Role of WASP/WIP Complex in T Cell Function

WASP/WIP 复合物在 T 细胞功能中的作用

• 批准号: 8321056
• 负责人: RAIF SALIM GEHA
• 金额: $ 37.52万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8321056
• 关键词: Actins; Autoimmunity; B-Lymphocytes; Binding; Blood Cells; Blood Platelets; CD3 Antigens; Cell physiology; Cells; Chemotaxis; Cleaved cell; Colitis; Complex; Cytoskeletal Modeling; Cytoskeleton; Data; Defect; Dendritic Cells; Disease; Drops; Eczema; Functional disorder; Genes; Genetically Engineered Mouse; Homing; Human; Immunologic Deficiency Syndromes; In Vitro; Inbred BALB C Mice; Individual; Infection; Interleukin-2; Knock-in Mouse; Lead; Link; Malignant lymphoid neoplasm; Mind; Mus; Mutation; Normal Range; Pathogenesis; Patients; Peptides; Peripheral; Phenotype; Play; Predisposition; Proliferating; Protein Binding; Proteins; Receptor Signaling; Recurrence; Role; Signal Transduction; Stromal Cell-Derived Factor 1; Structure; Surface; T-Lymphocyte; Testing; Thrombocytopenia; Transgenic Organisms; WASP protein; Wiskott-Aldrich Syndrome; in vivo; molecular pathology; monocyte; mouse Waspip protein; mutant; novel; protein complex; protein function; reconstitution; research study; response

T cell dysfunction contributes to the susceptibility of patients with the Wiskott-Aldrich syndrome (WAS) to infection, autoimmunity and lymphoid malignancy. This project probes the function of WASP and its partner WASP interacting protein WIP in T cells. Most of WASP is complexed with WIP in T cells. WASP levels of drop to ~10% of normal in T cells from WIP KO mice and from WAS patients with mutations that disrupt WIP binding, suggesting that WIP stabilizes WASP. Our Preliminary data show that the hierarchy of T cell dysfunction is WASP/WIP DKOWIP KOWASP KO, bearing in mind that since WASP levels are low in WIP KO T cells the individual contribution of WIP to the WIP KO phenotype is unknown. Our overall hypothesis is that WIP and WASP function in T cells both as a complex and independently. To test our hypothesis we propose to use genetically engineered mice which selectively lack WIP, WASP or both, or in which the WASP/WIP complex is disrupted. We will: 1. Compare T cells from WASP/WIP DKO mice to T cells from WT mice to define the combined role of WIP and WASP. In addition, comparison with WASP KO T cells would define WIP functions that may be independent of WASP. 2. Reconstitute T cells from WIP KO mice with a peptide that consists of the WASP binding domain (WBD) of WIP in order to restore WASP levels, and thus generate WIP deficient/WASP expressing T cells. We will compare T cells from these mice to WT T cells to define the role of WIP. 3. Disrupt the WASP/WIP complex expressing the WBD in WT T cells to test the hypothesis that there is a subset of signaling functions that is dependent on the WASP/WIP complex. 4. Examine the structure and function of T cells from knockin mice that express a WIP mutant that lacks the actin-binding region to define the role of WIP binding to actin in the subcellular localization and function of the WASP/WIP complex. The results of the proposed experiments in the mouse will aid in our understanding of the pathogenesis of WAS in humans and should lead to novel treatments for this disease.

T 细胞功能障碍导致 Wiskott-Aldrich 综合征 (WAS) 患者易感 感染、自身免疫和淋巴恶性肿瘤。该项目探讨了 WASP 及其合作伙伴的功能 T 细胞中的 WASP 相互作用蛋白 WIP。 大多数 WASP 与 T 细胞中的 WIP 复合。 T 细胞中 WASP 水平下降至正常值的约 10% WIP KO 小鼠和来自 WAS 患者的突变破坏了 WIP 结合，表明 WIP 稳定 WASP。我们的初步数据显示，T 细胞功能障碍的层次是 WASP/WIP DKOWIP KOWASP KO，请记住，由于 WIP KO T 细胞中的 WASP 水平较低，因此个体 WIP 对 WIP KO 表型的贡献尚不清楚。 我们的总体假设是 WIP 和 WASP 在 T 细胞中既作为一个复合体又独立地发挥作用。到 检验我们的假设，我们建议使用选择性缺乏 WIP、WASP 或 WIP 的基因工程小鼠 两者都有，或者 WASP/WIP 复合体被破坏。我们将： 1. 比较 WASP/WIP DKO 小鼠的 T 细胞与 WT 小鼠的 T 细胞，以确定 WIP 的联合作用 和黄蜂。此外，与 WASP KO T 细胞的比较将定义 WIP 功能，这些功能可能是 独立于 WASP。 2. 使用由 WASP 结合域 (WBD) 组成的肽重建 WIP KO 小鼠的 T 细胞 WIP 的目的是为了恢复 WASP 水平，从而产生 WIP 缺陷/WASP 表达 T 细胞。我们将 将这些小鼠的 T 细胞与 WT T 细胞进行比较，以确定 WIP 的作用。 3. 破坏 WT T 细胞中表达 WBD 的 WASP/WIP 复合物，以检验以下假设： 依赖于 WASP/WIP 复合体的信令功能子集。 4. 检查来自表达缺乏 WIP 突变体的敲入小鼠的 T 细胞的结构和功能 肌动蛋白结合区，用于定义 WIP 与肌动蛋白结合在亚细胞定位和功能中的作用 WASP/WIP 复合体。 所提出的小鼠实验结果将有助于我们了解疾病的发病机制 存在于人类中，应该会导致这种疾病的新治疗方法。