Development of Cell-Penetrating Monobodies

细胞穿透单体的开发

• 批准号: 2342675
• 负责人: Qing Lin
• 金额: $ 49.8万
• 依托单位: SUNY at Buffalo
• 依托单位国家: 美国
• 项目类别: Standard Grant
• 财政年份: 2024
• 资助国家: 美国
• 起止时间: 2024-03-01 至 2027-02-28
• 项目状态: 未结题
• 来源: https://www.nsf.gov/awardsearch/showAward?AWD_ID=2342675&HistoricalAwards=false
• 关键词: Development; Cell; Penetrating; Monobodies

With the support of the Chemistry of Life Processes (CLP) program in the Division of Chemistry, Professor Qing Lin of SUNY at Buffalo is developing new approaches for designing cell-permeable antibody-like small proteins called monobodies. While monobodies have provided a versatile research tool for dissecting the intracellular signaling pathways, it remains a challenge to design cell-penetrating monobodies for therapeutic applications. The proposed approaches introduce selective chemical crosslinking into monobodies to reinforce their structure, and together with surface supercharging endow cell permeability to the engineered monobodies. This project will provide multidisciplinary research opportunities to underrepresented undergraduate students through collaborations with several minority-focused programs at SUNY at Buffalo. The results of this project will also be integrated into the Experiential Learning Network for undergraduates to enhance their classroom learning.This research project seeks to optimize orthogonally crosslinked monobody scaffolds for the design of bioactive cell-penetrating monobodies targeting the intracellular proteins. By incorporating a genetically encoded noncanonical amino acid called beta-lactam-lysine into a monobody for selective chemical crosslinking, two complementary approaches will be explored to enhance the capability of the monobody scaffold: a computational approach based on deep learning; and an experimental approach based on bacterial surface display. The utility of the optimized monobody scaffolds will be evaluated by designing potent and selective cell-permeable monobodies targeting oncogenic KRAS mutants and examining efficacy. These studies are expected to provide insight into the relationship between protein topology and cytosolic transport, which is crucial for the use of protein-based ligands to study life processes.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.

在化学过程中的化学过程（CLP）方案的支持下，布法罗的纽约州立大学的辛尼教授正在开发新的方法，用于设计可渗透的细胞抗体样小蛋白，称为单体形成。虽然单几幅为剖析细胞内信号通路提供了一种多功能的研究工具，但对于治疗应用设计细胞渗透单体形成仍然是一个挑战。所提出的方法将选择性化学交联引入单基形成，以增强其结构，并与表面增压的endoder endow细胞渗透性一起对工程的单体形成。该项目将通过与布法罗纽约州立大学的几个以少数族裔为中心的计划合作，为代表性不足的本科生提供多学科的研究机会。该项目的结果还将集成到本科生的经验学习网络中，以增强其课堂学习。本研究项目旨在优化针对生物活性细胞渗透单体反应的靶向细胞内蛋白的正交交联的单试支架。通过将称为β-内酰胺 - 赖氨酸的遗传编码的非环氨酸纳入单体，以进行选择性化学交联，将探索两种互补方法，以增强单体支架的能力：基于深度学习的计算方法；以及基于细菌表面显示的实验方法。优化的单体支架的效用将通过设计靶向致癌KRAS突变体并检查功效的有效和可渗透性的单体形成来评估。预计这些研究将洞悉蛋白质拓扑与胞质转运之间的关系，这对于使用基于蛋白质的配体来研究生活过程至关重要。该奖项反映了NSF的法定任务，并被认为是通过基金会的知识分子优点和更广泛影响的审查标准来通过评估来通过评估来支持的。