Imaging tumor and T cell responses to metabolic and immune modulation

项目总结/摘要 项目的目的。该应用程序的重点是使用成像，以更好地了解，扭转， 监测侵袭性/转移性乳腺癌小鼠模型中的免疫抑制和代谢。高 LDH-A和单羧酸转运蛋白1和4（MCT-1和MCT-4）与预后不良有关， 和更大的转移潜力。基于GEO和MSKCC的cBio Portal的临床分析：i）肿瘤 与乳酸代谢相关的基因表达增加的人更具侵略性， 生存，和ii）糖酵解基因的表达与免疫相关的 基因.这些数据支持我们的假设：肿瘤糖酵解的高速率导致了富含乳酸的肿瘤 在排除T细胞的情况下，TME导致更具侵袭性的肿瘤， 形成转移。我们进一步假设，用代谢抑制剂逆转T细胞排斥， 使对使用检查点阻断的免疫调节（CTLA-4、PD-1）有抗性的肿瘤成为 由于肿瘤微环境内T细胞的再增殖，肿瘤细胞对治疗有反应。 在这里，我们建议使用多模态成像来探索代谢和免疫的机制， 调节疗法逆转临床相关鼠中细胞毒性T细胞的限制和失活 侵袭性乳腺癌的模型。我们计划：1）表征肿瘤乳酸和T细胞的体内变化 LDH-A和MCT-1/4抑制期间的浸润; 2）评价对佐剂和新佐剂单次给药的应答 和联合治疗（体内代谢抑制和“检查点”阻断）; 3）验证成像 结果通过使用离体方法评估糖酵解生物标志物和T细胞浸润之间的相关性来获得 免疫荧光（IF）、免疫组织化学（IHC）和荧光辅助细胞分选（FACS）;以及4） 评估临床翻译的潜力。翻译的目标是双重的：1）诱导原发性视网膜病变的消退， 和转移性乳腺癌，通过增加细胞毒性T细胞的肿瘤穿透和效应功能，和2） 通过非侵入性成像监测治疗反应。 实验策略。我们建立了几种侵袭性乳腺癌的小鼠模型， 有能力的宿主动物。在目标1中，体外和体内研究将确定肿瘤细胞代谢如何影响T 细胞功能，并确定优化T细胞再增殖的幅度和时间的“治疗窗口” LDH和MCT药理学抑制后侵袭性小鼠乳腺肿瘤的生长。在目标2中，代谢和 免疫PET成像将监测TME的变化（由抗LDH和抗MCT治疗诱导） 影响T细胞浸润和功能。基于目标1和2研究，“优化”乳酸抑制治疗 免疫策略（在目的1和2中建立）将与免疫检查点阻断（抗PD 1和抗PD 2）相结合。 CTLA 4），并在目标3中进行评估。成像将用于定量肿瘤乳酸盐， 超极化13 C-丙酮酸盐至乳酸盐、糖酵解（反映在FDG摄取中）和CD 8 + T细胞的运输。

PROJECT SUMMARY/ABSTRACT Purpose of the project. This application focuses on the use of imaging to better understand, reverse, and monitor immune suppression and metabolism in murine models of aggressive/metastatic breast cancer. High LDH-A and monocarboxylate transporters 1 and 4 (MCT-1 and MCT-4) have been linked to poor prognosis, and greater metastatic potential. Based on clinical analyses of GEO and MSKCC’s cBio Portal: i) tumors with increased expression of genes involved in lactate metabolism are more aggressive and have poor survival, and ii) there is an inverse correlation between the expression of glycolysis genes and immune-related genes. These data support our hypothesis: that high rates of tumor glycolysis leads to a lactic acid-rich tumor microenvironment (TME) with the exclusion of T cells, resulting in more aggressive tumors with a propensity to form metastases. We further hypothesize that reversal of the T cell exclusion with metabolic inhibitors will render tumors that are resistant to immune modulation with checkpoint blockade (CTLA-4, PD-1) to be responsive to the treatment, due to repopulation of T cells within the tumor microenvironment. Here, we propose to use multimodal imaging to explore the mechanisms by which metabolic and immune modulation therapy reverse the restriction and inactivation of cytotoxic T cells in clinically relevant murine models of aggressive breast cancer. We plan to: 1) characterize in vivo changes in tumor lactate and T cell infiltration during LDH-A and MCT-1/4 inhibition; 2) evaluate the responses to adjuvant and neo-adjuvant single and combination therapy (metabolic inhibition and “checkpoint” blockade in vivo); 3) validate the imaging results by assessing correlations between glycolytic biomarkers and T cell infiltration using ex vivo immunofluorescence (IF), immunohistochemistry (IHC), and fluorescence-assisted cell sorting (FACS); and 4) assess the potential for clinical translation. The translational goal is two-fold: 1) to induce regression in primary and metastatic breast cancer, by increasing tumor penetration and effector function of cytotoxic T cells, and 2) to monitor treatment response by non-invasive imaging. Experimental Strategy. We have established several murine models of aggressive breast cancer in immune competent host animals. In Aim 1, in vitro and in vivo studies will define how tumor-cell metabolism affects T cell function, and identify a “therapeutic window” for optimizing the magnitude and timing of T cell repopulation of aggressive murine breast tumors following LDH and MCT pharmacologic inhibition. In Aim 2, metabolic and immune-PET imaging will monitor how changes in the TME (induced by anti-LDH and anti-MCT treatment) affect T cell infiltration and function. Based on Aim 1 and 2 studies, an “optimized” lactate inhibition treatment strategy (established in Aims 1 and 2) will be combined with immune checkpoint blockade (anti-PD1 and anti- CTLA4) and assessed in Aim 3. Imaging will be used to quantitate tumor lactate, the conversion of hyperpolarized 13C-pyruvate to lactate, glycolysis (reflected in FDG uptake) and the trafficking of CD8+ T cells.