CAREER: Engineered multispecific antibodies to interrogate and manipulate immune checkpoint protein trafficking

癌症可以通过结合t细胞表面的特定蛋白质来抑制免疫反应。这种结合会关闭t细胞的肿瘤杀伤功能。被设计用来阻止这种结合的抗体被称为检查点抑制剂。检查点抑制剂允许正常的免疫反应，因此t细胞可以攻击癌细胞。不幸的是，检查点抑制剂治疗仅被批准用于大约12种癌症类型，并且在少数病例中有效。设计和测试通过独特机制阻断免疫检查点途径的抗体是该项目的重点。增强我们对免疫反应动力学的理解将有助于设计更有效的癌症治疗方法。研究项目与教育和推广项目交织在一起。巴尔的摩的高中生将参与互动研究体验，一个研讨会将让当地学生及其家人参与蛋白质工程。与巴尔的摩教师的合作将把工程学整合到K-12课程中，并激励学生在工程学、免疫学和医学的交叉领域追求职业。临床抗体药物竞争性地抑制表面结合免疫检查点蛋白与其激活配体之间的相互作用。这个项目将会针对检查点蛋白到达细胞表面之前的分子运输行为。在单个跨膜蛋白上加入多个表位可以诱导聚集和下调，消除细胞表面的免疫检查点蛋白。这调动了免疫反应。本项目的研究目标是发现和设计针对免疫检查点蛋白的多特异性下调抗体（MSDR Abs），并利用其探索分子运输，评估其对免疫系统的影响。MSDR抗体将使免疫检查点途径的分子表征成为可能，并有助于阐明对当前治疗的潜在耐药性机制。这项提议的工作也将为开发定制设计的蛋白质奠定基础，这些蛋白质制定了一种独特的基于贩运的机制，并具有作为癌症治疗的巨大潜力。此外，本文开发的模块化平台可以很容易地扩展到其他生物系统，用于一系列研究和医学应用。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

Cancers can suppress the immune response by binding specific proteins on the surface of T-cells. This binding turns off the tumor-killing function of the T-cells. Antibodies designed to block that binding are known as checkpoint inhibitors. Checkpoint inhibitors allow the normal immune response so T-cells can attack the cancerous cells. Unfortunately, checkpoint inhibitor treatment is only approved for about a dozen cancer types and is effective in a minority of cases. Designing and testing antibodies that block immune checkpoint pathways through a unique mechanism is the focus of this project. Enhancing our understanding of the dynamics of the immune response will aid in the design of more effective cancer therapeutics. The research program is interwoven with an education and outreach program. Baltimore high school students will engage in interactive research experiences, and a workshop will engage local students and their families in protein engineering. Collaboration with Baltimore teachers will integrate engineering into the K-12 curriculum and inspire students to pursue careers at the interface of engineering, immunology, and medicine.Clinical antibody drugs competitively inhibit interactions between surface-bound immune checkpoint proteins and their activating ligands. This project will instead target the molecular trafficking behavior of the checkpoint proteins before they reach the cell surface. Engaging multiple epitopes on a single transmembrane protein can induce clustering and down-regulation, eliminating immune checkpoint proteins from the cell surface. This mobilizes the immune response. The research objective of this project is to discover and design multi-specific down-regulating antibodies (MSDR Abs) targeting immune checkpoint proteins, use them to explore molecular trafficking, and evaluate their effects on the immune system. MSDR Abs will enable molecular characterization of immune checkpoint pathways and help elucidate the mechanisms underlying resistance to current therapeutics. The proposed work will also lay the foundation for development of custom-designed proteins that enact a distinctive trafficking-based mechanism and have great potential as cancer therapies. Moreover, the modular platform developed herein can be readily extended to other biological systems for a range of research and medical applications.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.