Characterization of WWOX, the Cancer Gene Spanning FREA16D

跨越 FREA16D 的癌症基因 WWOX 的表征

• 批准号: 7259527
• 负责人: C. Marcelo Aldaz
• 金额: $ 31.86万
• 依托单位: UNIVERSITY OF TX MD ANDERSON CAN CTR
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-01 至 2009-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7259527
• 关键词: 16q; Address; Adult; Affect; Alleles; Amino Acid Sequence; Amino Acids; Anchorage-Independent Growth; Antibodies; Binding; Binding Proteins; Biochemical; Biological; Breast; Breast Cancer Cell; Cancer cell line; Catalytic Domain; Cell physiology; Chimeric Proteins; Chromosome Arm; Chromosome Fragile Sites; Chromosomes; Complex; Cytogenetics; Development; Disruption; Embryonic Development; Epithelial; Esophageal carcinoma; Exons; Generations; Genes; Genetic Recombination; Genomics; Green Fluorescent Proteins; Immunohistochemistry; In Vitro; LacZ Genes; Malignant Neoplasms; Malignant neoplasm of esophagus; Mammary Neoplasms; Messenger RNA; Mouse Strains; Multiple Myeloma; Mus; Mutation; Nephroblastoma; Numbers; Oncogenes; Outcome; Ovarian; Ovarian Carcinoma; Oxidoreductase; Patients; Pattern; Primary carcinoma of the liver cells; Prognostic Marker; Promoter Regions; Prostate; Protein Array; Proteins; RNA Splicing; Reporting; Role; Sampling; Series; Source; Spectrum Analysis; Staging; System; Technology; Tertiary Protein Structure; Tissues; Translocation Breakpoint; Tumor Suppression; Tumorigenicity; Two-Dimensional Gel Electrophoresis; base; carcinogenesis; density; embryonic stem cell; in vivo; malignant breast neoplasm; mutant; ovarian neoplasm; polypeptide; prognostic; protein expression; protein protein interaction; recombinase; tool; tumor; tumor growth

DESCRIPTION (provided by applicant): Chromosomal and genomic abnormalities affecting chromosome 16q have frequently been reported in cytogenetic and allelotypic studies of various epithelial tumors. We recently cloned and described WWOX, a gene spanning a genomic region of more than 1 MB located in ch. 16q23.3-24.1. This specific region is the most frequent target for LOH affecting 16q in breast, ovarian, prostate and other tumors. Others and we demonstrated that this is the very same region as that of the common chromosomal fragile site 16D (FRA16D). Translocation breakpoints common in multiple myeloma are also found within the WWOX genomic region. We determined that one of the WWOX alleles is inactivated very early in breast carcinogenesis and the vast majority of primary breast tumors and breast cancer lines display this abnormality. Homozygous deletions affecting WWOX exons were detected in ovarian carcinoma, and mutations affecting WWOX were reported in esophageal carcinoma. Recently, the occurrence of aberrantly and other cancer lines spliced forms affecting WWOX mRNA have also been reported to occur in multiple tumor types. In summary, evidence from various sources indicate that WWOX is a gene commonly affected at the genomic and transcriptional level in various tumor types. Recently, we demonstrated that ectopic WWOX expression strongly inhibits anchorage-independent growth of breast cancer cell lines, and induces a dramatic inhibition of tumorigenicity of the most aggressive breast cancer cell lines. We speculate that the normal cellular function of WWOX may be affected by various different mechanisms. Our hypothesis is that WWOX is an important cancer gene that can behave as a suppressor of tumor growth and that abnormalities affecting this gene at the genomic and transcriptional level are relevant in carcinogenesis. Thus, we propose in Aim 1: To generate mouse strains containing WWOX disrupted alleles. We will A) generate a conditionally targeted WWOX mutant allelic series utilizing the cre-lox and tip-fit recombinase system and B) generate mice expressing a WWOX-LacZ fusion protein product of disruption of the endogenous WWOX alleles. This truncated WWOX form will be similar to that produce in various tumor types. Aim 2: To identify WWOX protein domains important for tumor inhibition and cellular localization. To this end mutant WWOX forms will be created affecting the WW domains and critical portions of the oxidoreductase enzymatic domain. Aim 3: To identify cellular proteins that interact with WWOX. Various candidate in vitro WWOX binding proteins containing the matching PPXP motifs have been already identified and cloned. Various approaches will be followed to define the true in vivo interacting proteins. Aim 4: To determine patterns and levels of WWOX protein expression in breast and ovarian carcinomas assessing its relevance as a prognostic tool.

描述（由申请人提供）：在各种上皮肿瘤的细胞遗传学和等位基因型研究中经常报道影响染色体 16q 的染色体和基因组异常。我们最近克隆并描述了 WWOX，这是一个跨越超过 1 MB 基因组区域的基因，位于 ch. 16q23.3-24.1。该特定区域是影响乳腺癌、卵巢癌、前列腺癌和其他肿瘤中 16q 的 LOH 最常见的靶点。其他人和我们证明，这是与常见染色体脆弱位点 16D (FRA16D) 完全相同的区域。多发性骨髓瘤中常见的易位断点也存在于 WWOX 基因组区域内。我们确定，其中一个 WWOX 等位基因在乳腺癌发生的早期就失活，并且绝大多数原发性乳腺肿瘤和乳腺癌系都表现出这种异常。在卵巢癌中检测到影响 WWOX 外显子的纯合缺失，在食道癌中报告了影响 WWOX 外显子的突变。最近，据报道，影响 WWOX mRNA 的异常剪接形式和其他癌细胞系剪接形式也发生在多种肿瘤类型中。总之，来自各种来源的证据表明，WWOX 是一种在各种肿瘤类型的基因组和转录水平上普遍受到影响的基因。最近，我们证明异位WWOX表达强烈抑制乳腺癌细胞系的贴壁独立生长，并诱导最具侵袭性乳腺癌细胞系的致瘤性的显着抑制。我们推测WWOX的正常细胞功能可能受到多种不同机制的影响。我们的假设是，WWOX 是一种重要的癌症基因，可以作为肿瘤生长的抑制因子，并且在基因组和转录水平上影响该基因的异常与癌发生相关。因此，我们在目标 1 中提出：生成含有 WWOX 破坏等位基因的小鼠品系。我们将A)利用cre-lox和tip-fit重组酶系统产生有条件靶向的WWOX突变等位基因系列，并且B)产生表达内源WWOX等位基因破坏的WWOX-LacZ融合蛋白产物的小鼠。这种截短的 WWOX 形式与各种肿瘤类型中产生的类似。目标 2：鉴定对于肿瘤抑制和细胞定位重要的 WWOX 蛋白结构域。为此，将产生影响WW结构域和氧化还原酶酶结构域的关键部分的突变WWOX形式。目标 3：鉴定与 WWOX 相互作用的细胞蛋白。含有匹配 PPXP 基序的各种候选体外 WWOX 结合蛋白已被鉴定和克隆。将采用各种方法来定义真正的体内相互作用蛋白。目标 4：确定乳腺癌和卵巢癌中 WWOX 蛋白表达的模式和水平，评估其作为预后工具的相关性。