CONTROL AND ACTIVATION OF THE TUMOR NECROSIS FACTOR RECEPTORS

肿瘤坏死因子受体的控制和激活

• 批准号: 10551737
• 负责人: GERHARD WAGNER
• 金额: $ 77.8万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 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/10551737
• 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; Proliferating; Property; Protocols documentation; Receptor Activation; Receptor Inhibition; Receptors, Tumor Necrosis Factor, Type II; 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; antagonist; cancer immunotherapy; dimer; high throughput technology; inhibitor; member; nanobodies; prevent; receptor; receptor function; screening; therapeutic development

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.

项目摘要 该提案的目的是确定肿瘤中受体激活的规则 坏死因子受体超级家族（TNFRSF），以帮助治疗抗体的发展 针对这些受体。有真正需要深入了解该机制的需求 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 伴随受体聚类的激活是TNFRSF受体的一般现象， 开发激动/拮抗抗体的现有方法主要依赖于反复试验，没有 已经观察到抗体亲和力与功能之间的相关性。拟议的研究是基于 我们最近的发现，对于TNFRSF的一些成员，受体跨膜螺旋（TMH）可以 介导高阶受体聚类以驱动下游信号传导，并表明 在没有配体的情况下，受体外生域正在防止TMH介导的受体激活。因此 开发特异性调节外生域自身抑制状态的抗体将产生更多 受体的选择性有效活化剂或抑制剂。调查此的一般适用性 机制，我们将对受体TMH低聚和领先的结构和功能研究进行结构和功能研究 TNFRSF其他成员协会。根据我们对受体机制的理解 自动抑制，我们将开发高通量技术，以发现激活或抑制的抗体 TNFRSF的成员以特定于构象的方式。具体来说，在AIM 1中，我们将进行彻底的 对受体TMH聚类的调查，以识别具有TMH的TNFRSF成员，这些成员能够形成更高的 订购交互网络和驾驶信号。在AIM 2中，我们将表征前配体协会 TNFRSF的少数代表成员的结构，以了解受体外生域如何物理 抑制TMH聚类。在AIM 3中，我们将广泛调查受体激活的基本要求，以测试我们的 提出的3-2受体激活规则。在AIM 4中，我们将利用自动抑制概念来实施 酵母展示技术以筛选纳米型/晶圆厂的筛选，这些技术专门破坏或稳定前凸台 外生域的关联分别作为激活或抑制受体的手段。提议 研究将大大提高我们对TNFRSF受体激活的机械理解，而 有可能针对多个TNFRSF成员（例如DR5）发现特定的激动剂或拮抗剂 TNFR1，TNFR2，OX40，4-1BB，CD40和GITR，所有这些都是重要的免疫疗法靶标。