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: 10240717
• 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/10240717
• 关键词: Address; Adopted; Affect; Amino Acids; Antibodies; Architecture; B-Cell Activation; Binding; Biochemical; Biological; Biological Assay; 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/antagonist; insight; member; mutant; neuron development; new therapeutic target; novel therapeutics; overexpression; polyglycine; pre-clinical; 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.

项目摘要 这项建议旨在解决激活非典型受体酪氨酸激酶(Rtk)的分子基础。 家庭-好吧。ALK家族由ALK和LTK两个受体组成。碱性磷酸酶参与神经细胞 和其他RTK一样，发育在许多癌症中都是至关重要的。特别是，ALK是 神经母细胞瘤的致癌驱动因素-一种侵袭性的、通常是致命的儿童癌症。值得注意的是，ALK 受体不同于其他19个RTK家族(共56个受体)。也许最突出的是，他们确实这样做了 不共享标准的RTK架构。通常，RTK由公共结构域的多个重复组成 (例如，Ig、纤维连接蛋白、富含半胱氨酸)在它们的胞外区。恰恰相反，碱性磷酸酶的细胞外“感官” 该区域高度富含甘氨酸(称为甘氨酸富集区，GRD)。就在最近，ALK GRD才 已显示为增强器(AUG)信号的接收器。增强剂是一种~50高度的小肽 保守的氨基酸，足以刺激碱性磷酸酶的活性。鉴于ALK的非凡分子 设计，我们希望ALK有一个同样可区分的受体控制机制-潜在地扩大 RTK监管的范例。尽管这个受体家族在癌症中很重要，但令人惊讶的是，几乎没有 已知受体如何发挥作用：1)GRD、AUG或其复合体的结构尚未报道， 2)配体的参与如何转化为受体激酶活性尚不清楚。这项研究 在此提出的重点是在8月之前了解ALK和LTK激活的分子机制。使用 在这个基础上，我们希望揭示致癌失调是如何篡夺了在 正常发育。我们在未来10年的目标是：1)发展一种完整的机械性理解 ALK受体如何感知增强子并传递其信号，2)了解受体突变是如何 而基质的变化改变了癌症中的ALK信号转导，以及3)以确定ALK的独特结构和 调控因素为治疗干预提供了新的目标。我们将研究配体-受体复合体 生物化学和结构技术，以详细了解Aug肽结合是如何导致 激活信号复合体中的ALKS。在继续研究碱性磷酸酶激活的过程中，我们将调查 临床前的单抗和已发现的FABs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所传递的信号提供必要的基本洞察力 规范的RTK架构。了解这一家族中受体激活的机制对于 设计和部署有效的抑制剂，需要不同于针对其他经过充分研究的RTK的抑制剂。 此外，我们的发现将揭示由常见癌症突变引起的信号差异 探讨ALK依赖型疾病的临床策略。