Identifying opportunities for extracellular inhibition of ALK signaling

确定细胞外抑制 ALK 信号传导的机会

• 批准号: 10407647
• 负责人: Daryl Ewald Klein
• 金额: $ 38.32万
• 依托单位: YALE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2020
• 资助国家: 美国
• 起止时间: 2020-09-01 至 2025-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10407647
• 关键词: Address; Adopted; Affect; Amino Acids; Antibodies; Architecture; B-Cell Activation; Binding; Biochemical; Biological Assay; Biological Products; Biophysics; Caenorhabditis elegans; Cellular Assay; Cessation of life; Childhood; Clinical; Complex; Complex Analysis; Cysteine; Development; Disease; Drosophila genus; Embryonic Development; Epitope Mapping; Epitopes; Family; Fibronectins; Foundations; Genetic; Genetic study; Glycine; Goals; Homologous Gene; Immunoglobulin Fragments; In Vitro; Invertebrates; Knowledge; LTK gene; Lead; Length; Ligand Binding; Ligands; Link; Logic; Malignant Childhood Neoplasm; Malignant Neoplasms; Maps; Mass Spectrum Analysis; Mediating; Molecular; Monoclonal Antibodies; Mutation; Neuroblastoma; Normal tissue morphology; Oncogenic; Outcome; Pathogenicity; Pathway interactions; Peptides; Peripheral Nervous System; Phage Display; Phosphotransferases; Play; Proteins; Receptor Activation; Receptor Protein-Tyrosine Kinases; Receptor Signaling; Regulation; Regulatory Element; Reporting; Research; Sensory; Signal Pathway; Signal Transduction; Structure; Sum; System; Techniques; Testing; Therapeutic; Therapeutic Intervention; Translating; Vertebrates; anaplastic lymphoma kinase; biophysical properties; design; dimer; disorder risk; extracellular; high risk; inhibitor; insight; member; mutant; neuron development; new therapeutic target; novel therapeutics; overexpression; polyglycine; pre-clinical; rational design; receptor; receptor expression; receptor function; relating to nervous system; response; targeted treatment; tool; tumor; virtual

Project Summary This proposal aims to resolve the molecular basis for activation of an atypical receptor tyrosine kinase (RTK) family – ALK. The ALK family consists of two receptors, ALK and LTK. ALKs are involved in neuronal development and, like other RTKs, are critically important in numerous cancers. In particular, ALK is the oncogenic driver in neuroblastoma – an aggressive and often lethal childhood cancer. Significantly, ALK receptors are distinct from the 19 other RTK families (56 receptors in total). Perhaps most prominent, they do not share the standard RTK architecture. Typically, RTKs are composed of multiple repeats of common domains (e.g. Ig, fibronectin, cysteine-rich) in their extracellular region. On the contrary, ALK’s extracellular ‘sensory’ region is highly enriched in glycines (termed the glycine rich domain, GRD). Only recently the ALK GRD has been shown to be a receiver for Augmentor (AUG) signals. Augmentors are small peptides with ~50 highly conserved amino acids that are sufficient for stimulating ALKs kinase activity. Given ALK’s remarkable molecular design, we expect ALK to have an equally distinguishable mechanism of receptor control – potentially broadening the paradigm for RTK regulation. Despite the importance of this receptor family in cancer, surprisingly little is known about how the receptor functions: 1) there are no reported structures for the GRD, AUG or their complex, and 2) it remains unknown how engagement of ligand translates into receptor kinase activity. The research proposed here focuses on understanding the molecular mechanism of ALK and LTK activation by AUG. With this foundation we hope to reveal how oncogenic dysregulation usurps the control mechanisms in place during normal development. Our goals over the next 10 years are: 1) To develop a complete mechanistic understanding of how ALK receptors perceive Augmentors and transmit their signals, 2) To understand how receptor mutations and matrix changes alter ALK signal transduction in cancer and 3) to determine if ALK’s unique structural and regulatory elements present novel targets for therapeutic interventions. We will study ligand-receptor complexes biochemically and with structural techniques, in order to understand in detail how AUG peptide binding leads to activation of ALKs in the signaling complex. In pursuing studies of ALK’s activation, we will investigate how preclinical monoclonal antibodies and discovered Fabs modulate and inhibit ALK function. In sum, our studies will provide necessary fundamental insight into signaling by this unique family of receptors that does not fit into the canonical RTK schema. Understanding the mechanism of receptor activation in this family will be crucial for designing and deploying effective inhibitors that will need to differ from those targeting other well-studied RTKs. In addition, our findings will reveal the signaling differences induced by common cancer mutations that will steer clinical strategies to approach ALK dependent diseases.

项目概要 该提案旨在解决非典型受体酪氨酸激酶（RTK）激活的分子基础 家庭——ALK。 ALK 家族由两种受体组成：ALK 和 LTK。 ALK 参与神经元 与其他 RTK 一样，它对许多癌症至关重要。特别是，ALK 是 神经母细胞瘤的致癌驱动因素——一种侵袭性且往往致命的儿童癌症。值得注意的是，ALK 受体与其他 19 个 RTK 家族（总共 56 个受体）不同。也许最突出的是，他们确实 不共享标准 RTK 架构。通常，RTK 由公共域的多个重复组成 （例如 Ig、纤连蛋白、富含半胱氨酸）位于细胞外区域。相反，ALK 的细胞外“感觉” 该区域富含甘氨酸（称为富含甘氨酸结构域，GRD）。直到最近 ALK GRD 才 已被证明是增强器（AUG）信号的接收器。增强剂是小肽，具有约 50 个高度 足以刺激 ALK 激酶活性的保守氨基酸。鉴于 ALK 非凡的分子 设计上，我们期望 ALK 具有同样独特的受体控制机制——可能会拓宽 RTK 监管范例。尽管该受体家族在癌症中很重要，但令人惊讶的是，它的作用却很少。 已知受体如何发挥作用：1）没有报道 GRD、AUG 或其复合物的结构， 2) 配体的结合如何转化为受体激酶活性仍然未知。研究 这里提出的重点是了解AUG激活ALK和LTK的分子机制。和 在此基础上，我们希望揭示致癌失调如何篡夺期间的控制机制 正常发育。我们未来 10 年的目标是： 1) 形成完整的机制理解 了解 ALK 受体如何感知增强器并传输其信号，2) 了解受体突变如何 和基质变化改变癌症中的 ALK 信号转导，3) 确定 ALK 的独特结构和 监管要素为治疗干预提供了新的目标。我们将研究配体-受体复合物 生物化学和结构技术，以便详细了解 AUG 肽结合如何导致 信号复合物中 ALK 的激活。在进行 ALK 激活研究时，我们将研究如何 临床前单克隆抗体和发现的 Fab 可调节和抑制 ALK 功能。总而言之，我们的研究 将为这一独特的受体家族的信号传导提供必要的基本见解，该受体不适合 规范的 RTK 模式。了解该家族中受体激活的机制对于 设计和部署有效的抑制剂，这些抑制剂需要不同于针对其他经过充分研究的 RTK 的抑制剂。 此外，我们的研究结果将揭示常见癌症突变引起的信号差异，这些差异将引导 治疗 ALK 依赖性疾病的临床策略。