Combinatorial Targeting of Epigenomic and Microenvironmental Pathways to Suppress MAPK Inhibitor Resistance in Melanoma

表观基因组和微环境途径的组合靶向抑制黑色素瘤中 MAPK 抑制剂耐药性

• 批准号: 10189526
• 负责人: ROGER S LO
• 金额: $ 37.05万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2013
• 资助国家: 美国
• 起止时间: 2013-08-07 至 2024-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/10189526
• 关键词: ATAC-seq; Address; Antibodies; Antigen Presentation; Automobile Driving; BRAF gene; CD8-Positive T-Lymphocytes; Catalogs; Cell Compartmentation; Cell Line; ChIP-seq; Clinical; DNA Methylation; Data; Dependence; Development; Disease Progression; Enhancers; Epigenetic Process; Evolution; Experimental Models; Foundations; Future; Genes; Genome; Genomics; Growth Factor; Human; Immune; Immunocompetent; Immunologic Memory; Immunologics; Immunotherapy; In Vitro; Inflammation; Life; Link; MAP Kinase Gene; MEKs; Melanoma Cell; Mitogen-Activated Protein Kinase Inhibitor; Modeling; Mus; Mutation; PD-L1 blockade; PTPRC gene; Pathway interactions; Patients; Phenotype; Process; Proto-Oncogene Proteins c-akt; Public Health; Publishing; RNA analysis; Recurrence; Resistance; Resistance development; Signal Pathway; Signal Transduction; Sustainable Development; T-Cell Development; T-Lymphocyte; T-cell inflamed; Testing; Time; Tissue Sample; Transforming Growth Factor beta; Translating; Translations; Tumor Tissue; VEGFA gene; Work; Xenograft procedure; addiction; anti-PD-1; anti-PD-L1; anti-PD1 therapy; anticancer research; base; bone morphogenetic protein receptors; cancer therapy; combinatorial; comparative; efficacy evaluation; epigenome; epigenomics; exhaustion; immune checkpoint; immune resistance; in vivo; inhibitor/antagonist; insight; melanoma; member; mutant; neoplastic cell; neutralizing antibody; next generation; non-genomic; patient derived xenograft model; pre-clinical; programmed cell death ligand 1; promoter; resistance mechanism; response; restoration; single-cell RNA sequencing; small molecular inhibitor; therapy resistant; transcription factor; transcriptome; transcriptome sequencing; transcriptomics; tumor; tumor microenvironment; tumor-immune system interactions; unpublished works

ABSTRACT Mutation-targeted small molecular inhibitors and immune checkpoint antibodies have extended the quality and quantity of life for patients with advanced MUTBRAF melanoma. Development of combinations based on these foundational therapies should yield further survival benefits in the near future. The combo of BRAF and MEK inhibitors specifically suppresses a form of resistance driven mainly by genetic alterations that result in MAPK- reactivation and restoration of MAPK-addiction. However, this clinically validated approach to suppress BRAF inhibitor resistance does not address epigenomic and immunologic alterations that drive resistance via MAPK- redundant and CD8 T-cell-depleted tumor states. We propose to uncover combinatorial targets in advanced V600BRAF mutant melanoma by understanding MAPK inhibitor-induced epigenomic and immune-suppressive mechanisms and their interplay. We hypothesize that MAPK inhibitor-induced tumor cell-intrinsic and extrinsic adaptations, which have been linked to innate anti-PD-1 resistance, converge on the elaboration and actions of TGFβ, BMP, or VEGFA in the tumor immune microenvironment. The Lo Lab has a track record of integrating analysis of clinical and experimental resistance evolution to derive translatable preclinical strategies to suppress resistance. We propose (Aim 1) to generate a landscape perspective of epigenome-directed tumor cell-intrinsic resistance evolution by comparative analysis of human melanoma cell lines, patient-derived xenografts and immune-competent murine melanoma models. This analysis seeks to identify master transcriptional factors regulating phenotypic transitions to provide insights into resistance-regulatory growth factors and signaling pathways. In Aim 2, we will test the hypothesis that MAPK inhibitors induce adaptive epigenomic and immune-suppressive processes via elaboration of specific TGFβs or BMPs. The action of these factors on the tumor cells and the tumor microenvironment will be analyzed, respectively, by characterization of cis-regulatory enhancer or super-enhancer modules involving TGFβ/BMP receptor-regulated SMADs and by single-cell RNA-seq of dissociated tumors. In Aim 3, we will evaluate the efficacy and mechanisms of blocking TGFβ, BMP, or VEGFA on top of the MAPKi+aPD-L1 foundation and dissect the influence of the most efficacious combination on single T-cell clonotypes and epigenetic states of activation or exhaustion. Together, these studies will advance translation of our understanding of multi-faceted resistance mechanisms into next-generation combinatorial therapies for advanced melanoma.

抽象的 针对突变的小分子抑制剂和免疫检查点抗体延长了治疗的质量和 晚期 MUTBRAF 黑色素瘤患者的生命质量。在此基础上开发组合 基础疗法应该会在不久的将来带来更多的生存益处。 BRAF 和 MEK 的组合 抑制剂专门抑制一种主要由基因改变驱动的耐药性，导致 MAPK- MAPK 成瘾的重新激活和恢复。然而，这种经过临床验证的抑制 BRAF 的方法 抑制剂耐药性并不能解决通过 MAPK- 驱动耐药性的表观基因组和免疫学改变 冗余和 CD8 T 细胞耗尽的肿瘤状态。我们建议提前发现组合目标 通过了解 MAPK 抑制剂诱导的表观基因组和免疫抑制来了解 V600BRAF 突变黑色素瘤 机制及其相互作用。我们假设 MAPK 抑制剂诱导的肿瘤细胞具有内在和外在的作用 与先天性抗 PD-1 耐药性相关的适应，集中于以下因素的阐述和行动： 肿瘤免疫微环境中的 TGFβ、BMP 或 VEGFA。 Lo 实验室拥有整合临床和实验耐药性进化分析的记录，以得出 抑制耐药性的可转化临床前策略。我们建议（目标 1）生成景观 通过比较分析人类表观基因组导向的肿瘤细胞内在耐药性进化的视角 黑色素瘤细胞系、患者来源的异种移植物和免疫活性小鼠黑色素瘤模型。本次分析 旨在识别调节表型转变的主要转录因子，以提供对以下方面的见解 耐药调节生长因子和信号通路。在目标 2 中，我们将检验 MAPK 的假设 抑制剂通过特定 TGFβ 的阐述诱导适应性表观基因组和免疫抑制过程或 BMP。将分析这些因素对肿瘤细胞和肿瘤微环境的作用， 分别通过涉及 TGFβ/BMP 的顺式调节增强子或超级增强子模块的表征 受体调节的 SMAD 和分离肿瘤的单细胞 RNA-seq。在目标 3 中，我们将评估 在 MAPKi+aPD-L1 基础上阻断 TGFβ、BMP 或 VEGFA 的功效和机制，以及 剖析最有效的组合对单个 T 细胞克隆型和表观遗传状态的影响 激活或耗尽。总之，这些研究将促进我们对多方面理解的转化 晚期黑色素瘤的下一代组合疗法的耐药机制。