Roswell Park Cancer Institute Corp

NIH Research Projects · FY 2026 · 2026-06

Project Summary Advanced EGFR mutant lung adenocarcinoma (LUAD) is treated with EGFR tyrosine kinase inhibitors (TKIs). While effective, virtually all patients progress on therapy. Mechanisms known to cause acquired EGFR TKI resistance include secondary EGFR mutations, activation of bypass signaling pathways, or histological transformation to lineage variants no longer dependent on EGFR signaling. Mechanisms of acquired EGFR TKI resistance have been identified in only about half of the cases examined, however, despite extensive DNA sequencing efforts. Transcriptional adaptations enabling LUAD cells to survive and proliferate during therapy are emerging as a prerequisite for many forms of acquired EGFR TKI resistance. The overarching hypothesis addressed in this grant proposal is that reversible and dynamic transcriptional adaptations accelerate acquired EGFR TKI resistance in LUAD, and blocking this transcriptional plasticity will improve EGFR TKI treatment outcomes. While transcriptional adaptations have been defined in experimental models and clinical samples, the pathways and mechanisms regulating transcriptional plasticity itself remain largely unknown. This is a significant gap because transcriptional plasticity controls how fast and how well LUAD cells adapt to EGFR TKI treatment. The primary goal of this application is to fill this gap. Aim 1 tests the novel concept that loss of the RB1 tumor suppressor gene (Rb) accelerates acquired EGFR TKI resistance by driving transcriptional plasticity. Rb loss is suggested to de-repress retroelements inducing chronic, cancer cell intrinsic inflammatory signaling. Inflammatory signaling then drives plasticity and EGFR TKI resistance. Aim 2 tests the innovative idea that initial adaptation to EGFR TKI is driven by YAP/TEAD/β-catenin mediated reprogramming of LUAD cells to a quiescent, alveolar transcriptional state. This alveolar state is extinguished in favor of other transcriptional adaptations as cells progress through therapy. While parallel and distinct, the aims are complementary and interconnected. Both aims address the role of Rb in altering the trajectory of LUAD transcriptional adaptations during EGFR TKI therapy. Both aims interrogate the impact of specific signaling pathways on transcriptional plasticity and EGFR TKI responses. Each aim employs cutting edge experimental models unique to this research team, including genetically engineered mouse models. Use of these animal models is required to experimentally assess the complex interactions between cancer cells and normal cells within a physiological tumor growth environment that can impact acquired therapeutic resistance and cancer progression. Each aim is led by a different MPI with complementary expertise. Collaboration between aims creates synergies by sharing experimental models, technical expertise, data, and patient specimens. Successful completion of the work will advance fundamental understanding of LUAD phenotypic plasticity and acquired EGFR TKI resistance while identifying targets for future therapeutic intervention.

NIH Research Projects · FY 2026 · 2026-06

High-grade serous ovarian carcinoma (HGSOC) is a quintessential example of a highly fatal malignancy that has not greatly benefited from therapeutic or screening advances in oncology. As symptoms are vague, 75% of patients are diagnosed at late stage, and most will recur with chemo-resistant disease within three years. Although it is well recognized that tumor immunity plays a critical role in the pathogenesis of HGSOC, nearly 85% of patients don’t respond to immunotherapy and comprehensive interrogation of the HGSOC tumor immune microenvironment (TIME) remains preliminary, wherein optimal patterns of immune infiltration and spatial relationships have not been well defined. At present, clinical characteristics (age at diagnosis, tumor stage, presence of ascites, and debulking status) are the only established prognostic factors and little is known about the impacts of modifiable lifestyle exposures, and related immune biomarkers, in improving HGSOC survival. Despite consistent recommendations from leading organizations to adhere to healthy lifestyles during cancer treatment and survivorship, which lifestyle factors, and whether they work together to improve treatment and survival outcomes remains a fundamental unanswered question. Moreover, whether lifestyle interventions during active therapy are associated with improved treatment and survival outcomes remains unknown. To this end, an emerging body of pre-clinical evidence from a variety of animal models suggests the TIME is highly plastic and modulated by unhealthy host exposures including obesity and inactivity via metabolic and immune dysregulation, while voluntary wheel running enhances anti-tumor immunity, increases infiltration of cytotoxic immune cells and mitigates entry of immune-suppressive cells into the tumor. Although compelling, our current understanding of how obesity and physical activity (PA) impact immunity in patients is mostly limited to circulating markers, which may not reflect the TIME, the most relevant site for tumor progression and prognosis. Herein, we contend that optimal body composition and regular PA in the peri-diagnosis period are associated with improved survival in advanced-stage HGSOC, and that these associations are mediated by enhanced immunity in the TIME. To test this hypothesis, we will use the high throughput PhenoCycler platform to assess 15 immune subsets in FFPE tumor sections from 500 patients diagnosed with advanced-stage HGSOC from Roswell Park and Moffitt Comprehensive Cancer Centers for linkage with lifestyle, epidemiological and clinical data. Leveraging multi-scale spatial analyses and formal mediation pathway analysis we propose the following three Specific Aims: 1) Define associations of body composition and physical activity in the peri-diagnosis period with immune cell abundance and spatial distributions (the immune contexture) in the HGSOC TIME; 2) Identify associations of immune cell abundance and spatial distributions in the TIME with HGSOC survival; and 3) Delineate the associations of body composition and physical activity with improved HGSOC survival into direct and indirect (mediating) pathways relayed through the tumor immune contexture.

NIH Research Projects · FY 2026 · 2026-05

Abstract This proposal addresses PAR-24-306 (Research Projects to Enhance Applicability of Mammalian Models for Translational Research) by developing and credentialling NeuroEndocrine Bladder Carcinoma (NEBC) models for cross-species comparison and evaluating innovative treatment approaches. Most bladder cancer (BC) are conventional urothelial carcinomas (UC) that originate from urothelial cells, the lining of the bladder. Histology variants/subtypes can be found in up to 30% of bladder cancer patients and often co-exist with UC tumors. NEBC significantly contributes to BC death because NEBC is incurable and often relapses after the standard etoposide and cisplatin (EP) chemotherapy. A clear unmet need is to understand the different pathogenesis of NEBC tumors and develop novel treatment strategies for overcoming chemotherapy resistance. However, no studies have systemically investigated NEBC, due to a lack of relevant genetically engineered mouse models (GEMMs) and patient derived organoid/allograft (PDO/PDX). Genomic studies of NEBC tumors have revealed 3 common altered pathways: TP53/RB1/PTEN(PIK3CA). Inactivating mutations of TP53 RB1, and PTEN are more enriched in NEBC tumors (n=47, TP53 83%, RB1 79%, PTEN 13%) than in muscle invasive BC (MIBC) tumors. Therefore, we developed two complementary triple knockout (TKO: Trp53-/-; Pten-/-; Rb1-/-) NEBC GEMMs. These include the UP2-Cre;TKO model, driven by the bladder-specific Uroplakin 2 promoter (UP2-Cre), and the Ad-Cre;TKO model, by intravesical injection of adenovirus-Cre (Ad-Cre) driven by a CMV promoter. The UP2-Cre;TKO model developed pure NEBC tumors, while the Ad-Cre;TKO model developed mixed NEBC/UC tumors. These mouse NEBC (mNEBC) tumors exhibited resistance to EP chemotherapy and a marked activation of the ATM pathway following EP treatment. Additionally, we established PDOs/PDXs from NEBC patients who exhibited resistance to EP chemotherapy. Based on these NEBC GEMMs and PDOs/PDXs, we are ideally positioned to test the hypothesis that NEBC GEMMs faithfully recapitulate human NEBC (hNEBC) biology, particularly in the molecular mechanism and their therapeutic resistance to EP chemotherapy. Our hypotheses will be tested in two specific aims: 1) Aim 1: Develop and credential novel syngeneic NEBC GEMMs and mixed NEBC/UC GEMMs. 2) Determine whether mNEBC GEMMs replicate hNEBC treatment responses and assess the efficacy of ATM inhibitors (ATMi) in enhancing EP chemotherapy responses. This proposal leverages our team expertise in the clinical management of BC and NEBC, as well as the GEMMs/PDXs pathological and bioinformatic characterization, and bladder cancer therapeutics. Our approach is innovative, utilizing novel immune-competent NEBC GEMMs, human NEBC PDXs, and incorporating advanced technologies such as single-cell ATAC/RNA- seq and CHIP-seq. This research is significant as it seeks to develop innovative treatment strategies to overcome chemotherapy resistance.

NIH Research Projects · FY 2026 · 2025-12

PROJECT ABSTRACT/SUMMARY (From the Parent Grant) Invasion is one of the most detrimental features of all cancers, including breast cancer, as it allows cells to escape the primary site and form metastases at distant organs. Despite progress in prevention and early lesions detection, the mortality associated with metastatic breast cancer is still extremely high. This is especially true for patients presenting with triple negative breast cancer (TNBC, characterized by lack of expression of ER, PR, and Her2), which is the most aggressive and deadliest subtype of breast cancer and the one that so far lack specific targets for therapeutic intervention. Understanding the mechanisms that facilitate the invasion of tumor cells will enable us to design more efficient therapeutic strategies to prevent or reduce metastasis. Our group has established a fundamental connection between GTP metabolism and tumor cell invasiveness; we have unveiled GTP and its metabolic enzymes (GME) as key players in tumor progression and metastatic potential. We have developed unique fluorescent reporters for intracellular GTP that have allowed us to determine that, in live cells, the intracellular GTP distribution is not uniform, and brought forward the hypothesis that local concentration of GTP can influence GTP-dependent processes. In particular, we have previously shown that genetic or pharmacological modulation of the GTP metabolic pathway deeply affected the activation status of small GTPases of the RHO-family and, with it, the tumor cells' invasive capability. Thus, in Aim 1 we will explore a novel mechanism of G-proteins activation based on GME subcellular localization. Our preliminary results showed that the rate-limiting enzyme for GTP de novo production, inositol monophosphate dehydrogenase 2 (IMPDH2) enriches at cell membrane sites that are critical for cell migration and invasion (namely focal adhesion, FA, and invadopodia). The role of IMPDH2 at these sites is virtually uncharacterized. Thus, in Aim 2 we will assess the catalytic and structural role of IMPDH2 in FA and invadopodia formation, as well as in focal adhesion kinase (FAK)-directed oncogenic motility. The understanding of GTP metabolic enzymes transcriptional regulation is far from complete. Identification of transcriptional master regulators of the GTP biosynthetic pathway that could be pharmacologically targeted would offer a more efficient way of suppressing this pathway. Our preliminary results suggest that Kruppel-like factor 9 (KLF9) and aryl hydrocarbon receptor (AHR) play antagonistic roles in the transcriptional regulation of GTP metabolic enzymes, with KLF9 suppressing, whereas AHR inducing GTP production. Thus, in Aim 3 we will elucidate this regulation and explore pharmacological treatments to regulate the activity of these transcription factors.

NIH Research Projects · FY 2025 · 2025-09

ABSTRACT Tobacco product brand/line extensions continue to proliferate as manufacturers seek to maintain or grow their market share. The Tobacco Control Act (TCA) introduced a number of potential pathways for marketing tobacco products, including a substantial equivalence (SE) process. Typically, SE determinations are made by comparing the “new” product to a pre-existing product in terms of design and emissions, with an eye toward whether the new product raises different questions of public health than what was previously on the market. The SE process raises important questions and concerns as to its impacts on the market and on public health. This understudied area of regulatory science could be leveraged by the tobacco industry to retain or attract consumers while avoiding the stricter regulatory standard applied to new products. Under the SE regulatory model in its present form, cigarettes and cigars can continue to evolve through accumulation of small changes and proliferation of line extensions that bear functional similarity to existing products. Notably, human studies are not explicitly required for SE determinations, which raises concerns as to whether a product that receives an SE Order on technical grounds is experienced in the same way as the predicate by consumers. Our Specific Aims are to evaluate the state of the market with respect to SE determinations and assess consumer product differentiation in two combustible tobacco product classes – cigarettes and cigars. These are the products of focus as they are clearly the most associated with health risks, and both of these markets were very well established prior to the TCA. In Aim 1 we will characterize the marketplace, leveraging Nielsen data and laboratory techniques to assess the design, contents, and emissions of 15 leading subbrands of cigarettes and cigars. In Aim 2 we will assess consumer behaviors with respect to substantial equivalence. In Study 2.1 we will leverage PATH data, which contains self-reported brands and sub-brands, to examine trends in the market share of brands. In Study 2.2, we will recruit a panel of adults (21+) who currently use one of the 15 leading subbrands of cigarettes and cigars as identified in Aim 1. The goal of this study is to examine whether consumers using these products can identify changes to their brand during the course of multiple year follow-up. In Aim 3, we assess industry behaviors with respect to substantial equivalence by compiling and integrating SE determinations and other orders posted on FDA’s website to examine trends in product characteristics and predicate product usage. A key goal is to determine if there are commonalities to what aspects of product design or emissions are changing, or if there are common predicate products employed. This proposal addresses the Product Composition and Design, Behavior, and Impact Analysis priority research domains.

NIH Research Projects · FY 2025 · 2025-09

This grant aims to support the career development of Lori Kern, a central developer of several facets of the NIH/NHGRI/NCI funded Bioconductor project: an open-source, open-development software project that develops, supports, and distributes software for genetic and genomic research, analysis, discovery and visualization. It will invest in the Research Software Engineer’s identified project goals to enhance and improve development and dissemination of Bioconductor. These goals include: Goal 1: Upgraded support for global computational genomic data science. This involves enhancing Bioconductor’s process for curating, documenting, and distributing Findable, Accessible, Interoperable and Reusable for Research Software (FAIR4RS) data and annotation for use in computational genomic data science. Experiment data, genomic reference information, and analytic algorithms are combined to produce statistical interpretations of novel experimental findings. Bioconductor strives to provide high quality data and up-to-date genomic annotations either through Bioconductor Hubs that reference remotely available data for download or traditional Bioconductor R packages. There are currently 71787 annotation resources from 2994 species available for download from the current AnnotationHub, 8332 Experiment data resources available in ExperimentHub, 926 standard Annotation Packages, and 430 standard Experiment Packages available in Bioconductor. Refinement and evaluation of currently available resources to ensure accuracy and relevance, and providing easier direct access in Bioconductor/R to other existing relevant genomic reference information, is essential for researchers to achieve novel experimental findings. Goal 2: Personal skill development that will enhance the sustainability of Bioconductor, with particular attention to concepts of cloud-scale research software, data, and genomic annotation production, management, and distribution as well as integration and collaboration with other relevant biological and bioinformatic communities. Goal 3: Project and Community leadership strengthening. Bioconductor leverages its extensive community to contribute and propel the project forward. With over 1192 active community developers, wrangling a highly technical and evolving ecosystem becomes vital. A balance of integrating cutting-edge, new technology and methodology with reliability and efficiency becomes essential to Bioconductor sustainability and relevance. Enhancements to the contribution process and learning opportunities for the community will help reach and connect the thousands of developers and millions of users. The outcomes will strengthen the well-established impact of Bioconductor in genome research in academia and industry.

NIH Research Projects · FY 2025 · 2025-09

This study addresses the unmet clinical needs for management of non-muscle invasive bladder cancer (NMIBC). NMIBC represents ~75% of bladder cancer cases and has a favorable five-year survival rate, but typically recurs (50-70% in 5 years) and progresses to muscle-invasive disease (MIBC, 10-30%). Intravesical Bacillus Calmette-Guerin (BCG) is the most effective therapy to prevent NMIBC recurrence and progression, yet BCG has a 30% failure rate, with various adverse effects leading to intolerance. Two critical needs exist: 1) a risk stratification tool to identify patients at high risk of recurrence and progression for early and aggressive treatment, and 2) a response prediction tool to identify patients who are unresponsive or intolerant to BCG for other treatment options. Our goal is to develop and validate risk-stratification and BCG-response tools by incorporating molecular signatures into the current pathological system for optimal clinical care of NMIBC. Transcriptome analysis is powerful to identify genes, and importantly, to define cancer molecular subtypes associated with different therapeutic and prognostic outcomes. This approach has been applied in bladder cancer but primarily focused on MIBC. Research on NMIBC is an unmet need. Building on the Bladder Cancer Epidemiology, Wellness, and Lifestyle Study (Be-Well), one of the largest prospective cohorts of NMIBC patients, we propose to conduct a comprehensive transcriptomic analysis of NMIBC and examine the utility of molecular subtypes with additional prognostic genes in predicting NMIBC treatment response and prognostic outcomes. Our central hypothesis is that molecular signatures (i.e., molecular subtypes and/or other genes) could unveil NMIBC heterogeneity and thus improve the current pathological classification system for tailored NMIBC care. We will: Aim 1. Develop a risk stratification tool for NMIBC prognostic outcomes by incorporating molecular subtypes, genes, clinicopathological and demographic factors. Primary outcomes will be disease recurrence and progression, with survival explored. We will define molecular subtypes and identify genes by NMIBC prognostic outcomes (1a), build risk prediction models in Be-Well (n=928) (1b), validate molecular signatures and the risk-stratification tool in an independent validation cohort (n=959) (1c), and explore the tool in sex and race specific groups in the pooled cohorts (1d). Aim 2. Develop a response prediction tool for BCG outcomes by incorporating molecular subtypes, immune signatures, genes, clinicopathological and demographic factors. Primary outcomes will be BCG unresponsive with BCG intolerant explored. Given the high immunogenic nature of bladder cancer and BCG therapy, we will develop the BCG- response prediction tool with further consideration of immune signatures in patients who received BCG in Be- Well (n=426) and a validation cohort (n=691). We will explore characterization of molecular subtypes by sex, race/ethnicity, and etiological risk factors. Clinical translation will be accelerated given the generation of NMIBC-specific molecular signatures using NanoString in a large health care delivery system setting.

NIH Research Projects · FY 2025 · 2025-09

Breast cancer remains the most commonly diagnosed malignancy and the second most common cause of cancer death in women, mostly from disease of aggressive features like estrogen receptor-negative and triple-negative status. The incidence of aggressive breast cancer has been linked to a prolonged pro-inflammatory immune response and dysfunction in the tumor microenvironment (TME). We hypothesize that genetic selection and adaptation to distinct local pathogenic environments during the evolution of different human populations from thousands of years ago have led to genetic differences in host immunity, which contribute to breast cancer disparities. This hypothesis is supported by previous studies that showed genetic estimates of African ancestry are strong predictors of systemic inflammatory response. Moreover, studies from our group have shown that the West African ancestry-linked Duffy-null allele, which confers malaria resistance, has a strong influence on circulating plasma chemokine levels. While these studies provide insight into systemic immunity, the ancestral and genetic determinants of immune phenotypes in the TME are largely unknown. Recent genome-wide association studies (GWAS) have shown that the immune response in the TME is shaped significantly by germline genetics; however, all these GWAS were performed in populations with lower genetic estimates of African ancestry. Thus, it is timely to conduct research to examine immunogenomic factors that shape the TME in patients of different genetic ancestries to better understand the causes of breast cancer disparities. We propose to leverage a rich body of genetic, pathological, and transcriptomic data from > 5,000 women with breast cancer in the context of three large studies. We will 1) perform the first local ancestry-informed GWAS of pathological tumor-infiltrating lymphocytes (TILs) in a group of breast cancer patients. Moreover, we will investigate the role of selection and adaptation in the presence and abundance of TILs by calculating population branch statistics and integrated haplotype scores for significantly associated SNPs. Moreover, we will 2) examine the role of germline genetics and ancestry in the inflammatory state of the TME through a cross-ancestry comparison of inferred immune cell composition, immune gene expression, and quantitative trait loci (QTL) with tumor RNA-seq and matched genotype data from > 1300 women. The results of this proposal will practically be used to decrease disparities in the following ways: 1) Generate the largest publicly available local ancestry-informed expression-QTL/splicing-QTL information for breast cancer tissue for the field. 2) Identify variation in immune response across ancestries that may function as targets for novel immunotherapies or vaccines. Our findings will advance our understanding of the multifaceted mechanisms of cancer health disparities from a novel perspective of genetic ancestry and tumor immunity and may inform clinical investigations of immunotherapy for breast cancer in all women.

NIH Research Projects · FY 2025 · 2025-09

Project Summary/Abstract The goal of our proposed Data and Resource Coordinating Center (DRCC) application is to catalyze collaboration across the Cancer Immunoprevention Network (CIP-Net) and to manage, integrate and disseminate the data and resources generated through the network. By leveraging cutting-edge, multidisciplinary team approaches, we will support CIP-Net in fostering a collaborative community of cancer immunoprevention researchers, collectively working to understand the basic mechanisms of immunoprevention, discover novel immunoprevention strategies, and develop and test preclinical interventions. Our strategy is to boost the productivity of CIP-Net investigators by fostering a collaborative research community, accelerate CIP-Net research by removing barriers to accessing analytics expertise, improve the interoperability and usability of CIP- Net data by deploying best practices for data standards and harmonization, and unlock the full potential of CIP- Net activities by sharing data and resources with the broader scientific community. First, we will establish an effective administrative infrastructure and organizational process to coordinate CIP-Net activities, building on our proven and continuously improving systems that currently support the coordination of NCI programs. Second, we will ensure that CIP-Net data are harmonized using standards interoperable with the broader cancer data ecosystem and findable through a centralized virtual repository, leveraging our extensive track record and robust systems for collaborative data sharing and dissemination in consortium studies. Third, we will provide multidisciplinary analytics expertise to support CIP-Net’s collaborative research, drawing on our collective strengths in biostatistics, bioinformatics, and biomedical informatics from Roswell Park’s Department of Biostatistics & Bioinformatics. Fourth, we will actively promote CIP-Net and engage diverse stakeholders to build an inclusive community that leverages a range of perspectives and approaches to address key scientific questions in immunoprevention. Taken together, we envision that our DRCC, in close collaboration with the CIP-Net stakeholders, will enable us to support highly effective and innovative translational studies that address the critical unmet needs in cancer immunoprevention. The main deliverables include: administrative and organizational support to coordinate network activities and facilitate communication and collaboration (Aim 1); policies, workflows, and infrastructure to ensure that all resources generated by CIP-Net are harmonized using standards interoperable with the broader cancer data ecosystem and findable through a centralized virtual repository; multidisciplinary analytics support to accelerate CIP-Net research progress, along with innovative solutions to address CIP-Net’s evolving needs (Aim 2); and community-building and outreach efforts to support immunoprevention research, integrate patient advocates into CIP-Net, and coordinate career development opportunities for early-career scientists (Aim 3).

NIH Research Projects · FY 2025 · 2025-09

Project Summary Innovative adoptive cellular therapies have emerged as revolutionary treatment options for patients with hematologic malignancies. One such promising strategy, Chimeric Antigen Receptor (CAR) T cell therapy, involves genetic modification of autologous T cells to express a CAR, redirecting the cytotoxic response of T cells towards hematologic tumors expressing human B cell maturation antigen (hBCMA) on multiple myeloma or CD19 on B cell lymphomas and leukemias. While response rates to CAR T cell therapy are strikingly high in patients with hematologic malignancies, most patients experience disease relapse, highlighting the urgent need to improve CAR T cell efficacy for a durable response. Preliminary data from our lab and published literature indicate that targeting endogenous CD28 on CAR T cells is a viable approach to enhance CAR T cell efficacy. The CAR contains a costimulatory domain (such as CD28), which undergoes activation upon ligation of the target antigen to the CAR, delivering costimulatory signals to contribute to CAR T cell activation, leading to subsequent CAR T cell effector functions. Importantly, as the CAR is integrated into T cells, T cells also express natural endogenous CD28, which can engage CD28 ligands in the tumor microenvironment or on other T cells themselves, providing yet another source of costimulatory signals that contribute to CAR T cell activation. Interestingly preliminary data from our lab show that deletion of endogenous CD28 enhances the functional persistence of CAR T cells in vivo. As such, we hypothesize that endogenous CD28 contributes to the overstimulation of CAR T cells which increases exhaustion, decreases persistence, and hampers efficacy. To test our hypothesis, will rigorously evaluate how knockout of endogenous CD28 using CRISPR/Cas9 technology affects CAR T cell function and efficacy using established preclinical in vitro and in vivo models. These studies have the potential to identify a novel role of endogenous CD28 in hindering CAR T cell efficacy, while highlighting a clinically relevant strategy to improve CAR T cell efficacy.

NIH Research Projects · FY 2025 · 2025-08

Summary Multiple Myeloma (MM) is the second most common hematologic malignancy and is considered incurable for most patients. There has been significant improvement in MM patient survival, mainly due to the use of novel treatments including immunomodulatory drugs (IMiDs), proteasome inhibitors (PIs), monoclonal antibodies (mabs), bispecific antibodies and cellular therapies. However, many of these treatments heavily rely on a healthy patient immune system to fully leverage their efficacy. Unfortunately, prior studies have shown that the phenotype and function of immune cells in the BM tumor microenvironment (TME) in MM is dysregulated. The accumulation of pathological myeloid cells has emerged as a major mechanism by which tumors evade anti- tumor immunity and are a primary obstacle to develop efficient cancer immunotherapies. To characterize the phenotype and transcriptional changes in myeloid cells in tumor, we used single-cell transcriptomics to dissect the differences between the myeloid cell populations in the BM and osteolytic lesions (OL) TME of MM patients (N = 13) and compared them with myeloid ME of BM of healthy controls (HDBM) (N = 3). OL are an aggressive manifestation of MM and can lead to pathologic fractures. We discovered a significant accumulation of CXCR2 positive myeloid cells in BM and OL of MM patients but not HDBM. Using murine preclinical models of MM, we found that targeting CXCR2 positive cells using anti-CXCR2 drugs alone or in combination with standard treatment (bortezomib plus dexamethasone) significantly increased the overall survival in tumor bearing mice. CXCR2 inhibitors are safe, effective, and commercially available (the CXCR1/CXCR2 inhibitor (SX-682) used in this trial is provided by Syntrix Pharmaceuticals). The goal of the clinical trial from which the samples for this correlative study are obtained is to explore safety and tolerability of CXCR2 blockade as a novel, yet practical approach that effectively mitigates myeloid cell dysregulation in MM patients to boost the response to standard treatments in a phase I clinical trial with two specific aims: Aim1) Evaluate the effect of SX-682 on the spatial architecture of the TME in MM on bone marrow samples at baseline and after 6 cycles of treatment. Aim 2) Evaluate the immunomodulatory effect of SX-682 on the BM TME and peripheral blood (PB).

NIH Research Projects · FY 2026 · 2025-04

Breast cancer (BCa) metastases are associated with extremely high mortality; hence, the development of effective prevention and treatment strategies for metastatic breast cancer is critical. Bone is typically the first and most frequent site of breast cancer metastatic expansion, and the resulting metastases can cause severe pain, immobility, fractures, and nerve damage. Additionally, our recent studies underscore the important role bone metastases can play in spreading cancer cells to other organs, thus making the bone metastatic process a very attractive target for the prevention and treatment of metastatic breast cancer. Presently, current treatments for bone metastases, such as bisphosphonates, have shown limited success, largely because these drugs predominantly target bone turnover and remodeling instead of directly eradicating cancer cells in the bone environment. Moreover, tumor cells can at times survive adjuvant therapies given to patients after surgery and eventually cause late-onset relapse in bone. It is therefore imperative to establish new therapeutic strategies to improve the outcome of patients with breast cancer bone metastases. Our preliminary studies are helping to unravel the transition of biological behaviors from the early phase of bone metastases (termed microscopic-bone metastasis or micro-BoM) to established BoM (termed macro-BoM). At the micro-BoM stage, the colonized metastatic tumor cells grow slowly and undergo amino acid and lipid metabolism to collect and accumulate necessary resources to sustain their survival. Our mechanistic studies have suggested that in the micro-BoM stage, the transcriptional coactivators YAP/TAZ are regulated by a bone- specific calcium-signaling-related mechanism, and YAP/TAZ activation orchestrates a repertoire of metabolic gene expression. Furthermore, we have developed delicate strategies that allow us to treat the microscopic bone lesions. The overall objectives of this proposal are to understand the underlying molecular mechanisms by which YAP/TAZ activation drives the advancing of BCa BoM. Our central hypothesize is that YAP/TAZ orchestrates an evolution of metabolic behaviors in micro-BoM and targeting of YAP/TAZ could potentially impede cellular adaptation processes and arrest the progression of micro-BoM. Our long-term goals are to decipher the crosstalk between YAP/TAZ activation in bone-colonized tumor cells and the bone microenvironment and uncover new therapeutic targets to treat both the microscopic and fully established BoM. To achieve these goals, we propose the following two aims: 1) determine the molecular mechanisms driving the dynamical activation of YAP/TAZ in BCa BoM; 2) determine the functional role of YAP/TAZ-activation-driven metabolic reprograming in BCa micro- BoM. A major impact of this study will be the identification of previously undescribed mechanisms underlying the adaptation and progression of microscopic BoM that can inform the development of new biomarkers and novel therapeutic interventions for advanced or metastatic BCa.

NIH Research Projects · FY 2026 · 2025-01

Bladder cancer is one of the top 10 cancers in the nation, and the 4th most common cancer in men. The majority (70-80%) of bladder cancer is diagnosed at early stages, known as non-muscle invasive bladder cancer (NMIBC). NMIBC can be removed, yet typically recurs (up to 75%), and some progress with poor prognosis. The majority of patients have no treatment available besides life-long active surveillance, rendering bladder cancer the most expensive cancer to treat. Compelling preclinical and epidemiological evidence show that dietary isothiocyanates (ITCs) from cruciferous vegetables (cruciferae) are particularly potent against bladder cancer given their exclusive excretion to the urine and exposure to the bladder epithelium. Our prospective cohort study also found that high intake of cruciferae, i.e., high ITCs, was associated with delayed bladder cancer recurrence and reduced progression. We translated our bench-side findings to develop a novel, dietary intervention to reduce bladder cancer recurrence and progression in NMIBC patients. Our intervention—POW-R-Health— significantly increased cruciferae intake and urinary ITC levels from baseline to 6 months, achieving urinary ITC levels sufficient to stop or kill at least 50% of bladder cancer cells in in vitro models. However, a considerable literature has documented the diminished effects of behavioral dietary interventions over time. To ensure POW- R Health’s impact on NMIBC outcomes, urinary ITC levels need to be sustained past 6 months because the majority of NMIBC recurrences occur within 24 months post-diagnosis. To sustain urinary ITC levels, we propose to establish the efficacy of POW-R Health with a maintenance component to maintain urinary ITC levels to at least 10 M through a 2-arm randomized controlled trial (RCT). We hypothesize that the POW-R Health + Maintenance arm will result in higher urinary ITC levels maintained compared to the POW-R Health Only arm. Aim 1: Use a systematic process to adapt maintenance strategies from evidence-based dietary interventions to develop an 18-month cruciferae-focused maintenance component for NMIBC survivors. We will expand POW- R Health by adding the adapted maintenance component. Aim 2: Assess the efficacy of POW-R Health + Maintenance compared to POW-R Health Only using a 2-group RCT design in 344 participants with the primary outcome of urinary ITC and the secondary outcome of cruciferae intake. Assessments will occur at baseline, 6, 12, 18, and 24 months. Exploratory Aim 3: Assess the preliminary efficacies of POW-R Health + Maintenance versus POW-R Healthy Only on bladder cancer recurrence and progression at 24 months through medical chart review. Recurrence and progression will be assessed as time to the event at 24 months post enrollment. Urinary proteomics will be conducted on urine samples to provide biological insights on ITC’s effects in the bladder. Our overall goal is to develop a new delivery of care model whereby our intervention can be implemented in healthcare institutions to reduce bladder cancer recurrence and progression for population-level impact on survivorship.

NIH Research Projects · FY 2026 · 2025-01

The tumor microenvironment (TME) of TNBC is extremely heterogenous, which plays critical roles in shaping tumor aggressiveness and response to therapies. Within the TME, tumor cells adapt to the selective pressures of limited nutrient and oxygen supplies, evade immune surveillance, and emerge as aggressive metastatic variants. We recently uncovered that the metabolic enzyme PFKFB4 is a potential driver of aggressive breast cancer. PFKFB4 is one of the rare bifunctional enzymes containing both a kinase and a phosphatase domain and controls the flow of glucose flux towards anabolic pathways. We discovered a non-canonical function of PFKFB4 acting as a protein kinase that phosphorylates the transcriptional coregulator SRC-3 to regulate gene activation. Our preliminary findings indicate that increased expression of PFKFB4 is significantly associated with poor survival in TNBC patients, and elevated levels were enriched in the metastatic lesions. Orthotopic implantation of human TNBC cells expressing inducible shRNA-PFKFB4 constructs significantly attenuated metastasis in xenograft models, whereas in syngeneic mouse TNBC models, depletion of PFKFB4 significantly increased CD8+ T cell infiltration and activation, which effectively cleared tumors. Photoacoustic imaging in live animals confirmed that hypoxia within mouse tumors robustly activated PFKFB4 expression. RNAseq analysis identified a pro-metastatic gene signature in the integrin αvβ3 pathway that was selectively activated by PFKFB4. We hypothesize that bifunctional activation of PFKFB4 in response to TME stressors accentuate TNBC progression to metastasis through intrinsic mechanisms that regulate metabolic and genetic plasticity, and extrinsic effects on the TME that enhance immune suppression. In Aim 1, we will investigate the bifunctional mechanisms of the PFKFB4 kinase and phosphatase activities regulating metabolic and genetic plasticity in the hypoxic TME and identify the mechanisms that promote selection of clones with metastatic competence. Aim 2 will interrogate the impact of PFKFB4-dependent tumor metabolic adaptations on the immune-TME and identify mechanisms of immunosuppression and trafficking that impair CD8+ T cell infiltration and/or their effector function leading to tumor-immune escape. Our proposed research will unmask the novel mechanisms in the hostile TME that accentuates the development and progression of metastatic TNBC progression by increasing tumor-immune escape.

NIH Research Projects · FY 2025 · 2024-09

Prostate cancer (PrCa) shows significant racial disparities, especially impacting African American (AA) men, who have a 1.4 times higher risk of PrCa diagnosis, and a double risk of PrCa-related mortality compared to European American (EA) men. AA men are also more frequently diagnosed with early-onset PrCa (≤55 years) than other racial/ethnic groups. Despite these disparities, we have limited knowledge about how ancestry/race influences epigenetic age and whether early-onset PrCa is linked to accelerated epigenetic aging due to genetic or environmental factors. This study builds on prior research that suggests ribosomal DNA (rDNA) methylation is a reliable marker of biological aging, with direct links to aging, longevity, and cancer We hypothesize that accelerated rDNA methylation (rDNAm) age is linked to early-onset PrCa (EO) in African American (AA) patients, potentially explaining their higher risk. By studying the rDNA methylation-EO PrCa relationship in AA men, we aim to uncover factors contributing to racial disparities. This insight may lead to targeted interventions for reducing the increased risk of EO PrCa in AA men. In the first aim, we investigate whether African American (AA) prostate cancer patients have a higher rDNA methylation age at diagnosis compared to age-matched European American (EA) prostate cancer patients. The second aim explores if AA men with prostate cancer exhibit an increased rDNA methylation age at diagnosis in comparison to age- and race-matched non-cancer controls. In the third aim, we assess the performance of the rDNA methylation clock in AA prostate cancer patients within their age range, relative to alternative metrics of epigenetic aging. Our hypothesis is that the rDNAm clock's age prediction will rival or surpass other measures like the Horvath, Hannum, PhenoAge, and GrimAge clocks in both prostate cancer patients and non-cancer controls. This cross-platform validation strengthens study rigor, advancing clock performance evaluation in the context of health disparities, ultimately progressing the field. Our main objective is to uncover the reasons behind racial disparities in prostate cancer among AA patients. In the short term, we expect the rDNA clock to emerge as a valuable tool for assessing the risk of early-onset prostate cancer in African American patients. In the long term, our research aims to provide strong evidence supporting the widespread adoption of the rDNA clock as a biomarker for monitoring age-related diseases in this unique group of cancer patients. Most importantly, this proposal lays the groundwork for future research to establish rDNA methylation epigenetic age as a fundamental baseline for epigenetic aging measurements in studies involving exercise, dietary, or other interventions for individuals at risk of prostate cancer, especially those of African descent.

NIH Research Projects · FY 2024 · 2024-09

PROJECT SUMMARY/ABSTRACT. As a Comprehensive Cancer Center with a mission to `understand, prevent and cure cancer', Roswell Park has long recognized the value of human biospecimens linked with clinical and outcomes data for driving research discoveries. As such, the institute has supported the collection and banking of remnant tumor tissue from surgical procedures in the Pathology Network Shared Resource (PNSR) and the collection and storage of peripheral blood samples in the Hematologic Procurement Shared Resource (HPSR) and the DataBank and BioRepository (DBBR). Furthermore, samples from specific research initiatives, including large multi-institutional team science grants, have also been banked for research use. Unfortunately, these efforts have all arisen independently of each other with no coordinated vision or standardized operating procedures, resulting in scores of -80 and liquid nitrogen (LN2) freezers spread across the Roswell Park campus. The haphazard and fragmented growth of these resources and the fact that some of these collections have inventories that cannot be electronically queried by the Roswell Park research community has made access to these samples difficult and has delayed advances in biomedical research. Importantly, with some epidemiological studies including thousands of participants with multiple aliquots of blood components for each, retrieval of samples for analysis is an extremely laborious and time-consuming process, and keeping freezer doors opened for long periods of time to retrieve samples jeopardizes sample integrity. Here we propose to consolidate sample procurement, processing and banking for the three existing shared resources (PNSR, HPSR, and DBBR) and external studies into ONE Biorepository – a comprehensive and unified biobanking system with accompanying services at Roswell Park. We will also construct a state-of- the-art biospecimen storage facility on the first floor of GBSB, centered around acquisition of a Hamilton BiOS M6, an ultra-low temperature automated storage system with robotic sample handling and retrieval, such that freezer doors are never opened, thereby maintaining the highest levels of biospecimen integrity during storage, tracking and accession. ONE Biorepository will be accompanied by a renovated laboratory space that can accommodate the operational needs of all three preexisting biobanks and their research services. The newly renovated Biorepository and laboratory facilities enable streamlined and efficient practices for the processing, banking and use of human samples to advance biomedical research, at Roswell and with our regional and national NIH-funded partners.

NIH Research Projects · FY 2025 · 2024-09

PROJECT SUMMARY / ABSTRACT The current application seeks salary support for Prashant K. Singh, PhD who serves as the Director of the Genomics Shared Resource (GSR) at Roswell Park Comprehensive Cancer Center (Roswell Park) and Assistant Professor in the Department of Cancer Genetics and Genomics. Dr. Singh works closely with Cancer Center Support Grant (CCSG) leadership and members to provide a centralized, efficient approach of supporting research, clinical and population based genomic endeavors, while facilitating peer-reviewed funding, publications, and recruitment efforts. Dr. Singh supports all genomics initiatives at Roswell Park by providing expertise in all aspects of assay design, sample preparation, QC, data generation, and analyses. Dr. Singh acts as a subject matter expert for CCSG members utilizing genomics approached to acquire and analyze genomic data sets across basic, translational, clinical and population studies. Dr. Singh plays a vital role in discovery- driven research of CCSG members as well as in diagnostic applications that complement traditional clinical evaluation of cancer samples. Dr. Singh is involved in the research and design of NGS assays that support the goals of developing personalized therapies based on the unique genetic makeup of a patient’s cancer. The GSR is an integral part of Roswell Park’s CCSG (P30CA016056) for which the cancer center Director, Dr. Candace Johnson, serves as the Principal Investigator. In the current CCSG reporting period (2018-present), as of October 2023, 128 NIH/NCI grant applications utilizing the GSR services and Dr. Singh’s expertise were awarded to Roswell Park faculty. In the same period, the GSR contributed to 210 peer reviewed publications. Dr. Singh has over 19 years of experience in the field of genomics and epigenomics with a strong focus in cancer research. Dr. Singh actively participates in meetings focused on correlative sciences related to clinical trials, multi-PI, and multi-institutional programs such as the Roswell Park Ovarian SPORE (P50CA159981-08), P01 Program grant (P01CA234212), and the Western New York Center for Research on Flavored Tobacco Products (CRoFT, U54CA228110). Dr. Singh’s leadership and expertise is critical for continued success of the CCSG program and Roswell Park’s research mission. As the PI of Roswell Park’s CCSG (P30CA016056), Dr. Johnson will serve as the Unit Director for this R50 Core-based scientist (PAR-22-188) application.

NIH Research Projects · FY 2025 · 2024-09

Until recently, the concept of the “oncology nurse” was limited to those having extra competency in administering chemotherapy, coordinating care delivery, or working with investigators to screen, enroll, and manage patients in clinical trials. With increasing recognition of the ability of PhD oncology nurse researchers to walk seamlessly between the “bedside and the bench” and acquire high value data, there is an urgent need for providing new state-of-the- art postdoctoral-level training in oncology for nurse-scientists. Nurses have an unduplicated role in patient care, monitoring overall well-being, managing symptoms, and developing a personal relationship over time with the patients and families. Research results obtained by nurse-scientists who can leverage these skills and interactions to conduct meaningful research on both the biological aspects of treatment response and the totality of the patient experience can provide important insights that cannot be obtained by other health care workers. And, as in the case with the success of the field of “symptom science”, they can change the way patient care is delivered as well as improving patient outcomes. However, despite increasing recognition of the unique and valuable role of the PhD nurse-scientist, there is an acknowledged shortage of these pivotal researchers in oncology. The Paul Calabresi Clinical Oncology Scholar Training Program provides a unique opportunity to address this gap by providing training opportunities for nurse-scientists with PhDs (or equivalent) in advanced cancer research skills and career-building opportunities needed to develop successful research careers in oncology. Indeed, with over 54-NCI-Designated Comprehensive Cancer Centers in the US, there is a significant need for a new generation of nurse scientists with the cutting-edge postdoctoral research training to lead essential patient-oriented research in oncology. While there are now 130+ programs leading to the PhD degree in Nursing, including one here at the University at Buffalo’s School of Nursing, training in oncology is significantly lacking and although the NIH Reporter lists 19 T32 training grants for nurse-scientists, at this point in time, few are funded through the NCI with a focus on cancer-related training. Moreover, for nurse-scientists to move up in their career, postdoctoral research training and career development and mentorship, such as afforded by the K12 mechanisms, has become the requirement for careers in either academia or clinical practice. Yet, there are no K12 programs for nurse scientists in oncology at this time in the US. Our new K12 program, led by a team of outstanding advisors and faculty, is designed to prepare a new generation of PhD educated nurse scientists in oncology, giving them front-line opportunities to conduct innovative, independent clinical and translational research and providing them with outstanding career development opportunities.

NIH Research Projects · FY 2025 · 2024-09

PROJECT ABSTRACT/SUMMARY Immunotherapy has revolutionized how oncologists treat cancer patients. One type of immunotherapy includes immune checkpoint inhibitors (ICIs), such as those that engage PD-1 on cytotoxic CD8+ T cells to bolster their effector functions. However, in several cancer types, single-agent therapy is moderately effective, but when combined with standard-of-care (SOC) chemotherapy, the combination regimen improves overall response rates. Therapeutic efficacy may be hampered by numerous barriers, including the immune suppressive tumor microenvironment (TME). One major cellular component of the TME includes myeloid-derived suppressor cells (MDSCs), which correlate with poorer survival outcomes and suppress antitumor activity of CD8+ T cells in response to these ICIs. To overcome these obstacles of immune suppression, we developed a novel approach to target MDSC ‘biogenesis’ in the bone marrow to mitigate their production and function, and boost ICI activity in mouse models of triple-negative breast cancer (TNBC), a cancer type that elicits a robust MDSC response. We identified a metabolic vulnerability in MDSCs and targeted that dependency using agents known as dihydroorotate dehydrogenase (DHODH) inhibitors. DHODH inhibitors block de novo pyrimidine metabolism and are being clinically tested as a therapy in acute myeloid leukemia. We found that combining the DHODH inhibitor, brequinar (BRQ), with PD-1 blockade significantly reduced tumor growth, and that therapeutic efficacy depended upon a reduction in MDSC suppressive activity. However, tumors still grew introducing two important gaps. First, it remains unclear how DHODH blockade ‘reprograms’ MDSC function and, secondly, it remains unclear how therapeutic efficacy can be further augmented. Regarding the first gap, in Aim 1 of the F99 phase, we will focus on the unfolded protein response (UPR) based on the rationale that we found that the UPR pathway is one of the top downregulated pathways in myeloid progenitors that give rise to MDSCs and that prior work in the field showed that the UPR is a key pathway by which MDSCs mediate their activities, particularly, via the transcription factor XBP1. Regarding the second gap, we propose to build on a SOC chemo-immunotherapy platform to augment antitumor responses. Therefore, in Aim 2 of the K00 phase, we will determine whether incorporating BRQ to a paclitaxel/anti-PD-1 monoclonal Ab regimen, which is clinically used in TNBC, bolsters CD8+ T cell responses and achieves durable antitumor immunity. Our central hypothesis is that DHODH blockade reprograms MDSCs toward less immune suppressive states through an XBP1-dependent mechanism in the UPR pathway. We further hypothesize that MDSC mitigation through this pathway will further sensitize TNBC to a SOC chemo-immunotherapy regimen. New advances gained from this research have the potential to inform the clinical design of novel, more effective combination immunotherapies.

NIH Research Projects · FY 2024 · 2024-08

ABSTRACT Some non-combustible tobacco products, such as various forms of smokeless tobacco (ST), can pose significantly fewer risks to individual users than traditional cigarettes, including those for several types of cancers. Although not safe, these products might offer the potential for harm reduction for adults who smoke cigarettes who completely switch to them. Recognizing these issues, the Food and Drug Administration (FDA) developed a new pathway by which tobacco companies can apply for modified-risk tobacco product (MRTP) status and to legally make modified risk claims about their products. In March 2023, the FDA announced that Copenhagen, the top-selling ST brand in the US, could legally make a modified risk claim in its advertising that complete switching from cigarettes to the product reduces risk of lung cancer. FDA had previously authorized a comparable claim for another ST brand, General Snus, about reduced risks for multiple diseases. The FDA’s authorization of these ST claims is notable given that previous implicit health claims and tactics used by the tobacco industry in marketing cigarettes (notably “light” cigarettes”) are now banned by the FDA. Given the relative recency of these ST claim authorizations, little is still known about how such claims may impact consumer behavior and health. Effects may depend on several factors including whether participants are exposed to the claims and find them to be believable and persuasive. Another open and innovative question is whether newly authorized claims for smokeless tobacco products, which make explicit health references, may improve expectancies about and subjective ratings for these smokeless products when using them. Indeed, previous studies show that tobacco marketing, such as packaging color and marketing descriptors (e.g., “smooth), can impact subjective ratings of those products when used. This is important given that negative product perceptions related to subjective use characteristics (such as taste, satisfaction, liking) have been noted as barriers/reasons why smokers have not switched to these products in the past. A negative sensory experience may undermine confidence in reduced risk claims and lead to reduced use intentions, whereas enjoyable sensory experiences may lead to a reevaluation of beliefs about harmfulness, which in turn may increase future intentions to use. Overall, the current study seeks to explore consumer responses to the announcement and implementation of these new modified risk claims using a series of complementary and innovative research activities, including an online survey and experiment, an experimental auction, and an in- person product use lab study. Two specific aims will explore 1) consumers’ exposure and reactions to authorized smokeless tobacco MRTP claims; and 2) whether the MRTP claims influence smokeless tobacco demand and sensory evaluations.

NIH Research Projects · FY 2025 · 2024-07

PROJECT SUMMARY/ABSTRACT This K08 mentored career development award proposal details a research and training plan that is designed to facilitate the career development and transition of Lisa Niswander, MD PhD to an independent physician-scientist investigator. Dr. Niswander is a pediatric oncologist and an Instructor of Pediatrics at the Children’s Hospital of Philadelphia (CHOP) and University of Pennsylvania (Penn). Her long-term goal is to lead an NIH-funded research program focused on the preclinical development and early bench-to-bedside translation of targeted therapies and immunotherapies to improve outcomes for children with leukemia. During the 5-year award period, critical technical and scientific training will be acquired in T cell and chimeric antigen receptor (CAR) T cell immunobiology, single cell transcriptomics, early-phase clinical trial design, leadership, and scientific communication. Under the primary mentorship of Dr. Sarah Tasian, a leader in the development of molecularly- targeted and CAR T cell therapies for high-risk pediatric leukemias, and co-mentorship of Dr. Martin Carroll, an expert in acute leukemia signaling, in tandem with a complementary advisory committee of senior scientists, this scientific research program and training plan will be bolstered by the resource-rich environment at CHOP/Penn. Despite high cure rates in most children with B-cell acute lymphoblastic leukemia (B-ALL), infants with B-ALL have dismal outcomes. Most cases of infant B-ALL are characterized by KMT2A rearrangements (KMT2A-R) with resulting fusion proteins that complex with critical adaptor protein menin to drive leukemogenesis and/or by frequent activating RAS pathway mutations. Promising preclinical and early clinical efforts have focused on small molecule disruption of KMT2A-menin binding (menin-i) and on MEK inhibitor (MEK-i) blockade of RAS signaling. CD19-directed CAR T cell immunotherapy (CD19CART) has achieved high response rates in children with B- ALL, including infants with KMT2A-R ALL. However, at least 50% of these patients will ultimately relapse. The central hypothesis of this proposal is that combining CD19CART with pharmacologic disruption of specific KMT2A-R ALL biologic vulnerabilities, including menin-binding or aberrant RAS signaling, will improve treatment efficacy and long-term remission in KMT2A-R ALL. To test this hypothesis, the combinatorial strategy of menin- i (Aim 1) and MEK-i (Aim 2) with CD19CART against KMT2A-R ALL will be investigated. The effects of these dual approaches will be elucidated for both anti-leukemia therapeutic activity and for CD19CART functionality and persistence. Successful completion of this career development award will identify promising co-therapeutic strategies with CD19CART poised for rapid clinical translation for children with KMT2A-R ALL. These research and training efforts will also propel Dr. Niswander’s transition to independence by establishing a robust scientific pipeline for future investigation of targeted inhibitors with immunotherapies in pediatric leukemias.

NIH Research Projects · FY 2025 · 2024-06

Despite notable improvements in childhood cancer survival rates, patients with relapsed or refractory disease face grim prospects, and childhood cancer remains the commonest cause of death from disease in children. This program aims to tackle this issue, focusing primarily on the challenging pediatric cancer, high-risk neuroblastoma (NB), a tumor of the sympathetic nervous system which accounts for up to 15% of childhood cancer deaths and is the commonest solid tumor of young children. Recent research by our international collaborative team has revealed a novel mechanism of cancer drug resistance linked to epigenetic activation of LINE1 (L1) retrotransposons. We have provided preliminary evidence that these retrotransposons can be effectively targeted by existing nucleoside reverse transcriptase inhibitors (NRTIs), offering a promising avenue for repurposing these safe and effective antiviral agents as potential treatments for refractory tumors. While the significance of retrotransposon activation has been established in various adult cancer types, a substantial knowledge gap exists concerning its role in pediatric malignancies. Closing this gap is imperative, as controlling retrotransposon activation with NRTIs, and potentially also with immunotherapeutic approaches, holds substantial potential for significantly improving treatment outcomes for pediatric cancers, including refractory NB. Our program seeks to explore the feasibility of retrobiome-targeted pharmacological and immunotherapeutic strategies in NB. This exploration will be guided by a comprehensive assessment of retrotransposons’ activities and their impact on tumor characteristics, in NB and also in all other solid tumors of childhood, employing a unique and extensive range of preclinical models and clinical samples with associated comprehensive clinical and molecular data. Our program capitalizes on a longstanding, fruitful collaboration between US and Australian teams, drawing upon our complementary expertise in basic and translational science. We also build upon our teams' foundational research on the epigenetic control of retrotransposons in cancer, their recognition by humoral and T cell immunity, and the potential druggability of retrotransposon-induced malignant traits, particularly drug resistance. Our specific aims are as follows: (SA1) to comprehensively characterize retrotransposon activity in NB and other solid tumors of childhood, and assess its potential therapeutic and diagnostic significance, (SA2) to investigate the impact of retrotransposon derepression on NB treatment resistance and the potential of reverse transcriptase inhibitors to enhance refractory NB treatment outcomes in preclinical models and (SA3) to explore immune responses to L1 retrotransposon antigens and evaluate the efficacy of anti-L1 immunotherapy in preclinical NB models. The outcomes of the proposed program will guide the rational design of future pharmacological and immunotherapeutic interventions for refractory NB and subsequently other poorly curable pediatric cancers.

NIH Research Projects · FY 2026 · 2024-04

Pediatric nonadherence to medication is a significant public health problem, and rigorous research repeatedly documents that nonadherence increases risk for hospitalization, healthcare cost, disease progression, and death. Participants in our previous studies have described medication adherence as “one of the most stressful parts” of pediatric cancer caregiving, and we found that 93% of parents experience barrier(s) to medication administration. We have designed MedSupport, a theory-based multilevel intervention that is designed to address root barriers to medication adherence. In Aim 1 we will determine if the MedSupport intervention increases the proportion of patients with chemotherapy adherence 95% or higher. Our study design leverages a unique opportunity as a companion study for an upcoming therapeutic trial and will enroll at 8 pediatric cancer programs. This will enhance methodological rigor through leveraging robust clinical trial infrastructure to achieve multi-site recruitment in diverse geographic and clinical sites. We will recruit families of pediatric patients with ALL (N = 150) on home-based chemotherapy. Families will be randomized 1:1 to (1) the MedSupport intervention and (2) usual care with standardized education control. We will use both MEMS electronic medication monitoring and innovative biomarkers of drug metabolites to measure adherence. In Aim 2 we will test a theory-based mechanism of intervention effectiveness. These results will increase conceptual significance through rigorous examination of mechanisms of action of the MedSupport intervention to inform future intervention optimization and translation. In Aim 3 we will use the Implementation Outcomes Framework to examine implementation effectiveness to enhance future dissemination of the MedSupport intervention. We will examine the relationship between implementation quality on intervention effectiveness (Aim 3a) and strategies that may hinder or support uptake within routine care to inform future implementation strategies (Aim 3b). Our design incorporates numerous elements that substantially increase rigor and reproducibility including leveraging a large therapeutic trial for multi-site participant recruitment and rigorous measurement of adherence through objective behavioral measures (electronic medication monitoring) and pharmacological measures (validated biomarkers of drug metabolites). Findings will make significant conceptual contributions through examining how the MedSupport intervention works to pave the way for future intervention optimization. Findings will have significant translational impact through examining implementation effectiveness to enhance future dissemination of the MedSupport intervention. Upon completion of this research, we will have an innovative, low-cost, and scalable intervention to enhance home-based medication adherence in pediatric cancer that is ready to be translated to clinical practice. While this study focuses on pediatric ALL, the MedSupport intervention has potential for high impact in numerous pediatric diseases.

NIH Research Projects · FY 2026 · 2024-04

A. Project Summary/Abstract Ovarian cancer is the second most common cause of gynecologic cancer death in women around the world. High-grade serous ovarian cancer (HGSOC) accounts for more than 70% of all ovarian cancer deaths. Approximately 50% of HGSOCs harbor alterations in the genes involved in homologous recombination (HR) DNA repair pathway. These tumors tend to respond well to PARP inhibitors and chemotherapy. CCNE1 (Cyclin E1) gene amplification and overexpression is frequently found in HR-proficient ovarian cancer and is associated with primary chemoresistance and poor clinical outcomes. CCNE1 gene amplification is present in 15-20% ovarian tumors, and overexpression of Cyclin E1 is detected in over 40% of ovarian cancer specimens. Currently, there are no effective treatment approaches for this type of HGSOC. By using Cyclin E1-overexpressing syngeneic and humanized ovarian patient-derived xenograft (PDX) models of HGSOC, we will identify novel therapies for the treatment of HGSOC cancers with amplification or high expression of Cyclin E1. Our preliminary data suggests that Cyclin E1 downregulates IRF1 and type I interferon (IFN) expression in a CDK2 kinase-independent manner in ovarian cancer cells. Our preliminary data also reveal that overexpression of Cyclin E1 in ovarian cancer cells promotes pro-tumor polarization of tumor-associated macrophages (TAMs) in the tumor microenvironment (TME). Expression of IRF1 and type I IFNs has been implicated in pro-tumor polarization of TAMs in cancers. We plan to investigate the molecular mechanisms underlying the regulation of Cyclin E1 on the type I IFN signaling pathway and TAMs in Cyclin E1-amplified/overexpressing HGSOC tumors and to develop therapeutic approaches for this type of ovarian cancer. We hypothesize that downregulation of IRF1 and type I IFNs by Cyclin E1 in tumor cells fosters an immunosuppressive TME in HGSOC with Cyclin E1- amplification/overexpression by promoting pro-tumor macrophage polarization. We also hypothesize that targeting the conventional tumor cell-intrinsic oncogenic function of Cyclin E1 with CDK2i (CDK2 inhibitors) or ATRi (Ataxia telangiectasia, and Rad3-related inhibitors), in combination with remodeling the pro-tumor TAMs that induced by expression of Cyclin E1 in the TME with immunomodulators, such as STING agonists, will provide promising therapeutics for ovarian cancer patients with Cyclin E1-amplification/overexpression. In Aim 1, we will determine how cyclin E1 regulates IRF1 and type I IFNs in both human and mouse ovarian cancer cell lines. In Aim 2, we will determine how the Cyclin E1-regulated type I IFN signaling pathway promotes pro-tumor polarization of TAMs and immunosuppression in the TME of HGSOC tumors with Cyclin E1- amplification/overexpression both in vitro and in vivo. In Aim 3, based on our recent findings that STING agonism activates type I IFN signaling pathway and repolarizes pro-tumor macrophages to an anti-tumor status, we will assess the therapeutic efficacy and toxicity of STING agonists in combination with CDK2i or ATRi, in both syngeneic mouse models and humanized PDX models.

NIH Research Projects · FY 2025 · 2024-03

Project Summary/Abstract Given the high mortality of castration-resistant prostate cancer (CRPC) as a result of relapse after androgen deprivation therapy (ADT), novel treatment strategies are urgently needed. Tumor suppressor gene- phosphatase and tensin homolog ( PTEN) is mutated in approximately 20% of primary prostate cancers, and in as many as 40~60% of CRPC. In addition to activation of PI3K oncogenic signaling pathway, loss of PTEN is also associated with cytokine and chemokine signaling that creates an immunosuppressive microenvironment. Both defects in antigen presentation in tumor cells and the immunosuppressive tumor microenvironment (TME) have been implicated in CRPC progression after ADT and therapeutic resistance in prostate cancer in the clinic. Genetic or pharmacologic inactivation of β isoform of PI3K (p110β) in PTEN-deficient prostate cancer has been reported to significantly inhibit tumor growth and progression. Ataxia telangiectasia and Rad3-related (ATR) exhibits essential functions in controlling DNA replication stress and DNA damage response in cancer cells. PTEN-deficient cells have been reported to be more sensitive to ATR inhibition. PI3Kβ is also functionally associated with DNA damage response and genomic integrity in tumor growth. By using a syngeneic genetically engineered mouse model of CRPC driven by co-deletion of PTEN and TP53 (PP-CRPC), we found that addition of p110β inhibitor significantly reduces the viability of ATR inhibitor-treated PP-CRPC cells. Moreover, our preliminary data show that inhibition of p110β or ATR alone significantly increases MHC class I antigen presentation in mouse or human CRPC cells. Based on these novel preliminary data, we hypothesize that p110β inhibitor in combination with ATR inhibitor will show promising anti-tumor activity in PTEN-deficient CRPC via inducing cell death in cancer cells and anti-tumor immunity in the tumor microenvironment. We also hypothesize that inhibition of p110β will increase DNA replication stress and sensitize PTEN-deficient CRPC cells to ATR inhibition. In this study, we will 1) evaluate the therapeutic efficacy and antitumor immune responses of combined ATR inhibition and p110β inhibition i n syngeneic PTEN-deficient CRPC mouse models; 2) investigate the molecular mechanisms of the combined effect of ATR inhibitor and p110β inhibitor in replication stress and DNA damage-induced cell death in PTEN-deficient CRPC cells. Our study will not only provide fundamental information for translational research of p110β inhibitors and ATR inhibitors in cancer treatment, but also provide a novel treatment strategy for patients with PTEN-deficient advanced CRPC.