PHORBOL ESTER-INDUCED DIFFERENTIATION OF LEUKEMIC CELLS

佛波酯诱导白血病细胞分化

基本信息

  • 批准号:
    3186192
  • 负责人:
  • 金额:
    $ 15.68万
  • 依托单位:
  • 依托单位国家:
    美国
  • 项目类别:
  • 财政年份:
    1987
  • 资助国家:
    美国
  • 起止时间:
    1987-05-01 至 1995-06-30
  • 项目状态:
    已结题

项目摘要

Neoplasia is a disorder characterized by abnormalities in proliferation and differentiation. Preliminary clinical trials using therapies designed to stimulate leukemic cell differentiation have yielded promising results. Phorbol esters are a group of agents which induce in vitro differentiation of human leukemic cells more potently than agents currently being evaluated for clinical use. However, mitogenic effects in other tissues preclude their use in a "clinical setting. Phorbol esters (TPA) exert their effects by activating members of the protein kinase C (PKC) gene family. Because of differences among the seven PKC isoforms, it is hypothesized that individual isoforms may selectively mediate TPA-induced events. Thus, studies are warranted to directly assess the involvement of the individual isoforms in mediating the effects of TPA. Using the human monoblastoid U937 leukemic cell line which differentiates into a macrophage-like cell after exposure to TPA, we have: (1) demonstrated the existence of a previously uncharacterized PKC isoform; (2) observed that PKC zeta is contained in the U-937 cell and does not translocate or down regulate in response to TPA; (3) found that TPA treatment functionally induces a partial down regulation PKC a activity by altering its substrate specificity and (4) using PKC mutants prepared by recombinant polymerase chain reaction techniques that are constituitively activated in the absence of TPA determined that individual isoforms elicit qualitatively different effects on gene expression. This data form the basis for this revision of the previously submitted proposal. The goal of the current proposal is to determine the mechanism(s) by which TPA stimulates U937 differentiation. The main thrust of the proposal is to determine the role of individual isoforms in mediating TPA stimulated differentiation and other TPA-induced responses. Pursuant to this goal, we will: (1) directly examine the role of individual isoforms by assessing the ability of PKC mutants constituitively activated in the absence to TPA to elicit U937 differentiation and other TPA-induced responses; (2) analyze the role of individual isoforms on TPA-stimulated differentiation by selectively depleting isoforms with antisense DNA; (3) determine the factors activating PKC-zeta, the cellular responses elicited by activation of this isoform and examine the role of the second cysteine-rich repeat in the Cl domain in conferring TPA-responsiveness to PKC; (4) analyze the mechanisms responsible for TPA-induced alterations in PKC a substrate specificity; (5) identify and characterize a newly recognized PKC activity in the U937 cell; (6) determine the effects of altering endogenous PKC isoform expression on TPA-responsiveness in the U937 cell; and (7) to analyze the role of the DNA region 5' to the beta and gamma isoforms in regulating transcriptional activity of these genes in the U937 cell using CAT constructs containing these regions. By determining the role of individual isoforms in mediating TPA-induced differentiation and other phorbol ester stimulated responses, information obtained from these studies could have direct clinical implications in the treatment of leukemias. These results would focus the development of agonists selectively activating the involved isoform and could form the basis for designing PKC mutants which could be used as a selective form of gene therapy for certain leukemias.
肿瘤是一种以增殖异常为特征的疾病 和差异化。 使用疗法的初步临床试验 旨在刺激白血病细胞分化已产生 有希望的结果。 佛波酯是一组诱导剂 人类白血病细胞的体外分化能力比药物更有效 目前正在评估其临床用途。 然而,促有丝分裂作用 在其他组织中的存在妨碍了它们在“临床环境中的使用。佛波醇 酯类 (TPA) 通过激活蛋白质成员发挥作用 激酶 C (PKC) 基因家族。 由于七个PKC之间的差异 同种型,假设单个同种型可以选择性地 介导 TPA 诱发的事件。 因此,研究有必要直接 评估各个亚型在介导中的参与 TPA 的影响。 使用可分化的人单母细胞 U937 白血病细胞系 暴露于 TPA 后进入巨噬细胞样细胞,我们有: (1) 证明了以前未表征的 PKC 同工型的存在; (2)观察到U-937细胞中含有PKC zeta,并且不存在 响应 TPA 进行易位或下调; (3)发现TPA 治疗功能性诱导部分下调 PKC a 活性 通过改变其底物特异性和(4)使用制备的 PKC 突变体 通过重组聚合酶链反应技术 在没有 TPA 的情况下组成型激活确定 各个亚型对基因产生不同质的影响 表达。 该数据构成了本次修订的基础 提交的提案。 当前提案的目标是通过以下方式确定机制: TPA 刺激 U937 分化。 该计划的主要推力是 提议是确定各个亚型在介导 TPA 中的作用 刺激分化和其他 TPA 诱导的反应。 根据 为了实现这一目标,我们将:(1)直接检查各个亚型的作用 通过评估 PKC 突变体组成型激活的能力 缺乏 TPA 来引发 U937 分化和其他 TPA 诱导的分化 回应; (2)分析各个异构体对TPA刺激的作用 通过用反义 DNA 选择性地消除同种型来进行分化; (3) 确定激活 PKC-zeta 的因素,即细胞反应 通过激活该亚型引起并检查第二个亚型的作用 Cl 结构域中富含半胱氨酸的重复序列赋予 TPA 响应性 PKC; (4)分析TPA引起的改变的机制 PKC 中具有底物特异性; (5) 识别并表征新的 识别 U937 细胞中的 PKC 活性; (6) 确定效果 改变内源性 PKC 亚型表达对 TPA 反应性的影响 U937细胞; (7) 分析 DNA 区域 5' 对 β 的作用 和 γ 亚型调节这些基因的转录活性 在 U937 细胞中使用包含这些区域的 CAT 构建体。 经过 确定各个异构体在介导 TPA 诱导的过程中的作用 分化和其他佛波酯刺激反应、信息 从这些研究中获得的结果可能具有直接的临床意义 白血病的治疗。 这些结果将集中于发展 激动剂选择性地激活所涉及的亚型并可以形成 设计可用作选择性形式的 PKC 突变体的基础 某些白血病的基因治疗。

项目成果

期刊论文数量(0)
专著数量(0)
科研奖励数量(0)
会议论文数量(0)
专利数量(0)

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

数据更新时间:{{ journalArticles.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ monograph.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ sciAawards.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ conferencePapers.updateTime }}

{{ item.title }}
  • 作者:
    {{ item.author }}

数据更新时间:{{ patent.updateTime }}

DOUGLAS K. WAYS其他文献

DOUGLAS K. WAYS的其他文献

{{ item.title }}
{{ item.translation_title }}
  • DOI:
    {{ item.doi }}
  • 发表时间:
    {{ item.publish_year }}
  • 期刊:
  • 影响因子:
    {{ item.factor }}
  • 作者:
    {{ item.authors }}
  • 通讯作者:
    {{ item.author }}

{{ truncateString('DOUGLAS K. WAYS', 18)}}的其他基金

PHORBOL ESTER-INDUCED DIFFERENTIATION OF LEUKEMIC CELLS
佛波酯诱导白血病细胞分化
  • 批准号:
    3186193
  • 财政年份:
    1994
  • 资助金额:
    $ 15.68万
  • 项目类别:
PROTEIN KINASE C INHIBITION OF HEPATOMA GROWTH
蛋白激酶 C 抑制肝癌生长
  • 批准号:
    2291668
  • 财政年份:
    1993
  • 资助金额:
    $ 15.68万
  • 项目类别:
CHARACTERIZATION OF PROTEIN KINASE C GENE RESPONSE ELEME
蛋白激酶 C 基因反应元件的表征
  • 批准号:
    3023319
  • 财政年份:
    1991
  • 资助金额:
    $ 15.68万
  • 项目类别:
PHORBOL ESTER INDUCED DIFFERENTIATION OF LEUKEMIC CELLS
佛波酯诱导白血病细胞分化
  • 批准号:
    3457923
  • 财政年份:
    1987
  • 资助金额:
    $ 15.68万
  • 项目类别:
PHORBOL ESTER-INDUCED DIFFERENTIATION OF LEUKEMIC CELLS
佛波酯诱导白血病细胞分化
  • 批准号:
    3186194
  • 财政年份:
    1987
  • 资助金额:
    $ 15.68万
  • 项目类别:
PHORBOL ESTER INDUCED DIFFERENTIATION OF LEUKEMIC CELLS
佛波酯诱导白血病细胞分化
  • 批准号:
    3457925
  • 财政年份:
    1987
  • 资助金额:
    $ 15.68万
  • 项目类别:
PHORBOL ESTER INDUCED DIFFERENTIATION OF LEUKEMIC CELLS
佛波酯诱导白血病细胞分化
  • 批准号:
    3447011
  • 财政年份:
    1987
  • 资助金额:
    $ 15.68万
  • 项目类别:
PHORBOL ESTER INDUCED DIFFERENTIATION OF LEUKEMIC CELLS
佛波酯诱导白血病细胞分化
  • 批准号:
    3457922
  • 财政年份:
    1987
  • 资助金额:
    $ 15.68万
  • 项目类别:
PHORBOL ESTER INDUCED DIFFERENTIATION OF LEUKEMIC CELLS
佛波酯诱导白血病细胞分化
  • 批准号:
    3457926
  • 财政年份:
    1987
  • 资助金额:
    $ 15.68万
  • 项目类别:
PHORBOL ESTER INDUCED DIFFERENTIATION OF LEUKEMIC CELLS
佛波酯诱导白血病细胞分化
  • 批准号:
    3457924
  • 财政年份:
    1987
  • 资助金额:
    $ 15.68万
  • 项目类别:

相似海外基金

Development of a method for preserving transplanted lung function using Gapmer-type antisense nucleic acid
开发利用Gapmer型反义核酸保存移植肺功能的方法
  • 批准号:
    22K09003
  • 财政年份:
    2022
  • 资助金额:
    $ 15.68万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Myostatin antisense nucleic acid therapy for rhabdomyosarcoma
肌肉生长抑制素反义核酸治疗横纹肌肉瘤
  • 批准号:
    21K07762
  • 财政年份:
    2021
  • 资助金额:
    $ 15.68万
  • 项目类别:
    Grant-in-Aid for Scientific Research (C)
Suppression of PHOX2B (+7Ala mutant) expression by antisense nucleic acid
反义核酸抑制 PHOX2B(7Ala 突变体)表达
  • 批准号:
    20K16927
  • 财政年份:
    2020
  • 资助金额:
    $ 15.68万
  • 项目类别:
    Grant-in-Aid for Early-Career Scientists
Pathogenesis and Antisense nucleic acid, glycosylation supplementation, and AAV therapy development forFukuyama muscular dystrophy and related diseases
福山性肌营养不良症及相关疾病的发病机制和反义核酸、糖基化补充以及 AAV 疗法的开发
  • 批准号:
    20H00526
  • 财政年份:
    2020
  • 资助金额:
    $ 15.68万
  • 项目类别:
    Grant-in-Aid for Scientific Research (A)
Synthesis of antisense nucleic acid incorporating cyclic sulfonamide backbone
掺入环状磺酰胺主链的反义核酸的合成
  • 批准号:
    20K21245
  • 财政年份:
    2020
  • 资助金额:
    $ 15.68万
  • 项目类别:
    Grant-in-Aid for Challenging Research (Exploratory)
Antisense nucleic acid splice correction therapy for Duchenne muscular dystrophy and related disorders
杜氏肌营养不良症及相关疾病的反义核酸剪接校正疗法
  • 批准号:
    G0900887/1
  • 财政年份:
    2011
  • 资助金额:
    $ 15.68万
  • 项目类别:
    Research Grant
CHEMICAL SYNTHESIS OF A NEW MATERIAL OF ANTISENSE NUCLEIC ACID "2'-PHOSPHORYLATED RNAS" -DIRECTED TOWARD ITS BASIC STRUCTURAL STUDIES AND REGULATION OF EXPRESSION OF HIV VIRUS-
反义核酸新材料“2-磷酸化RNAS”的化学合成-针对其基础结构研究和HIV病毒表达调控-
  • 批准号:
    05558090
  • 财政年份:
    1993
  • 资助金额:
    $ 15.68万
  • 项目类别:
    Grant-in-Aid for Developmental Scientific Research (B)
CHEMICAL SYNTHESIS OF A NEW MATERIAL OF ANTISENSE NUCLEIC ACID"2"PHOSTHORYLATEDRNAS" DIRETED TOWARD IIS BASIC STRUCTRAL STUDIES AND REGULATION OF EXPRESSION OF HIV VIRUS-
针对 IIS 基础结构研究和 HIV 病毒表达调控的反义核酸新材料“2”磷酸化 RNA 的化学合成-
  • 批准号:
    04453031
  • 财政年份:
    1992
  • 资助金额:
    $ 15.68万
  • 项目类别:
    Grant-in-Aid for General Scientific Research (B)
{{ showInfoDetail.title }}

作者:{{ showInfoDetail.author }}

知道了