Microenvironment-Leukemia Communication Through Lectins

描述(申请人提供)：Ph阳性的急性淋巴细胞白血病(ALL)预后仍然很差，因为经常治疗失败和迅速复发。所有细胞在其微环境中都受到基质细胞的显著保护，但介导这种保护的分子信号并不明确。所有的细胞，特别是免疫系统的细胞，都被碳水化合物修饰的蛋白质和糖脂覆盖着一层致密的层，其功能尚未被探索。然而，所有细胞和基质之间沟通的第一步是通过细胞表面结构进行的，其中碳水化合物可能扮演关键角色。我们的初步数据表明，PolyLacNAC结合凝集素Galectin-3(Gal3)是基质细胞创造的微环境的一部分，并确定细胞外Gal3是所有细胞和基质之间的通讯分子，保护所有细胞免受药物治疗。我们推测，在ALL微环境中，Gal3是一种分子通信器，通过糖基化细胞表面蛋白的胞外晶格形成和与bcr/Abl的细胞内b相互作用促进Ph阳性ALL细胞的存活。这一假设的推论是：Galectin-3的去除将1)中断这些细胞类型之间的重要信号，2)将产生对药物治疗更敏感的所有细胞。以下目标将与同时研究的独立实验一起验证这些预测：目标1将通过使用不同的互补方法鉴定Ph阳性ALL细胞上Gal3的细胞表面配体/结合伙伴来确定信号的胞外成分。目的2将通过检测bcr/abl和Gal3之间的分子相互作用及其对所有细胞存活的影响来研究信号的细胞内成分。目标3将确定细胞外的意义 通过比较Gal3-/-和Gal3+/+Pre-B ALL细胞的基因表达，外源性Gal3刺激和尼洛替尼处理这些细胞，以及将Gal3-/-和Gal3+/+Pre-B ALL细胞移植到Gal3-/-和Gal3+/+Pre-B ALL细胞体内，比较归巢、增殖和长春新碱和尼洛替尼治疗的效果。这些实验将首次全面描述Gal3在任何类型癌症中的重要性，特别是将梳理这种碳水化合物结合蛋白在其细胞外位置作为间质与所有细胞之间的通信信使以及在其细胞内位置作为抗凋亡分子的功能。

DESCRIPTION (provided by applicant): Prognosis of Ph-positive acute lymphoblastic leukemia (ALL) remains poor because of frequent treatment failure and rapid relapse. ALL cells are significantly protected against drug treatment by stromal cells in their microenvironment, but the molecular signals that mediate the protection are not well-defined. All cells, and in particula cells of the immune system, are covered by a dense layer of carbohydrate-modified proteins and glycolipids, of which the function has not been explored. However, the first step in the communication between ALL cells and the stroma takes place through cell surface structures, of which carbohydrates are likely to play key roles. Our preliminary data show that the polyLacNAC-binding lectin Galectin-3 (Gal3) is part of the microenvironment created by stromal cells and identify extracellular Gal3 as a communication molecule between ALL cells and the stroma that protects ALL cells against drug treatment. We hypothesize that Gal3 in the ALL microenvironment is a molecular communicator that promotes Ph-positive ALL cell survival through extracellular lattice formation of glycosylated cell surface proteins and intracellularly b interactions with Bcr/Abl. Corollaries of this hypothesis are: Removal of Galectin-3 will 1) interrupt an important signal between these cell types and 2) will generate ALL cells that are more sensitive to drug treatment. The following Aims, with independent experiments that will be studied concurrently, will test these predictions: Aim 1 will determine the extracellular component of the signal by identification of the cell surface ligands/binding partners of Gal3 on Ph-positive ALL cells using different complementary approaches. Aim 2 will investigate the intracellular component of the signal by examining molecular interactions between Bcr/Abl and Gal3 and their effects on ALL cell survival. Aim 3 will determine the significance of extracellular and intracellular Gal3 to pre-B ALL cell function both in vitro, by comparing gene expression in gal3-/- and gal3+/+ pre-B ALL cells, in these cells stimulated with exogenous Gal3, and treated with nilotinib and in vivo through transplant of gal3-/- and gal3+/+ pre-B ALL cells into gal3 -/- and +/+ recipients and comparing homing, proliferation and the effect of vincristine and nilotinib treatment. These experiments together will be the first to fully characterize the significance of Gal3 in any type of cancer, and specifically will tease apart the function of this carbohydrate-binding protein in its extracellular location as communication messenger between stroma and ALL cells and in its intracellular location as an anti-apoptotic molecule.