Roswell Park Cancer Institute Corp

NIH Research Projects · FY 2025 · 2021-09

Mitochondrial protein homeostasis (proteostasis) has been implicated in cancer and is regulated by mitochondrial unfolded protein response (UPRmt). However, it is unknown whether UPRmt promotes tumorigenesis and whether it could be targeted for therapeutic benefit in prostate cancer (PCa). This proposal will define how heat shock protein 60 (HSP60), a key component of UPRmt, promotes aggressive and resistant PCa. Using genetically-engineered triple knockout (TKO: deletion of Pten, Trp53, and Rb1) tumors, we observed that HSP60 is upregulated in aggressive tumors and castration-resistant prostate cancer (CRPC) compared to WT prostatic tissues. TCGA data analysis and our preliminary data using human PCa-specific TMAs demonstrated that HSP60 is upregulated in prostate tumors with higher Gleason Scores. HSP60-silencing induced caspase activation and inhibited cellular proliferation whereas HSP60 overexpression promoted cancer cell survival and proliferation. We provide the first evidence that, genetic deletion of HSP60 in TKO mouse and inhibition of HSP60 oligomerization by introducing HSP60D3G KI during prostate tumorigenesis, reduced tumor burden in vivo. We observed that activating transcription factor 5 (ATF5), specific for HSP60 expression and UPRmt activation, was upregulated with higher Gleason Scores, and ATF5 was translocated to nucleus during stress. Using in silico analysis, we have identified a novel UPRmt inhibitor (referred to as DCEM1), which induced robust apoptosis in PCa cells and blocked tumor growth in vivo. Based on these findings, we hypothesized that HSP60- dependent mitochondrial unfolded protein response promotes cancer cell adaptation during tumor progression and therapeutic resistance in PCa. Identification of UPRmt inhibitor provides alternative treatment option for patients with PCa. We propose the following Specific Aims to test this hypothesis. Aim 1. Define the role of transcription factor ATF5 in activating mitochondrial unfolded protein response. Aim 2: Evaluate whether HSP60 oligomerization maintains functional mitochondria and inhibits apoptosis to develop aggressive PCa. Aim 3. Explore the clinical relevance of HSP60 inhibition using patient-derived xenografts (PDXs) and primary tumor cells. Impact: The findings will provide fundamental understanding on how UPRmt is activated and how persistent mitochondrial stress is attenuated by UPRmt leading to development of aggressive and lethal PCa. Identification of unique UPRmt inhibitor represents a new therapeutic vulnerability in PCa that does not rely on androgen modulation. Therefore, UPRmt inhibition by DCEM1 will have greater therapeutic benefits for patients with androgen-dependent and androgen-independent CRPC.

NIH Research Projects · FY 2025 · 2021-08

Project Summary Blood and marrow transplant (BMT) is an effective cure for many life-threatening hematologic diseases. Survival after BMT has improved dramatically over the past two decades, however up to 40% of patients still die within one year after HLA-matched unrelated donor allogeneic BMT. This project will build upon our prior genome-wide (GWAS) and exome-wide association studies (ExWAS) involving ~2,900 patients named Determining the Influence of Susceptibility COnveying Variants Related to one-Year mortality after BMT (DISCOVeRY-BMT). Our GWAS identified several donor loci that significantly increased recipient’s risk of disease-related mortality (DRM) and donor-recipient genotype mismatches significantly increased risk of transplant-related mortality (TRM) in European Americans (EAs). Our ExWAS discovered a rare nonsynonymous coding variant, where a donor-recipient genotype mismatch correlated with TRM and additional novel genes (e.g. TEX38, OR51D1, and NT5E) correlated with overall survival, TRM and DRM. Our goal for this proposal is two-fold: to deepen our understanding of non-HLA genetic contributors to BMT mortality, and to build the clinical-genomic prognostic models to translate such understanding into clinical practice. The first goal will be fulfilled in two directions. 1) We will systematically survey both rare and common variants using whole-exome sequencing (WES) and meta- GWAS in EAs as well as under-studied diverse populations in an effort to bridge the BMT survival disparity between EAs, African Americans, Asians and Hispanics. Our prior ExWAS in EAs demonstrated the important roles played by rare coding variants in BMT mortality, however, only 2% of rare variants are in the exome array. Therefore, we will use WES to assay all exonic variants in 5,598 multi-ethnic donor-recipient pairs. As the variants/genes we identify are direct candidates for causality, functional validation will be performed to investigate such relationships. In parallel, we will perform the largest meta-GWAS of BMT mortality to date. Through our collaborations, we have assembled all BMT GWAS data available in the US (8,576 donor-recipient pairs including 1,978 minority pairs). 2) We will interrogate WES and GWAS data to further reveal the biological networks contributing to BMT mortality. To meet the second goal, we will leverage our unique and powerful GWAS resource to develop prognostic models to predict patients’ personalized mortality risk. This is the first study to use next-generation sequencing technology to analyze the contribution of non-HLA coding variants on post-BMT mortality. GWAS data on 8,576 donor-recipient pairs, of which 5,598 pairs also have WES data, will make this the largest genetic study ever undertaken, and provide a real opportunity to understand the genetics of BMT mortality across diverse populations. The prognostic models we develop will provide a valuable tool to help reduce BMT mortality and enhance donor-recipient matching in routine clinical practice. Importantly, the data generated by this project will be shared publicly to serve as a resource for additional research to improve survivorship after BMT and enhance the public investment in this project.

NIH Research Projects · FY 2025 · 2021-08

PROJECT SUMMARY Despite advances in treatment, such as targeted and immune therapies, lung cancer remains a deadly malignancy with five-year survival below 25%. About two-thirds of NSCLC diagnoses are made in former tobacco smokers, who are at 6-fold higher risk for the disease compared to non-smokers. Unfortunately, attempts to identify chemopreventive agents that reduce the risk of cancer in ex-smokers have been unsuccessful. Currently in the US, about 60% of ex-smokers are either overweight or obese. We have observed that for lung cancer, the well-known anti-cancer effect of the common diabetes drug metformin is restricted to patients who are overweight or obese. In investigations that followed this novel finding, we have found that in both humans and mice, obesity is associated with changes in the lung tumor immune microenvironment that promote disease progression, and that these changes are susceptible to reversal by metformin. Prominent among these changes is the impact of metformin on activation of immunosuppressive regulatory T cells (Tregs), which is known to be an important immunological event in carcinogenesis. We hypothesize that the obesity-specific immunomodulatory action of metformin also occurs in obese/overweight ex-smokers at high risk of lung cancer. If true, this concept will establish a basis for metformin's chemopreventive potential to abate lung cancer development in a major fraction of the population at high risk for the cancer. To examine this preventive potential of metformin, we will conduct a small phase II trial with at- high-risk obese/overweight subjects to establish that months-long oral metformin treatment diminishes markers of immunosuppressive Tregs in lungs and enhances markers local pulmonary and systemic immunosurveillance activity (Specific Aim 1). To identify mechanisms that underlie the obesity-specific immunomodulatory effects of metformin, we will study the impact of this drug in obese and non-obese mice of two distinct but complementary mouse lung cancer models (Specific Aim 2).

NIH Research Projects · FY 2025 · 2021-07

Project Summary: Pancreatic ductal adenocarcinoma (PDAC) is an aggressive disease that remains largely incurable. Although the cause for this profound therapeutic resistance is poorly understood, it is however partly blamed on signaling factors present in the tumor microenvironment (TME), which supports the proliferation and survival of neoplastic cells. Apart from being stroma rich, PDAC TME is associated with a distinctive tumor immune infiltrate. Paradoxically, most immunotherapy trials using immune checkpoint inhibitors, either as monotherapy or combination, failed to increase patient survival motivating exploration of new therapeutic strategies. To that end, the cytokine mediated heterotypic interactions between cancer cells and immune cells remain largely unexplored. In a recent study, we demonstrated that cytokines, IL4 and IL13, secreted by TH2 cells (a subtype of CD4+ T cells), provide trophic support for PDAC development. Mechanistically, inhibiting this cytokine mediated crosstalk between cancer-TH2 cells either genetically or pharmacologically drastically reduces tumor growth and increases survival in a preclinical model. Our subsequent preliminary work identified a potent inflammatory cytokine, IL33 which is overexpressed and released by PDAC cells that attract and activate TH2 and other immune cells such as innate lymphoid cells 2 (ILC2) and Tregs. Importantly, we found that the release of IL33 by PDAC cells is mediated by intratumor mycobiome. Inhibition of IL33 or anti-fungal treatment leads to a decrease in the infiltration and activation of type 2 immune cells (TH2 and ILC2) and Treg cells, accompanied by significant PDAC tumor regression. Taking these observations together, we hypothesize that type 2 immune response plays an important role in PDAC tumorigenesis and intratumor mycobiome is key to the IL33 secretion. The major objective of this proposal is to elucidate the role of mycobiome in the IL33 mediated type 2 immune response and provide pre-clinical evidence to guide future clinical studies with an anti-IL33 monoclonal antibody in PDAC patients. To that end, we will determine the molecular mechanism of mycobiome mediated IL33 release in cell and organoid models of PDAC. Further, to conduct a clinically relevant study, we will analyze IL33, intratumor mycobiome and type 2 immunocytes in the PDAC patient tumor and serum samples. While our preliminary studies using the syngeneic orthotopic model have shown a significant tumor regression upon IL33 deletion or anti-fungal treatment, synergistic combination strategies are expected to be even superior in efficacy. So, we propose to use an anti-IL33 antibody in combination with anti-fungal treatment for superior efficacy. Finally, to block the IL33-TH2/ILC2 axis we have three genetically engineered mouse models that will allow rigorous testing of the function of IL33 in PDAC tumorigenesis. In conclusion, our study is poised to identify a novel strategy to target PDAC patients and provide mechanistic insights for future clinical development of anti- IL33 therapy.

NIH Research Projects · FY 2025 · 2021-06

ABSTRACT Immune checkpoint inhibitors (ICIs) are a powerful and innovative mode of cancer therapy, believed to be partially responsible for the largest single-year drop in cancer mortality from 2016 to 2017. Their use has increased dramatically over the past 3 years. However, little data has been collected about ICI treatment response among patients of African ancestry (AA). In addition, little is known about the toxicities, treatment patterns, long-term outcomes, and post-treatment quality of life associated with ICIs outside the clinical trials setting. A prospective cohort study with a focus on racial differences between AA patients and patients of European ancestry (EA) in community oncology settings could address these knowledge gaps. Focusing on racial differences in ICI impact is important for three reasons. First, at the population level, AA patients are more likely than EA patients to have advanced cancers, an important disease group ICIs are intended to treat. Second, due to racial differences in host immunity, AA individuals tend to have a stronger pro-inflammatory response than EAs. This could lead to a higher risk of immune-related adverse events (irAEs) while on ICIs. Third, as a result of immune differences, AA patients who manage irAEs and continue ICI treatment may be more likely to benefit than EA patients. However, AA populations may experience multiple barriers while accessing healthcare (e.g., discrimination, financial toxicity) that may lead to discontinuing ICIs. We have a unique opportunity to assess the treatment, disease, individual, lifestyle, and quality of life factors that contribute to differential experiences of AA patients on ICIs, by accruing a prospective cohort through the nationwide NCI Community Oncology Research Program (NCORP) network. We will include all patients receiving anti-PD-1/-L1 therapy regardless of cancer site and enroll a total of 600 AA and 1,200 EA patients, with 1:2 match of AA to EA patients on cancer type within NCORP site. Our Specific Aims are: 1. To examine racial differences and predictors of irAEs, comparing AA and EA patients on incidence and severity of irAEs and assessing disease, individual, and lifestyle factors as predictors of these differences. 2. To examine treatment delay and discontinuation between AA and EA patients and assess racial differences in irAEs, healthcare barriers, and other factors as potential causes of treatment interruptions. 3. To examine short- and long-term treatment outcomes, comparing AA and EA patients on objective response rate (ORR), recurrence, death, and HRQOL after ICIs, and assessing treatment, disease, individual, and lifestyle factors as predictors of patient outcomes and potential causes of racial differences. We envision this to be the first large cohort study of diverse AA and EA patients treated with ICIs. We will gain valuable knowledge of the usage, effects, and challenges of ICIs in community oncology settings. Our findings may inform use of ICIs, management of irAEs and reduction of healthcare barriers across populations.

NIH Research Projects · FY 2025 · 2021-04

Project Abstract/Summary. T cells are unique in their requirement for two activation signals to become functional. This separation of powers between the T cell receptor (TCR) and co-receptors, such as CD28, allows exquisite regulation of T cell responses. In recent years, T cell co-receptors have emerged as valuable targets of immunotherapy for autoimmune diseases and cancer. In autoimmunity blocking activating co-receptors can reduce the destructive effects of T cells on normal tissue, while in cancer blocking inhibitory co-receptors can activate a T cell response against malignant cells. However, co-receptor targeted immunotherapy often fails to produce desired results. Moreover, there remain large gaps in knowledge of how co-receptors coordinate the myriad cellular changes that allow T-cells to gain functional properties. The project proposed in this grant application will explore a novel mechanism by which T cell co-receptors coordinate changes in RNA maturation important for T-cells to generate sufficient numbers and make key molecules that allow them to kill other cells. These killer properties underlie the ability of T cells to contain infectious diseases and control tumor growth or to damage healthy tissue in autoimmune diseases. Preliminary studies provide evidence that CD28 signaling coordinates many changes in alternative splicing of newly made RNAs through effects on the RNA binding protein ARS2. Alternative splicing allows one gene to code for several different proteins by changing how the RNA produced from that gene is assembled. We find that in activated T cells CD28-ARS2 dependent alternative splicing of the mRNA coding for metabolic enzyme pyruvate kinase favors production of an isoform, known as PKM2, with known proliferation promoting properties. Proposed studies seek to 1) determine if alternative splicing to PKM2 induces changes in how T cells use nutrients to fuel proliferation, 2) examine how ARS2 regulates alternative splicing in T cells, and 3) establish CD28 regulated changes in alternative splicing as potential modulators of immunotherapy. The long-term goals of these studies are to understand how RNA binding proteins and RNA maturation shapes gene expression during the process of T-cell activation and to determine if such mechanisms of gene regulation can be therapeutically targeted to alter T-cell function in patients with cancer or autoimmune diseases.

NIH Research Projects · FY 2025 · 2021-04

Metastatic prostate cancer (PCa) remains a major clinical challenge. Although androgen deprivation therapy (ADT) is effective in treating PCa, majority of the patients quickly develop resistance to therapy and the tumor relapses as hormone refractory castration-resistant prostate cancer (CRPC). Men with CRPC frequently progress to an aggressive lethal disease that metastasizes to bones and other visceral organs accounting for high morbidity and mortality. Transcriptional activation of steroid receptor coactivator-2 (SRC-2; also known as NCOA2/TIF2/GRIP1) plays a critical role in the pathogenesis of PCa by driving a metabolic switch towards de novo fatty acid biosynthesis. Although increased lipogenesis is a known hallmark of hormone refractory PCa progression, it is less clear how mitochondrial enzymes communicate with nuclear receptor coregulators to rapidly fuel and support fat biosynthesis. Our preliminary findings indicate that sustained activity of mitochondrial aconitase (ACO2) enzyme is critical for regulating citrate synthesis. We found that acetylation of ACO2 is essential for enzyme functions, which is negatively regulated by sirtuin-3 (SIRT3). In human prostate cancer patients, SIRT3 expression is repressed and increased expression of SRC-2 with concomitant reduction of SIRT3 was found to be a genetic hallmark in metastatic PCa. Based on these findings, we hypothesize that the transcriptional coregulator SRC-2 drives the nuclear-mitochondrial regulatory axis by repressing tumor suppressor SIRT3 thus promoting prostate tumor survival and metastasis competence. So our objectives in this proposal are (1) to investigate the mechanisms regulating sustained activation of mitochondrial ACO2 to promote lipogenesis, (2) define the role of nuclear receptor coregulator SRC-2 regulating SIRT3 expression, and (3) evaluate the impact of this nuclear-mitochondrial regulatory axis on prostate tumor survival and adaptation leading to bone colonization and growth. Our study will uncover molecular links between mitochondrial metabolism and transcriptional regulation that enables hormone refractory PCa adaptation, survival and ultimately metastatic competency.

NIH Research Projects · FY 2025 · 2021-04

ABSTRACT A patient's own T cells can be modified using gene therapy technology to express receptors, termed chimeric antigen receptors or CARs, which allow these immune T cells to recognize proteins on the tumor cell surface, and in turn allow these CAR modified T cells to recognize and kill the patient's own tumor cells. This approach has been successful in some hematological malignancies, however, it has not been successful to date in solid tumors including small cell lung cancer (SCLC). Two mechanisms by which SCLC may evade T cell-mediated killing are loss of expression of antigens, and suppression of T cell function in the tumor microenvironment. In this proposal, we will attempt to overcome these barriers by designing CAR T cells that target two SCLC antigens simultaneously, and that produce multiple factors (“armors”) that enhance T cell activity in solid tumors. We hypothesize that these dual-armored, dual targeted (DADT) CAR T cells will be more effective against SCLC than previous T cell-mediated and immune therapies. We have previously shown that CAR T cells targeted to either the antigen GD3 or to the antigen DLL3, both of which are expressed on the majority of small cell lung cancers, are capable of killing SCLC cells in preclinical systems. Additionally, we have developed multiple armored CAR T cells that secrete factors such as IL-18, or an antibody-derived single-chain variable fragment (scFv) that blocks the immune checkpoint receptor PD-1, or an scFv blocking the phagocytosis-inhibitory signal CD47 on tumor cells. All of these armors enhance CAR T cell activity in our in vivo model systems through different mechanisms. In Aim 1 of this proposal, we will generate CAR T cells targeting DLL3 and GD3 simultaneously, to overcome antigen heterogeneity and antigen loss in tumors as a means of escape from T cell-mediated killing. Simultaneously, in Aim 2, we will test pairs of armors to identify the pair that is the most effective at enhancing the activity of single antigen-targeted CAR T cells against SCLC in vivo in immunocompetent systems. We then analyze the immune cells in the SCLC tumor microenvironment following CAR T cell treatment to assess changes mediated by the armored CAR T cells. Ultimately, in Aim 3, we will combine these approaches to generate CAR T cells that recognize GD3 and DLL3 and produce multiple armors. These DADT CAR T cells for SCLC may be suitable for further preclinical testing in preparation for clinical trials beyond the scope of this proposal, representing a novel therapeutic approach to SCLC. Given our robust track record in CAR T cell clinical translation, we fully anticipate having new CAR T cells suitable for clinical trials at the conclusion of funding. Additionally, these novel CAR T cells may be used as tools to explore the interactions between T cells and the SCLC microenvironment. The analysis of changes in SCLC tumors induced by the armored CAR T cells proposed here may reveal novel aspects of SCLC biology and illuminate mechanisms of immune escape and treatment failure in SCLC.

NIH Research Projects · FY 2025 · 2021-03

Abstract This proposal describes a 5-year research career development program focused on a non-canonical role of RB1 loss in bladder cancer. Dr. Qiang Li is an Assistant Professor of Oncology at Roswell Park Comprehensive Cancer Center in the Department of Urology. The proposal builds on the candidate’s previous experience and current research projects using genetically engineered mouse models (GEMMs) and organoids. The proposed experiments and training will enable his transition to independence as a physician scientist in bladder cancer translational research. He will be mentored primarily by Dr. David Goodrich. Dr. Goodrich is an expert in RB1 cancer biology, genetically engineered mouse models, acquired drug resistance and cancer cell plasticity. The training plan includes the following goals: (1) Enhance expertise in preclinical cancer modeling (GEMM and organoids); (2) Probe the molecular mechanisms of bladder cancer cellular plasticity and drug resistance; and (3) Gain expertise in bioinformatic analysis. RB1 mutations are predictive of pathologic response after neoadjuvant chemotherapy in bladder cancer. Other clinical observations suggest that the basal type of bladder cancer is more likely to respond to chemotherapy than the luminal type. However, the biological impact of RB1 loss on molecular subtypes of bladder cancer pathogenesis and chemotherapy response has not been investigated. Newly discovered features of the RB1 pathway in other cancer types suggest that RB1 loss promotes lineage plasticity and acquired therapy resistance. Thus, we hypothesize that RB1 loss promotes bladder cancer progression, metastasis, and cellular plasticity (luminal to basal, and therapeutic resistance). We use two transgenic mouse systems (Uroplakin II driven reverse tetracycline trans-activator, TRE-Cre) to investigate the role of RB1 loss in bladder urothelium by facilitating deletion of tumor suppressor genes (Trp53, Pten, Rb1) under control of doxycycline administration. We engineered doxycycline inducible triple knockout mice Trp53-/-: Pten-/-: Rb1-/- (referred as TKO) and double knockout mice Trp53-/-: Pten-/- (referred as DKO). We propose the following Specific Aims: (1) Define the function of RB1 loss in accelerating tumor progression, metastasis, and cellular plasticity in bladder cancer GEMMs; (2) Dissect the impact of cell-of-origin on bladder tumorigenesis, metastasis and response to chemotherapy in TKO tumors derived from basal cells versus luminal cells. Successful completion of this proposal will allow the candidate to gain valuable technical knowledge and expertise in preclinical modeling of advanced bladder cancer and further his development as an independent physician scientist. The work will also establish experimental models and analysis pipelines that will provide the foundation for the candidate’s independent research program.

NIH Research Projects · FY 2025 · 2021-03

Triple negative breast cancers (TNBCs) do not express estrogen receptor-α (ERα), progesterone receptor (PR), or human epidermal growth factor receptor 2 (HER2), and therefore, none of the targeted drugs currently in use for breast cancer are effective against them. Approximately 60-80% of TNBCs express estrogen receptor-β (ERβ). However, pro- versus anti-tumorigenic capabilities of ERβ remains controversial. Another key molecular characteristic of TNBC is the high frequency (80%) of p53 mutation. In addition to losing tumor suppressor properties and exerting dominant-negative regulation over any remaining wild type p53 (WTp53), mutant p53 also acquires oncogenic gain-of-function. Increasing evidence suggests that not all mutant p53s function similarly. Although ERβ and p53 have been implicated in TNBC pathology, whether p53 has a role in the pro- versus anti-proliferative functional duality of ERβ remains an open question. The long-term goal is to understand and exploit the role of ERβ-p53 crosstalk in breast cancer for the development of better therapeutic strategies. The objective is to study how specific mutations in p53 impinges upon ERβ function in TNBC, with the prediction that specific p53 mutation will determine its role in the ERβ-mutant p53-p73 signaling axis impacting multiple aspects of tumor progression and metastasis. The hypothesis is that ERβ binds to and inhibits both WTp53 and mutant p53, leading to opposite effects on progression and therapeutic response of TNBC to agents such as Tamoxifen (Tam). The rationale for the proposed research is that understanding how ERβ elicits opposite functions in a p53 status-dependent manner will be critical to stratify TNBC patients to repurpose established therapeutic agents such as Tam to treat large percentage of TNBC patients. The specific aims are: (1 Determine the interaction of different p53 mutants with p73 and ERβ in TNBC cells; (2) Analyze the differential effects of p53 mutants on tumor progression, metastasis and therapeutic response in vivo; and (3) Evaluate the clinical significance of the ERβ-p53-p73 signaling axis. In specific aim 1, Isogenic TNBC cells expressing different combinations of ERβ and WT and p53 mutants generated using CRISPR technology and shRNA-mediated conditional knockdown will be used for analyzing the mechanisms underlying the interaction and its impact on cellular functions in vitro and tumor progression in vivo. In specific aim 2, the effect of different p53 mutations on tumor growth and metastasis will be analyzed in vivo. The clinical relevance of these studies will be evaluated using well-characterized patient derived xenografts (PDXs); patient tumor-derived organoids (PDOs); and patient tumor tissues with linked clinical database (specific aim 3). The contribution of this research is expected to be better understanding of the mechanisms by which ERβ-p53-p73 axis in the context of different p53 mutations affects the disease progression and therapeutic response. The proposal is innovative because analyzing the differential effects of different p53 mutations as part of an integrated ERβ-mutant p53-p73 signaling axis is a departure from the status quo and has the potential of developing novel therapeutic strategies against TNBC.

NIH Research Projects · FY 2025 · 2021-02

1 Squamous cell skin cancer (SCC) is the second most common cancer in the US. There are methods available 2 to prevent SCC but are not appropriately used because we lack methods of evaluating their effectiveness in a 3 timely manner. Ultraviolet light (UV) from the sun induces genomic damage which is the most important cause 4 of skin cancer. Early in the process of cancer formation UV causes mutations in cells which result in small 5 clones, clusters of mutated cells. The early mutations that result in the growth of these clones are called 6 clonogenic mutations (CM). CMs are early changes during SCC formation, which appear decades before 7 clinically detectable cancer. Based on previous evidence CMs may signal skin cancer risk and evaluate the 8 efficacy of preventative treatment strategies and sun protection. CM are in low abundance in the skin which 9 make them challenging to detect. However, recent advances in genomic sequencing technology and 10 computational tools allow accurate identification and quantitation of CMs in the skin. Preliminary data has shown 11 that CMS can be accurately detected and used to evaluate sun damaged skin areas. Many of the CMs found in 12 normal sun exposed skin are also common in SCC. The central hypothesis for this application is that CMs are 13 biomarkers of sun induced skin damaged and that CMs can measure how well strategies for skin cancer 14 prevention and preventative treatment work. In the first set of studies we will refine the previously developed 15 panel of sun induced CMs by identifying the most common CMs in sun exposed versus non-sun exposed skin. 16 Subsequent studies will examine the impact of UV exposure on changes in the CM panel and development of 17 skin cancer. These studies will evaluate patterns of CMs and the risk of developing skin cancer. Next, the 18 refined panel of CMs will be used to examine how well treatments designed to prevent skin cancer in heavily 19 sun damaged skin areas reduce CMs and skin cancer formation. In the final set of studies, CMs will be used to 20 evaluate the efficacy of sun protection strategies, such as sunscreens. Sun protection factor (SPF) is widely 21 used to evaluate sunscreens. However, SPF measures reduction in redness of the skin instead of the actual 22 DNA damage. Genomic DNA damage contributes to skin cancer, not “redness” in the skin. Genomic damage 23 can be caused by long term sun damage that does not cause a sunburn. In the final set of studies, CMs are used 24 to evaluate the effectiveness of sunscreens to protect against genomic damage and skin cancer. These studies 25 will change how we evaluate a patient’s risk of developing skin cancer and how we determine the effect of skin 26 cancer prevention. These studies have the potential to shift the focus from treating cancer to preventing the 27 occurrence of skin cancer. This would result in an improvement in cancer care outcomes, improve treatment 28 strategies and ultimately improve the life of individual with a history of sun damage and pre-cancerous lesions. 29 This work focuses on skin cancer but as CMs play a crucial first step in cancer growth in most human cancers 30 our findings and the framework of this study will have implications for the wider field of preventative oncology.

NIH Research Projects · FY 2025 · 2021-01

ABSTRACT This application is designed to address the scientific goals of FOA-PA-19-112. Coronary microvascular disease (CMD) is major sequelae of chest radiotherapy in cancer survivors. Blockade of the larger coronary arteries can be treated by stents or surgical bypass; however, there are no effective therapies currently available to target CMD. This project aims to investigate the novel and previously unexplored mechanisms of ionizing radiation (IR)-induced coronary microvascular injury, and test the beneficial effects of a small molecule, N-acetyl- ser-asp-lys-pro (Ac-SDKP), to counteract these effects. The scientific premise of this proposal is based on our recent studies demonstrating profound endothelial cell injury with marked increase in coronary vascular permeability, and fibrosis, after thoracic radiation exposure in rodents. We also found that radiation-induced CMD was dose-dependently associated with the transcriptional inhibition of claudin-1 (cldn1) expression. Importantly, administration of Ac-SDKP, a thymosin β4-derived endogenous peptide, normalized endothelial cell permeability, reconstituted cldn1, and reduced cardiac fibrosis. Despite its cardioprotective potential, therapeutic application of Ac-SDKP has been challenging due to its short half-life (T1/2 of 4.5 mins) in serum. Therefore, we have developed a stable, liposomal Ac-SDKP (Lip- Ac-SDKP) formulation, which we intend to test for sustained systemic effects. We hypothesize that Ac-SDKP mitigates radiation-induced coronary endothelial damage, and prevents microvascular leakage by inhibiting IR-mediated cldn1 loss. In Aim I, we will examine the uptake efficiency and bioactivity of Lip-Ac- SDKP in the heart and in coronary microvascular endothelial cells. In Aim II, we will examine the effects of Ac- SDKP on endothelial barrier integrity after radiation and study the role of cldn1 in this process. In Aim III, we will determine the effects of Ac-SDKP treatment on radiation-induced coronary blood flow and regional and global cardiac function. We will accomplish these aims by using advanced molecular biology and imaging approaches. We have developed a novel genetically engineered mouse model of endothelial cell-specific cldn1 gain-of- function. We have also developed a cldn1 loss-of-function model using a next generation in vivo siRNA delivery technology. Additionally, we will utilize tumor-bearing syngeneic and xenograft models to examine Ac-SDKP effects after multi-dose thoracic irradiation. This project will provide mechanistic insight on the protective effects of Ac-SDKP against radiation-induced CMD, and will have important therapeutic implications for timely and targeted interventions in cancer patients susceptible to radiotherapy-induced CMD and cardiac ischemia.

NIH Research Projects · FY 2025 · 2021-01

Durable outcomes in subsets of solid cancer patients treated with immune checkpoint inhibitors (ICI) or adoptive cell transfer (ACT) immunotherapy has driven interest in gaining a better understanding of resistance mechanisms that could identify novel druggable targets. Myeloid-derived suppressor cells (MDSC) have emerged as one such barrier based on their ability to inhibit innate and adaptive immunity. While elevated blood MDSC are recognized as a poor prognostic indicator in cancer patients, it is widely thought that the main effector site for MDSC is within the tumor microenvironment (TME). This is in line with the well-documented contact- dependent mechanisms involving short-lived intermediates that underlie known mechanisms of T cell suppression by MDSC. Our published and preliminary studies enlarge on this view, showing that MDSC also function outside the TME through an unprecedented mechanism of intravascular immune suppression. The proposed study builds on our discovery that circulating MDSC initiate contact-dependent cleavage of the L- selectin homing receptor on target T cells that substantially reduces antigen-driven expansion of cytotoxic T cells in lymph nodes. We further found that L-selectin loss coincides with the formation of stable MDSC clusters in the blood of murine tumor models and advanced cancer patients. We term these new structures circulating myeloid cell (CMC) clusters. These observations led us to hypothesize that CMC clusters are an unrecognized functional niche for systemic immune suppression in cancer. To test this hypothesis, we will first determine if blood-borne MDSC target not only naïve T cells, but more broadly attack stem cell memory and central memory T cells and natural killer cells that each require L-selectin for their antitumor activity. Secondly, we will determine if CMC clusters are the active site of L-selectin cleavage by using a multipronged genetic approach to examine L-selectin fate following disruption of MDSC-T cell conjugate formation in vivo. These mechanistic studies center on β2 integrins that are highly expressed by MDSC but are normally inactive on leukocytes in fast-flowing blood under non-pathological conditions. Thirdly, we will examine the translational relevance of CMC clusters during ICI or ACT therapy in a preclinical model in which blood is the primary effector site for MDSC due to their exclusion from the TME (by blocking chemokine-directed trafficking) and spleen (by splenectomy). We will deplete circulating MDSC in this model using antibodies or a clinically relevant liver-X-receptor agonist that induces MDSC-intrinsic apoptosis to establish if blood-borne MDSC contribute to therapeutic resistance. Complementary studies will test the hypothesis that combining the analysis of circulating MDSC with CMC clusters and/or T cell L-selectin will formulate an immunosuppressive signature that predicts response to first-line therapy in metastatic cancer patients. The proposed studies will provide new insights into an unprecedented function of circulating myeloid cells and could lead to the consideration of CMC clusters as a functional biomarker for prognostication or preselection of patients that would benefit from MDSC-depleting regimens during cancer immunotherapy.

NIH Research Projects · FY 2025 · 2020-12

Abstract Head and neck squamous cell carcinomas (HNSCC) are aggressive neoplasms that result in debilitating changes in speech, appearance, and quality of life in humans. Response rates in HNSCC patients have remained relatively unchanged over the years, especially in patients with human papillomavirus (HPV) negative HNSCC highlighting the critical need for novel strategies to meet the therapeutic needs of this patient population. As recognized by PAR-17-245, a critical step in discovering novel therapies for HNSCC patients is the development of tumor models that can reliably recapitulate human disease biology, heterogeneity and therapeutic response. The overall goal of this application is to validate and credential a panel of patient- derived and immunocompetent models of HNSCC. Systematic and in-depth comparison of histopathologic, genomic, and therapeutic response profiles will be performed across multiple preclinical platforms in vitro (organoids) and in vivo (allografts/xenografts). Paired in vitro and in vivo models across these platforms will be used to assess their response to standard of care chemoradiation and immune checkpoint blockade. The models will also be used to screen the activity of novel and FDA-approved agents (‘drug repurposing’) targeting critical pathways implicated in the pathogenesis of HNSCC. The application builds on an existing collaboration between several investigators at Roswell Park Comprehensive Cancer Center with extensive experience and expertise in mammalian models, head and neck cancer, cancer imaging, tumor immunology, genomics, bioinformatics and cancer therapeutics. The project will employ innovative multimodal functional imaging methods to better define and enhance the true translational utility of mammalian models. The proposal will establish a robust panel of credentialed mammalian models of HNSCC and enable development of an integrated preclinical pipeline to assess efficacy of novel therapeutics, identify resistance mechanisms and enable biomarker discovery in HNSCC.

NIH Research Projects · FY 2024 · 2020-07

ABSTRACT Breast cancer rates among African-American (AA) women continue to rise and may further widen breast cancer disparities experienced by AA women, who are more likely to develop aggressive tumor types with a worse prognosis. The biological reasons for these differences remain largely unknown. Recent genome-wide, high-throughput studies highlight an emerging role of long noncoding RNAs (lncRNAs) as a novel class of regulatory molecules in cancer. LncRNAs form an important regulatory layer in global gene expression, and increasing evidence indicates that abnormal expression of specific lncRNAs can contribute to breast cancer carcinogenesis and progression. Studies to date, however, are focused exclusively on EA women, have not commonly used high-throughput next generation sequencing (NGS) to provide unbiased comprehensive profiling, and mostly do not incorporate rigorous normal tissue controls. Motivated by these research gaps and limitations, we recently completed a pilot study of genome-wide lncRNA expression profiling in normal and tumor breast tissues from AA and EA women. LncRNA expression data showed clear tissue- and subtype- specific expression patterns. Importantly, we noted a number of differentially abundant lncRNAs between AA and EA women by estrogen receptor (ER) status. These results indicate that there are unique lncRNA expression patterns in AA tumors, which we hypothesize contributes to aggressive tumor biology and high breast cancer-related mortality. We propose a cost-effective study in a well-characterized cohort of AA breast cancer patients in the Women’s Circle of Health Study (WCHS), which has available tumor tissue blocks, and extensive data on tumor characteristics, clinical outcomes, treatments received, lifestyle factors, and genome- wide DNA methylation. As such, our Specific Aims are: 1) Perform tissue lncRNA expression profiling using total RNA sequencing (1181 AA cases from WCHS and 100 AA controls from Komen Tissue Bank) to determine lncRNAs that are breast cancer- and ER subtype- specific (tumor, ER+, ER- vs. normal) and those associated with clinico-pathological factors (e.g., grade); 2) Examine associations of lncRNA expression levels with breast cancer survival, and use a machine learning approach to identify a combined panel of lncRNAs associated with breast cancer survival; and further perform computational prediction and in vitro functional assays to determine their biological relevance; and 3) Integrate paired data on lncRNA expression and DNA methylation to determine which of these cancer- and prognosis-relevant lncRNAs are regulated by DNA methylation, and explore whether diet, obesity and other lifestyle-related factors are associated with aberrant DNA methylation. This work is novel and findings are anticipated to advance our understanding of molecular mechanisms contributing to aggressive tumor biology and poor cancer prognosis observed in AA women that can be translated into the development of targeted strategies for prevention and therapeutics.

NIH Research Projects · FY 2024 · 2020-07

Project Summary Immune checkpoint receptor (ICR) blockade as a cancer treatment strategy has yielded favorable response rates in some solid tumor types, yet cellular mechanisms of immune escape have resulted in limited efficacy in other tumors, including high grade serous carcinoma (HGSC). Our long-term goal is to understand the link between DNA damage repair (DDR) and HGSC immunogenicity, in order to develop combinatorial treatments for HGSC through DDR inhibition that turn hypoimmunogenic or “cold” HGSC into “hot” tumors with improved responses to immunotherapy. The current knowledge regarding DDR in HGSC has focused on the homologous recombination (HR) pathway. There is a fundamental gap in knowledge regarding the link between other DDR pathways and immune response; the objective of this proposal is to bridge that gap so that we can address the unmet need of treatment strategies for HGSC patients for whom immunotherapy or PARP inhibitor therapy, either alone or in combination, is ineffective. Our central hypothesis is that the inhibition of the non-homologous end joining (NHEJ) pathway is a novel immune priming strategy to sensitize cold HGSC to immune checkpoint blockade. Through the characterization of the role of NHEJ protein DEK in the pathogenesis of HGSC, we have identified a downstream targetable kinase, aurora kinase A (AURKA), that serves as a novel and potent checkpoint against DNA damage and type-I interferon (IFN-I) signaling. To test our central hypothesis, we will focus on three specific aims. Aim 1. To test the hypothesis that the inhibition of NHEJ results in robust IFN-I signaling. Aim 2. To characterize the mechanism by which the inhibition of NHEJ induces immune cell activation. Aim 3. To test the hypothesis that NHEJ inhibition sensitizes cold HGSC to immune checkpoint blockade. The experiments for these aims will be carried out using human HGSC cell lines with known BRCA mutation status and HR-proficiency or HR-deficiency as well as primary, patient derived tumor cells established in cell culture. To characterize the immune changes following NHEJ inhibition in tumor cells and the interplay with host immune cells, we will employ the ID8/p53-/- and ID8/p53-/- /BRCA1-/- mouse cell lines on the C57BL/6 background to allow for studies utilizing established genetic knockouts of critical IFN-I signaling proteins. A second, genetically engineered mouse model of HGSC will also be studied. As outcomes of our work, we expect our findings to contribute to our understanding of the link between NHEJ and tumor immunogenicity. Furthermore, this work will provide critical insights into treatment strategies for HR proficient and BRCA wild-type cancers in which current PARP inhibitor-based therapies are less effective. These contributions will be significant, as they will define the underlying mechanisms and lead to effective combinatorial strategies for the treatment of HGSC and other solid malignancies, allowing us to move forward with immediate translation to clinical trials.

NIH Research Projects · FY 2024 · 2020-07

PROJECT SUMMARY This application describes my research on cell signaling in acute myeloid leukemia (AML) to be performed within the context of a 5-year mentored career development plan. My ultimate goal is to become an independent physician-scientist in the area of laboratory-based academic Hematology/Oncology. Under the guidance of my primary research mentor, Dr. Martin Carroll, at the University of Pennsylvania (UPENN), I have developed a structured training plan consisting of intensive laboratory research, didactics, and oversight by an experienced faculty advisory committee. The proposed research will focus on mechanisms of resistance to FLT3 targeted therapy in AML based on key insights from the clinical trials of these agents conducted at UPENN. Early generation FLT3 inhibitors (FLT3i) were met with lukewarm enthusiasm due to poor target specificity and limited bioavailability. Newer agents have recently been developed with improved activity against FLT3 and clinical efficacy as evidenced by clearance of leukemic blast cells. However, these agents are not curative and many patients respond with differentiation rather than eradication of the leukemic clone. This raises important questions about how FLT3 regulates the differentiation state of leukemia cells. In preliminary studies, I identified a novel pathway downstream of FLT3 inhibition that leads to rapid downregulation of the histone methyltransferase, EZH2. EZH2 is the catalytic component of the PRC2 transcriptional repressor. This research represents the first demonstration of FLT3 regulation of an epigenetic modifier. Loss of EZH2 activity has been linked to increased myeloid differentiation and decreased leukemogenicity, making it an attractive target to study as a potential mechanism for FLT3i-induced differentiation. This proposal aims to demonstrate that PRC2 is necessary for FLT3-ITD leukemogenesis (Aim 1), demonstrate that FLT3i functionally inhibit PRC2 activity (Aim 2), and determine the mechanism of EZH2 downregulation after FLT3 inhibition (Aim 3). These findings will provide insight into the biology of FLT3 signaling and identify improved approaches to induce terminal differentiation of FLT3-ITD leukemia cells. In undertaking the proposed studies and training plan, I will develop the skills and expertise necessary to establish an independent career in translational research.

NIH Research Projects · FY 2024 · 2020-03

The revised P01CA234212 tests novel strategies to promote selective CTL entry into tumor micro- environments (TME) and sensitize “cold” tumors to immunotherapy. Our preclinical and early clinical data demonstrate that the chemokine-modulating (CKM) regimen targeting toll-like receptor-3 (TLR3), type-1 interferons (IFN) and the PGE2 system, selectively enhances CTL numbers but reduces regulatory T(reg) cells in TME, uniformly sensitizing tumors for the therapeutic effectiveness of PD-1 blockers and specialized dendritic cell vaccines (αDC1) in melanoma, colorectal cancer (CRC) and ovarian cancer (OvCa). We will now: 1) Determine local immunologic efficacy of systemically- or locally applied CKMs in cancer patients; 2) Identify the most effective ways of using CKM to enhance antitumor effects DC therapies and PD-1 blockade; and 3) Evaluate the clinical activity of the resulting therapies in PD-1-resistant cancer patients, and identify the most relevant TME correlates of clinical benefit. Project 1 Combinatorial adjuvants promote uniform and selective intratumoral CTL infiltration will test in a Phase IIa trial NCT03403634 whether systemic administration of CKM composed of rintatolimod (TLR3- ligand) IFNα and celecoxib promotes local CTL accumulation in TME of metastatic colorectal cancer (CRC). Magnitude of effects, tumor-selectivity (vs surrounding tissues) and mouse studies will guide the design of the second trial which will evaluate the clinical efficacy of sequential CMK/anti-PD-1 application in CRC patients. Project 2 Local immunotherapy corrects chemokine patterns in OvCa will complete the phase II portion of trial NCT02432378 to test the specificity of local CKM in attracting CTLs (rather than Tregs) to the TME of OvCa patients vaccinated with αDC1 loaded with own tumor cells (αDC1[tumor]) and identify “secondary” mechanisms or treatment resistance. The results will inform preclinical studies and the design of the second trial to determine the clinical activity of sequential treatment with DC[tumor]/CKM followed with PD-1 blockade. Project 3 Chemokine modulation to enhance CD8+ TIL recruitment and cross-priming in the TME is based on our latest observations (NCT01876212) of 57% objective response rate (ORR) to αDC1 vaccine targeting tumor blood vessels (αDC1[TBVA] in the 4 of 7 melanoma patients with primary PD1 resistance and 46% objective clinical benefit overall (6/13 patients). We will now perform phase II trial to evaluate the clinical activity of αDC1[DBVA] combined with systemic CKM (BB-IND16,704) in stage IV melanoma patients with primary PD1 resistance. Using correlative studies and mouse in vivo models, we will develop optimized and potentially simplified vaccines to complement CKM and PD-1 blockade for durable therapeutic benefit. Impact: We will test widely-applicable complementary approaches to promote selective entry of therapeutic CTLs into tumors. Since intratumoral CTL numbers predict survival and therapeutic advantage of checkpoint blockers in multiple cancer types, the results are likely to benefit a broad range of cancer patients.

NIH Research Projects · FY 2026 · 2019-06

In this renewal R01A1 application (parent R01CA240290; “Novel Therapeutic Strategies to Co-Target Undifferentiated Prostate Cancer (PCa) Stem Cells and Bulk PCa Cells”), we continue to develop novel strategies to tackle a pervasive and intertwined problem of clinical significance in PCa, i.e., PCa cell heterogeneity and plasticity. PCa continues to claim a high mortality with >35,000 American men estimated to die from metastatic castration-resistant PCa (mCRPC) in 2024. This high mortality and increased diagnosis of advanced PCa have been linked to PCa cell heterogeneity (e.g., in androgen receptor; AR) and treatment- induced plasticity. It has become clear that the entire spectrum of primary PCa, CRPC and metastases harbors not only AR-expressing (AR+) cells but also a population of PCa cells with little/no AR (AR-/lo), the latter of which often become expanded in mCRPC. Clinically, since introduction of the new generation of antiandrogens such as enzalutamide (Enza), there has been a dramatic increase (up to 20-40%) in AR-/lo mCRPC. Compared to AR+ cells, the AR-/lo PCa cells are understudied and poorly understood. Our focused work over the past 22 years has advanced our understanding of AR-/lo PCa cells and the dynamic relationship between AR-/lo and AR+ cells in PCa ecosystem and during PCa progression. For example, we have demonstrated that the AR-/lo PCa cell population in treatment-naïve tumors harbors PCa stem cells (PCSCs), and we have identified novel targets in AR-/lo PCa cells that have been translated to clinical trials (e.g., NCT03751436). Herein, we have made new and unexpected findings that AR-/lo PCa cells preferentially express genes involved in homologous recombination (HR) based DNA damage response/repair (DDR) including ATR, ATM and BRCA2. Remarkably, many of the same HR genes are also highly expressed in AR-/lo genetically engineered mouse models (GEMMs) of PCa (Pten-/-;Rb1-/-; DKO and Pten-/-Rb1-/-p53-/-; TKO) as well as in patient AR- mCRPC that have undergone Enza-induced plasticity. In contrast, AR+ PCa cells mainly express PARP1 and NHEJ genes and utilize NHEJ for DDR. These exciting preliminary data led us to hypothesize that the AR-/lo PCa cells primarily employ the HR pathway to repair the DNA damage which makes these cells highly vulnerable to HR pathway inhibition. Therefore, combination of ATR inhibitor (ATRi) and ADT/Enza will elicit synthetic lethality and represents a novel therapeutic regimen to holistically target both AR-/lo and AR+ PCa cells. We test this hypothesis with 3 Aims: Aim 1: Test in PCa GEMMs that ATRi combined with castration/Enza will inhibit CRPC and ATRi alone will be highly effective against AR-lo CRPC/NEPC; Aim 2: Evaluate the therapeutic effects of ATRi alone on AR-/lo and ATRi/Enza combination on AR+ human CRPC/PDX models; and Aim 3: Define the DDR pathway dynamics and functions in AR+ and AR-/lo PCa cells using cell line and organoid models as well as patient tumor samples. These aims will be accomplished by therapeutic studies in the DKO/TKO GEMMs and human xenograft/PDX/organoid models combined with mechanistic investigations.

NIH Research Projects · FY 2025 · 2019-03

PROJECT SUMMARY The Roswell Park Comprehensive Cancer Center (Roswell Park) has had a long tradition of contributing to the national cooperative groups over many decades. Those contributions have been scientific, administrative, and participatory (enrollment on clinical trials). Contributions have been consistent and numerous over the years. This extension briefly summarizes the intent of its 4 PD/PI’s (Levine/Puzanov/Lele/Singh) and the membership of Roswell Park to sustain its commitment to the many strengths inherent to cooperative group research: therapeutic advances; a better understanding of the biology of cancer; cancer prevention; means to improve the quality of life of cancer patients; piloting of new drugs and radiologic and radiation and surgical techniques; establishing the relevance of new cellular and molecular advances to the predictive, prognostic, and therapeutic approaches to patients; and the advancement of patient advocacy. Investigators at Roswell Park and its supporting staff fully recognize the essential importance of the timely and accurate submission of data and specimens to achieve these aims

NIH Research Projects · FY 2025 · 2016-09

The current application seeks continued salary support for Hans Minderman, PhD who serves as the Associate Director of the Flow and Image Cytometry Shared Resource (FICSR) of the Roswell Park Comprehensive Cancer Center (Roswell Park). This position was created in 2007 to oversee the research applications of the various imaging (confocal, live cell imaging) and flow cytometry platforms available in the facility and to apply his extensive experience in these areas in collaborations and consultations with the FICSR investigator user base. With over 35 years of experience in clinical and research applications of flow and image cytometry (30 years at Roswell Park), Dr Minderman throughout his career has collaborated with many investigators at the Cancer Center on NIH funded projects. The FICSR is an integral part of Roswell Park’s Cancer Center Support grant (P30CA016056) for which the cancer center Director, Dr Johnson serves as the Principal Investigator. The Tumor Immunology & Immunotherapy (TII) CCSG program, previously co-led by the Unit Director on the original application, Dr Odunsi, is the major user base of the FICSR; but numerous NIH-funded researchers across all 5 CCSG programs actively collaborate and consult with Dr Minderman commensurate with the purpose of the PAR 21-286 funding opportunity. During his first R50 tenure (2016-2021), Dr Minderman’s contributions to Roswell Parks research enterprise have been significant and impactful specifically with regards to the NIH-funded research. As of October 2021, 137 extramural grant applications dependent on FICSR were awarded to Roswell Park faculty, 59 were awarded by NIH, 22 of which to members of the TII program. The cancer center has also made significant investments in the FICSR with the acquisition of a new confocal microscope, two spectral flow cytometry analyzers and a 3rd Fortessa analyzer. With the changes in the R50 program now making the distinction between Laboratory (PAR-20-288)- and Core (PAR-20-287)-based scientists and with the FICSR being an integral part of the Roswell Park CCSG infrastructure, Dr Johnson as the PI of the CCSG grant, will now serve as the Unit Director for the renewal application.

NIH Research Projects · FY 2025 · 2016-01

PROJECT SUMMARY/ABSTRACT Radiation therapy is utilized to treat approximately half of all cancer patients. For some patients, radiation therapy can achieve local tumor control and cure. For other patients, radiation therapy palliates symptoms and alleviates suffering. However, radiation therapy can also cause acute toxicity and late effects that diminish quality of life. The goal of our research program is to widen the therapeutic window of radiation therapy by dissecting mechanisms of normal tissue radiation injury and tumor response to radiotherapy. As I am a sarcoma radiation oncologist, my research group not only studies sarcoma response to radiation therapy, but also sarcoma development and metastasis. During the prior funding period, we adapted CRISPR/Cas9 technology to develop novel genetically engineered mouse models of sarcoma that facilitated lineage tracing of sarcoma clones during tumor progression. This approach identified novel regulators of metastasis, which are potential targets for new cancer therapies. We also used new mouse and in vitro models to discover a novel mechanism for the exquisite radiosensitivity of myxoid liposarcoma: DNA-damage induced phosphorylation of a prion-like domain in the FUS-CHOP fusion protein disrupts interactions with chromatin remodeling complexes that are required for tumor maintenance. We initiated new projects with a novel sarcoma model with high tumor mutation burden that revealed tumor-intrinsic and immune-mediated mechanisms of response and resistance to radiotherapy and immunotherapy. Our findings provided the rationale for a randomized clinical trial in sarcoma patients testing radiation therapy with immune checkpoint inhibition. We also used our genetically engineered mice to uncover mechanisms regulating acute toxicity and late effects of radiation, such as radiation carcinogenesis. Our proposed research will build on the success of our recent work. We will use innovative model systems and sophisticated approaches to make discoveries in three broad areas: 1. Response of sarcomas to radiotherapy 2. Response and resistance of sarcomas to radiation and immunotherapy 3. Normal tissue injury from radiation The knowledge gained from the proposed studies will not only deepen our understanding of radiation and sarcoma biology, but will also inform the design of future clinical trials that aim to widen the therapeutic ratio of radiation therapy to improve the outcome for patients with sarcomas and other cancers.

NIH Research Projects · FY 2025 · 2014-09

Roswell Park is the only NCI-designated Comprehensive Cancer Center in Upstate New York. As a free-standing cancer center, Roswell Park has a long-standing (70+ years) tradition of developing and implementing rigorous educational programs across the training continuum, including the NCI-R25 (R25CA181003, currently in Year 10) supported program that has provided immersive research experiences in cancer sciences and oncology to undergraduate students and clinical trainees (medical/dental/nursing). Since its initial award in 2015, over 300 interns have successfully completed this program. Trainee research productivity during the previous award period has been excellent with R25 interns serving as primary authors or co-authors on a total of 35 publications in cancer-specific journals including 11 publications in high-impact journals. Leveraging these successes over the previous funding period and to meet the evolving needs of cancer research, this R25 renewal application seeks to provide research experiences to college and medical students focused on quantitative sciences and emerging technologies. Specifically, the program will train students on quantitative physical sciences including cancer imaging, genomics/bioinformatics, computational oncology, and emerging technologies including artificial intelligence (AI) based methods. The program leverages the expertise of over 50 faculty members at Roswell Park and their innovative research programs and benefits from access to state-of-the-art technologies in our CCSG-supported resources. Key metrics of programmatic reach (enrollment trends) and outcomes (knowledge gains, career choices) will be continually measured and opportunities for improvement identified based on trainee and mentor feedback to ensure successful training experiences. These experiences will allow interns to appreciate how mathematics, physics, and engineering are integrated with biology in cancer research and allow them to explore how quantitative approaches can accelerate discoveries in oncology and improve outcomes in cancer patients.

NIH Research Projects · FY 2025 · 2013-09

OVERALL SUMMARY – Roswell Park-University of Chicago Ovarian Cancer SPORE This is the revised competing renewal application of the Roswell Park-University of Chicago Ovarian Cancer SPORE. We have made significant progress on the translational objectives and human endpoints of each Individual Research Project (IRP). Our highly successful Developmental Research Program (DRP) and Career Enhancement Program (CEP) have catalyzed new translational ovarian cancer research projects, collaborations, and extramural funding for awardees. Our overarching goal remains unchanged: to conduct multidisciplinary, mechanism-based and collaborative translational research that will have the highest possible impact for patients with ovarian cancer. Because immunotherapies have met with only modest success in ovarian cancer patients, we continue to uniquely focus on novel strategies for generating effective anti-tumor immunity by unraveling immune-resistance mechanisms and identifying novel proteogenomic biomarkers of responsiveness. After significant planning and guidance by our Internal and External Advisory Boards, and Patient Advocate Committee, we have leveraged our highly successful DRP and CEP to propose two bi-directional translational IRPs addressing basic and clinical research questions of importance in ovarian cancer. The new IRP1 evolved as a result of a DRP award and the new IRP2 developed from a CEP award. IRP1 will test an oncolytic virus armed with a CXCR4 antagonist in combination with PDL1 blockade to abrogate tumor immune suppression and limit T cell exhaustion in a randomized Phase I/II clinical trial. IRP2 addresses the completely novel concept of identifying mismatch between immunopeptidomes of ovarian cancer cells versus dendritic cells and leveraging a computational approach of bypassing such mismatch. IRP1 commences with a planned clinical trial; the Phase I/II clinical trial in IRP2 will commence in year 3 following preclinical, IND-enabling translational studies. The program also continues to expand opportunities for new avenues of ovarian cancer translational research via its successful DRP and CEP. The four highly integrated, interconnected shared resource cores – Administration, Biospecimen & Pathology, Biostatistics & Bioinformatics, and Immunogenomics – bring innovative technology and resources to the SPORE and do not duplicate pre-existing shared resources available at Roswell Park or the University of Chicago. This application is strongly supported by over $3.6MM of institutional commitment to ensure its success of conducting highly innovative translational research that changes the clinical practice paradigm in ovarian cancer.

NIH Research Projects · FY 2025 · 2001-07

PROJECT SUMMARY/ABSTRACT The overarching goals of the Tumor Immunology Training Program (TITP) at Roswell Park Comprehensive Cancer Center Graduate Division are to educate, train, and prepare talented pre-doctoral trainees for a profession in cancer immunology research. Driven by significant advances in understanding how the immune system can be weaponized against cancer, future leaders in tumor immunology will not only be well-trained in the discipline but also will have a strong appreciation of the potential translational impact to human cancer biology and therapy. The unique, cancer-focus of the TITP introduces trainees to an advanced didactic and conceptual cancer immunology educational paradigm that encapsulates knowledge spanning the continuum of basic science to clinical application. The uniqueness of this TITP is further enhanced by an academic environment comprised of faculty with multi-disciplinary scientific expertise in basic, translational, and clinical research in tumor immunology that are concentrated within a prestigious NCI-designated Comprehensive Cancer Center. Throughout the program, trainees are continually exposed to a comprehensive portfolio of themes and concepts in tumor immunology, responsible conduct of research, and the principles of rigor and reproducibility. Trainees conduct their doctoral research with TITP mentors with expertise in basic, translational, and clinical tumor immunology. The primary mission of the TITP is to support competitively selected trainees during their third and fourth years of study. The funds requested cover stipends and tuition for 4 pre-doctoral trainees per year. This funding is crucial to continue the upward trajectory experienced for nearly 20 years of NRSA support in the careers of pre-doctoral trainees focused on complex and challenging immunologic questions in cancer. NRSA-supported trainees are prepared for a competitive biomedical career through didactic lectures and concept-driven learning in tumor immunology and biology, grant writing, responsible conduct of research, and rigor and reproducibility. After trainees complete all formal course work and pass a qualifying exam, degree conferral is dependent upon fulfilling a first-authored publication requirement and passing the dissertation defense reflecting the novel and original findings of the trainee’s body-of-work. Trainees who complete this TITP will be well-versed in all major facets of tumor immunology and will have the solid foundation upon which to build cancer-focused careers, guided by a clear vision of its impact on human cancer biology and treatment.