CaM Kinase Cascade in Myeloid Differentiation/Function

骨髓分化/功能中的 CaM 激酶级联

基本信息

批准号：
6784616
负责人：
PETER C GAINES
金额：
$ 9.8万
依托单位：
YALE UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2002
资助国家：
美国
起止时间：
2002-09-30 至 2006-03-31
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/6784616
关键词：
RNase protection assay calmodulin dependent protein kinase cell cycle cell differentiation cell growth regulation cell line colony stimulating factor enzyme activity granulocyte laboratory rabbit leukocyte activation /transformation myelocyte neutrophil northern blottings phosphorylation polymerase chain reaction postdoctoral investigator protein structure function

项目摘要

DESCRIPTION (provided by applicant) The molecular mechanisms that mediate myeloid differentiation from multipotent hematopoietic progenitors have yet to be clearly defined. The retinoic acid (RA) signaling pathway has been suggested to play a critical role in myeloid differentiation, and several direct targets for RA signaling have been recently identified. One such target, CaM Kinase Kinase alpha (CaMKKa), has been identified in our laboratory. CaMKKa shows rapid upregulation in promyelocytic MPRO cells in response to RA-induced myeloid differentiation. To further characterize the role of the CaM kinase cascade in myeloid differentiation, we have ectopically expressed a constitutively active form of CaMKKa in a myeloid cell line (32Dcl3) that differentiates into mature neutrophils in response to G-CSF. Our preliminary results suggest deregulated activation of CaMKKa disrupts proliferative and/or survival signaling mediated by G-CSF. We aim to further characterize this phenotype and investigate potential roles of CaM kinase signaling in regulating neutrophil differentiation and function. Specifically, we aim to: 1) characterize the expression of CaM kinase family members, including a recently identified granuloctye-specific CaM kinase I-like kinase, in normal and leukemic models of neutophil differentiation; 2) determine which component of the cell cycle machinery and/or pro-apoptotic pathway is disrupted by constitutive activation of CaM kinase cascade, and 3) characterize the phenotypic effect of inducible expression of constitutively active and dominant negative forms of CaM kinase family members on myeloid differentiation and functional activation. This work will both increase our understanding of the signaling pathways that control myelopoiesis, and will complete the training of the Principle Investigator, Dr. Peter Gaines, as a fully prepared independent investigator in the field of molecular hematopoiesis.

描述（由申请人提供）介导髓样分化与多能的分子机制造血祖细胞尚未明确定义。视黄酸（RA）已建议信号通路在髓样中发挥关键作用最近已经有分化和RA信号的几个直接目标确定。一个这样的目标是CAM激酶激酶α（Camkka）在我们的实验室中确定。 Camkka显示出临时细胞的快速上调 MPRO细胞响应RA诱导的髓样分化。进一步表征CAM激酶级联反应在髓样分化中的作用，我们在异位表达了Camkka的组成型活性形式细胞系（32DCL3），该细胞系因响应于成熟的中性粒细胞而分化为 G-CSF。我们的初步结果表明camkka的激活失调破坏由G-CSF介导的增殖和/或存活信号传导。我们的目标进一步表征这种表型并研究CAM的潜在作用激酶信号在调节中性粒细胞分化和功能中。具体而言，我们的目的是：1）表征CAM激酶家族的表达成员，包括最近确定的颗粒特异性凸轮激酶I样的成员激酶，在正常和白血病模型的中性粒细胞分化模型中； 2）确定细胞周期机械和/或促凋亡的哪个组件 CAM激酶级联反应的组成型激活会破坏途径，而3）表征组成性诱导表达的表型效应 CAM激酶家族成员在髓样中的主动和主导的负面形式分化和功能激活。这项工作都会增加我们的了解控制髓鞘的信号通路，并将完成主要研究者彼得·盖恩斯博士的培训分子领域的完全准备的独立研究者造血。