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transcriptome sequencing; transcriptomics; tumor; tumor microenvironment; tumor-immune system interactions; unpublished works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 靶向突变的小分子抑制剂和免疫切解点抗体已扩大质量和 晚期Mutbraf黑色素瘤患者的寿命数量。基于这些的组合开发 基础疗法应在不久的将来产生进一步的生存益处。布拉夫和梅克的组合 抑制剂特别抑制了一种主要由遗传改变驱动的抗性形式，这导致MAPK- MAPK添加的重新激活和恢复。但是，这种抑制BRAF的临床验证方法 抑制剂耐药性不能解决表观基因组和免疫学改变，这些改变通过MAPK-驱动抗性 冗余和CD8 T细胞缺失的肿瘤状态。我们建议在高级中发现组合靶标 通过了解MAPK抑制剂诱导的表观基因组和免疫抑制，V600BRAF突变体黑色素瘤 机制及其相互作用。我们假设MAPK抑制剂诱导的肿瘤细胞中性和外部 适应与先天性抗PD-1抗性有关的适应性，汇聚 肿瘤免疫微环境中的TGFβ，BMP或VEGFA。 LO实验室具有整合临床和实验抗性演化分析的记录，以得出 可翻译的临床前策略以抑制抵抗力。我们建议（目标1）生成景观 通过比较人的比较分析，表观基因组指导的肿瘤细胞中性抗性演变的透视图 黑色素瘤细胞系，患者衍生的Xenographics和免疫能力的鼠类黑色素瘤模型。这个分析 试图确定调查表型过渡的主转录因素，以提供洞察力 电阻调节生长因子和信号通路。在AIM 2中，我们将测试MAPK的假设 抑制剂通过阐述特定的TGFβ或 BMP。这些因素对肿瘤细胞和肿瘤微环境的作用将进行分析， 分别通过表征顺式调节增强子或超增强器模块涉及TGFβ/BMP 受体调节的SMAD和通过分离肿瘤的单细胞RNA-seq。在AIM 3中，我们将评估 在MAPKI+APD-L1基础上阻塞TGFβ，BMP或VEGFA的功效和机制 剖析最有效组合对单个T细胞克隆型和表观遗传态的影响 激活或精疲力尽。这些研究将共同​​推动我们对多方面的理解的翻译 用于晚期黑色素瘤的下一代组合疗法的抗性机制。