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

通过使用重现人类胶质瘤分子特征的基因工程小鼠整合免疫疗法和护理标准来提高恶性胶质瘤的治疗效果

• 批准号: 10377182
• 负责人: VASSILIKI A BOUSSIOTIS
• 金额: $ 4.53万
• 依托单位: BETH ISRAEL DEACONESS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2018
• 资助国家: 美国
• 起止时间: 2018-07-01 至 2023-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10377182
• 关键词: Ablation; Administrative Supplement; Affect; Animals; Cell physiology; Cells; Collaborations; Funding; Genes; Genetic; Genetically Engineered Mouse; Genome; Glioblastoma; Glioma; Human; Immune; Immunotherapy; Knock-out; Knockout Mice; Knowledge; Learning; Ligands; Malignant Glioma; Malignant Neoplasms; Molecular; Molecular Biology; Mouse Strains; Mus; Resistance; Stains; Techniques; Transgenic Organisms; Treatment outcome; Tumor-infiltrating immune cells; anti-PD-1; cancer cell; conditional knockout; embryonic stem cell; immune checkpoint blockade; immune function; interest; parent grant; programmed cell death ligand 1; programmed cell death protein 1; response; single-cell RNA sequencing; standard of care; tumor; tumor growth; vector

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.

项目总结/摘要 在这份行政补充材料中，候选人沃恩·罗杰斯先生将产生一个检查点配体， PD-L2条件性基因敲除小鼠品系。在父母补助金的第一个供资期内， 通过单细胞RNA测序，我们发现PD-L1和PD-L2都主要表达在 免疫细胞而不是GBM癌细胞，我们的小鼠GBM肿瘤对抗 PD-1检查点阻断治疗。假设抗PD-1免疫疗法的抗性是 与肿瘤免疫细胞中PD-L1和PD-L2的高水平表达有关。PD-L1和PD-L2是 PD-1的配体和许多癌症中的研究表明，这些配体的高表达赋予了 抗PD-1免疫疗法的抗性。在这里罗杰斯先生感兴趣的是研究基因的影响， PD-L1和PD-L2的消融。通过与Boussiotis实验室的合作，Charest实验室已经获得了 并表征了条件性PD-L1敲除菌株。在这里，罗杰斯先生的项目是产生一个PD- L2条件性基因敲除小鼠，以研究先天性免疫缺陷病毒中PD-L1和PD-L2失活的后果。 肿瘤内免疫细胞对抗PD-1免疫疗法的功效的影响。在这样做的过程中，他将学习基础知识 通过产生Cre/lox条件敲除小鼠， PD-L2检查点配体的菌株。他将利用他的分子技术知识 构建靶向载体，将其导入小鼠胚胎干细胞（mES细胞），选择， 分离克隆、筛选克隆并产生嵌合动物和种系传递的敲除染色。 然后将该小鼠与免疫细胞特异性Cre转基因物结合使用，以选择性地 消除特定免疫细胞中PD-L2和PD-L1的表达，并确定对 GBM肿瘤生长及其对抗PD-1检查点阻断疗法的响应。