Dissecting the role and mechanism of EML4-ALK condensates in oncogenic signaling and tumor growth

剖析 EML4-ALK 缩合物在致癌信号和肿瘤生长中的作用和机制

• 批准号: 10634392
• 负责人: Trever G Bivona
• 金额: $ 67.02万
• 依托单位: UNIVERSITY OF CALIFORNIA, SAN FRANCISCO
• 依托单位国家: 美国
• 财政年份: 2023
• 资助国家: 美国
• 起止时间: 2023-04-11 至 2028-03-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10634392
• 关键词: ALK gene; Adaptor Signaling Protein; Applications Grants; Architecture; Biological; Biology; Cancer Control; Cancer Etiology; Cancer Patient; Cells; Chimeric Proteins; Clinical; Complement; Complex; Cytoplasmic Granules; Cytoplasmic Protein; Data; Development; Drug resistance; Epidermal Growth Factor Receptor; Foundations; Fusion Oncogene Proteins; Future; Gene Rearrangement; Genetic; Goals; Growth; Histologic; Lung Adenocarcinoma; MAP Kinase Gene; Malignant Neoplasms; Malignant neoplasm of lung; Membrane; Membrane Lipids; Methodology; Molecular; Mutagenesis; Non-Small-Cell Lung Carcinoma; Oncogenic; Oncoproteins; Outcome; Output; Pathogenicity; Pathway interactions; Patients; Phase; Phenotype; Phosphotransferases; Physical condensation; Proteins; Proteomics; Receptor Protein-Tyrosine Kinases; Recurrence; Role; Set protein; Signal Transduction; Testing; Therapeutic; Variant; Work; anaplastic lymphoma kinase; cancer cell; clinically relevant; design; genetic approach; growth factor receptor-bound protein 2; improved; innovation; insight; kinase inhibitor; mortality; multidisciplinary; mutant; precision medicine; protein expression; recruit; response; tool; treatment strategy; tumor growth

PROJECT ABSTRACT. Lung cancer is the leading cause of cancer mortality worldwide, with non-small cell lung cancer (NSCLC) the predominant histologic subtype of lung cancer and lung adenocarcinoma the major subset of NSCLC. ALK gene rearrangements (e.g., EML4-ALK fusions) are validated targets in NSCLC and current ALK kinase inhibitors yield impressive responses. Despite this clinical progress drug resistance remains a problem that limits patient survival. Improved therapeutic strategies are critical to identify to improve clinical outcomes. We propose an innovative, multidisciplinary, and collaborative project to hopefully improve the survival of NSCLC patients by defining a new mechanism of oncogenic signaling that we uncovered by studying ALK fusion oncoproteins. We aim to capitalize on our discovery of membraneless cytoplasmic protein granules (condensates) as a distinct mechanism of oncogenic kinase signaling in cancer. Our data suggest an emerging paradigm in which certain ALK fusion oncoproteins, as well as other clinically-relevant oncoprotein kinase fusions such as RET fusions, form de novo their own phase separated protein-based subcellular compartment devoid of lipid membranes and utilize higher-order protein assembly as distinguishing principles underlying oncogenic output. These membraneless cytoplasmic protein granules comprise a mode of oncogenic signaling that is different from that of native receptor tyrosine kinase (RTK) signaling and oncogenic, mutant forms of other RTKs such as EGFR, which use classical lipid membrane-based signaling. The pathogenic biomolecular condensates formed by ALK (and other RTK) fusion oncoproteins locally concentrate the RAS activating complex GRB2/SOS1 and activate RAS in a lipid membrane-independent manner. RTK protein granule formation is critical for oncogenic RAS/MAPK signaling output in cells. We identified a set of protein granule signaling components and established structural rules that define ALK protein granule formation. For instance, protein granule formation requires the adaptor proteins GRB2 and SHC, in addition to the ALK fusion oncoprotein. Our findings reveal membraneless, higher-order cytoplasmic protein assembly as a distinct subcellular platform for organizing oncogenic RTK and RAS signaling in cancer. We propose 2 complementary Specific Aims using innovative methodologies to probe condensate biology to understand the role of phase separation in ALK fusion oncogenic signaling. We further define the protein architecture of ALK fusion protein granules and identify the key interacting proteins required for ALK fusion protein granule formation, oncogenic signaling and tumor growth. The proposed studies will establish a mechanistic understanding of RTK fusion condensate biology to lay a firm foundation for the future design of mechanism-based therapeutic strategies to interfere with ALK protein granule assembly per se and that complement conventional ALK-targeted clinical agents, which are ALK kinase inhibitors. This project will provide insight into this distinct form of oncogenic signaling with a focus on ALK, with broader implications for the understanding of condensate and RTK fusion biology and the design of differentiated treatment strategies.

项目摘要。肺癌是世界范围内癌症死亡率的主要原因，非小细胞肺癌（NSCLC） 肺癌（NSCLC）是肺癌的主要组织学亚型，肺腺癌是主要亚型 的NSCLC。ALK基因重排（例如，EML4-ALK融合）是NSCLC和当前ALK的经验证靶点 激酶抑制剂产生令人印象深刻的反应。尽管临床取得了这些进展，耐药性仍然是一个问题 限制了病人的生存改进的治疗策略对于确定改善临床结果至关重要。 我们提出了一个创新的，多学科的，合作的项目，希望提高非小细胞肺癌的生存 通过定义我们通过研究ALK融合发现的致癌信号传导的新机制， 癌蛋白我们的目标是利用我们发现的无膜细胞质蛋白颗粒 （缩合物）作为癌症中致癌激酶信号传导的独特机制。我们的数据显示 某些ALK融合癌蛋白以及其他临床相关癌蛋白激酶融合的范例 例如RET融合物，从头形成它们自己相分离的基于蛋白质的亚细胞区室， 并利用高阶蛋白质组装作为区分致癌的基础原则 输出.这些无膜细胞质蛋白颗粒包含致癌信号传导模式， 不同于天然受体酪氨酸激酶（RTK）信号传导和致癌的其它RTK的突变形式 例如EGFR，其使用经典基于脂质膜的信号传导。病原性生物分子凝聚物 由ALK（和其他RTK）融合癌蛋白形成的RAS激活复合物GRB 2/SOS 1局部集中 并以不依赖于脂质膜的方式激活RAS。RTK蛋白颗粒的形成对于 致癌RAS/MAPK信号输出。我们鉴定了一组蛋白质颗粒信号成分， 建立了定义ALK蛋白颗粒形成的结构规则。例如，蛋白质颗粒的形成 除了ALK融合癌蛋白外，还需要衔接蛋白GRB 2和SHC。我们的发现揭示了 无膜，高阶胞质蛋白组装作为一个独特的亚细胞平台， 致癌RTK和RAS信号在癌症中的作用。我们提出了两个互补的具体目标，利用创新的 探索冷凝物生物学的方法，以了解相分离在ALK融合致癌中的作用 信号我们进一步定义了ALK融合蛋白颗粒的蛋白质结构，并确定了关键的相互作用机制。 ALK融合蛋白颗粒形成、致癌信号传导和肿瘤生长所需的蛋白质。拟议 研究将建立对RTK融合冷凝物生物学的机械理解，为 未来设计基于机制的治疗策略来干扰ALK蛋白颗粒组装， 其补充了作为ALK激酶抑制剂常规ALK靶向临床药物。这个项目 将深入了解这种独特的致癌信号传导形式，重点关注ALK，具有更广泛的意义 了解冷凝物和RTK融合生物学以及设计差异化处理策略。