Role of polycystin-1 in centrosome duplication and cell cycle control

Polycystin-1 在中心体复制和细胞周期控制中的作用

• 批准号: 7921098
• 负责人: Gabriele Luca GUSELLA
• 金额: $ 10.17万
• 依托单位: ICAHN SCHOOL OF MEDICINE AT MOUNT SINAI
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-09-18 至 2011-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7921098
• 关键词: Accounting; Adult; Affect; Autosomal Dominant Polycystic Kidney; Cell Cycle; Cell Cycle Checkpoint; Cell Cycle Progression; Cell Cycle Regulation; Centrosome; Chromosome abnormality; Chromosomes, Human, Pair 1; Cyst; DNA Damage; Data; Disease; Epithelial Cells; Event; Genetic; Genetic Heterogeneity; Genomic Instability; Germ-Line Mutation; Goals; Human; In Vitro; Individual; Kidney; Lead; Licensing; Mediating; Mitotic; Modeling; Molecular; Mus; Normal Cell; Patients; Phase; Phenotype; Phosphotransferases; Process; Proteins; Renal Tissue; Role; Small Interfering RNA; Stress; System; TP53 gene; Testing; Therapeutic; Transgenic Mice; drug development; in vivo; inhibitor/antagonist; mouse model; novel; polycystic kidney disease 1 protein; public health relevance; renal epithelium; response; separase; therapeutic target

DESCRIPTION (provided by applicant): To date, the molecular mechanisms responsible for the remarkable genetic heterogeneity found in ADPKD cysts remain unknown. Similar karyotypic alterations have been described consequently to genomic instability. Our overall hypothesis is that PC1 disregulation contributes to the cystogenic hyperproliferative process by altering normal centrosomal functions and cell cycle progression, leading to genomic instability. Our preliminary results have uncovered strong evidence in support of this proposition. Specific siRNA- mediated inhibition of PC1 altered cell cycle progression and centrosome amplification in vitro, a finding corroborated in human ADPKD renal tissues. Loss of centrosome integrity was associated with dramatic genomic instability. The experimental focus of this application is to elucidate the molecular mechanisms underlying these novel findings. The first specific Aim is to determine whether PC1 disregulation causes an abnormal modulation of the proteins involved in the centrosome duplication process. The second specific Aim is to evaluate molecular mechanisms by which PC1 knockdown may cause interference with the normal cell division cycle progression. The third specific Aim is to characterize in vivo the role of PC1 knockdown in cystic hyperproliferation and centrosome amplification, using a transgenic mouse model containing a doxycyclin inducible siRNA specific for murine PKD1. Taken together, these novel studies will expand therapeutic cell cycle targets for drug development, as well as our understanding of the disease process. PUBLIC HEALTH RELEVANCE The elucidation of mechanisms underlying ADPKD hyperproliferative processes will help the understanding of the events triggering cystogenesis. Our study will contribute to ongoing efforts assessing the therapeutic potential for cell cycle inhibitors in the treatment of ADPKD.

描述(申请人提供)：到目前为止，在ADPKD包囊中发现的显著遗传异质性的分子机制仍不清楚。类似的核型改变也被描述为基因组不稳定的结果。我们的总体假设是，PC1的失调通过改变正常的中心体功能和细胞周期进程，导致基因组不稳定，从而促进了囊性过度增殖过程。我们的初步结果发现了支持这一主张的强有力的证据。特异性siRNA介导的PC1抑制在体外改变了细胞周期进展和中心体放大，这一发现在人类ADPKD肾组织中得到证实。中心体完整性的丧失与基因组的剧烈不稳定有关。这一应用的实验重点是阐明这些新发现背后的分子机制。第一个特定的目的是确定PC1的失调是否会导致中心体复制过程中涉及的蛋白质的异常调制。第二个具体目的是评估PC1基因敲除可能导致干扰正常细胞分裂周期进程的分子机制。第三个特定的目的是利用含有多西环素诱导的小鼠PKD1特异性siRNA的转基因小鼠模型，在体内表征PC1基因敲除在囊状细胞过度增殖和中心体扩增中的作用。综上所述，这些新颖的研究将扩大药物开发的治疗细胞周期目标，以及我们对疾病过程的理解。 公共卫生相关性 阐明ADPKD过度增殖过程的机制将有助于理解触发囊变的事件。我们的研究将有助于评估细胞周期抑制剂在ADPKD治疗中的治疗潜力。