CONTROL AND ACTIVATION OF THE TUMOR NECROSIS FACTOR RECEPTORS

肿瘤坏死因子受体的控制和激活

基本信息

批准号：
10092951
负责人：
JAMES Jeiwen CHOU
金额：
$ 79.69万
依托单位：
HARVARD MEDICAL SCHOOL
依托单位国家：
美国
项目类别：
财政年份：
2020
资助国家：
美国
起止时间：
2020-02-01 至 2025-01-31
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10092951
关键词：
Address Agonist Antibodies Antibody Affinity Apoptosis Autoimmune Autoimmune Diseases Automobile Driving Back Binding Biochemistry Cells Cessation of life Cryoelectron Microscopy Cytoplasmic Tail Death Domain Environment Excision Family Family member Goals Immune Immunotherapy Ligands Lipid Bilayers Malignant Neoplasms Mediating Membrane Molecular Conformation Mutagenesis Negative Staining OX40 Property Protocols documentation Receptor Activation Research Role Signal Transduction Structure Surface Surveys TNFRSF10B gene TNFRSF1A gene TNFRSF1B gene TNFRSF5 gene TNFRSF8 gene Technology Testing Therapeutic antibodies Tumor Necrosis Factor Activation Tumor Necrosis Factor Receptor Yeasts base cancer immunotherapy dimer family structure high throughput technology inhibitor/antagonist member nanobodies prevent receptor receptor function screening therapeutic development

项目摘要

PROJECT SUMMARY The goal of this proposal is to determine the rules governing the activation of the receptors in the tumor necrosis factor receptors super family (TNFRSF), for aiding the development of therapeutic antibodies targeting these receptors. There are genuine needs for in-depth understanding of the mechanism by which these receptors are activated, as many of them are targets for antibody-based immunotherapy. While receptor activation accompanying receptor clustering is a general phenomenon observed for receptors of the TNFRSF, existing approaches of developing agonistic/antagonistic antibodies rely largely on trial and error, and no correlation between antibody affinity and functionality has been observed. The proposed research is based on our recent finding that, for a few members of the TNFRSF, the receptor transmembrane helix (TMH) alone can mediate higher-order receptor clustering to drive downstream signaling and that an important role of the receptor ectodomain in the absence of ligand is preventing the TMH-mediated receptor activation. Thus developing antibodies that specifically modulate the auto-inhibitory state of the ectodomain would yield more selective and efficient activators or inhibitors of the receptors. To investigate the general applicability of this mechanism, we will perform structural and functional studies of receptor TMH oligomerization and pre-ligand association for other members of the TNFRSF. Based on our understanding of the mechanism of receptor autoinhibition, we will develop high-throughput technology for discovering antibodies that activate or inhibit members of the TNFRSF in conformation-specific manner. Specifically, in Aim 1, we will perform a thorough survey of receptor TMH clustering to identify TNFRSF members with TMHs that are capable of forming higher- order interaction network and driving signaling. In Aim 2, we will characterize the pre-ligand association structures for a few representative members of the TNFRSF to understand how receptor ectodomain physically inhibit TMH clustering. In Aim 3, we will broadly survey the basic requirements of receptor activation to test our proposed 3-2 rule of receptor activation. In Aim 4, we will exploit the autoinhibition concept to implement a yeast display technology to screen for nanobodies/Fabs that specifically break or stabilize the pre-ligand ectodomain association, as means of activating or inhibiting the receptors, respectively. The proposed research will significantly advance our mechanistic understanding of receptor activation for the TNFRSF while potentially discovering specific agonistic or antagonistic agents for several TNFRSF members such as DR5, TNFR1, TNFR2, OX40, 4-1BB, CD40, and GITR, all of which are important immunotherapy targets.

项目概要该提案的目标是确定肿瘤中受体激活的规则坏死因子受体超家族 (TNFRSF)，用于帮助开发治疗性抗体靶向这些受体。确实需要深入了解其机制这些受体被激活，因为其中许多是基于抗体的免疫疗法的靶标。而受体伴随受体聚集的激活是 TNFRSF 受体观察到的普遍现象，开发激动/拮抗抗体的现有方法很大程度上依赖于反复试验，并且没有已经观察到抗体亲和力和功能之间的相关性。拟议的研究基于我们最近发现，对于 TNFRSF 的一些成员，仅受体跨膜螺旋 (TMH) 就可以介导高阶受体聚集以驱动下游信号传导，并且在没有配体的情况下，受体胞外域会阻止 TMH 介导的受体激活。因此开发特异性调节胞外域自身抑制状态的抗体将产生更多选择性且有效的受体激活剂或抑制剂。调查本条的普遍适用性机制，我们将进行受体TMH寡聚化和预配体的结构和功能研究 TNFRSF 其他成员协会。基于我们对受体机制的理解自抑制，我们将开发高通量技术来发现激活或抑制的抗体 TNFRSF 的成员以构象特异性的方式。具体来说，在目标1中，我们将彻底执行对受体 TMH 聚类的调查，以识别具有能够形成更高级的 TMH 的 TNFRSF 成员秩序交互网络和驱动信号。在目标 2 中，我们将描述预配体缔合的特征 TNFRSF 的一些代表性成员的结构，以了解受体胞外域的物理结构抑制 TMH 聚类。在目标 3 中，我们将广泛调查受体激活的基本要求，以测试我们的提出了受体激活的3-2规则。在目标 4 中，我们将利用自动抑制概念来实现酵母展示技术，用于筛选特异性破坏或稳定前配体的纳米抗体/Fab 胞外域联合，分别作为激活或抑制受体的手段。拟议的研究将显着推进我们对 TNFRSF 受体激活机制的理解，同时可能会发现几个 TNFRSF 成员（例如 DR5）的特定激动剂或拮抗剂， TNFR1、TNFR2、OX40、4-1BB、CD40 和 GITR，所有这些都是重要的免疫治疗靶点。