MODULATION OF HOST CELL APOPTOTIC RESPONSES BY HPVE7

HPVE7 对宿主细胞凋亡反应的调节

• 批准号: 6497542
• 负责人: Karl Munger
• 金额: $ 25.63万
• 依托单位: HARVARD MEDICAL SCHOOL
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-02-15 至 2005-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6497542
• 关键词: apoptosis; biological signal transduction; cell transformation; cysteine endopeptidases; growth factor receptors; human papillomavirus; human tissue; immunofluorescence technique; immunoprecipitation; intermolecular interaction; oncoproteins; p53 gene /protein; retinoblastoma protein; tumor necrosis factor alpha; virus protein

In normal cells, the processes of cellular growth, differentiation, and death are intricately coupled. In contrast, these processes are imbalanced in cancer cells. During differentiation of squamous epithelia, differentiation is tightly linked to cellular proliferation in that the keratinocytes withdraw permanently from the cell division cycle while they undergo differentiation. The terminally differentiated keratinocytes undergo cell death and are sloughed off. Human papillomaviruses (HPVs) infect keratinocytes and their life cycle is connected to the differentiation program of the host epithelial cells. To allow their replication, the HPVs have to maintain a replication-competent milieu in the infected host cell. Hence, the HPVs encode proteins that allow replication in differentiating keratinocytes. During carcinogenic progression the viral genome is often integrated into the cellular genome. This results in the dysregulated expression of these viral regulatory proteins. The two viral proteins E6 and E7 are consistently expressed in HPV-positive cancers. We have previously shown that the HPV E7 oncoprotein can inactivate cell cycle checkpoints by inactivating the retinoblastoma tumor suppressor protein, pRB, and the cyclin dependent kinase inhibitor p21. Here we present preliminary results that demonstrate, that E7 also has the capacity to modulate cell death pathways. Depending on the stimulus, HPV E7 can either enhance or inhibit the apoptotic program. HPV E7-expressing cells are predisposed to undergo apoptosis in response to growth factor deprivation while they are protected from undergoing apoptosis in response to the cytokine tumor necrosis factor alpha (TNF). The capacity of E7 to modulate cell death pathways correlates with ability to destabilize the retinoblastoma tumor suppressor protein, pRB and the stabilization of the p53 tumor suppressor. Here I propose an integrated plan to determine whether the regulation of the stabilities of pRB and p53 are coupled, and to analyze the relative importance of p53 and/or pRB in these apoptosis pathways. Furthermore I will determine the point of action of E7 in the TNF pathway. Our preliminary results indicate that this may be at the level of coupling of caspase 8 (FLICE) to the TNF-receptor. In concert, these two aims will provide insight into critical regulatory circuits of cell death. Since they may also be dysfunctional in other, non-HPV associated cancers they should provide useful targets for anticancer therapy.

在正常细胞中，细胞生长、分化和死亡的过程是错综复杂的。 相反，这些过程在癌细胞中是不平衡的。 在鳞状上皮细胞的分化过程中，分化与细胞增殖紧密相关，因为角质形成细胞在经历分化时永久地退出细胞分裂周期。 终末分化的角质形成细胞经历细胞死亡并脱落。 人乳头瘤病毒（HPV）感染角质形成细胞，其生命周期与宿主上皮细胞的分化程序有关。 为了允许它们的复制，HPV必须在感染的宿主细胞中维持有复制能力的环境。 因此，HPV编码允许在分化的角质形成细胞中复制的蛋白质。 在致癌进展过程中，病毒基因组通常整合到细胞基因组中。这导致这些病毒调节蛋白的表达失调。 两种病毒蛋白E6和E7在HPV阳性癌症中一致表达。 我们以前已经证明，HPV E7癌蛋白可以通过使视网膜母细胞瘤肿瘤抑制蛋白pRB和细胞周期蛋白依赖性激酶抑制剂p21失活来阻断细胞周期检查点。 在这里，我们提出的初步结果表明，E7也有能力调节细胞死亡途径。 根据刺激，HPV E7可以增强或抑制凋亡程序。 表达HPV E7的细胞倾向于响应于生长因子剥夺而经历凋亡，而它们响应于细胞因子肿瘤坏死因子α（TNF）而被保护免于经历凋亡。 E7调节细胞死亡途径的能力与破坏视网膜母细胞瘤肿瘤抑制蛋白pRB的稳定性和p53肿瘤抑制蛋白的稳定性相关。在这里，我提出了一个综合计划，以确定是否pRB和p53的稳定性的调节耦合，并分析p53和/或pRB在这些凋亡途径的相对重要性。 此外，我将确定E7在TNF途径中的作用点。 我们的初步结果表明，这可能是在耦合的caspase 8（FLICE）的TNF受体的水平。 这两个目标将共同提供对细胞死亡关键调控回路的深入了解。由于它们在其他非HPV相关癌症中也可能功能失调，因此它们应该为抗癌治疗提供有用的靶点。