Nitric Oxide as a Modifier of Oxidative Stress

Division Of Clinical Sciences - Nci

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Abstract

NOS2 and COX2 shape the tumor microenvironment that leads to poor outcome. We have lead the science for the development of the tumor Altas for NOS2 and COX2. in numerous cancers express elevated levels of NOS2 and COX2 and associated with poor outcome. Our research has uncovered that in p53 mutated cells related to the most aggressive cancers, result is "the major driver of advancement of cancer progression to clinically challenging tumors" leading to most clinically challenging cancers. Our focus initially was on spatial breast cancer, but we have begun to expand liver, lung, ovarian, and prostate cancer. A remarkable finding was that homogenous cells in vitro treated cancer cells with IFNG/IL1 TNF and or LPS tumor and macrophage show that NOS2 and COX2 are expressed in different cells (PMID: 31683257). Furthermore, in tumor tissue NOS2 and COX2 occupied different cellular neighborhoods in the TME (PMID: 37169743PMID: 39356141). This spatial orthogonality implies important intercellular and regional communication in these cellular niches with unique defined roles in progression of the tumor. To address the TME at a single cellular level, we have begun to develop with Optical Microscopy Analysis Laboratory, members of Cancer Innovation Laboratory, Laboratory of Pathology, Human Carcinogenesis, Women Malignancy Branch, Division of Cancer Therapeutic and Diagnostics and CBITT in the NCI as well as external collaborators from Houston Methodist, UPMC, Oxford University; University of Galway, to develop a functional platform to decipher the specific role of these cellular niches and their role in cancer progression and poor outcome. This platform utilizes several multiplexing immunofluorescent techniques (antibodies), different transcriptomics strategies, novel spatial analysis supervised and unsupervised methods that to define cellular neighborhoods that are associated specifically for metastasis, cancer stem cellness, chemoresistance and immunosuppression. The intertwining of this "Redox. Inflammatory Landscape" with progression of cancer has prompted us to explore this complex intercellular communication network in 2-3 dimensions. The use of these spatial platforms has provided unique insight into the science of spatial orientation as a factor that drives poor outcome. Over the last several years we have uncovered novel cellular neighborhoods that are related to tumor-COX2 and tumor NOS2. Using the TCGA and GEO data bases comparing patients with high NOS2/COX2 and poor outcome with low reveal that the microenvironment was unexpected dominated by Type II interferon, IL1 and TNF suggesting an inflamed area. In vitro treatment of breast cancer cell lines shows in single cell RNAseq reveal that tumor NOS and tumor COX2 were strongly associated with only IFNG/TNF/IL1. Furthermore, IL6, IL8, and IL1 as well CD44 were increased only where NOS2/COX2 were high in the T-nse plot. IFNTNF induction of NOS2 and COX2 lead to increase metastasis. But there is a conundrum where cytokines are associated with a positive outcome but is required for induction of NOS2 and COX2 such powerful drivers of poor outcome. The answer is revealed from the spatial orientation of immune response to tumor. The number and location of CD8 Teff cells are considered a major factor in determining outcome in triple negative cancer. Restriction or elimination from the tumor nest is an indication of poor responding tumor. NOS2 driven metastatic niche are proximal to restricted extra tumoral CD8 and lymphoid cells that produce IFN/cytokines inducing in the tumor NOS2 and COX2. This spatial orientation CD8 and tumor NOS2 and COX2 are important in the formation of cancer stem cells and metastasis. Further examination in patient with poor outcome reveal the development of immune deserts which are regions develop lymphocytes often referred as cold tumors. These lymphoid devoid regions from the tumor nest. As the elimination and maturation of these desert, tumor-COX2 has predominate role leading to elimination of leukocytes extratumoral (within 1 mm) revealing tumor nest with no proximal lymphocytes and sporadic tumor COX2. These three types of restricted regions of restricted CD8 cells can be classified in three categorized as I) restricted inflamed (tumor-NOS2/tumor-COX2) II) developing immune desert tumor-COX2 and III) mature immune desert tumor-COX2+ sporadic. These specific regions that are associated with poor outcome suggest a temporal progression from inflamed to immune cold desert. Furthermore, Type I is associated with metastasis, while types II and III are indication of treatment resistance with very low lymphocytes. Verifying these on larger transcriptomic data bases it was found that NOS2/CD8 and NOS2/ IFN ratio show a powerful relationship with survival demonstrating the importance of the antagonism between NOS2 and CD8. Overall, this reveals that spatial orientation and defining specific cellular niches provide novel insight on how tumor microenvironment impacts patient outcome opening up novel approaches to diagnosis and insight into novel therapies. The chemical biology of NO, A Map for Identification of Prevention/post treatment strategies to reduce cancer progression and morality from Cancer. One approach to cancer for limiting the mortality of cancer is to develop prevention that reduces the risk of higher-grade tumors often associated with metastasis as well as prevent recurrence that often involves the development metastasis. The map provided by NO concentration and oncogenic signaling as well as the spatial imaging shows an evolving highly interactive map that can be applied to inflammation landscapes to identity new agents and application of old agents. The oncogenic map shows several pathways that can be targeted to limit metastasis, angiogenesis, tumor growth, and chemoresistance. However, immune system response and polarization has a significant impact on in particular at the time of treatment where efficacy can actually be dampened by strategies involved in prevention. Utilizing these maps formed from NOS2 and COX2 provides a map to discover different agents and strategies. One of the interesting aspects of PP2A was that the phase II compounds found in foods like broccoli and wasabi have potential pharmaceutical uses. Exploring different ADT-NSAID, we show that they had antiangiogenic and antitumor proliferation effect while being 40 times safer than aspirin. APOE mimetics were shown to have a powerful anti-inflammatory impact suggesting the lipid processing APOE phenotype may have a role in the nature regulation. We have developed a 5-phase map in the development of compounds that can be used at different phases (PMID: 34694129). The two basic strategies of prevention and treatment. Where Prevention strategies (not diagnosis) could benefit most patient are faced with a cancer diagnosis. There are 4 phases and different phases in treatment: Pretreatment, at the time of treatment, Early post treatment and late post treatment. We can now more broadly organize agents conventional and alternative in timing strategy for maximum positive patient result. It is important to understand the biology at these stages to proper devise optimal strategies that don't cross purposes. But this opens the idea of new, standard, repurposing and alternative compounds as derived from nutrients that may play a role.