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: 7952613
• 负责人: Richard Goff James
• 金额: $ 12.71万
• 依托单位: UNIVERSITY OF WASHINGTON
• 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 2012-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7952613
• 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; base; cancer therapy; chemical genetics; gain of function; improved; in vitro Model; in vivo; interest; kinase inhibitor; loss of function; post-doctoral training; public health relevance; reconstitution; research study; small molecule; tissue culture

DESCRIPTION (provided by applicant): Currently I am using chemical genetics, siRNA screens and mass spectrometry- based proteomics to probe the Wnt/b-catenin signaling pathway. Using these techniques we identified Tec kinases as negative regulators of Wnt/b-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/b-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. PUBLIC HEALTH RELEVANCE: In addition to testing an interesting basic science hypothesis, the results of this proposal are likely to be relevant to the mission of the NHLBI in two areas: 1) informing clinicians treating patients diagnosed with X-linked agammaglobulinemia to focus attention to Wnt/b-catenin related symptoms such as colorectal carcinoma, and 2) using Tec kinase inhibition to increase the efficacy of human cord blood in clinical engraftment.

描述(申请人提供)：目前我正在使用化学遗传学、siRNA筛选和基于质谱学的蛋白质组学来探索Wnt/b-catenin信号通路。利用这些技术，我们发现Tec激酶是Wnt/b-catenin信号的负调节因子。由于Tec激酶BTK的突变是X连锁无丙种球蛋白血症的原因，我们试图证实我们在B细胞上的最初发现。我们发现，在培养的B细胞中，Tec激酶也负向调节Wnt/b-catenin信号。这项工作使我产生了这样的假设，即Tec激酶和Wnt信号的相互作用将在体内的造血过程中发挥重要作用。我正在申请独立之路奖，以延长我的博士后培训，以便了解疾病和造血的动物模型，并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用流式细胞仪分析这些实验的实用知识。正如提案中所概述的，我将使用所有这些方法来检验我的假设。作为一名独立的研究者，我计划利用我在蛋白质组学和造血学的交叉点上的独特地位，探索细胞分化中信号转导的分子机制。这个方向不仅可以让我充分利用我迄今为止的培训，还可以让我进入一个具有重要临床意义的领域，如脐带血移植、骨髓移植和癌症治疗。 公共卫生相关性：除了检验一项有趣的基础科学假说外，这项提议的结果可能与NHLBI在两个领域的使命相关：1)告知治疗被诊断为X连锁无丙种球蛋白血症的临床医生将注意力集中在Wnt/b-catenin相关症状，如结直肠癌，以及2)使用Tec激酶抑制来提高人脐血在临床移植中的疗效。