MAPPING AND CLONING THE 17Q-LINKED BREAST CANCER LOCUS

绘制和克隆 17Q 连锁乳腺癌基因座

基本信息

批准号：
2097008
负责人：
MARK H SKOLNICK
金额：
$ 26.27万
依托单位：
UNIVERSITY OF UTAH
依托单位国家：
美国
项目类别：
财政年份：
1993
资助国家：
美国
起止时间：
1993-02-01 至 1996-01-31
项目状态：
已结题

项目摘要

The localization of a susceptibility allele for breast cancer to chromosome 17q is the first successful mapping of a common cancer. The goal of this proposal is to clone and genetically characterize this 17q breast cancer susceptibility locus. Just as localizing common diseases to a specific chromosome has proven to be difficult and error-prone, the subsequent fine structure mapping and gene isolation will be more difficult than for rare mendelian traits. The unique characteristics of the Utah population for genetic analysis will be an essential asset for this project. Our project will be one of many tied together in a scientific consortium jointly providing the resources necessary for the analysis of the 17q susceptibility locus. A key to the success of this project is the ability to identify large informative kindreds for mapping studies. Highly informative kindreds with a high likelihood of segregating a breast cancer susceptibility will be studied. Linkage analysis will determine whether a kindred's susceptibility is linked to 17q. A map of highly informative markers will be developed around the susceptibility locus. We have identified recombinants which flank the breast cancer susceptibility locus and characterized a hybrid panel created by K. Fournier with breakpoints surrounding the gene. A cosmid library will be constructed from a Fournier hybrid which contains 10 to 15 megabases of human DNA including the region containing the gene. Clones which map genetically and physically within the candidate gene region are used as nucleation points to develop a contig of the region. The final product will be a YAC contig, a pulsed field map of rare cutters, and a cosmid contig of the region with genetic boundaries defining the candidate region where the susceptibility locus must lie. We have eliminated seven potential candidate genes (EDBH17, ERB B2, HOX2, NM23, WNT3, RARA, and Prohibitin) from the region; therefore, it is virtually certain that the susceptibility locus is a novel gene. Clones derived from cDNA libraries will be selected by subtractive hybridization, hybridization of cDNAs to an affinity column, and direct hybridization of cosmid clones to cDNA clones. All cDNAs will be screened for DNA sequence differences between cases and controls once intron-exon boundaries are defined. We will characterize the phenotypic effect of the 17q-linked susceptibility locus in terms of its site and age-specific penetrance. We will examine questions of laterality of disease and try to identify any histopathologic features characteristic of 17q-linked breast cancer. Data on reproductive history will be gathered and interaction with the susceptibility locus will be assessed. If, during the course of this study the 17q gene is identified, we will screen for mutations in each family and stratify the analyses described on a mutation-specific basis.

乳腺癌易感性等位基因的定位 17q 染色体首次成功绘制了常见癌症的图谱。这该提案的目标是克隆该 17q 并对其进行遗传表征乳腺癌易感位点。就像定位常见疾病一样事实证明，对特定染色体进行定位是困难且容易出错的，后续的精细结构图谱和基因分离将会更加比罕见的孟德尔特征更困难。独特的特点用于遗传分析的犹他州人口将成为重要资产这个项目。我们的项目将是众多项目之一科学联盟共同提供必要的资源 17q 易感位点分析。此次活动成功的一个关键项目是识别大量信息丰富的亲属以进行绘图的能力研究。信息丰富的亲属，很有可能将研究分离乳腺癌易感性。连锁分析将确定亲属的易感性是否与 17q。将围绕该区域开发一张信息丰富的标记地图易感位点。我们已经鉴定出侧翼的重组体乳腺癌易感性位点和混合组的特征由 K. Fournier 创建，在基因周围设有断点。粘粒文库将由 Fournier 混合体构建，其中包含 10 至 15 兆碱基的人类 DNA，包括含有该基因的区域。在候选基因内进行遗传和物理定位的克隆区域被用作成核点以形成该区域的重叠群。最终产品将是 YAC 重叠群，这是稀有物质的脉冲场图切割器和具有遗传边界的区域的粘粒重叠群定义易感性位点必须位于的候选区域。我们已经消除了七个潜在的候选基因（EDBH17、ERB B2、HOX2、来自该地区的 NM23、WNT3、RARA 和 Prohibitin）；因此，它是几乎可以肯定，易感位点是一个新基因。克隆来自 cDNA 文库的衍生品将通过消减法进行选择杂交，cDNA 与亲和柱杂交，以及直接杂交粘粒克隆与cDNA克隆的杂交。所有 cDNA 将筛查病例和对照之间的 DNA 序列差异一次定义了内含子-外显子边界。我们将表征表型 17q连锁易感位点对其位点的影响特定年龄的外显率。我们将研究偏侧性问题疾病并尝试识别任何组织病理学特征 17q连锁乳腺癌。有关生殖史的数据将收集并评估与易感性位点的相互作用。如果在本研究过程中鉴定出 17q 基因，我们将筛选每个家族中的突变并对描述的分析进行分层在特定突变的基础上。