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: 10525194
• 负责人: EMILY H CHENG
• 金额: $ 53.4万
• 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/10525194
• 关键词: APAF1 gene; Abscopal effect; Affect; Allografting; Antigens; Apoptosis; BAX gene; BCL2 gene; CASP9 gene; CD4 Positive T Lymphocytes; CD8-Positive T-Lymphocytes; CD8B1 gene; Caspase; Cell Death; Cell Line; Cell Nucleus; Cells; Clear cell renal cell carcinoma; Computer Models; Cytosol; 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; Process; Protein Family; RNA; Recurrence; 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; base; 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; 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.

项目三。黑色素瘤和肾细胞癌中的工程免疫原性细胞死亡 实验负责人：程 计算主管：Leslie 实验共同研究员：李 项目概要 最近批准的免疫检查点阻断（ICB）治疗多种癌症类型彻底改变了癌症治疗， 但只有一小部分患者获得持久的完全缓解。因此，迫切需要 基于免疫疗法的新型治疗策略可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此类策略之一。坏死性凋亡和 细胞焦亡是程序性坏死的两种形式，具有促炎性并且可能具有免疫原性。相比之下， 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 线粒体凋亡的调节因子。 BAX 和 BAK 一旦被 BH3-only 分子激活，就会触发 线粒体外膜透化 (MOMP)，释放细胞色素 c 以激活 凋亡体和半胱天冬酶。最近的范式转变发现表明，BAX/BAK 激活 半胱天冬酶的缺失会触发线粒体 DNA 通过一个称为胞质溶胶的过程释放到细胞质中 “线粒体内膜透化”（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 依赖性、半胱天冬酶非依赖性细胞 死亡，我们称之为“mimptosis”，是高度炎症性的。在这里，我们设计了诱导细胞死亡的模型 小鼠黑色素瘤和透明细胞肾细胞癌基因工程小鼠模型 (GEMM) (ccRCC) 比较体内对细胞凋亡、有丝分裂、细胞焦亡和坏死性凋亡的免疫反应。我们的 目标是确定最具免疫原性的细胞死亡，不仅杀死癌细胞，还指导免疫系统 系统增强 ICB 反应并使用综合的方法阐明潜在的分子机制 系统生物学方法。在目标 1 中，我们将描述不同类型工程的影响 使用集成单细胞对肿瘤免疫生态系统动力学黑色素瘤进行免疫原性细胞死亡（ICD） 多组学、配对单细胞 RNA 和 T 细胞受体 (TCR) 测序以及多重免疫荧光。 计算模型将定义响应 ICD 的免疫细胞基因表达和调控程序 为改善 ICB 的治疗策略提供信息。在目标 2 中，我们将确定免疫原性细胞是否以及如何 消除肿瘤进展和转移，并产生针对肿瘤再攻击的免疫记忆 在 ccRCC 的 GEMM 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针对不同环境的 TCR 库 细胞死亡的类型将揭示促进抗肿瘤免疫的机制。 TCGA 分析显示，低 凋亡体成分 APAF1 和 Caspase-9 的表达与更好的生存结果相关， 表明不依赖 caspase 的 mimptosis 可能激活抗肿瘤免疫并赋予生存益处 在ccRCC。我们将通过对 ccRCC 患者进行单核多组学和 MxIF 来评估这一假设 样本来表征肿瘤免疫串扰，比较具有低凋亡体表达和高凋亡体表达的 ccRCC。