Project III: Engineering immunogenic cell death in melanoma and renal cell carcinoma.

项目 III：工程化黑色素瘤和肾细胞癌中的免疫原性细胞死亡。

• 批准号: 10705788
• 负责人: EMILY H CHENG
• 金额: $ 49.56万
• 依托单位: SLOAN-KETTERING INST CAN RESEARCH
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-09-16 至 2027-08-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10705788
• 关键词: APAF1 gene; Abscopal effect; Affect; Allografting; Antigens; Apoptosis; BAX gene; BCL1 Oncogene; CASP9 gene; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CD8B1 gene; Caspase; Cell Death; Cell Line; Cell Membrane Permeability; Cell Nucleus; Cells; Clear cell renal cell carcinoma; Computer Models; Cytosol; Dimensions; Distant; Ecosystem; Engineering; Epigenetic Process; Evolution; Gene Expression; Genetic Engineering; Genetically Engineered Mouse; Goals; Human; Immune; Immune response; Immune system; Immunofluorescence Immunologic; Immunologic Memory; Immunologics; Immunotherapy; In complete remission; Inflammatory; Inner mitochondrial membrane; Interferon Type I; Label; Lead; Luciferases; Melanoma Cell; Memorial Sloan-Kettering Cancer Center; Mitochondria; Mitochondrial DNA; Modeling; Molecular; Mus; Mutation; Necrosis; Neoplasm Metastasis; Outer Mitochondrial Membrane; Pathway interactions; Patients; Probability; Process; Protein Family; RNA; Recurrent tumor; Renal Cell Carcinoma; Research Personnel; Resolution; Sampling; Stimulator of Interferon Genes; System; Systems Biology; T cell receptor repertoire sequencing; T-Lymphocyte; T-cell receptor repertoire; The Cancer Genome Atlas; Therapeutic; Transplantation; Tumor Immunity; adaptive immune response; anti-CTLA-4 therapy; anti-PD-1; cancer care; cancer cell; cancer type; cell type; cytochrome c; draining lymph node; immune checkpoint blockade; immunogenic; immunogenic cell death; immunogenicity; improved; in vivo; insight; melanoma; mouse model; multiple omics; novel; programs; response; success; survival outcome; tumor; tumor heterogeneity; tumor progression

Project III. Engineering immunogenic cell death in melanoma and renal cell carcinoma Experimental Lead: Cheng Computational Lead: Leslie Experimental Co-Investigator: Li PROJECT SUMMARY Recent approval of immune checkpoint blockade (ICB) in multiple cancer types has revolutionized cancer care, but only a small fraction of patients achieve a durable complete response. Hence, there is an urgent need for novel immunotherapy-based therapeutic strategies to enhance response and offer long-term survival benefits. We hypothesize that induction of immunogenic cancer cell death is one of such strategy. Necroptosis and pyroptosis, two forms of programmed necrosis, are pro-inflammatory and probably immunogenic. By contrast, mitochondrial apoptosis is generally considered immunologically silent. The BCL-2 family proteins are central regulators of mitochondrial apoptosis. BAX and BAK, once activated by BH3-only molecules, trigger mitochondrial outer membrane permeabilization (MOMP), which releases cytochrome c to activate the apoptosome and caspases. Recent paradigm-shifting discoveries have shown that BAX/BAK activation in the absence of caspases can trigger the release of mitochondrial DNA to the cytosol through a process called “mitochondrial inner membrane permeabilization” (MIMP), which activates the cGAS/STING pathway and type I interferon response. These findings indicate that this form of BAX/BAK-dependent, caspase-independent cell death, which we termed “mimptosis”, is highly inflammatory. Here, we engineer models of inducible cell death in murine melanoma and genetically engineered mouse models (GEMMs) of clear cell renal cell carcinoma (ccRCC) to compare the immune response to apoptosis, mimptosis, pyroptosis, and necroptosis in vivo. Our goal is to identify the most immunogenic cell death that not only kills cancer cells but also instructs the immune system to enhance ICB response and elucidate the underlying molecular mechanisms using a comprehensive systems biology approach. In Aim 1, we will characterize the impact of different kinds of engineered immunogenic cell death (ICD) on tumor-immune ecosystem dynamics melanoma using integrated single-cell multiome, paired single-cell RNA and T cell receptor (TCR) sequencing, and multiplexed immunofluorescence. Computational modeling will define immune cell gene expression and regulatory programs in response to ICD to inform therapeutic strategies to improve ICB. In Aim 2, we will determine whether and how immunogenic cell abrogates tumor progression and metastasis and generates immunological memory against tumor rechallenge in GEMMs of ccRCC. A comprehensive multidimensional assessment and computational modeling of tumor- immune ecosystem evolution, T cell epigenetic and functional states, and TCR repertoire in response to different types of cell death will unravel mechanisms that promote anti-tumor immunity. TCGA analysis revealed that low expression of apoptosome components, APAF1 and Caspase-9, is associated with better survival outcomes, suggesting that caspase-independent mimptosis may activate anti-tumor immunity and confer survival benefits in ccRCC. We will assess this hypothesis by performing single-nucleus multiome and MxIF in patient ccRCC samples to characterize tumor-immune cross-talk comparing ccRCC with low vs high apoptosome expression.

项目III黑色素瘤和肾癌的工程化免疫原性细胞死亡 实验领队：程 计算主管：莱斯利 实验联合调查员：李 项目总结 最近批准免疫检查点阻断(ICB)治疗多种癌症类型，使癌症治疗发生了革命性的变化， 但只有一小部分患者获得了持久的完全缓解。因此，迫切需要 以免疫治疗为基础的新治疗策略，以增强反应并提供长期生存益处。 我们假设诱导免疫原性癌细胞死亡是其中一种策略。坏死性下垂和 上睑下垂是程序性坏死的两种形式，是促炎的，可能是免疫原性的。相反， 线粒体凋亡通常被认为是免疫沉默的。BCL-2家族蛋白是核心蛋白 线粒体凋亡的调节者。BAX和BAK一旦被仅BH3分子激活，就会触发 线粒体外膜通透性(MOMP)，释放细胞色素c激活 凋亡体和半胱氨酸天冬氨酸酶。最近的范式转换发现表明BAX/BAK在脑内的激活 半胱氨酸氨基转移酶的缺失可以触发线粒体DNA释放到细胞质中，这个过程被称为 “线粒体内膜通透性”(MIMP)，激活cGAS/STING途径和I型 干扰素反应。这些发现表明，这种形式的Bax/BAK依赖于caspase的细胞 死亡，我们称之为“小儿麻痹症”，具有高度的煽动性。在这里，我们设计了诱导细胞死亡的模型 小鼠黑色素瘤和基因工程小鼠肾透明细胞癌模型 (CcRCC)比较体内对细胞凋亡、小儿麻痹、下垂和坏死性下垂的免疫反应。我们的 目标是识别免疫原性最强的细胞死亡，这种细胞死亡不仅能杀死癌细胞，还能指导免疫 系统，以增强ICB反应和阐明潜在的分子机制使用全面 系统生物学方法。在目标1中，我们将描述不同类型的工程技术的影响 免疫原性细胞死亡(ICD)对黑色素瘤肿瘤免疫生态系统动力学的影响 多组、配对单细胞RNA和T细胞受体(TCR)测序以及多重免疫荧光。 计算模型将定义免疫细胞基因表达和应对ICD的调节程序 为改善ICB的治疗策略提供信息。在目标2中，我们将确定免疫原细胞是否以及如何 阻断肿瘤的进展和转移，并产生免疫记忆以抵御肿瘤的再攻击 在CCRCC的GEMMS中。肿瘤的综合多维评估和计算模型- 免疫生态系统进化，T细胞表观遗传和功能状态，以及TCR谱对不同 细胞死亡的类型将解开促进抗肿瘤免疫的机制。TCGA分析显示，Low 凋亡体成分APAF1和Caspase-9的表达与更好的生存结果有关， 提示caspase非依赖的mimtosis可能激活抗肿瘤免疫并提供生存益处。 在ccrcc。我们将通过对慢性肾细胞癌患者进行单核多组体和MxIF来评估这一假说。 比较ccRCC低表达与高凋亡体表达的样本以表征肿瘤免疫串扰。