Fhit modulation of cell cycle progression and DNA damage response

Fhit 调节细胞周期进程和 DNA 损伤反应

• 批准号: 7673532
• 负责人: KAY HUEBNER
• 金额: $ 31.59万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-09-01 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7673532
• 关键词: Allelic Imbalance; Apoptosis; Cell Cycle; Cell Cycle Progression; Cell Death; Cell Proliferation; Cell physiology; Cells; Chromosome Fragile Sites; Cyclin D1; DNA Damage; DNA biosynthesis; DNA damage checkpoint; Data; Defect; Exposure to; FHIT gene; Genomic Instability; Goals; Growth; Hyperplasia; Induction of Apoptosis; Infection; Lesion; Malignant Neoplasms; Mus; Mutation; Nature; Oral; Oxidative Stress; PPP3CA gene; Pathway interactions; Process; Proteins; Research; Role; Stress; Upper digestive tract structure; Virus; base; cell transformation; gene therapy; mutant; prevent; response; survivin; tumor; tumor progression

Fhit modulation of cell cycle progression and DMA damage response It has recently been proposedthat in early preneoplastic lesions, DNA replication stress leads tc the DNA Damage Response (DDR), which elicits growth arrest and cell death. Thus, before occurrence o-' genomic instability and transformation, cells activate DNA damage response networks that delay or prevent cancer; mutations that compromise the checkpoints, such as defects in Atm or Atr pathway proteins, allcw escape from the checkpoint block, leading to tumor progression. Importantly, for the research proposed here, the early hyperplasias showed allelic imbalance at common fragile sites, particularly the FRA3B/FH1T locus. "Fragile site LOH was targeted during the period of maximal DDR and may be considered a "signature" of stalled replicationforks" (Gorgoulis et at, Nature 434:907-913, 2005). The researchproposed is based on the hypothesis that loss of the Fhit tumor suppressorvery early in the preneoplastic process, coincident with activation of the DNA damage response (DDR) checkpoint, effects critical cellular processes, cell proliferation, DNA damage response and apoptosis, that can tip the tBalance toward genome instability and tumor progression. The hypothesis is based on preliminary data demonstrating that Fhit-deficient cells show altered expression of cell,cycle, DNA damage response andapoptosis associated proteins and that Fhit over-expression modulates expression of Cyclin D1, Hus1, Chk1 jind Akt/mTor/Survivin. ¿; ¿ The goals of this project are to understand the mechanisms underlying Fhit modulation of these pathways by the following aims: 1) Examine the mechanism of Fhit down-modulation of Cyclin D1 level after infection of Fhit negsitive cells with AdenoFHIT and AdenoFHIT mutant viruses. ¿ 2) Determine the mechanisms through which Fhit modulates" the level of Hus1, pChkl and the CNA damage response pathway by manipulation of Fhit .expression in Fhit-deficient cells. 3) Examine the role of Fhit in induction of apoptosis after AdenoFHIT and AdenoFHIT mutant viius infection of Fhit-deficient cells and exposure to oxidative stress conditions. 4) Define the sequenceof modulation of expression of cell.cycle, DDR and apoptosis associated proteins in early oral and upper gastrointestinal tract lesions of Fhit-deficient mice before and after FHIT replacement by AdenoFHIT oral gene therapy. ,

细胞周期进程和DMA损伤响应的调制 最近有人提出，在早期肿瘤病变中，DNA复制应力导致TC DNA 损坏反应（DDR），引起逮捕和细胞死亡。在发生之前，O-'基因组 细胞不稳定和转化，激活DNA损伤反应网络，这些反应网络延迟或预防癌症。 损害检查点的突变，例如ATM或ATR途径蛋白中的缺陷，AllcW逃脱 从检查点块，导致肿瘤进展。重要的是，对于这里提出的研究， 早期增生性在常见的脆弱部位，尤其是fra3b/fh1t基因座，表现出垂体失衡。 “在最大DDR期间针对脆弱的现场LOH，可以被视为一个“签名” 停滞的replicationforks”（Gorgoulis et，Nature 434：907-913，2005）。 研究规模是基于以下假设。 与DNA损伤响应（DDR）检查点的激活相一致的肿瘤塑性过程，效果 关键的细胞过程，细胞增殖，DNA损伤反应和凋亡，可以使Tbalance尖锐 迈向基因组不稳定性和肿瘤进展。该假设基于初步数据证明 缺陷型细胞显示细胞表达改变，循环，DNA损伤反应和凋亡 相关蛋白质，并且过表达调节Cyclin D1，HUS1，CHK1 Jind的表达 Akt/mtor/survivin。 �; » 该项目的目标是了解这些途径的FHIT调制的机制 以以下目的： 1）检查感染FHIT阴性细胞后细胞周期蛋白D1水平的FHIT下调机制 带有腺体和腺体突变病毒。 » 2）确定FHIT调节“ HUS1，PCHKL和CNA的水平”的机制 通过操纵FHIT的损害响应途径。 3）检查adenofhit和adenofhit突变体Viius后凋亡诱导的作用 缺乏FHIT的细胞感染和暴露于氧化物应力条件。 4）定义细胞表达的调节顺序。循环，DDR和凋亡相关蛋白 在FHIT之前和之后，在早期口服和上胃肠道病变中 用adenofhit口服基因疗法替代。 ，，，，