SEARCH FOR SELECTIVE THERAPY OF CML

描述(由申请人提供)： 我们的总体目标是设计出更具选择性的慢性粒细胞白血病治疗方法。我们之前报道了12个pTyr蛋白在CML中组成酪氨酸磷酸化，但在正常祖细胞中不是。所有这些蛋白都已被鉴定，包括3个新的蛋白p62dok-1、p56dok-2和SHIP2。识别和启动由特定细胞因子或其他分子诱导的特定信号所涉及的蛋白质-蛋白质和蛋白质-磷脂相互作用非常复杂，调节这些信号的传递和调节适当的细胞反应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良好的进展。在项目1中，Clarkson博士和Resh博士及其同事正在研究Dok、Ship和其他蛋白质在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。最近发现了一种化合物(PD 173955)，它对bcr-Abl的抑制作用大约是ST1571的100倍，目前正在进行研究，以表征这种化合物，并设计和合成更具选择性的抑制剂。在项目2中，“DOK蛋白在个体发生和白血病发生中的作用”是项目#2的新标题。Pandolfi博士和他的同事已经在小鼠中灭活了DOK-1、2和3基因，目前正在检测这些小鼠以及交叉双基因和三基因敲除小鼠的表型变化，目的是确定这些蛋白在个体发生、造血和白血病发生中的作用，并确定对它们的功能和CML发病机制至关重要的基因。在项目3中，Van Aelst博士和同事正在研究p62dok在p210bcr-abl和PDGFR信号转导中的功能作用。与Pandolfi博士一起，他们发现p62dok在PDGF诱导的细胞增殖和p210bcr-abl介导的转化中发挥负面调节作用，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，并正在表征与p62dok和p210bcr-abl相关的蛋白质复合体。与克拉克森博士的团队一起，他们将通过微阵列分析，在细胞系以及原代和CML祖细胞中识别和表征受p210bcr-abl影响的基因和信号通路。与M.Myers博士一起，他们还将绘制p210bcr-abl和PDGF触发的p62dok的酪氨酸磷酸化位点图，并尝试应用质量分析和蛋白质测序技术识别更多与p62dok相互作用的蛋白质。

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.