C-N-, and L-MYC Effects on Hematopoietic Cells

C-N- 和 L-MYC 对造血细胞的影响

• 批准号: 7613498
• 负责人: Edward Victor Prochownik
• 金额: $ 24.42万
• 依托单位: UNIVERSITY OF PITTSBURGH AT PITTSBURGH
• 依托单位国家: 美国
• 财政年份: 1999
• 资助国家: 美国
• 起止时间: 1999-04-01 至 2010-04-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7613498
• 关键词: Apoptosis; Apoptotic; Behavior; Cell Cycle Progression; Cell physiology; Cells; Cessation of life; Complement; DNA Microarray Chip; Evolution; Family; Gene Targeting; Genes; Genomic Instability; Goals; Growth; Growth Factor; Hematopoietic; Human; IL3 Signaling Pathway; Individual; Malignant Neoplasms; Molecular; Morphology; Myeloid Cells; Normal Cell; Normal tissue morphology; Oncogene Proteins; Oncogenes; Pathogenesis; Pathway interactions; Phenotype; Population; Property; Proteins; Retroviral Vector; Route; Series; Staphylococcal Protein A; Transgenic Mice; Transgenic Model; Tumor Cell Invasion; Work; base; beta-Chemokines; c-myc Genes; cancer therapy; cell transformation; cellular engineering; chemokine; expression vector; insight; member; novel; transcription factor; tumor; vector

DESCRIPTION (provided by applicant): Myc oncoproteins (c-, N-, and L-Myc) are transcription factors, which are involved in the pathogenesis of many human cancers. In addition to being transforming, c-Myc, the most intensely studied member also promotes apoptosis, alters morphology, inhibits differentiation, accelerates cell cycle progression, and promotes genomic instability. A major goal is to identify the genes regulated by c-Myc, since this would provide significant insight into the molecular basis of Myc's phenotypes. Another goal is to determine whether different Myc members regulate the same target genes. Several c-Myc target genes can mimic a limited number of c-Myc properties. However, the cells in which they have been shown to do so all express endogenous c-Myc, thus making it impossible to determine whether these targets behave in a truly c-Myc-independent manner. We have identified a c-Myc target gene, MT-MC1, which has the unique property of imparting multiple c-Myc phenotypes in cells engineered to not express c-Myc (c-Myc "K.O" cells). This suggests that MT-MC1, and a carefully selected group of other complementing genes, might be used to reconstruct the c-Myc phenotype in KO cells, thus defining a minimum, although not necessarily unique, functional target gene population. We have therefore devised a novel retroviral vector (pRetroFLOX-GFP) that permits an unlimited number of genes to be sequentially transduced and stably expressed. This will allow us in Specific Aim 1, to construct and characterize a series of pRetroFLOX-GFP vectors for a defined subset of c-Myc target genes. In Specific Aim 2, we will determine whether over-expression of individual c-Myc target genes in KO cells can mimic the same c-Myc phenotypes as they do in parental cells. In Specific Aim 3, we will consecutively express c-Myc target genes in KO cells or transgenic mice in order to recapitulate the "complete" c-Myc phenotype. In related studies, we have shown that the CCL6 chemokine is regulated oppositely by c-Myc and L-Myc. Together with c-Myc, CCL6 imparts growth factor independence and a transformed phenotype to IL-3-dependent myeloid cells and enhances the invasive and metastatic behavior of established tumor lines. This appears to be a result of CCL6's ability to induce apoptotic death in adjacent normal cells, thus destroying the normal tissue barriers that limit tumor spread. Therefore, in Specific Aim 4, we will determine whether CCL6 must be secreted in order to impart IL-3-independent growth and more aggressive tumor behavior. In Specific Aim 5, we will determine the mechanism(s) by which CCL6 and c-Myc subvert the IL-3 signaling pathway. In Specific Aim 6, we will determine whether related chemokines impart CCL6-like properties. Finally, in Specific Aim 7, we will develop a transgenic model of CCL-6-dependent tumor invasion. Together, these studies will provide new insights into the mechanisms by which c-Myc and its transcriptional targets subvert normal cellular pathways en route to establishing a transformed cell.

描述（由申请人提供）：Myc 癌蛋白（c-、N- 和 L-Myc）是转录因子，参与许多人类癌症的发病机制。除了转化之外，c-Myc 作为研究最深入的成员，还可以促进细胞凋亡、改变形态、抑制分化、加速细胞周期进程并促进基因组不稳定。主要目标是鉴定受 c-Myc 调节的基因，因为这将为了解 Myc 表型的分子基础提供重要的见解。另一个目标是确定不同的 Myc 成员是否调节相同的靶基因。一些 c-Myc 靶基因可以模拟有限数量的 c-Myc 特性。然而，已证明它们这样做的细胞都表达内源性 c-Myc，因此无法确定这些靶标是否以真正不依赖于 c-Myc 的方式发挥作用。我们已经鉴定了一个 c-Myc 靶基因 MT-MC1，它具有在经工程改造后不表达 c-Myc 的细胞（c-Myc“K.O”细胞）中赋予多种 c-Myc 表型的独特特性。这表明MT-MC1和一组精心挑选的其他互补基因可用于重建KO细胞中的c-Myc表型，从而定义最小但不一定是唯一的功能性靶基因群。因此，我们设计了一种新型逆转录病毒载体（pRetroFLOX-GFP），它允许连续转导和稳定表达无限数量的基因。这将使我们能够在特定目标 1 中为 c-Myc 靶基因的定义子集构建并表征一系列 pRetroFLOX-GFP 载体。在具体目标 2 中，我们将确定 KO 细胞中单个 c-Myc 靶基因的过度表达是否可以模仿与亲本细胞中相同的 c-Myc 表型。在具体目标 3 中，我们将在 KO 细胞或转基因小鼠中连续表达 c-Myc 靶基因，以重现“完整”c-Myc 表型。在相关研究中，我们发现CCL6趋化因子受到c-Myc和L-Myc相反的调节。 CCL6 与 c-Myc 一起赋予 IL-3 依赖性骨髓细胞生长因子独立性和转化表型，并增强已建立肿瘤系的侵袭和转移行为。这似乎是由于 CCL6 能够诱导邻近正常细胞凋亡，从而破坏限制肿瘤扩散的正常组织屏障。因此，在具体目标 4 中，我们将确定是否必须分泌 CCL6，以赋予不依赖于 IL-3 的生长和更具侵袭性的肿瘤行为。在具体目标 5 中，我们将确定 CCL6 和 c-Myc 破坏 IL-3 信号通路的机制。在具体目标 6 中，我们将确定相关趋化因子是否具有 CCL6 样特性。最后，在具体目标 7 中，我们将开发 CCL-6 依赖性肿瘤侵袭的转基因模型。总之，这些研究将为 c-Myc 及其转录靶标在建立转化细胞过程中颠覆正常细胞途径的机制提供新的见解。

项目成果

Dynamic in vivo interactions among Myc network members.

Myc 网络成员之间的动态体内相互作用。

• DOI: 10.1038/sj.onc.1204606
• 发表时间: 2001
• 期刊: Oncogene
• 影响因子: 8
• 作者: Yin,X;Landay,MF;Han,W;Levitan,ES;Watkins,SC;Levenson,RM;Farkas,DL;Prochownik,EV
• 通讯作者: Prochownik,EV

c-Myc as a therapeutic target in cancer.

• DOI: 10.1586/14737140.4.2.289
• 发表时间: 2004-04-01
• 期刊: Expert review of anticancer therapy
• 影响因子: 3.3
• 作者: Prochownik, Edward V

Inverse regulation of cyclin B1 by c-Myc and p53 and induction of tetraploidy by cyclin B1 overexpression.

• 发表时间: 2001-09
• 期刊: Cancer research
• 影响因子: 11.2
• 作者: Xiao-ying Yin;L. Grove;N. Datta;K. Katula;M. Long;M. Long;E. Prochownik;E. Prochownik

Modularity of the oncoprotein-like properties of platelet glycoprotein Ibalpha.

血小板糖蛋白 Ibalpha 的癌蛋白样特性的模块化。

• DOI: 10.1074/jbc.m806222200
• 发表时间: 2009
• 期刊: The Journal of biological chemistry
• 作者: Li,Youjun;Lu,Jie;Prochownik,EdwardV
• 通讯作者: Prochownik,EdwardV

Deregulation of common genes by c-Myc and its direct target, MT-MC1.

• DOI: 10.1073/pnas.0507902102
• 发表时间: 2005-12
• 期刊: Proceedings of the National Academy of Sciences of the United States of America
• 影响因子: 11.1
• 作者: K. Rogulski;D. Cohen;D. Corcoran;P. Benos;E. Prochownik