Advancing Treatment Outcomes in Malignant Glioma by Integrating Immunotherapy and Standard of Care using Genetically Engineered Mice that Recapitulate Molecular Feature of Human Glioma

项目摘要/摘要 在这份行政副刊中，候选人沃恩·罗杰斯先生将生成一个检查点配体 PD-L2条件性基因敲除小鼠品系。在母基金的第一个资助期内，使用 单细胞RNA序列，我们发现PD-L1和PD-L2主要表达在 免疫细胞而不是GBM癌细胞，我们的小鼠GBM肿瘤对抗生素具有抵抗力 PD-1关卡封锁治疗。假设对抗PD-1免疫疗法的抵抗力是 与肿瘤免疫细胞高水平表达PD-L1和PD-L2有关。PD-L1和PD-L2是 PD-1的配体和对许多癌症的研究表明，这些配体的高表达使 对抗PD-1免疫治疗的抵抗。在这里，罗杰斯先生对研究基因的影响很感兴趣 消融Pd-L1和Pd-L2。在与Boussiotis实验室的合作下，Charest实验室已经获得 并鉴定为条件PD-L1基因敲除株。在这里，罗杰斯先生的项目是产生一个PD- L2条件基因敲除小鼠研究先天PD-L1和PD-L2失活的后果 瘤内免疫细胞对抗PD-1免疫治疗效果的影响在这样做的过程中，他将学习基础知识 通过产生Cre/lox条件基因敲除小鼠来研究分子生物学和基因组操作 为PD-L2检查点配基的菌株。他将运用他的分子技术知识并学习 构建靶向载体，将其导入小鼠胚胎干细胞，筛选， 分离克隆，筛选克隆，产生嵌合动物和生殖系传递的基因敲除染色。 然后，这种小鼠将与免疫细胞特异性Cre转基因基因一起使用，选择性地 阻断特定免疫细胞中PD-L2和PD-L1的表达，并确定其后果 GBM肿瘤生长及其对抗PD-1检查点阻断治疗的反应

Project Summary/Abstract In this Administrative Supplement, the candidate, Mr. Vaughn Rogers will generate a checkpoint ligand PD-L2 conditional knockout mouse strain. During the first funding periods of the parent grant, using single cell RNA seq, we discovered that both PD-L1 and PD-L2 are predominantly expressed on immune cells rather than on GBM cancer cells and that our mouse GBM tumors are resistant to anti PD-1 checkpoint blockade treatment. The hypothesis is that resistance to anti PD-1 immunotherapy is related to the high levels of PD-L1 and PD-L2 expression in tumor immune cells. PD-L1 and PD-L2 are ligands of PD-1 and studies in many cancers have shown that high expression of these ligands confers resistance to anti PD-1 immunotherapy. Here Mr. Rogers is interested in studying the effects of genetic ablation of PD-L1 and PD-L2. In collaboration with the Boussiotis lab, the Charest lab already obtained and characterized a conditional PD-L1 knockout strain. Here, Mr. Rogers’ project is to generate a PD- L2 conditional knockout mouse to study the consequences of inactivating PD-L1 and PD-L2 in innate intratumoral immune cells on efficacy of anti-PD-1 immunotherapy. In doing so, he will learn the basics of molecular biology and genome manipulation by generating a Cre/lox conditional knockout mouse strain for the PD-L2 checkpoint ligand. He will used his knowledge of molecular techniques and learn new ones to construct a targeting vector, introduce it into mouse embryonic stem (mES) cells, select, isolate clones, screen clones and generate chimeric animals and germline transmitted knockout stain. This mouse will then be used in conjunction with immune cell-specific Cre transgenics to selectively ablate expression of PD- L2 and PD-L1 in specific immune cells and ascertain the consequences on GBM tumor growth and its response to anti PD-1 checkpoint blockade therapy.