Chemical Inhibition PTPN22 to boost anti-viral immunity

化学抑制 PTPN22 可增强抗病毒免疫力

• 批准号: 10844076
• 负责人: Robin C. Orozco
• 金额: $ 20.99万
• 依托单位: UNIVERSITY OF KANSAS LAWRENCE
• 依托单位国家: 美国
• 财政年份: 2022
• 资助国家: 美国
• 起止时间: 2022-11-29 至 2026-05-31
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/10844076
• 关键词: Address; Alleles; Amino Acid Substitution; Arginine; Binding; Biological; Biology; C-terminal; Cells; Chemicals; Chronic; Communicable Diseases; Computer Analysis; Data; Disease; Enzymes; Flow Cytometry; Funding; Future; Goals; Health; Immune; Immunology; Individual; Kansas; Length; Libraries; Molecular; Mus; Mutation; Myeloid Cells; N-terminal; PTPN22 gene; Peptides; Phenotype; Production; Protein Tyrosine Phosphatase; Proteins; Role; Specificity; Structure; Tryptophan; Universities; Viral Physiology; Virus Diseases; Work; antiviral immunity; assay development; cytokine; experimental study; in vivo; inhibitor; interest; novel; novel therapeutics; rational design; therapeutic target; tool

The overall goal of this project is to boost anti-viral immunity by inhibiting the protein tyrosine phosphatase non receptor 22 (PTPN22/Ptpn22) encoded enzyme Lyp (PEP in mice). Our hypothesis is that targeting Lyp/PEP C-terminal binding domain rather than the N-terminal enzymatic pocket will; 1- increase specificity and 2- chemically replicate the altered function caused by a commonly expressed PTPN22 allele (rs24766010), which causes an arginine (Arg) to tryptophan (Trp) amino acid substitution in the C-terminal region of Lyp/PEP and disrupts binding. In myeloid cells, N-terminal mutations rendering eenzymatically deadf Lyp/PEP compared to the health-relevant mutation in the C-terminal binding domain (rs24760010) can have different biological impact. We have previously shown that mice harboring this PEP Arg¨Trp Cterminal mutation have enhanced anti-viral immunity and can clear persistent virus infection. Now, we aim to therapeutically target PEP to achieve a comparable phenotype. This project utilizes the use of multiple CoBRE funded and non-CoBRE funded cores at the University of Kansas (KU). In brief, we will employ the protein production group to purify full length, N-terminal, and C- Terminal region of PEP. Then, the Infectious Disease Assay Development (IDAD) core will screen a compound library against these various PEP constructs. After identifying the compound of interest, this data will be submitted to the Computational Chemical Biology (CCB) core to computationally define these potential inhibitors. Further, the CCB core will work to rationally design a peptide based on the available structure of the P1 binding domain of PEP to binding partner Csk. Alongside these computational analyses, my lab will biologically validate the compounds evaluating anti-viral function in various immune cells, such as cytokine production, using spectral flow cytometry in the new Immunology Core. We will also molecularly assess the binding capacity of PEP to common protein partners. Future studies will submit the top compound(s) to the Synthetic Chemical Biology (SCB) core to be chemically stabilized and modified for in vivo experiments addressing anti-viral immunity. Discovery of such compounds results in a novel tool to further understand the role of Lyp/PEP during disease and potentially lead to a new therapeutic to better treat individualfs suffering from chronic infectious diseases.

这个项目的总体目标是通过抑制蛋白酪氨酸磷酸酶来增强抗病毒免疫力。 非受体22(PTPN22/PTPN22)编码酶LYP(小鼠的PEP)。我们的假设是，瞄准 LYP/PEP C-末端结合结构域而不是N-末端酶口袋Will；1-增加特异性 和2-化学复制由通常表达的PTPN22等位基因引起的改变的功能 (Rs24766010)，导致C-末端精氨酸(Arg)到色氨酸(Trp)的氨基酸取代 LYP/PEP区域并破坏结合。在髓系细胞中，N端突变以酶促方式呈现 Deadf LYP/PEP与健康相关的C末端结合域突变(Rs24760010)的比较 会产生不同的生物影响。我们之前已经证明，携带这种PEP Arg？Trp CTr的小鼠 突变可以增强抗病毒免疫，并可以清除持续的病毒感染。现在，我们的目标 在治疗上以PEP为靶点，以获得可比的表型。这个项目使用了多个 科布雷资助和非科布雷资助的核心在堪萨斯大学(KU)。简而言之，我们将采用 蛋白质生产小组提纯PEP全长、N-末端和C-末端区域。那么， 传染病分析开发(IDAD)核心将筛选针对这些不同类型的化合物库 PEP构造。确定感兴趣的化合物后，此数据将提交给 化学生物学(CCB)核心，通过计算定义这些潜在的抑制剂。此外，建行的核心将 致力于根据PEP的P1结合结构域的可用结构合理地设计多肽 绑定合作伙伴CSK。除了这些计算分析，我的实验室将从生物学上验证 评估各种免疫细胞中的抗病毒功能的化合物，如细胞因子的产生，使用 新的免疫学核心中的光谱流式细胞术。我们还将从分子水平评估其结合能力 将PEP提供给普通蛋白质伙伴。未来的研究将把顶级化合物(S)提交给合成的 化学生物学(SCB)核心将进行化学稳定和修改，以用于体内实验解决 抗病毒免疫。这类化合物的发现为进一步了解其作用提供了一种新的工具 LYP/PEP在疾病中的作用，并可能导致一种新的治疗方法，以更好地治疗患有 慢性传染病。