In vivo role of BTK-mediated inhibition of Wnt/b-catenin signaling during hematop

BTK 介导的 Wnt/b-catenin 信号抑制在 hematop 过程中的体内作用

• 批准号: 8514128
• 负责人: Richard Goff James
• 金额: $ 24.9万
• 依托单位: SEATTLE CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-08-14 至 2015-05-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8514128
• 关键词: Adult; Amino Acids; Animal Disease Models; Area; Attention; Award; B-Lymphocytes; Basic Science; Biological Models; Blood Cells; Bone Marrow; Bone Marrow Transplantation; Cancer cell line; Cell Culture Techniques; Cell Line; Cells; Chemicals; Clinical; Colorectal Cancer; Data; Development; Diagnosis; Embryo; Engraftment; Event; Flow Cytometry; Galactosidase; Gene Targeting; Genetic Suppression; Hematopoiesis; Hematopoietic System; Hematopoietic stem cells; Home environment; Human; Investigation; Knowledge; Large Intestine Carcinoma; Learning; Mass Spectrum Analysis; Mediating; Methods; Mission; Modeling; Molecular; Monitor; Mus; Mutation; National Heart, Lung, and Blood Institute; Pathway interactions; Patients; Peptides; Phosphorylation; Positioning Attribute; Proteomics; Regulation; Reporter; Research Personnel; Role; Sampling; Signal Pathway; Signal Transduction; Small Interfering RNA; Sorting - Cell Movement; Spleen; Stable Isotope Labeling; Stem cells; Symptoms; TEC Protein Tyrosine Kinase; Techniques; Testing; Training; Umbilical Cord Blood; Umbilical cord structure; Work; X-Linked Agammaglobulinemia; Zebrafish; abstracting; base; cancer therapy; chemical genetics; gain of function; improved; in vitro Model; in vivo; interest; kinase inhibitor; loss of function; post-doctoral training; reconstitution; research study; small molecule; tissue culture

Project Abstract Currently I am using chemical genetics, siRNA screens and mass spectrometry- based proteomics to probe the Wnt/ -catenin signaling pathway. Using these techniques we identified Tec kinases as negative regulators of Wnt/ -catenin signaling. Because mutations in the Tec kinase BTK are responsible for X-linked agammaglobulinemia, we sought to corroborate our original findings in B cells. We found that Tec kinases also negatively regulate Wnt/ -catenin signaling in B cells in culture. This work has led me to the hypothesis that the interplay of Tec kinases and Wnt signaling will have a significant role in hematopoiesis in vivo. I am applying for the Pathway to Independence Award in order to extend my postdoctoral training so that I can learn about animal models of disease and hematopoiesis and gain practical knowledge of how to dissect mice and collect bone marrow, how to perform murine bone marrow transplantation experiments and how to analyze these experiments by flow cytometry. As outlined in the proposal, I will use all of these methods in order to test my hypothesis. As an independent investigator I plan to exploit my unique position at the intersection of proteomics and hematopoiesis to explore the molecular mechanisms of signal transduction in cellular differentiation. Not only would this direction allow me to fully utilize my training to date, it would allow me to enter a field that has important clinical implications, such as cord blood engraftment, bone marrow transplant and cancer treatments.

项目摘要 目前，我正在使用化学遗传学，siRNA筛选和质谱- 基于蛋白质组学来探测Wnt/β-catenin信号通路。使用这些 技术，我们确定Tec激酶作为Wnt/β-连环蛋白的负调节因子 发信号。因为Tec激酶BTK中的突变负责X连锁的 在无丙种球蛋白血症中，我们试图证实我们在B细胞中的原始发现。 我们发现Tec激酶在B细胞中也负调节Wnt/β-catenin信号传导 细胞培养。这项工作使我提出了一个假设，即Tec 激酶和Wnt信号传导在体内造血中具有重要作用。我 我申请独立之路奖，以延长我的 博士后培训，这样我就可以了解疾病的动物模型， 造血和获得如何解剖小鼠和收集 骨髓，如何进行小鼠骨髓移植实验 以及如何通过流式细胞术分析这些实验。中概述的 我将使用所有这些方法来验证我的假设。作为 独立调查员我计划利用我在 蛋白质组学和造血，探讨信号的分子机制 细胞分化中的转导。这个方向不仅能让我 充分利用我迄今为止的训练，这将使我进入一个领域， 重要的临床意义，如脐带血移植，骨髓 移植和癌症治疗。