array-based screening for the expression and regulation of tumor supressor genes

基于芯片的抑癌基因表达和调控筛选

基本信息

批准号：
8509664
负责人：
Patrick Kyongmin Ha
金额：
$ 11.81万
依托单位：
JOHNS HOPKINS UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-07-13 至 2014-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/8509664
关键词：
Adenoid Cystic Carcinoma Affect Aggressive behavior Azacitidine Behavior Bioinformatics Biological Assay Candidate Disease Gene Cell Line Characteristics Clinical Data Data Set Development Disease Distant Metastasis Dose Epigenetic Process Evaluation Gene Silencing Generations Genes Genome Goals Growth Harvest Human Hypermethylation Immunodeficient Mouse In Vitro Indolent Investigation Lead Life Lung Malignant Epithelial Cell Malignant Neoplasms Methods Methylation Modeling Molecular Mus Neoplasm Metastasis Nude Mice Operative Surgical Procedures Pathway interactions Patients Pattern Pharmaceutical Preparations Postoperative Period Primary Neoplasm Process Radiation therapy Regulation Research Resources Role Running Salivary Gland Adenoid Cystic Carcinoma Salivary Glands Sampling Source Stable Disease Surveys Techniques Therapeutic Tissues Tumor Cell Line Tumor Suppressor Genes Tumor Tissue Up-Regulation Validation base bisulfite bone carcinogenesis cohort cytotoxic demethylation experience gene discovery genome wide association study genome wide methylation genome-wide implantation novel novel strategies perineural promoter public health relevance research study screening therapy development tumor

项目摘要

DESCRIPTION (provided by applicant): Adenoid cystic carcinoma of the salivary glands (ACC) is a rare malignancy with an unusual clinical behavior, marked by aggressive local growth which is often controlled with surgery and postoperative radiation therapy, followed by a dormant period that may exist for years. However, up to 50% of patients experience distant metastasis, most often to lung or bone, which may remain indolent for years as well. Not much is known about the molecular underpinnings of ACC and thus cytotoxic or targeted therapy has been empiric and inconsistent. Promoter methylation has emerged as a prominent epigenetic mechanism of tumor suppressor gene silencing in ACC and most other tumor models. By capitalizing on the ability of 5-aza-cytidine treatment to produce global demethylation, we are able to pharmacologically unmask tumor suppressor genes, previously silenced in primary ACCs. This exposure will lead to gene upregulation, which can be detected with expression microarrays, and promoter demethylation, which can be detected with newer generation methylation arrays. While this discovery assay is typically performed in vitro, there presently are no reliable ACC cell line models. We thus propose performing a novel approach of 5-aza-cytidine treatment in heterotransplanted ACC tumors from primary patients and treating these nude mice with daily, moderate-dose 5-aza-cytidine for 28 days. Once performed, the tissue may be harvested, arrays run, and the bioinformatic integration and analysis completed. The gene list will then be validated for its methylation status in a subset of normal salivary gland an primary ACC samples through bisulfite sequencing. At the conclusion of this project, we will have genome-wide characterization of candidate tumor suppressor genes under the control of promoter methylation. This powerful technique involving integration across array platforms allows for a wider range of gene discovery than by the use of methylation arrays alone, and will provide us with a more streamlined validation process. The candidate genes identified will then provide more focused targets for further investigation into the basic mechanisms of ACC carcinogenesis.

描述（由申请人提供）：唾液腺（ACC）的腺样性囊性癌是一种罕见的恶性肿瘤，具有不寻常的临床行为，其特征是侵略性局部生长，通常由手术和术后放射治疗控制，随后是可能存在多年的休眠期。但是，多达50％的患者经历了遥远的转移，最常见于肺或骨骼，这种转移可能会持续多年。关于ACC的分子基础知之甚少，因此细胞毒性或靶向治疗是经验性和不一致的。启动子甲基化已成为ACC和大多数其他肿瘤模型中肿瘤抑制基因沉默的显着表观遗传机制。通过利用5-氮杂 - 胞苷处理产生全球脱甲基化的能力，我们能够在药理学上揭示抑制肿瘤基因，以前在主要ACC中沉默。这种暴露将导致基因上调，可以用表达微阵列检测到，启动子脱甲基化，可以用新的甲基化阵列检测到。虽然该发现测定法通常是在体外进行的，但目前有没有可靠的ACC单元线模型。因此，我们提出了一种新的方法，该方法是从初级患者的异质移植ACC肿瘤中进行5-氮杂胞苷治疗的新方法，并用每天的，中等剂量的5-aza-cytidine治疗这些裸鼠28天。一旦执行，就可以收获组织，运行阵列，并完成生物信息学的整合和分析。然后，将通过亚硫酸盐测序在正常唾液腺的一部分中对其甲基化状态的甲基化状态进行验证。在该项目的结尾，我们将在启动子甲基化的控制下对候选肿瘤抑制基因的全基因组表征。这种涉及跨数组平台集成的功能强大的技术可以比单独使用甲基化阵列进行更广泛的基因发现，并将为我们提供更简化的验证过程。然后，所鉴定的候选基因将提供更多的重点靶标，以进一步研究ACC癌变的基本机制。

项目成果

期刊论文数量（1）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Identification of methylated genes in salivary gland adenoid cystic carcinoma xenografts using global demethylation and methylation microarray screening.

DOI：
10.3892/ijo.2016.3532
发表时间：
2016-07
期刊：
International journal of oncology
影响因子：
5.2
作者：
Ling S;Rettig EM;Tan M;Chang X;Wang Z;Brait M;Bishop JA;Fertig EJ;Considine M;Wick MJ;Ha PK
通讯作者：
Ha PK