PROTEIN KINASE C BASED ANTILEUKEMIC THERAPY

基于蛋白激酶 C 的抗白血病治疗

• 批准号: 2895354
• 负责人: RICHARD M STONE
• 金额: $ 12.54万
• 依托单位: DANA-FARBER CANCER INST
• 依托单位国家: 美国
• 财政年份: 1995
• 资助国家: 美国
• 起止时间: 1995-09-12 至 2001-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/2895354
• 关键词: SCID mouse; acute myelogenous leukemia; antileukemic agent; antineoplastics; blood /lymphatic neoplasm; bryostatin; cell differentiation; clinical trials; combination cancer therapy; disease /disorder model; drug interactions; enzyme activity; human subject; human therapy evaluation; isozymes; neoplasm /cancer chemotherapy; neoplastic cell; phorbols; protein kinase C; retinoate; tissue /cell culture; transfection

DESCRIPTION: (Applicant's Abstract) The objective of this application is to determine whether activation of protein kinase C (PKC) is a viable strategy for anti-leukemic therapy. Acute myeloid leukemia (AML) and myelodysplasia (MDS) are manifested by bone marrow failure resulting from a partial or complete block in maturation of hematopoietic elements. The protein kinase C (PKC) family of calcium and phospholipid-dependent serine-threonine kinases plays a critical role in transducing extracellular signals involved in monocytic differentiation. The phorbol ester 12-0-tetradecanoyl-phorbol-13- acetate (TPA), a potent inducer of maturation along the monocytic lineage, binds to and activates PKC. The applicant demonstrated that all-trans retinoic acid (ATRA) treatment of the TPA-resistant HL-60 cell subline (HL- 525) markedly increases the relatively low constitutive levels of PKCb mRNA and protein found in this variant. The ATRA- mediated increase in PKC activity and PKCb gene product is associated with the acquisition of TPA responsiveness characterized by adherence, non-specific esterase staining, and c-fms gene expression. He has demonstrated that bryostatin 1 (a non-tumor promoting compound which activates PKC at nanomolar concentrations) treatment of ATRA- primed HL-60 and U-937 cells also leads to a marked augmentation of features of monocytic differentiation in these cells. Therefore, the combination of ATRA and bryostatin 1 may be useful in diseases characterized by a block in myeloid differentiation. This application will build upon the applicant's preliminary observations by defining the PKC isoform(s) which are critically increased by retinoic acid and subsequently activated by TPA or bryostatin 1 in several leukemia cell lines and human leukemic cells (Specific Aim 1). Downstream elements directly interacting with the relevant PKC isoform will be determined by means of glutathione-S-transferase fusion constructs (Specific Aim 2). To insure that his findings with leukemic cell lines are generalizable, he will test the retinoic acid/bryostatin 1 combination against human leukemias propagated in immunodeficient mice (Specific Aim 3). Finally, he will conduct a phase II trial of all trans retinoic acid in combination with bryostatin 1 in patients with MDS and refractory AML, correlating therapeutic response with studies to assess biologic effects of therapy including the ability of sera from treated patients to induce differentiation in vitro and in vivo (Specific Aim 4).

描述：（申请人摘要）本申请的目的 是为了确定蛋白激酶 C (PKC) 的激活是否可行 抗白血病治疗策略。急性髓系白血病 (AML) 和 骨髓增生异常（MDS）表现为骨髓衰竭导致 造血成熟部分或完全受阻 元素。钙和蛋白激酶 C (PKC) 家族 磷脂依赖性丝氨酸-苏氨酸激酶起着关键作用 转导参与单核细胞的细胞外信号 差异化。佛波酯 12-0-十四烷酰基-佛波醇-13- 醋酸盐（TPA），单核细胞成熟的有效诱导剂 谱系，结合并激活 PKC。申请人证明 全反式视黄酸 (ATRA) 处理 TPA 抗性 HL-60 细胞 亚系（HL-525）显着增加了相对较低的本构 在此变体中发现的 PKCb mRNA 和蛋白质水平。 ATRA- 介导的 PKC 活性和 PKCb 基因产物的增加相关 获得以坚持为特征的 TPA 响应能力， 非特异性酯酶染色和 c-fms 基因表达。他有 证明苔藓抑素 1（一种非肿瘤促进化合物， 在纳摩尔浓度下激活 PKC）ATRA 引发的处理 HL-60 和 U-937 细胞也导致特征显着增强 这些细胞中的单核细胞分化。因此，组合 ATRA 和苔藓抑素 1 可能有助于治疗以以下特征为特征的疾病： 骨髓分化受阻。该应用程序将建立在 申请人通过定义 PKC 同工型得出的初步意见 视黄酸会显着增加，随后 在几种白血病细胞系中被 TPA 或苔藓抑素 1 激活 人类白血病细胞（具体目标 1）。直接下游元件 与相关 PKC 同工型的相互作用将通过以下方式确定 谷胱甘肽-S-转移酶融合构建体（具体目标 2）。到 为了确保他对白血病细胞系的发现具有普遍性，他 将针对人类测试视黄酸/苔藓抑素 1 组合 白血病在免疫缺陷小鼠中繁殖（具体目标 3）。最后， 他将对所有反式视黄酸进行 II 期试验 与苔藓抑素 1 联合治疗 MDS 和难治性 AML 患者， 将治疗反应与评估生物学效应的研究相关联 治疗的效果，包括来自接受治疗的患者的血清诱导 体外和体内分化（具体目标 4）。