Chromosome 22q genetic studies in the colon

结肠染色体 22q 遗传学研究

• 批准号: 7238488
• 负责人: Anil K Rustgi
• 金额: $ 32.31万
• 依托单位: UNIVERSITY OF PENNSYLVANIA
• 依托单位国家: 美国
• 财政年份: 2000
• 资助国家: 美国
• 起止时间: 2000-03-01 至 2008-09-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7238488
• 关键词: 17p; 18q; 22q; 22q13; Adenomatous Polyposis Coli; Adenomatous Polyps; Allelic Imbalance; Animal Model; Bacterial Artificial Chromosomes; Bacteriophage P1; Behavior; Biochemical; Biological; Biological Assay; Breast; Candidate Disease Gene; Cell Line; Cell physiology; Cessation of life; Chromosomes; Chromosomes, Artificial, Yeast; Clinical; Cloning; Colon; Colon Carcinoma; Colonic Neoplasms; Colorectal; Colorectal Cancer; Complement; Cosmids; Custom; DNA Markers; DNA Sequence; Data; Databases; Development; Disease; Embryonic Development; Epigenetic Process; Epithelial; Epithelial Cell Proliferation; Epithelial Cells; Expressed Sequence Tags; Gene Expression; Gene Mutation; Genes; Genetic; Genomics; Genus Cola; Homeostasis; Human; Human Genome Project; Immunohistochemistry; In Situ Hybridization; In Vitro; Individual; Intestines; Length; Link; Loss of Heterozygosity; Maintenance; Malignant - descriptor; Malignant Neoplasms; Messenger RNA; Methylation; Microsatellite Repeats; Modeling; Molecular; Mutation; Nucleotides; Oral; Ovarian; Pathogenesis; Pattern; Physical Chromosome Mapping; Premalignant; Property; Proteins; Relative (related person); Research Personnel; Reverse Transcriptase Polymerase Chain Reaction; Role; SSCP Analysis; Single-Stranded Conformational Polymorphism; Small Interfering RNA; Source; TP53 gene; Technology; Tissues; Tumor Suppressor Genes; United States; Variant; Villus; Work; Xenograft Model; base; cDNA Arrays; cancer genetics; carcinogenesis; chromosome 5q loss; colon cancer cell line; innovation; insight; mRNA Expression; malignant state; novel; outcome forecast; positional cloning; programs; tumor

DESCRIPTION (provided by applicant): There are approximately 140,000 new cases of colorectal cancer annually in the United States, with nearly 60,000 deaths related to complications of disease. The model of colorectal cancer development and progression, espoused by Volgelstein and colleagues, represents a paradigm in which to understand cancer genetics. The identification of novel genes involved in proliferation and malignant transformation has been greatly enhanced through cloning after localization of chromosomal regions that are deleted (designated as loss of heterozygosity-LOH- or allelic deletion) during malignant transformation. The colon is no exception and indeed, identification of critical genes such as APC for adenomatous polyposis coli (chromosome 5q), p53 (chromosome 17p), and Smad4 (chromosome 18q). Recognizing that other chromosomal regions are frequently deleted in the progression premalignant to malignant states in the colon, we have meticulously identified a new target region of allelic loss on chromosome 22q that is involved in human colorectal carcinogenesis. Fine genetic and physical mapping with microsatellite DNA markers demonstrates that the interval on chromosome 22q13.31 is less than 1 MB and contains 14 genes (including ESTs). We hypothesize that a tumor suppressor gene(s) exist (s) whose encoded proteins harbor critical genetic, biological and biochemical properties that ultimately is important in the maintenance of intestinal epithelial cell homeostasis and linked to progression to cancer. Therefore, to achieve the identification of the gene through well established technologies and attain its subsequent molecular characterization, we will pursue well-integrated Specific Aims: (1) To determine the mRNA expression of the 14 genes in colon tumors relative to normal colon using a custom made cDNA array; and to narrow the region of LOH using PCR-SNP technology; (2) To identify the chromosome 22q13 candidate gene(s) from among other genes in the region, each candidate gene will be studied for tumor-specific alterations (mutations) using PCRSSCP followed by DNA sequencing of SSCP variants, and to investigate for the possibility of epigenetic inactivation employing PCR-methylation assays; and (3) To evaluate gene expression at the mRNA and protein levels during development, states of differentiation and proliferation, and assess correlations with clinical parameters; to evaluate the functional properties of the tumor suppressor gene in vitro and in animal models. These innovative and cohesive studies will permit important mechanistic insights into molecular pathogenesis and create a platform for translational applications.

描述（由申请人提供）：在美国，每年大约有14万例结直肠癌的新病例，近60,000例与疾病并发症有关。 由Volgelstein及其同事拥护的结直肠癌发展和进展模型代表了一种了解癌症遗传学的范式。 通过克隆在恶性转化期间被删除的染色体区域的定位（指定为杂合性-LOH或等位基因缺失）后，通过克隆进行克隆，可以通过克隆进行克隆，从而极大地增强了参与增殖和恶性转化的新基因的鉴定。 结肠也不例外，实际上，鉴定关键基因（例如APC）用于腺瘤性息肉病（5q染色体），p53（17p染色体）和SMAD4（染色体18Q）。 认识到其他染色体区域经常在结肠前对恶性状态的进展中被删除，因此我们精心识别了与人类结直肠癌发生有关的新的等位基因损失的新目标区域。 用微卫星DNA标记物的细遗传和物理映射表明，染色体22q13.31的间隔小于1 Mb，并包含14个基因（包括ESTS）。 我们假设存在肿瘤抑制基因（S），其编码蛋白具有关键的遗传，生物学和生化特性，最终对于维持肠上皮细胞稳态而言至关重要，并且与癌症的进展有关。 因此，为了通过良好的技术实现基因的鉴定并获得其随后的分子表征，我们将追求良好的集成特定目的：（1）确定使用A相对于正常结肠的14个基因的mRNA表达定制制作的cDNA阵列；并使用PCR-SNP技术缩小LOH的区域； （2）为了从该地区的其他基因中识别候选基因22q13染色体，将研究每个候选基因的基因，以进行肿瘤特异性改变（突变），然后使用PCRSSCP进行SSCP变体的DNA测序，并研究该基因的DNA测序使用PCR-甲基化测定法进行表观遗传失活的可能性； （3）在发育过程中评估在mRNA和蛋白水平上的基因表达，分化和增殖状态，并评估与临床参数的相关性；评估体外和动物模型中肿瘤抑制基因的功能特性。这些创新和凝聚力的研究将允许对分子发病机理的重要机理见解，并为翻译应用创建平台。