Chemical Inhibition PTPN22 to boost anti-viral immunity

该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（PTPN 22/Ptpn 22）编码的酶Lyp（小鼠中的PEP）。我们的假设是， Lyp/PEP C-末端结合结构域而不是N-末端酶口袋将：1-增加特异性 和2-化学复制由通常表达的PTPN 22等位基因引起的改变的功能 （rs 24766010），其导致C-末端中精氨酸（Arg）至色氨酸（Trp）氨基酸取代 Lyp/PEP区域并破坏结合。在髓系细胞中，N-末端突变使酶促 deadf Lyp/PEP与C-末端结合结构域（rs 24760010）中的健康相关突变相比 会产生不同的生物学影响。我们先前已经证明，携带这种PEP Arg？Trp C末端的小鼠， 突变增强了抗病毒免疫力，可以清除持续性病毒感染。现在，我们的目标是 以PEP为治疗靶点，以获得可比较的表型。该项目利用多个 堪萨斯大学（KU）的CobRE资助和非CobRE资助的核心。简而言之，我们将采用 蛋白生产组纯化PEP的全长、N-末端和C-末端区域。然后 传染病检测开发（IDAD）核心将针对这些不同的化合物库筛选化合物库。 PEP构建体。在确定感兴趣的化合物后，该数据将提交给计算 化学生物学（CCB）的核心计算定义这些潜在的抑制剂。此外，建行核心将 基于PEP的P1结合结构域的可用结构， 结合伴侣Csk.除了这些计算分析，我的实验室还将从生物学上验证 评价在各种免疫细胞中的抗病毒功能如细胞因子产生的化合物， 光谱流式细胞术在新的免疫学核心。我们还将从分子水平评估 PEP与普通蛋白质伴侣的结合未来的研究将提交顶级化合物给合成 化学生物学（SCB）核心是化学稳定和修改的体内实验， 抗病毒免疫这些化合物的发现为进一步了解药物的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

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.