Imaging tumor and T cell responses to metabolic and immune modulation therapy

项目摘要/摘要 项目的目的。此应用程序侧重于使用成像来更好地理解、反转和 监测侵袭性/转移性乳腺癌小鼠模型的免疫抑制和代谢。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， 以及更大的转移潜能。基于GEO和MSKCC cBio门脉的临床分析：I)肿瘤 随着参与乳酸新陈代谢的基因表达增加，它们更具侵略性， 存活，以及ii)糖酵解基因的表达与免疫相关 基因。这些数据支持我们的假设：高比率的肿瘤糖酵解导致富含乳酸的肿瘤。 排除T细胞的微环境(TME)，导致更具侵袭性的肿瘤 形成转移。我们进一步假设，代谢抑制剂逆转T细胞排斥反应将 通过检查点阻断(CTLA-4，PD-1)使免疫耐受的肿瘤成为 对治疗的反应，由于T细胞在肿瘤微环境中的重新繁殖。 在这里，我们建议使用多模式成像来探索代谢和免疫 调制疗法逆转临床相关小鼠细胞毒性T细胞的限制和失活 侵袭性乳腺癌的模型。我们计划：1)表征体内肿瘤乳酸和T细胞的变化 LDH-A和MCT-1/4抑制过程中的渗透；2)评估单一佐剂和新佐剂的反应 联合治疗(体内代谢抑制和“关卡”阻断)；3)验证成像 利用体外实验评估糖酵解生物标志物与T细胞浸润的相关性 免疫荧光(IF)、免疫组织化学(IHC)和荧光辅助细胞分选(FACS)； 评估临床翻译的潜力。翻译的目标有两个：1)在小学阶段诱导回归 和转移性乳腺癌，通过增加肿瘤穿透性和细胞毒性T细胞的效应功能，以及2) 通过无创性影像监测治疗反应。 实验战略。我们已经建立了几种侵袭性乳腺癌的免疫小鼠模型 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细胞再繁殖的大小和时机的“治疗窗口” LDH和MCT药物抑制后的侵袭性小鼠乳腺肿瘤。在目标2中，代谢和 免疫-PET成像将监测TME(由抗LDH和抗MCT治疗引起)的变化 影响T细胞的浸润和功能。基于目标1和目标2的研究，一种“优化的”乳酸抑制治疗 战略(在目标1和2中建立)将与免疫检查点封锁(抗PD1和抗PD1)相结合 CTLA4)，并在AIM 3中进行评估。成像将用于定量肿瘤乳酸，转化为 超极化13C-丙酮酸转化为乳酸，糖酵解(反映在FDG摄取)和CD8+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.