generation styrene maleic acid copolymers. Membrane lipids influence integrin activation and ligand binding and are key to forming stable complexes. These polymers will afford a stream- lined method to extract and purify nanodiscs embedded with pre- formed integrin-ligand complexes in their native environment. I will use this system to study integrin in complex with talin, the central integrin- activator protein that binds a conserved motif on the cytoplasmic region on integrin. This will reveal in molecular detail how integrins are activated to relay signals allosterically across the plasma membrane and define a molecular basis for bidirectional signaling as well as provide a framework for designing biochemical, biophysical, and mechano- sensitive experiments to study larger complexes and gain comprehensive insight into integrin function. Ultimately, this work will provide a structural blueprint for the rational design of therapeutics for autoimmune diseases, which is a long-term goal of the lab.

NIH Research Projects · FY 2025 · 2022-09

The field of cancer diagnostics is in a rapidly expanding growth phase that goes hand in glove with the precision medicine revolution. However, the rapid pace at which new technologies are entering the marketplace makes rigorous evaluation via controlled studies infeasible for all but a relative few. This means that while we typically have some data about diagnostic test performance, we frequently lack evidence regarding the outcomes that drive clinical and policy decisions. The Research Program outlined in this application will tackle this data- evidence divide using the tools of modeling and analytics. Modeling is an increasingly accepted discipline for integrating knowledge about the process by which diagnostic performance drives outcomes. Analytics is the use of statistical learning techniques to fill in the knowledge gaps and to propagate uncertainty from model inputs to outcomes. The Principal Investigator has built a leading research program in modeling and analytics for evidence generation in cancer policy. Among many methodologic and substantive contributions, her work has informed prostate cancer screening guidelines from national policy panels, established best practices for estimation of overdiagnosis, and produced specific directions for screening high-risk populations including Black men. The Research Program outlined in this application will harness the modeling and analytics skillset developed by the Principal Investigator over nearly three decades to build a framework and tools for evidence generation around cancer diagnostics. The application details a sequence of projects for two technologies that are generating intense current interest with wide-ranging practice implications and serious evidence gaps: Multi-cancer early detection testing, and PSMA-PET/CT for newly diagnosed and recurrent prostate cancer. The MCED work will deepen our understanding of performance characteristics, provide guidance regarding a defensible test confirmation strategy, project benefits and harms of different MCED strategies and offer new ideas for shortcutting the typically lengthy process of cancer screening trials. The PSMA-PET/CT work will develop an approach for updating treatment benefit estimated derived from trials that included a mixture of patients with unknown PSMA status and will project lives saved of treatment reallocation on the basis of PSMA-PET.CT result. The tools and processes developed for modeling these technologies will be applicable to other new diagnostics that emerge during the lifetime of the Research Program. The modeling work will be accompanied by a sequence of real-world analytics projects to assess dissemination of and disparities in uptake of novel diagnostics and their consequences for healthcare utilization and costs. This work will establish collaborations with new real-world data partners and materially expand the Principal Investigator’s skillset to encompass a greater competency in medical informatics. The successful execution of the Research Program will improve our understanding of how novel cancer diagnostics impact clinical and policy relevant outcomes so that these technologies can be used wisely and equitably to improve care for all cancer patients.

NIH Research Projects · FY 2025 · 2022-09

Abstract For the last 30 years, the 5-year survival rate of small cell lung cancer (SCLC) has been less than 7% despite the addition of immune checkpoint inhibitors as treatment options. Therapies like immune checkpoint inhibitors that aim to reengage an immune response may not succeed for SCLC as previous studies have shown downregulation of MHC molecules, low PD-L1 expression and limited immune infiltration. However, SCLC is often associated with autoantibody-driven Paraneoplastic Syndromes, providing evidence for the immunogenicity of SCLC. We propose that chimeric antigen receptor T cells (CAR-Ts) as a novel approach for SCLC immunotherapy that overcomes impediments to endogenous immunity. CAR-Ts are synthetically engineered to fuse antibody ligand binding domains with costimulatory components that activate T cells after engagement of cell surface antigens, and have had considerable success in leukemia, lymphoma, and multiple myeloma. The microenvironment of SCLC is phenotypically closer to CAR-T responsive lymphoma than many solid tumors where CAR-Ts have thus far had limited success. A challenge for CAR-T cells in many solid tumors is the identification of target antigens that are tumor-specific. We have identified 13 novel cell surface antigen and here will prioritize 3 with high prevalence in SCLC. Each of these antigens have post-translational modifications that act as neoantigens and lead to autoantibody production in a high percentage of SCLC cases. We will capture these neoantigen-autoantibodies from SCLC patient-derived B cells, sequence the tumor specific binding sequences, and design and test CARs constructed from the single chain variable fragments (scFvs). The benefit of isolating autoantibodies from SCLC patients to detect tumor-specific neoantigens is three-fold: 1. The antigens identified have already proven to be immunogenic; 2. The variable regions of these human autoantibodies can be directly engineered into ligand binding domains of CAR-T cells; and 3. Autoantibodies can be detected in the blood of patients and serve as tissue surrogate biomarkers to guide CAR-T cell target selection. The CAR-T cells we develop will be rigorously tested in multiple preclinical models that address complementary but non-overlapping therapeutic barriers. These include testing CAR-T cell tumor infiltration, efficacy and toxicity in a library of genetically diverse SCLC patient derived xenografts and identifying, then overcoming, immunosuppressive mechanisms in the immune competent Rb/p53 genetically engineered mouse model. Our team of experts in lung cancer, autoantibody biomarkers, immunology and CAR-T cells is well equipped to execute the development of novel immunotherapies that are desperately needed in SCLC.

NIH Research Projects · FY 2025 · 2022-08

Project Summary The intestine serves both as a conduit for the uptake of food-derived nutrients and as a barrier that prevents host invasion by microorganisms. This barrier function is important to maintain intestinal integrity and it is promoted by immune cells, which can quickly respond to microbial presence in intestinal lumen coordinating protective functions. Alterations in timing of food intake and diet composition have been associated with the development of immune-mediated intestinal dysfunctions (e.g. irritable bowel syndrome). However, despite its profound biological and clinical relevance, there is a major gap in our understanding of how intestinal immune responses are modulated by food presence in the intestinal tract. The long-term goal of this proposal is to determine how, during feeding, dietary-derived signals are sensed in the intestine and promote alterations in intestinal immune responses. Recently, we uncovered a neuroimmune circuit that coordinates intestinal immune- mediated barrier functions in response to food consumption. This neuroimmune circuit is formed by the interaction of vasoactive intestinal peptide-producing enteric neurons (VIPens) and type 3 innate lymphoid cells (ILC3s). VIPens are activated by the presence of food in the intestinal tract, they directly inhibit ILC3 functions. Although VIPen-mediated inhibition of ILC3 during feeding reduces intestinal barrier functions, it also increases efficiency of fat absorption from the diet (immune-nutritional trade-off). Importantly, in experimental mouse models, perturbations in this neuroimmune circuit alters host resistance to enteropathogens and host-microbiota interactions. We propose to study the mechanism of activation of VIPens by dietary signals as an entry point to understand how feeding promotes alterations in intestinal immunity. A combination of cutting-edge technologies to measure neuronal activation in vivo (genetically encoded calcium indicators and intravital imaging), manipulate neuronal activity (chemogenetic tools and AAV-assisted CRISPR/Cas9-based genetic manipulation), dissect molecular profiles of cellular circuits (monosynaptic viral tracing and single cell genomics), and control ingestion of specific dietary signals (diet engineering), will allow us to acquire a mechanistic understanding of how food consumption can affect intestinal immunity through neuroimmune circuits. The Specific Aims of this proposal are: 1) to determine the nature of food-derived signals that, by triggering activation of VIPens, coordinate intestinal immune-nutritional trade-offs, and 2) to dissect the cellular and molecular pathways of VIPens activation by food-derived signals. These studies will provide the molecular underpinnings of how intestinal immune responses are being modulated by food consumption, as well as provide new insights of the intestinal mechanisms for sensing food-derived signals and orchestrating immune-nutritional trade-offs. These studies will also advance the development of dietary-based therapies to boost immune-mediated barrier functions and the mitigation of intestinal infectious and inflammatory diseases.

NIH Research Projects · FY 2025 · 2022-08

Project Summary Gastrointestinal (GI) cancers are a major cause of mortality and morbidity in the U.S. and their treatment uses a substantial proportion of healthcare resources. Of the GI cancers, colorectal cancer (CRC) and esophageal cancer (EAC) account for a majority of the cancer related deaths, and both are preventable by screening and surveillance. The current screening tests are suboptimal and have variable success. A major goal of CRC screening tests is to identify advanced tubular and serrated adenomas, which are high-risk for becoming CRC, as well as early stage CRC. The risk for CRC is variable with some people being at high risk because of family histories of CRC, hereditary cancer syndromes, or a personal history of adenomas. High risk people are placed on aggressive colonoscopy based surveillance programs and low-risk people are placed on minimal surveillance programs. Unfortunately, our current system for identifying high and low CRC risk is suboptimal resulting in under and over surveillance and preventable interval CRCs. Better risk markers for CRC to are needed to prevent interval CRCs and improve the overall effectiveness of CRC screening. Analogous to CRC, EAC arises from a precancerous condition of the esophagus called Barretts esophagus (BE), which is a specialized intestinal metaplasia of the esophagus and the highest risk factor for EAC. It is present in 5% of the US population. BE progresses to EAC through successive histologic steps of low grade dysplasia (LGD), high grade dysplasia (HGD) and then EAC. Screening and surveillance for BE is recommended using serial upper endoscopy, which is controversial in its effectiveness for preventing deaths from EAC. This is in part because, as with CRC, BE patients have variable risk of EAC and are placed on high- risk and low-risk screening programs. However, the current system for assigning risk is not accurate and the current screening test is expensive. More cost effective and accurate EAC and HGD screening/surveillance assays and accurate BE risk biomarkers are needed. We propose to develop an EDRN BCC that is integrated into the EDRN consortium and, through collaborations within and outside the EDRN, will develop effective GI cancer screening biomarkers. We propose to identify, validate, and develop accurate CLIA compliant risk biomarkers for CRC and for EAC in order to prevent EAC and CRC missed under current screening protocols. Moreover, the accurate risk stratification of patients for CRC and EAC will reduce the financial impact of current CRC and EAC prevention programs. We also propose to identify and validate accurate CLIA compliant early detection markers for HGD and early stage EAC that can be used in an inexpensive, non-endoscopic surveillance test.

NIH Research Projects · FY 2025 · 2022-08

PROJECT SUMMARY/ABSTRACT The recent SARS-CoV-2 pandemic has caused global catastrophe and needs little introduction. One of seven coronaviruses known to infect humans (HCoVs), SARS-CoV-2 infection leads to a range of pathogenic outcomes ranging from asymptomatic infection to severe pneumonia and death. Conversely, four HCoVs are endemic, circulate globally and typically cause only mild illness. The differences in pathogenesis between more lethal HCoVs and endemic HCoVs raises the possibility that there are differences in the innate immune response upon infection with the different HCoV species. The Type I interferon (IFN) response is the first line of innate immune defense against viruses and involves activation of a suite of IFN-stimulated genes (ISGs) that maintain IFN production and generate a cellular antiviral state. Several high-throughput screening studies have identified ISGs that contribute to the protective IFN response against SARS-CoV-2 infection, several of which did not overlap, suggesting much more remains to be learned. A smaller subset of ISGs is known to participate in IFN-mediated signaling in cells infected with endemic HCoVs such as HCoV-OC43, but the studies have not been comprehensive, and the role for IFN remains complex and understudied. Therefore, comprehensive approaches are needed to identify ISGs that are restrictive against HCoVs, particularly endemic HCoVs. Identifying ISGs that are active against endemic HCoVs will pave the way for comparative studies of ISG activity between HCoVs that are more pathogenic. Identifying the mechanisms that govern differential ISG activity against broad HCoV species may inform broad antiviral therapeutic strategies targeting current HCoVs and those that are likely to emerge in the future. In preliminary studies, we identified H1299 cells as permissible to infection with HCoV-OC43, and found a 92-fold protective effect against HCoV-OC43 infection in the presence of IFNβ. Here, we propose to identify IFN-mediated restriction factors against HCoV-OC43. To address the hypothesis that HCoVs with different pathogenic features are regulated by different features of the innate immune response, we will do a comparative study of ISG activity between SARS-CoV-2 and HCOV-OC43 and will seek to identify mechanistic explanations for differences we identify. During the mentored K99 phase: we will (1) develop a customized CRISPR-Cas9 knockout library for use with HCoV-OC43 in H1299 cells, (2) screen for ISGs responsible for the 92-fold protective IFNβ effect, (3) validate hits with single gene knockout cell lines and test for activity against SARS-CoV-2 and (4) initiate experiments to determine if these ISGs reflect direct viral protein-host protein interactions, if time allows. During the independent R00 phase: we will (1) comparatively test viral protein-host ISG interactions identified by co-immunoprecipitation and mass spectrometry, (2) complete expansion of CRISPR screening to other respiratory cell lines, and (3) initiate preliminary structural studies to characterize ISG-viral protein interfaces.

NIH Research Projects · FY 2026 · 2022-08

PROJECT SUMMARY Cell metabolism is the collection of biochemical processes that support the bioenergetic, biosynthetic, and signaling demands of life. While the biochemical composition of most major human metabolic pathways have been defined, we do not yet have a strong understanding of how metabolic pathways are differentially utilized to support the diverse needs of cells across cell states. Activation of cell proliferation is one such state that comes with substantial changes to metabolic pathway activities, however many of the mechanisms by which these metabolic changes support cell proliferation, and the consequences of their disruption, remain unknown. At the heart of cell metabolism is the mitochondrion, a double membrane bound organelle that serves as a metabolic hub by providing a separate biochemical compartment from the cytosol and through the unique metabolic capabilities afforded by the electron transport chain (ETC). While most famous for its role in ATP synthesis, studies from us and others have determined that mitochondrial metabolism is essential for supporting cell proliferation independent of ATP production. These findings have upended the traditional view of mitochondria as mere “powerhouses” and underscore the need for a new, holistic understanding of how mitochondria support cell functions. Our work has identified that complex I of the ETC is critical for cell proliferation by regenerating electron acceptors, which support the synthesis of the amino acid aspartate. In addition, our preliminary data uncover that the metabolic effects of impairments to complex II of the ETC are distinct from those of complex I, and we identify a novel, redox-driven mitochondrial metabolic pathway necessary for cell proliferation upon complex II dysfunction. Nevertheless, a comprehensive understanding of the metabolic contributions of mitochondrial processes to cell proliferation remains lacking. My research program uses state-of-the-art approaches to delineate the metabolic and functional consequences of disruptions to mitochondrial processes to gain a new, systems level understanding of how the interconnected metabolic pathways in mitochondria support cell proliferation. Notably, disruptions to mitochondrial function in humans have highly diverse clinical manifestations and so mechanistic understanding of the consequences of impairments to different mitochondrial processes will support the development of novel, targeted therapeutic approaches for the many diseases associated with mitochondrial dysregulation, including inborn errors of metabolism, cancer, neurodegeneration, aging, and others.

NIH Research Projects · FY 2026 · 2022-07

PROJECT SUMMARY The thymus, which is the primary site of T cell generation, is extremely sensitive to insult, but also has a remarkable capacity for endogenous repair. Even though there is likely continual thymic involution and regeneration in response to everyday insults like stress and infection, profound thymic damage caused by radiation injury leads to prolonged T cell lymphopenia. Consequently, identification of therapies that can boost T cell reconstitution in recipients after a dose of radiation is a clear priority. We have previously identified two distinct pathways of endogenous thymic regeneration, centered on the production of the regeneration factors IL-22 by innate lymphoid cells (ILCs), and BMP4 by endothelial cells (ECs); both of which mediate their regenerative effects by targeting thymic epithelial cells. More recently we have found that the trigger for these distinct regenerative pathways hinge on the balance between forms of cell death, with immunologically silent apoptosis (which is abundant in thymocytes during steady-state) suppressive to the regenerative program. On the other hand, after thymic damage caused by radiation injury, we found a switch toward immunogenic cell death, with the resulting release of damage-associated molecular patterns (DAMPs) sufficient to promote regeneration. Specifically, we identified that intracellular Zn was released after radiation injury, where it could signal through the G-protein coupled receptor 39 (GPR39) to stimulate production of BMP4 and IL-23, a key upstream regulator of IL-22 production. Separately, we also found that the release of the prototypical DAMP, ATP, was able to signal directly on thymic epithelial cells through purinergic (P2) receptors and promote their expression of Foxn1, key microenvironmental drivers of T cell development. Importantly, our preliminary data also suggests that each of these pathways can be therapeutically targeted to improve thymic recovery following radiation damage in young mice. Based on our preliminary data, we hypothesize that modulation of pathways associated with these DAMPs can be used as a countermeasure to improve immune function after radiation injury by stimulating the generation of new T cells in the thymus. Specifically, our proposal has the following aims: (1) To validate and optimize the targeting of GPR39 or P2 receptors to improve the production of new T cells and immune function after radiation injury; (2) to examine the ability of the thymus to respond to regenerative signals across mouse lifespan and sex; and (3) to comprehensively evaluate the potential for targeting GPR39 or P2 receptors to improve thymic function after radiation damage across mouse lifespan and sex. The studies outlined in this proposal not only have the potential to define important pathways underlying tissue regeneration across lifespan but could also result in innovative approaches to enhance T cell recovery after radiation damage.

NIH Research Projects · FY 2025 · 2022-07

ABSTRACT Antigen-specific therapies have long been pursued to improve outcomes in acute myeloid leukemia (AML). So far most exploited are monoclonal antibodies (mAbs) targeting CD33, a glycoprotein displayed on the cell surface of leukemic blasts in almost all cases and possibly leukemia stem cells in some. Longer survival of some patients treated with the CD33 antibody-drug conjugate gemtuzumab ozogamicin (GO) validates this approach, but GO is often ineffective, prompting efforts to develop improved, more potent CD33-directed therapeutics. Because AML cells are exquisitely sensitive to radiation in a dose-dependent fashion, radionuclides are ideal to arm anti- CD33 mAbs. Indeed, early phase clinical trials demonstrated substantial anti-AML efficacy of the anti-CD33 mAb lintuzumab (HuM195, SGN-33) when coupled with the a-emitter actinium-225 (225Ac). a-emitters deliver a very high amount of radiation over just a few cell diameters, thereby enabling precise and efficient target cell kill, rendering them particularly interesting for specific targeting of AML with radioimmunoconjugates (“RIT”). However, even with 225Ac-lintuzumab, an important shortcoming is CD33 expression on normal myeloid cells, which leads to “on-target, off-tumor cell” toxicities that manifest as severe and prolonged myelosuppression with life-threatening sequelae (e.g. infection). Thus, clinical use of CD33-directed RIT without immediate stem cell rescue is currently limited to suboptimal drug doses. We have recently demonstrated in mice and nonhuman primates that CRISPR/Cas9 nuclease-based editing of CD33 results in functionally normal hematopoiesis that expresses reduced levels of CD33 and is protected from GO and CD33-directed T cell-engaging therapeutics. We hypothesize CD33-edited normal hematopoietic stem and progenitor cells (HSPCs) will resist CD33-directed RIT with a-particle-emitting radionuclides and enable their safe use at maximally effective drug doses. However, the CRISPR/Cas9-based CD33 gene editing strategy suffers from significant off-target activity, and DNA double strand breaks (DSBs) can generate larger deletions and complex chromosomal rearrangements and cause TP53-dependent DNA damage response and cell cycle arrest. To address this limitation, we will optimize and characterize a novel gene-editing strategy to protect normal hematopoiesis from highly potent CD33-directed RIT by utilizing the recently described base editor (BE) technology. BEs induce precise nucleotide modifications without intentional introduction of DSBs, making them an attractive strategy to generate CD33null “normal” hematopoietic cells. We have assembled a multidisciplinary team of investigators with complementary expertise in CD33-directed therapies, preclinical optimization of RIT, and radiopharmaceutics to conduct well-controlled preclinical IND-enabling studies to develop BE-based CD33 engineering of normal human HSPCs for clinical use with a-emitter CD33-directed RIT for patients with AML and other CD33-expressing disorders.

NIH Research Projects · FY 2025 · 2022-07

SUMMARY/ABSTRACT The AR may be the earliest known example of a lineage oncogene: a master regulator of cell survival and growth to which neoplastic cells derived from prostate epithelium are addicted. Recognizing this unique feature, concerted efforts have focused on developing therapeutics capable of suppressing AR signaling. Androgen deprivation therapy (ADT) and AR pathway signaling inhibitors (ARSI) produce dramatic responses in the vast majority of patients with metastatic PC (mPC). Unfortunately, these responses are not accompanied by cures, with near universal development of treatment resistance. Studies from our group and others have determined that an increasing fraction of mPCs resisting AR pathway inhibition lose AR activity and gain a spectrum of new phenotypes, each of which exhibit an aggressive clinical course with limited treatment options. The processes by which tumor cells switch lineages under treatment pressure is not well understood. Determining the mechanisms that permit or drive this lineage plasticity may identify new treatment strategies. This proposal is designed to address a major clinical problem whereby AR pathway inhibition promotes tumor cell plasticity. We will test the hypothesis that targeting permissive epigenetic factors or lineage determinants together with AR pathway inhibition will prevent lineage-redirection, prolong response rates overall, and cure a subset of advanced prostate cancers. AIM 1. Identify the key determinants and permissive factors that promote a lineage switch from conventional AR- driven prostate cancer to new phenotypes following AR-directed treatment. AIM 2. Determine if modulating factors that drive or permit lineage specification can prevent, delay, or reverse resistance to AR pathway inhibition. AIM 3. Determine if co-targeting characteristics of re-directed lineages that emerge in the context of lineage switching will prolong responses to AR pathway inhibition. In order for effective therapeutics to be developed that can adequately address this new class of malignancy, the pathways permitting or driving lineage conversion must first be clearly defined; this project aims to elucidate those underlying mechanisms.

NIH Research Projects · FY 2025 · 2022-07

PROJECT SUMMARY/ABSTRACT The 20 years since publication of the draft human genome sequence in 2001 have been a truly transformational era in the field of cancer research marked by ever more significant scientific milestones and a steadily accelerating rate of progress. Global scientific initiatives focused on comprehensive characterization of tumor genomes – such as The Cancer Genome Atlas (TCGA) and the Pan-Cancer Analysis of Whole Genomes (PCAWG) – are radically transforming our approach to the prevention, diagnosis, and treatment of cancer. Surprisingly, although both TCGA and PCAWG included tumors from patients on many continents, few tumors from Africa were included in their analyses. Moreover, the number of African scientists involved in the two projects was limited. The paucity of African tumors analyzed by TCGA and PCAWG, and the dearth of African scientists involved in their analysis, is concerning given that Africa is home to 1.4 of the world’s 7.9 billion inhabitants, and much of human genetic variation is found only on the African continent. The profound under- representation of African genomes and tumors – and of African scientists – in cancer genomics studies is attributable to many factors, but the lack of requisite infrastructure for genomic research in Africa and the limited number of genomics professionals in Africa who are trained in both the generation and analysis of genomic data are major contributors. To ensure that the transformative benefits of recent advances in cancer genomics to the prevention, diagnosis, and treatment of cancer will be realized in Africa, dedicated initiatives must be undertaken to increase the number of trained genomics professionals in Africa. In this application, the Fred Hutchinson Cancer Research Center in Seattle, WA and the Uganda Cancer Institute in Kampala, Uganda – two premier institutions that have been collaborating since 2004 to conduct cancer research and to increase the capacity for high-quality cancer research, training, and clinical care in Uganda and East Africa – propose to extend their well- established collaboration to address the critical shortage of African genomics professionals who are trained in the generation and the analysis of cancer genomic data. We hypothesize that improving the supply of African professionals who can both generate cancer genomic data of reproducibly high quality and analyze those data rigorously and comprehensively will accelerate the development of an Africa-based cancer genomics community and ensure that the advances of the cancer genomics revolution will maximally benefit African cancer patients. Over a period of 5 years, and in conjunction with the Infectious Diseases Institute of the College of Health Sciences, Makerere University, the Fred Hutch and UCI will support 4 Ugandan or East African individuals who will complete master’s degrees, and 2 who will complete doctoral degrees, in Bioinformatics with a focus on cancer genomics and genomic data science. We anticipate that graduates of the program will become the leaders of the cancer genomics community in Uganda and East Africa.

NIH Research Projects · FY 2025 · 2022-06

Project Summary/Abstract Melanoma is the deadliest form of skin cancer. Its incidence is on the rise with 106,000 new cases expected in the U.S. in 2021. Immune checkpoint inhibitors (ICIs) have revolutionized the treatment of early and advanced melanoma, with concurrent anti-CTLA-4 and anti-PD-1 monoclonal antibodies demonstrating a response in ~50% of patients, including highly durable responses. Unfortunately, there are no adequate biomarkers to predict response to single agent or combination ICI, and dual checkpoint blockade is associated with significant grade 3/4 immune-related adverse events (irAEs) in ~55% of patients. The goals of our PTRC are designed to address two unmet clinical needs: (i) improve our understanding of mechanisms of resistance to ICIs to design more effective immunotherapies and combinations, and (ii) identify potential biomarkers to select patients appropriately for single agent vs combination immunotherapies and to predict and monitor irAEs. In our Preclinical Arm, we will perform integrated proteogenomic analysis of clinically annotated, pre-treatment biopsies from melanoma patients who received ICI. The data will be analyzed in the context of clinical annotations to refine an existing signature of melanoma ICI response identified by our team and to further elucidate mechanisms of ICI response/resistance and signatures associated with irAEs. In our Clinical Arm, we will analyze clinical trial biospecimens using MRM-based assays to confirm & extend findings generated in the Preclinical Arm.

NIH Research Projects · FY 2025 · 2022-05

ABSTRACT Hematopoietic stem cells (HSCs) maintain the adult blood and immune systems throughout the lifetime of an organism. While HSC transplants are used clinically for the curative treatment of patients with leukemias, lymphomas, and immune disorders, the success of HSC transplants varies considerably, with some patients having to undergo multiple transplants due to inefficient HSC engraftment. The goal of the proposed project is to identify how the heparan sulfate proteoglycan, Syndecan-2, regulates HSC maintenance and regeneration. Aim 1 will utilize newly generated tissue-specific knockout mice to analyze the contribution of Syndecan-2 in regulating HSC engraftment, while performing a structure-function analysis of the molecular motifs of Syndecan- 2 in mediating engraftment and niche regeneration. Aim 2 will analyze the role of Syndecan-2 in regulating hematopoietic recovery from stress by combining radiation injury and myelosuppression in vivo models of HSC self-renewal. These experiments will demonstrate the role of Syndecan-2 in mediating HSCs maintenance and regeneration, providing foundational knowledge to enable further study of how proteoglycans regulate HSCs. The knowledge procured from the completion of the proposed aims is expect to have broad potential contributions to patients undergoing chemotherapy and radiation therapy as a part of their treatment regimen for variety of cancers, immune diseases, leukemias and anemias and therapeutic value from a public health standpoint for treating patients that experience accidental radiation exposure. My career goals are to become an assistant professor at a top-tier academic research institution. I aim to lead a research program that investigates how proteoglycans regulate HSCs and the HSC microenvironment at homeostasis and during stress. To achieve these goals, the proposed project has embedded training in modeling HSC recovery from injury, while expanding my knowledge of glycobiology and growth factor signaling. Under this award, I will pursue the proposed training with the mentorship of Dr. John Chute (UCLA), who has substantial experience in translational modeling of HSC and niche regeneration, while supported by an Advisory Committee with expertise in stem cell biology, growth factor signaling and proteoglycan biology. As a postdoctoral fellow, I will present my work at international conferences and to my mentoring committee, which consists of experts in the field. As junior faculty, I will receive training in laboratory management/leadership, grant writing, negotiation, and mentoring to help me lead a successful research team. The proposed project will allow me to transition to an independent research position while developing my skillset in HSC biology. UCLA has superb research facilities, professional development resources and administrative support, which will provide the necessary infrastructure to support my research and career development as a mentored postdoctoral fellow and ultimately, my transition to independence.

NIH Research Projects · FY 2026 · 2022-04

Project summary There is a crisis of reproducibility and replicability of scientific results. This crisis is an increasing source of concern both in the scientific and popular press. The crisis is so acute that the United States Congress is currently investigating reproducibility of the scientific process. At the heart of this crisis is a collection of problems including small-sample sizes, under-powered studies, under-trained data analysts and an inability to directly leverage prior results in the statistical analysis of smaller, hypothesis-driven experiments using high-throughput technologies. Advances in technology have dramatically reduced the cost and difficulty of collecting high-throughput molecular data. Large collections of raw data are increasingly publicly available but are usually incorporated into individual analyses by NIGMS and other investigators on an ad-hoc basis. Meanwhile, the other costs of running a designed, hypothesis-driven study have not decreased at the same speed with technological advances. It is still expensive to identify, recruit, collect, and follow up samples even if the high-throughput measurements themselves are cheap. Despite the incredible amount of available public data, it is still common practice to perform statistical inference in these hypothesis-driven experiments study-by-study, only indirectly including previous data, estimates, and results. So findings from these studies may be highly variable, unreliable, or unreplicable. Our group has focused on developing statistical methods, data resources, and software and training that allow researchers to borrow strength empirically from public repositories, large-scale data generation projects, and crowd-sourced data to improve inference in individual, hypothesis driven studies. We propose to build on our work in developing statistical data sources, methods, software and training that facilitate and speed the work of our biological and medical collaborators. The result will be a research community that can take advantage of public data already collected at a large cost to the NIH to improve power, reduce required sample sizes, and improve replication in many new hypothesis driven molecular studies of development and disorder.

NIH Research Projects · FY 2024 · 2022-04

Project Summary Tumor-associated macrophages (TAMs) usually express an M2 phenotype which enables them to perform immunosuppressive and tumor-promoting functions. Reprogramming these TAMs toward an M1 phenotype could thwart their pro-cancer activities and unleash anti-tumor immunity, but current efforts to accomplish this are nonspecific and elicit systemic inflammation. Our group at Fred Hutchinson Cancer Research Center has developed a targeted nanocarrier that can deliver in vitro-transcribed mRNA encoding M1-polarizing transcription factors to reprogram TAMs without causing systemic toxicity. With the goal of designing the first clinical trial for treating chemotherapy-resistant ovarian cancer patients with this nanodrug, we propose here research that will generate the data required for an IND application. Specifically, we will (1) develop a robust protocol for the scaled-up production of genetic macrophage-programming nanoparticles under GMP- conditions so they can be carried forward into large primate and human studies, (2) identify potential infusion reaction risks, with reference to FDA regulations for nanomedicines, and (3) confirm safety of the nanoparticles for clinical use in a large-animal species. We expect the outcome of the proposed research will help propel this approach into clinical practice for the treatment of advanced ovarian cancer, and provide knowledge to design a broad repertoire of nanotherapeutics that genetically reprogram TAMs as a strategy to treat other tumor types.

NIH Research Projects · FY 2026 · 2022-02

PROJECT SUMMARY Influenza virus evolves rapidly to escape immunity elicited by prior infections and vaccinations. To combat this evolution, the strains in the influenza vaccine are updated every season to keep pace with viral evolution. However, it is currently difficult to accurately forecast which viral strain will dominate the coming season, and vaccines are less effective when the wrong strain is chosen for the vaccine. Here we propose a combined experimental / computational approach that will overcome some of the factors that currently make vaccine strain selection so difficult. First, we will use a new deep mutational scanning approach to directly map the genotypes of the viral surface proteins to their antigenic phenotype with respect to human antibody immunity. Second, we will use this new experimental approach to reconstruct the heterogeneous human immune landscape over which influenza virus evolves. Finally, we will build computational models that forecast how influenza evolves year-to- year in response to human immunity. All of our data and forecasts will be integrated into easy-to-use interactive visualizations. Overall, this work will improve the capacity to accurately identify which viral strains should be selected for the seasonal influenza vaccine.

NIH Research Projects · FY 2024 · 2022-01

Project Summary Protein synthesis begins via a multi-step and highly-regulated process that culminates with a ribosome poised at a start site on a messenger RNA (mRNA). Loss of control is broadly implicated in human disease, including cancers, developmental disorders, neurological diseases, and viral infections. Since translation initiation is rate limiting, an important regulatory strategy involves multi-protein complexes recruited to the opposite end of the mRNA. Recruited regulatory proteins directly enhance or inhibit assembly of the initiation machinery on the mRNA, thereby tuning protein production up or down. Many molecular mechanisms that underlie translation initiation and its long-range control remain unclear. Current paradigms rely on analyses of complexes that are stable for minutes to hours, as the intrinsic dynamics challenge approaches in bulk solutions. Here, I build off my initial postdoctoral research to track the human translation initiation machinery, mRNA, and regulatory complexes as they interact using single-molecule spectroscopy and purified components in vitro, which I complement with structural analyses. In Aim 1 (K99), I focus on how the ribosomal subunits are recruited to and load onto an mRNA, and determine how these landmark initiation events are dictated by mRNA features. In Aim 2 (K99), I examine how the final initiation steps and the transition into active protein synthesis are coordinated and governed by a universally-conserved GTPase, eIF5B. In Aim 3 (R00), I leverage the obtained training and expertise to examine how translation initiation is controlled via the 3’-end of the mRNA by the CCR4-NOT complex, a major regulator with human-health relevance. As my preliminary data demonstrate, my strategy will overcome previous roadblocks to provide a dynamic view of key molecular branchpoints that underlie translation initiation, reveal how they are targeted for control, and may define molecular bases of disease. Aided by strong collaborations and my mentoring team, the proposed research and training plan will provide me with new conceptual and experimental expertise in structural biology and biophysics and enhance my professional development. Together, this proposal will serve as a strong foundation as I transition into independence and continue my investigation of translational control and how it goes awry in human disease.

NIH Research Projects · FY 2026 · 2021-12

ABSTRACT HIV-specific gene therapies are a powerful and promising means to achieve HIV cure/stable remission in the absence of antiretroviral therapy (ART). Broadly neutralizing antibodies (bNAbs) and analogous molecules such as eCD4-Ig offer one of the clearest paths to a cure, but are hindered by three key obstacles. First, passive administration of bNAb/eCD4-Ig proteins is by definition a transient therapy; when circulating levels of these potent anti-HIV factors decline, virus replication is able to resume. Second, gene therapy vector-based approaches including adeno-associated virus (AAV) support prolonged expression of bNAbs and other antiviral transgenes, but are frequently limited by host immune responses. Third, potent ART regimens suppress viral replication to extremely low levels, rendering engineered HIV-specific lymphocytes unable to recognize and clear persistently infected cells. We have generated an exciting set of tools and preliminary data that directly addresses each of these barriers. To overcome the transient nature of bNAbs and associated immunogenicity of vectored delivery approaches, we have performed an in vivo screen in nonhuman primates (NHP) and identified engineered AAV variants that persist long term (consistent with a lack of recognition by the host immune system), and specifically target B cells. B cell tropic vectors will be packaged with CRISPR-Cas9 gene editing machinery, applying highly innovative covalent linkage methodology to double our vectors’ packaging capacity. We refer to our novel in vivo delivery approach as Non-Immunogenic, Cargo-Enhanced (NICE) AAV: in a single dose, NICE- AAV vectors will specifically reprogram B cells with bNAb or eCD4-Ig sequences targeted to the native IgG locus. Finally, we will overcome the significant problem of insufficient viral antigen by supplying cell-associated HIV-1 Env in trans. Our recent publication in the NHP model demonstrates the immense success of this strategy to stimulate HIV-1-specific chimeric antigen receptor (CAR) T cells and should similarly boost and trigger expansion of our gene-edited B cells. The central goals of our proposal are to validate the efficiency and specificity of B cell-targeted NICE-AAV (AIM 1), to demonstrate that this in vivo delivery approach enables persistent bNAb/eCD4-Ig expression in HIV anatomical compartments and reservoir sites (AIM 2), and most importantly, to achieve a therapeutic impact in humanized mouse and NHP models of HIV persistence (AIM 3). We will merge one of the most promising therapeutic modalities for HIV cure (bNAbs/eCD4-Ig) with our extremely unique in vivo delivery platform (NICE-AAV). Importantly, this approach will be applicable not only for HIV-1, but for the broad range of pathologies where monoclonal antibody therapies offer clinical benefit.

NIH Research Projects · FY 2026 · 2021-12

PROJECT SUMMARY/ABSTRACT Bronchiolitis obliterans syndrome (BOS) is the most severe manifestation of chronic graft-versus-host disease (cGVHD) in survivors of allogeneic hematopoietic cell transplant (alloHCT), leading to irreversible pulmonary impairment, poor quality of life, and 5-year survival of 40%. Fundamental gaps in knowledge of the pathogenic events that contribute to progressive lung dysfunction in BOS have not been well characterized, hampering our ability to intervene effectively. Our preliminary data suggest that respiratory viruses, including respiratory syncytial virus (RSV), parainfluenza (PIV), human metapneumovirus (HMPV), and influenza (FLU), are independent risk factors for the development of BOS. Additionally, we show that asymptomatic respiratory viral infections (RVI) are common posttransplant. We have shown that mobile wireless home spirometry is feasible in patients with cGVHD and can enable early diagnosis and a granular understanding of the trajectory of lung function decline. Our overarching hypothesis is that cumulative respiratory viral exposure leads to the development of BOS and poor outcomes in the context of alloimmunity. The overall aim of this proposal is to establish the temporal relationship between RVI along the continuum of disease presentations, from asymptomatic to symptomatic upper respiratory tract to lower tract disease, and the lung function trajectory of BOS. We propose to conduct a multicenter prospective longitudinal study of the natural history of RVI and lung function with an innovative home monitoring approach that overcomes the barriers to understanding clinical events that lead to BOS and severe BOS phenotypes. Aim 1 investigates the role of RVI as triggers BOS. We will enroll alloHCT recipients at risk for BOS (Cohort 1, n=200), including those with a diagnosis of cGVHD or a history of high-risk RVI (RSV/PIV/HMPV/Flu/SARS-CoV2). Patient will perform weekly home spirometry and protocolized surveillance and symptom-prompted self-collected nasal swab viral PCR. In addition, serum will be collected quarterly via a needle-less home blood collection kit and assayed with VirScan, a novel comprehensive serosurvey that detects epitopes of >1000 virus strains, in order to assess the impact of cumulative respiratory viral burden on BOS outcomes. Aim 2 examines the role of RVI on pulmonary exacerbations in BOS, as well as the association of cumulative RVI exposure (as determined by VirScan) on accelerated FEV1 decline in patients with a severe BOS phenotype. Patients with a clinical diagnosis of BOS (Cohort 2, n=80), will perform the same procedures as Cohort 1. For both aims, viral PCR and VirsScan results will be compared and analyzed as predictors for BOS development or accelerated FEV1 decline. The critical data generated by this study will improve recognition of early BOS in the context of RVI, risk stratify patients at highest risk for intensive monitoring, and identify tangible endpoints and biologic rationale for testing early interventions and novel therapies. Importantly, this proposal will also establish a unique adult and pediatric multicenter Consortium with the specific goal of addressing lung disease in HCT recipients, an area of significant and urgent unmet need.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY/ABSTRACT Advanced age is the single most significant risk factor for cancer. Although there are many plausible age-related mechanisms that mediate the increased risk for cancer in the elderly, functional evidence is lacking for the majority of these mechanisms. This proposal will focus on the role of the aged microenvironment, particularly senescence, in promoting the formation of colorectal cancer (CRC), which is a common age-related cancer in the U.S. It is well established that the majority of CRCs arise through the serial accumulation of mutations and epigenetic alterations in oncogenes and tumor suppressor genes in colon epithelial stem cells. However, our group and others have found cells with cancer-causing DNA alterations (e.g. oncogenic KRAS and TP53) in the histologically normal colon mucosa of people, which suggests that the DNA alterations alone are not sufficient to drive the complete CRC formation process. Our group has recently shown that a senescent normal colon tissue microenvironment can promote the neoplastic behavior of colon epithelial cells ex vivo. We found increased senescent fibroblasts in the colons of the elderly and of people at high risk for CRC. We found that certain senescence associated secretory phenotype (SASP) factors produced by senescent fibroblasts can induce cancer hallmark behaviors through the activation of oncogenic signaling pathways. Based on our preliminary data we hypothesize that senescent fibroblasts in older people and the accompanying specific Senescence Associated Secretory Phenotype (SASP) secreted proteins promote the tumorigenesis of colon epithelial cells that have acquired age-related oncogenic mutations, which we have defined as cancer-initiating cells. To provide direct functional evidence to test this hypothesis, we propose the following Specific Aims: AIM 1A. To determine whether senescent fibroblasts can promote the survival and/or transformation of colon cancer initiating cells in ex vivo and in vivo organoid model systems. AIM 1B. To determine the necessary and sufficient role of specific cancer-associated SASP factors produced by the senescent fibroblasts in cancer formation. AIM 2. To determine whether a senescent tissue microenvironment is necessary for the survival and clonal expansion of mutant APC and mutant KRAS cancer-initiating cells using a well characterized CRC mouse model, the inducible Villin-Cre;Apcflx/flx; KrasLSL-G12D (AK) mice, with two approaches: AIM 2A: a pharmacological approach using the senomorphic drug rapamycin and the senolytic drug treatment DNQ (dasatinib and quercetin) AIM 2B: a genetic approach using the p16-3MR transgenic model, which allows the inducible ablation of senescent cells.

NIH Research Projects · FY 2025 · 2021-09

Abstract My lab has been the leader in the field of mouse modeling for brain tumors over the past 15 years. We have developed a suite of genetically engineered mouse models that are demonstrably representative of human gliomas and other tumor types. These models have been used to inform treatment options for clinical agents, and this now enables us to propose these models are suitable testbeds for testing potential major improvements to how these diseases are treated. We have three projects. 1) We are understanding the central role of specific splice variants of TrkB in embryonic development and oncogenesis throughout the body. The RCAS modeling system has been used here to show that forced expression of the embryonic splice variant in adult tissues leads to cancer formation broadly. In this project we will investigate the mechanisms of oncogenesis for this splice variant and determine if it could be a good diagnostic or therapeutic target. 2) We are now using the modeling system developed for glioma to address the biology of rare tumors driven by gene fusions. In this grant we propose to understand the mechanisms of oncogenesis for YAP1 gene fusions in the rare tumors ependymoma, porocarcinoma and aggressive meningioma (all for which we have YAP1 gene fusion driven models currently). 3) And, we will use these mouse models to study therapeutic response and identify therapeutic strategies for these fusion driven tumors including identification of FDA approved drugs that would intervene downstream of the action of the gene fusion.

NIH Research Projects · FY 2025 · 2021-09

PROJECT SUMMARY / ABSTRACT Children and adolescents diagnosed with cancer now have, on average, nearly 85% 5-year survival. However, premature cardiovascular (CV) disease has become the leading non-cancer cause of late mortality among childhood cancer survivors. There is a robust body of evidence from the general population, and increasingly, among cancer survivors (even those exposed to cardiotoxic cancer therapies), that greater physical activity (PA) and improved diet quality can reduce future CV-related morbidity. However, while many general population and cancer-specific intervention studies have focused on a single lifestyle factor (e.g., PA or diet alone), given the interplay between PA and dietary factors in influencing CV health, a multi-faceted approach may result in overall better long-term CV health profiles. Research on lifestyle interventions in cancer survivors also has been predominantly conducted in women with breast cancer, and the evidence for survivors of childhood cancer is limited. To accomplish our aims, we will use the largest prospectively followed childhood cancer survivor cohort in the world, the Childhood Cancer Survivor Study (CCSS; n>24,000), to recruit adult-aged participants at increased risk of early CV disease (n=403) for a remotely conducted 12-month randomized controlled trial testing a multi-faceted approach at improving PA and diet quality. Specifically, the study will use a sequential multiple assignment randomized trial (SMART) design, where participants with low PA or poor diet will first be randomized between intervention and control conditions. Intervention participants will be further randomized to receive either clinician-led telehealth sessions focused on risk factor self-management, or weekly mobile health (mHealth) supported individualized PA and dietary goal-setting with social media peer support. The adaptive SMART design will allow further tailoring of the intervention experience based on initial response, which may increase overall intervention efficacy. Participants not initially responsive to their assigned intervention will be further randomized to receive an alternate intervention. The study will use consumer-grade mHealth applications that track PA and diet, thereby increasing future dissemination capacity. The study’s primary analyses will determine the overall intervention efficacy and whether specific intervention strategies and sequence of strategies are associated with optimal outcomes. Secondary analyses will examine potential predictors, mediators, and moderating factors associated with PA and dietary changes over time, as well as changes in participants’ cardiometabolic profiles. In summary, lifestyle change represents one of the few available strategies to mitigate CV risk in childhood cancer survivors. Significant barriers (e.g., time, training, resources) limit the ability of healthcare systems to facilitate such change. To fill this void, remote-based, personalized, and easily disseminated multi-faceted mHealth-supported interventions may play a transformative role.