Illuminating B7-H3 Protein Dimerization and Function

阐明 B7-H3 蛋白二聚化和功能

基本信息

批准号：
10179335
负责人：
Margie Nicole Sutton
金额：
$ 7.11万
依托单位：
UNIVERSITY OF TX MD ANDERSON CAN CTR
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：
2020-04-01 至 2023-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10179335
关键词：
Ablation Alleles Amino Acids Attenuated Binding Binding Proteins Biochemical Biological Assay Biological Process Biological Response Modifiers Bioluminescence Biosensor Bladder CD276 gene CD28 gene CTLA4 gene Candidate Disease Gene Cell Surface Proteins Cell Survival Cell membrane Cell surface Cells Colorectal Complement Complex Development Diffuse Dimerization EGF gene Energy Transfer Enterobacteria phage P1 Cre recombinase Epidermal Growth Factor Receptor Family Family member Fluorescence Homeostasis Immune Immune Evasion Immune response Immunomodulators Immunotherapy Integral Membrane Protein KRAS2 gene Kidney Kinetics Ligands Luciferases Lung Lymphocyte Malignant - descriptor Malignant Neoplasms Malignant neoplasm of prostate Methods Molecular Monitor Monoclonal Antibodies Oncogenic Optics Ovarian Pancreas Pathogenesis Pathogenicity Pathway interactions Phenotype Phosphotransferases Physiological Property Protein Analysis Protein Family Proteins Receptor Cell Reporter Research Roentgen Rays Role Signal Transduction Signaling Protein Solid Neoplasm Structure Structure-Activity Relationship Surface System T cell response T-Lymphocyte Techniques Technology Testing Therapeutic Time Training VTCN1 gene Work assay development base cancer cell cancer therapy differential expression dimer extracellular genetic regulatory protein imaging agent immune checkpoint immune checkpoint blockade immunological synapse immunoregulation improved in vivo inhibiting antibody innovation interest luminescence melanoma member mutant novel novel strategies optical imaging overexpression programmed cell death ligand 1 programmed cell death protein 1 protein complex protein function protein protein interaction receptor recombinase-mediated cassette exchange response spatiotemporal targeted treatment tool tumor tumorigenesis

项目摘要

SUMMARY/ABSTRACT Understanding the protein-protein interactions (PPIs) between the B7-family of immune regulatory proteins and their cognate binding partners on the T-cell have led to a new era in cancer treatment, where checkpoint blockade by monoclonal antibodies inhibiting CTLA-4 and PD-1 on the T-cell has revolutionized immunotherapy. These PPIs provide both stimulatory secondary signals to promote and sustain T cell responses, and they also contribute to negative secondary signals that downregulate T cell responses. PD-L1, PD-L2, B7-H3, B7-H4 and HHLA-2 can be expressed on non-hematopoetic cells and tumors, but the role of temporal, as well as, spatial differences in ligand expression and their contributions to pathogenic and protective immune responses have not been fully elucidated. Many B7 family members, similar to other families of cell surface signaling proteins, are known to exist as dimers, related to their signaling state. Thus, characterization of B7 family interactions and oligomerization is essential to understanding the molecular mechanisms by which they alter the cancer cell to confer an oncogenic phenotype. From a candidate gene approach, an interesting and novel target protein is B7- H3, a member of the B7-family, which is overexpressed on the surface of many solid tumors. This proposal aims to develop a bioluminescence-based, sequential resonance energy transfer (BLI-SRET) reporter of PPIs that can be used to define structural features that contribute to B7-H3 dimerization and oligomerization and determine the mechanism of how these interactions promote cancer cell survival and immune evasion. In aim 1, I will develop a BLI-SRET system which can distinguish dimeric or multimeric protein complexes in a spatiotemporal manner. This system will allow us to understand how signals are integrated at a molecular level through protein dimerization and oligomerization, and provide a platform to aid characterizing those interactions that contribute to pathogenesis. In aim 2, I will test the hypothesis that defined regions of B7-H3 contribute to protein clustering and that ablation of these interacting domains will disrupt protein function, both intracellularly and extracellularly, through the complemented immune synapse. As each interacting domain may serve a different downstream function, understanding the similar or different mechanisms that regulate B7-H3-driven tumorigenesis may better enable pathway-specific targeting of therapeutics and improve our understanding of B7-H3 oligomerization and how this relates to its function. The research proposed herein will support my training and development as an independent and productive cancer biologist, allowing me to gain a greater understanding of assay development, genetically encoded optical biosensors, and the application of these tools to understand complex biological processes. Using the longstanding practical expertise and development of optical imaging agents in the Piwnica- Worms group and my detailed understanding of KRAS protein clustering as it relates to cancer development, we aim to develop this system that will allow us to better understand how signals are integrated at a molecular level, and use it to aid in developing therapeutics to target the B7-family of oncogenic immune regulatory proteins.

摘要/摘要了解免疫调节蛋白的B7家庭之间的蛋白质 - 蛋白质相互作用（PPI）和他们在T细胞上的同源绑定伙伴导致了癌症治疗的新时代，检查点封锁了通过T-Cell上抑制CTLA-4和PD-1的单克隆抗体彻底改变了免疫疗法。这些 PPI提供了刺激性的次级信号，以促进和维持T细胞反应，它们也有助于下调T细胞反应的负次级信号。 PD-L1，PD-L2，B7-H3，B7-H4和 HHLA-2可以在非杂质细胞和肿瘤上表达，但时间和空间的作用配体表达的差异及其对病原和保护性免疫反应的贡献没有完全阐明。许多B7家庭成员，类似于其他细胞表面信号蛋白家族，被称为二聚体，与它们的信号状态有关。因此，B7家族互动的表征和低聚对于理解它们将癌细胞改变为分子机制至关重要赋予致癌表型。从候选基因方法中，有趣而新颖的靶蛋白是B7- H3是B7家庭的成员，在许多实体瘤的表面过表达。该提议的目的为了开发PPI的基于生物发光的顺序共振能量传递（BLI-SRET）报道。可用于定义有助于B7-H3二聚体和低聚的结构特征并确定这些相互作用如何促进癌细胞存活和免疫逃避的机制。在AIM 1中，我会开发一个可以区分时空中二聚体或多聚体蛋白复合物的Bli-Sret系统方式。该系统将使我们能够了解如何通过蛋白质在分子水平集成信号二聚体和寡聚化，并提供一个平台来帮助表征那些有助于的相互作用发病机理。在AIM 2中，我将检验以下假设，即定义的B7-H3区域有助于蛋白质聚类这些相互作用的结构域的消融将在细胞内和细胞外破坏蛋白质功能通过补充的免疫突触。由于每个相互作用的域可能会在下游服务功能，了解调节B7-H3驱动肿瘤发生的类似或不同的机制可能会更好启用特定途径的治疗剂，并提高我们对B7-H3低聚和的理解这与其功能如何相关。本文提出的研究将支持我的培训和发展独立和生产性癌症生物学家，使我能够对测定开发有更多的了解，遗传编码的光学生物传感器以及这些工具的应用来理解复杂的生物学过程。使用长期实用的专业知识和Piwnica-中光学成像剂的开发蠕虫小组和我对KRAS蛋白聚类的详细理解与癌症的发展有关，我们旨在开发该系统，以使我们能够更好地了解信号如何在分子级别集成，并使用它有助于开发治疗剂，以靶向致癌免疫调节蛋白的B7家庭。