SEARCH FOR SELECTIVE THERAPY OF CML

描述（由申请人提供）： 我们的总体目标是设计更有选择性的慢性粒细胞白血病治疗方法。 我们以前报道了12 pTyr蛋白组成型酪氨酸磷酸化的CML，但不是在正常的祖细胞。 所有这些蛋白质现已被鉴定，包括3种新蛋白质p62 dok-1、p56 dok-2和SHIP 2。 参与识别和启动由特定细胞因子或其他分子诱导的特定信号的蛋白质-蛋白质和蛋白质-磷脂相互作用是非常复杂的，参与调节这些信号的传递和控制适当的细胞应答的分子相互作用也是如此。 尽管它们很复杂，但我们在确定CML发病机制中涉及的几种蛋白质的相互作用方面取得了很好的进展。 在项目1中，克拉克森博士和Resh博士及其同事正在研究Dok、SHIP和其他蛋白在CML发病机制中的作用，并与Bornmann博士和Kuriyan博士合作，研究新化合物选择性抑制Abl激酶和其他分子靶点的能力。 最近已经鉴定出一种化合物（PD 173955），其对Bcr-Abl的抑制作用比ST 1571高约100倍，并且正在进行研究以表征该化合物并设计和合成更具选择性的抑制剂。 在项目2中，“个体发生和白血病发生中的Dok蛋白”是项目#2的新标题。 Pandolfi博士及其同事已经在小鼠中灭活了Dok-1，2和3基因，目前正在研究这些小鼠以及交叉双和三敲除小鼠的表型变化，目的是确定这些蛋白质在个体发育，造血和白血病发生中的作用，并确定对其功能和CML发病机制至关重要的基因。 在项目3中，货车Aelst博士及其同事正在研究p62 dok在p210 bcr-abl和PDGFR信号传导中的功能作用。 他们与Pandolfi博士一起发现，p62 dok作为PDGF诱导的细胞增殖和p210 bcr-abl介导的转化的负调节剂发挥作用，至少部分通过负面影响Ras/MAPK信号通路发挥作用。 他们目前正在研究抑制机制，并对与p62 dok和p210 bcr-abl相关的蛋白复合物进行表征。他们将与克拉克森博士的小组一起，通过微阵列分析细胞系和原代及CML祖细胞，寻求鉴定和表征受p210 bcr-abl影响的基因和信号通路。 与M博士Myers他们还将绘制p210 bcr-abl和PDGF触发的p62 dok中的酪氨酸磷酸化位点，并尝试应用质量分析和蛋白质测序技术鉴定其他p62 dok相互作用蛋白。

DESCRIPTION (provided by applicant): Our overall objective is to devise more selective treatment of CML. We previously reported 12 pTyr proteins constitutively tyrosine phosphorylated in CML but not in normal progenitors. All of these proteins have now been identified, including 3 novel proteins p62dok-1, p56dok-2, and SHIP2. The protein-protein and protein-phospholipid interactions involved in recognizing and initiating specific signals induced by specific cytokines or other molecules are extraordinarily complex, as are the molecular interactions involved in regulating the transmission of these signals and governing the appropriate cellular responses. Despite their complexity, we're making good progress in defining the interactions of several of the proteins involved in the pathogenesis of CML. In Project 1, Drs. Clarkson and Resh and coworkers are studying the roles of the Dok, SHIP and other proteins in the pathogenesis of CML, and in collaboration with Drs. Bornmann and Kuriyan, they are examining new compounds for their ability to selectively inhibit Abl kinase and other molecular targets. One compound (PD 173955) has recently been identified that is approximately 100-fold more inhibitory to Bcr-Abl than ST1571 and studies are underway to characterize this compound and to design and synthesize even more selective inhibitors. In Project 2, "Dok proteins in ontogenesis and leukemogenesis" is the new title of Project #2. Dr. Pandolfi and coworkers have inactivated Dok-1, 2, & 3 genes in mice and are currently examining phenotypic changes in these mice as well as in intercrossed double and triple knockout mice with the goals of defining the roles of these proteins in ontogenesis, hematopoiesis, and leukemogenesis and to identify the genes critical for their function and for the pathogenesis of CML. In Project 3, Dr. Van Aelst and coworkers are studying the functional role of p62dok in p210bcr-abl and PDGFR signaling. Together with Dr. Pandolfi, they found that p62dok functions as a negative regulator of PDGF-induced cell proliferation and p210bcr-abl-mediated transformation, acting at least in part by negatively influencing the Ras/MAPK signaling pathway. They are currently studying the mechanism of inhibition and are characterizing the protein complexes associated with p62dok and p210bcr-abl. Together with Dr. Clarkson's group they will seek to identify and characterize the genes and signaling pathways affected by p210bcr-abl by microarray analysis both in cell lines and primary and CML progenitors. With Dr. M. Myers they will also map tyrosine phosphorylation sites in p62dok triggered by p210bcr-abl and PDGF, and try to identify additional p62dok-interacting proteins applying techniques of mass analysis and protein sequencing.