Establishment of array-based CGH technology and its use for-diagnosis of genetic diseases in clinical setting

基于芯片的CGH技术的建立及其在临床遗传疾病诊断中的应用

• 批准号: 15390112
• 负责人: INAZAWA Johji
• 金额: $ 10.18万
• 依托单位: Tokyo Medical and Dental University
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2003
• 资助国家: 日本
• 起止时间: 2003 至 2004
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-15390112/
• 关键词: CGH; Genetic disease; Chromosome aberration; BAC; PAC; DNA methylation; Autism; contiguous gene syndrome

Comparative genomic hybridization (CGH) has already made a significant impact on cancer cytogenetics. However, CGH to metaphase chromosomes can provide only limited resolution at 5-10 Mb level. To circumvent this limitation, array-based CGH has been devised. We have constructed different types of BAC-based CGH-arrays. The first consisted of 〜4500 BACs for genome-wide analysis (named MCG Whole Genome Array-4500) ; this array provided a resolution of 〜0.7Mb. The second consisted of 〜800 BACs harboring 800 known cancer-relaed genes, intended for diagnosis of cancer-specific copy-number aberrations (MCG Cancer Array-800, see URL : http://www.cghtmd.jp/cghdatabase/index.html). The third of our arrays contains 212 contiguous BACs spanning a 〜20-Mb region at 1p36 (MCG 1p36 Contig Array). Furthermore, to accomplish high-throughput screening for methylated sites in the whole genome, we have combined array-CGH with MCA, and our preliminary data show that this "BAC array-based MCA (BAMCA)" can discriminate BAC clones that harbor methylated CpGs on an array platform. Our custom-made CGH-arrays paves the way for identification of disease-related genetic aberrations that have not yet been detected by existing technologies, and our array-based CGH technology should soon be practical for diagnosis of cancer or genetic diseases in clinical settings.

比较基因组杂交（CGH）已经对癌症细胞遗传学产生了重大影响。然而，中期染色体的CGH只能提供5-10 Mb水平的有限分辨率。为了克服这一限制，基于阵列的计算全息图已经被设计出来。我们已经构建了不同类型的BAC为基础的CGH阵列。第一个由4500个BAC组成，用于全基因组分析（命名为MCG全基因组阵列-4500）;该阵列提供了10.7Mb的分辨率。第二组由10800个BAC组成，其含有800个已知的癌症相关基因，旨在诊断癌症特异性拷贝数畸变（MCG Cancer Array-800，参见URL：http：//www.cghtmd.jp/cghdatabase/index.html）。我们的第三个阵列包含212个连续的BAC，跨越1 p36处的20-Mb区域（MCG 1 p36 Contig阵列）。此外，为了实现全基因组中甲基化位点的高通量筛选，我们将阵列CGH与MCA相结合，我们的初步数据表明，这种“基于BAC阵列的MCA（BAMCA）”可以在阵列平台上区分携带甲基化CpG的BAC克隆。我们定制的CGH阵列为识别现有技术尚未检测到的疾病相关遗传畸变铺平了道路，我们基于阵列的CGH技术应该很快就能在临床环境中用于癌症或遗传疾病的诊断。

项目成果

学術月報

学术月报

• 发表时间: 2004
• 作者: Inoue J;Inazawa J et al.;稲澤 譲治
• 通讯作者: 稲澤 譲治

Hirasawa A, Inazawa J et al.: "Association of 17q21-q24 Gain in Ovarian Clear Cell Adenocarcinomas with Poor Prognosis and Identification of PPM1D and APPBP2 as Likely Amplification Targets"Clinical Cancer Research. 9. 1995-2004 (2003)

Hirasawa A、Inazawa J 等人：“卵巢透明细胞腺癌中 17q21-q24 增益与不良预后的关联以及 PPM1D 和 APPBP2 作为可能扩增靶点的识别”临床癌症研究。

Yokoi S, Inazawa J et al.: "TERC Identified as a probable target within the 3q26 amplicon that is detected frequently in non-small cell lung cancers"Clinical Cancer Research. 15. 4705-4713 (2003)

Yokoi S、Inazawa J 等人：“TERC 被确定为 3q26 扩增子中的一个可能靶标，该扩增子在非小细胞肺癌中经常检测到”临床癌症研究。

Imoto I, Inazawa J, et al.: "Identification of ZASC1 encoding a Kruppel-like zinc finger protein as a novel target for 3q26 amplification in esophageal squamous-cell carcinomas."Cancer Research. 63. 5691-5696 (2003)

Imoto I、Inazawa J 等人：“鉴定编码 Kruppel 样锌指蛋白的 ZASC1 作为食管鳞状细胞癌 3q26 扩增的新靶标。”癌症研究。

Okamoto H, Inazawa J et al.: "PTK2 and EIF3S3 Genes May be Amplification Targets at 8q23-q24 and Are Associated with Large Hepatocellular Carcinomas"Hepatology. 38. 1242-1249 (2003)

Okamoto H、Inazawa J 等人：“PTK2 和 EIF3S3 基因可能是 8q23-q24 处的扩增目标，并且与大肝细胞癌相关”肝病学。