Targeting iNOS to inhibit myeloid-derived suppressor cells (MDSC) in melanoma

靶向 iNOS 抑制黑色素瘤中的骨髓源性抑制细胞 (MDSC)

• 批准号: 8507618
• 负责人: Andrew Sikora
• 金额: $ 13.49万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-08 至 2014-05-15
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8507618
• 关键词: Ablation; Address; Affect; Animals; Area; Biometry; Bone Marrow; CD8B1 gene; Cancer Biology; Cancer Immunology Science; Cancer Patient; Cell Count; Cell Culture System; Cell Differentiation process; Cells; Chemicals; Clinical Research; Clinical Trials; Critiques; Cutaneous Melanoma; Data; Data Analyses; Development; Development Plans; Down-Regulation; Environment; Enzymes; Experimental Designs; Funding; Generations; Genetic; Goals; Grant; Growth Factor; Homing; Human; ITGAM gene; Immune; Immune response; Immunization; Immunology; Immunosuppression; Immunosuppressive Agents; Immunotherapy; In Vitro; Infection; Infiltration; Inflammation; Inflammatory; Institution; Knockout Mice; Lead; Leukocytes; Lysine; Maintenance; Malignant Neoplasms; Mediating; Mediator of activation protein; Melanoma Cell; Mentors; Mentorship; Modeling; Molecular; Mus; Myelogenous; Myeloid Cells; Myeloproliferative disease; Natural Killer Cells; Nitric Oxide; Peptides; Physicians; Play; Production; Publications; Publishing; Recruitment Activity; Relative (related person); Research; Resistance; Role; STAT3 gene; Scientist; Secure; Serum; Shock; Skin Cancer; Solid Neoplasm; Spleen; Splenocyte; Suppressor-Effector T-Lymphocytes; T cell response; T-Lymphocyte; Testing; Transgenic Organisms; Translations; Vaccination; Vascular Endothelial Growth Factors; Work; Writing; base; cancer immunotherapy; career; career development; chemokine; clinical material; cytokine; design; experience; human NOS2A protein; improved; in vitro testing; inhibitor/antagonist; lymph nodes; melanoma; migration; mouse model; neoplastic cell; novel; novel strategies; patient population; programs; response; skills; small molecule; sound; trafficking; tumor; tumor progression; vaccine efficacy

DESCRIPTION (provided by applicant): Myeloid-derived suppressor cells (MDSC) are immature myeloid cells described in both tumor-bearing mice and human cancer patients, which play an increasingly recognized role in cancer maintenance, progression, and resistance to immunotherapy. MDSC infiltrate solid tumors, including coetaneous melanoma, and potently inhibit anti-tumor T cell responses through a variety of mechanisms including production of nitric oxide (NO) by inducible nitric oxide synthase (iNOS). While iNOS/NO have not previously been shown to regulate MDSC development, persistence, or localization within the tumor-bearing host, the well-known associations of iNOS with cancer, infection, shock, and other inflammatory states associated with MDSC accumulation make this an attractive hypothesis. Our preliminary data demonstrate that mice bearing syngeneic B16 or MT-RET melanomas accumulate splenic and tumor-infiltrating GR-1+CD11b+ MDSC, and experience a decline in splenic CD4+ and CD8+ T cells. Treatment with the small molecule iNOS inhibitor N6(1-iminoethyl)-L-lysine-dihydrochloride (L-NIL) decreases MDSC infiltration in tumor and spleen, reverses the tumor-mediated decrease in CD4+ and CD8+ splenocytes, and enhances the number of tumor-infiltrating CD4+ and CD8+ T cells. L-nil treatment was also associated with down regulation of activated STAT3 in the tumor, and normalization of elevated levels of VEGF and other inflammatory cytokines observed in sera of tumor-bearing mice. Serum VEGF levels were also suppressed, intratumoral MDSC infiltration reduced, and T cell numbers normalized in tumor-bearing iNOS-/- "knockout" mice, suggesting that host-derived iNOS plays a role in MDSC recruitment and migration. Together, these data suggest that MDSC recruitment and trafficking are controlled by cross-talk in which iNOS expression by tumor-infiltrating myeloid cells, melanoma cells, or both is required for release of soluble factors from the tumor which recruit additional MDSC from the bone marrow and drive their infiltration into the tumor. We further hypothesize that targeted inhibition of iNOS with small-molecule chemical antagonists will enhance the response to anti-tumor vaccination by suppressing the recruitment and suppressive capacity of intratumoral MDSC. In this proposal we aim to characterize the effect of iNOS inhibition on tumor-secreted factors responsible for recruitment and intratumoral homing of MDSC in syngeneic transplantable and spontaneous tumor models. We will seek to determine whether the effects of iNOS inhibition on production, by tumor and myeloid cells, of cytokines known to regulate MDSC are mediated by changes in STAT3 activation. We will determine whether iNOS inhibition decreases intratumoral MDSC numbers by affecting their trafficking, survival, or differentiation, or through a combination of these mechanisms. Finally, we will use a well-established model of adoptive CD8+ T cell transfer and anti-tumor vaccination to determine whether targeted iNOS inhibition boosts the number, effector function, and anti-tumor efficacy of vaccine-induced CD8+ CTL. The above project will be carried out in the context of a comprehensive, mentored career development plan leading to scientific independence. A team of outstanding mentors will provide critical guidance in every aspect of the candidate's scientific development, from hands-on critique of experimental design and data analysis, to publication of results, to acquisition of lab management and grant-writing skills which will lead to independent funding. These personal interactions will be supplemented by formal class work addressing specific areas targeted for improvement (biostatistics, clinical research skills and translation of basic findings to clinical trials). As the candidate is a physician-scientist with access to unique patient populations and clinical material, special emphasis has been placed on providing didactic background and supplemental mentorship in clinical research. The candidate has the benefit of a first class research environment in which immunology and cancer biology are centers of excellence within the institution, and already has secured the confidence and support of his department and the institution. His immediate goal of carrying out and publishing the research proposed in this application will significantly enhance our understanding of the relationship between inflammation and tumor-mediated immunosuppression, and provide a sound platform for establishing an independent research career in cancer immunology and translational cancer biology.

描述(申请人提供)：髓系衍生抑制细胞(MDSC)是在荷瘤小鼠和人类癌症患者中描述的未成熟髓系细胞，在癌症的维持、进展和免疫治疗的抵抗中发挥着越来越多的公认作用。MDSC可侵袭实体瘤，包括皮肤黑色素瘤，并通过多种机制有效地抑制抗肿瘤T细胞反应，包括诱导型一氧化氮合酶(INOS)产生一氧化氮(NO)。虽然iNOS/NO以前没有被证明调节MDSC的发展、持久性或在荷瘤宿主中的定位，但众所周知，iNOS与癌症、感染、休克和其他与MDSC聚集相关的炎症状态的关联使这一假说很有吸引力。我们的初步数据表明，携带同基因B16或MT-RET黑色素瘤的小鼠脾组织和肿瘤浸润性GR-1+CD11b+MDSC积聚，并经历了脾中CD4+和CD8+T细胞的下降。小分子诱导型一氧化氮合酶抑制剂N6(1-亚氨基乙基)-L-赖氨酸-二盐酸盐(L-NIL)治疗可减少肿瘤和脾组织中骨髓间充质干细胞的浸润，逆转肿瘤介导的CD_4~+和CD_8~+T细胞的减少，增加肿瘤侵袭的CD_4~+和CD_8~+T细胞的数量。L-NIL治疗还与下调肿瘤中激活的STAT3，并使荷瘤小鼠血清中升高的血管内皮生长因子和其他炎性细胞因子水平正常化有关。携带iNOS-/-“基因敲除”小鼠的血清VEGF水平也受到抑制，肿瘤内MDSC的浸润减少，T细胞数量恢复正常，表明宿主来源的iNOS在MDSC的募集和迁移中发挥了作用。综上所述，这些数据表明MDSC的募集和运输是由串扰控制的，在串扰中，肿瘤浸润性髓系细胞和/或黑色素瘤细胞需要iNOS的表达，以从肿瘤中释放可溶性因子，从骨髓中招募额外的MDSC，并将其渗透到肿瘤中。我们进一步假设，用小分子化学拮抗剂靶向抑制iNOS将通过抑制肿瘤内MDSC的募集和抑制能力来增强抗肿瘤疫苗的反应。在这项研究中，我们旨在研究iNOS抑制对肿瘤分泌因子的影响，这些因子负责MDSC在同基因移植和自发肿瘤模型中的募集和瘤内归巢。我们将寻求确定iNOS抑制肿瘤和髓系细胞产生已知的调节MDSC的细胞因子的效果是否通过STAT3激活的变化来调节。我们将确定iNOS抑制是否通过影响MDSC的贩运、存活或分化，或通过这些机制的组合来减少肿瘤内MDSC的数量。最后，我们将使用一个成熟的过继CD8+T细胞转移和抗肿瘤疫苗接种模型来确定靶向iNOS抑制是否增加疫苗诱导的CD8+CTL的数量、效应功能和抗肿瘤效果。上述项目将在导致科学独立的全面、有指导的职业发展计划的背景下进行。一支优秀的导师团队将在候选人科学发展的各个方面提供关键指导，从亲身实践对实验设计和数据分析的批评，到结果的发布，再到获得实验室管理和拨款撰写技能，这将导致独立资助。这些个人互动将被正式的课堂作业补充，涉及特定的目标改进领域(生物统计学、临床研究技能和将基本发现转化为临床试验)。由于候选人是一名内科科学家，能够接触到独特的患者群体和临床材料，因此在临床研究中特别强调提供教学背景和补充指导。候选人拥有一流的研究环境，其中免疫学和癌症生物学是该机构的卓越中心，并已获得他所在部门和该机构的信任和支持。他在本申请中提出的开展和发表这项研究的近期目标将显著增强我们对炎症和肿瘤介导的免疫抑制之间关系的理解，并为在癌症免疫学和翻译癌症生物学领域建立独立的研究生涯提供一个良好的平台。