Wwox and Fhit loss and the DNA damage response in breast cancer subtypes

Wwox 和 Fhit 丢失以及乳腺癌亚型中的 DNA 损伤反应

• 批准号: 8521109
• 负责人: KAY HUEBNER
• 金额: $ 15.02万
• 依托单位: OHIO STATE UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-09-18 至 2015-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8521109
• 关键词: African; Agreement; BRCA1 gene; Binding; Breast; Breast Cancer Cell; Cancer cell line; Carboplatin; Cell Line; Cell Nucleus; Cells; Chromosome Fragile Sites; Cisplatin; Clinical; Complex; Cytoplasm; Cytotoxic agent; DNA Damage; DNA Repair; DNA Repair Pathway; DNA damage checkpoint; Data; Defect; Development; Disseminated Malignant Neoplasm; ERBB2 gene; Epidermal Growth Factor Receptor; ErbB4 gene; FHIT gene; Funding; Genes; Genome; Gland; Goals; In Vitro; Infection; Knock-out; Knockout Mice; Laboratories; Link; Malignant Neoplasms; Mammary Neoplasms; Mammary gland; Mass Spectrum Analysis; Membrane; Modeling; Molecular; Multivariate Analysis; Mus; Neoplasms; Nickel; Nuclear; Nude Mice; Paclitaxel; Pathogenesis; Patients; Pharmaceutical Preparations; Phenotype; Predisposition; Proteins; Resistance; Risk; Role; Signal Pathway; Tamoxifen; Testing; Tissues; Transcription Factor AP-2 Alpha; Treatment outcome; Universities; Woman; Xenograft procedure; base; cancer therapy; chemotherapy; cytotoxic; cytotoxicity; high risk; in vivo; inhibitor/antagonist; killings; malignant breast neoplasm; mutant; neoplastic; promoter; research study; response; tissue culture; tumor; young woman

PROJECT SUMMARY / ABSTRACT In the last two years we have shown that: 1) Wwox, Ap2¿, and ErbB4 are independent markers of tamoxifen resistance. Reduced Wwox expression was better than PR in prediction of resistance, especially in high-risk patients, and nuclear Ap2¿ expression was better than Her2, especially in low-risk patients; 2) ErbB4 loss was an independent marker of tamoxifen resistance when adjusted for other significant predictors; 3) in vitro studies of tamoxifen resistant cells confirmed that Ap2¿ was bound by Wwox in the cytoplasm, released into the nucleus in Wwox negative, tamoxifen resistant cells and was a nuclear activator of HER2; 4) >800 invasive breast cancers on Tissue Micro Arrays (TMAs) were classified into specific subtypes: triple negative tumors (TN: ER, PR, HER2 negative, basal-like tumors) showed frequent expression of EGFR, CK5/6 and AP2¿ and frequent loss of Fhit and Wwox, suggesting that reduced Fhit and Wwox expression have roles in pathogenesis of basal differentiation in breast cancer. Alteration of expression of Fhit and Wwox occured in ~90% of the basal-like/TN breast cancers and may contribute to defects in DNA repair, as observed in BRCA1- deficient cancers. DNA damage response (DDR) proteins (¿H2AX, pChk2, p53) were expressed highly significantly more in TN and basal-like tumors. Thus, DDR checkpoint proteins could be targets for treatment of these cancers. Specific subtype breast cancer cell lines, for mechanistic studies, and TMAs with cores from primary and metastatic cancers with linked treatment and outcome data, for in vivo studies, will be used in experiments outlined in the following Aims: 1. Wwox effectors in breast cancer in vitro: a) using breast cancer cells of luminal A, HER2+ and TN/Basal subtypes, identify Wwox interactor proteins in specific subtypes, after infection with AdenoWWOX, isolation of the Wwox complex, followed by mass spectrometry identification of proteins in the complex; b) examine expression of candidate interactors in specific breast cancer subtypes in cell lines and tissues; 2. DNA damage response checkpoint inhibitors in breast cancer in vitro: a) Chk1 and Parp1 inhibitors will be tested for effect on specific breast cancer subtypes in tissue culture; b) Chk1 and Parp1 inhibitors, in combination with cytotoxic chemotherapeutic drugs, will be tested for effect on cytotoxicity of specific breast cancer subtypes; 3. Association of activated DDR checkpoint in breast cancers with treatment outcome: divide tumors on TMAs into subtypes based on ER, PR, HER2, EGFR, CK5,6 status; assess status of the DDR checkpoint proteins in tumors of each subtype and correlate markers with treatment and outcome; 4. Conditional mouse Wwox knockdown X Fhit knockout. The mouse cross will be examined for susceptibility to mammary gland cancer and mechanisms involved in development of normal and neoplastic glands. 1

项目摘要 /摘要 在过去的两年中 在抗药性预测中，WWOX表达的降低大于PR，尤其是在高危中 患者和核AP2»表达比HER2好，尤其是在低危患者中。 2）ERBB4损失是 调整其他重要预测因子时，他莫昔芬抵抗的独立标记； 3）体外研究 他莫昔芬抗药细胞证实，AP2曾在细胞质中被WWOX结合，释放到 WWOX阴性，他莫昔芬的耐药细胞中的核，是HER2的核激活剂； 4）> 800侵入性 将组织微阵列（TMA）上的乳腺癌分类为特定的亚型：三重阴性肿瘤 （TN：ER，PR，HER2阴性，基本肿瘤）经常表现出EGFR，CK5/6和AP2¿的表达 FHIT和WWOX经常丧失，表明降低的FHIT和WWOX表达在 乳腺癌基本分化的发病机理。 FHIT和WWOX表达的改变发生在 〜90％的低音状/TN乳腺癌，可能导致DNA修复的缺陷，如BRCA1-所观察到的那样 不足的癌症。 DNA损伤反应（DDR）蛋白（H2AX，PCHK2，P53）高度表达 在TN和基本肿瘤中明显更多。那就是DDR检查点蛋白可能是治疗的靶标 这些癌症。特定的亚型乳腺癌细胞系，用于机械研究和带有来自核心的TMA 用于体内研究的原始和转移性癌症将使用链接的治疗数据和结果数据 以下目的概述的实验：1。体外乳腺癌的WWOX效应：a）使用乳房 腔A，HER2+和TN/基础亚型的癌细胞鉴定特异性的WWOX相互作用蛋白 亚型，在感染Adenowwox之后，分离WWOX复合物，然后进行质谱法 鉴定复合物中的蛋白质； b）检查特定乳房中候选者相互作用的表达 细胞系和组织中的癌症亚型； 2。乳腺癌中的DNA损伤响应检查点抑制剂 体外：A）CHK1和PARP1抑制剂将对组织中特定乳腺癌亚型的影响进行测试 文化; b）将测试CHK1和PARP1抑制剂，结合细胞毒性化学治疗药物 对特定乳腺癌亚型细胞毒性的影响； 3。激活的DDR检查点的关联 具有治疗结果的乳腺癌：将TMA上的肿瘤分为基于ER，PR，HER2， EGFR，CK5,6状态； DDR检查点蛋白在每个亚型肿瘤中的评估状态并相关 带有治疗和结果的标记； 4。有条件的鼠标WWOX敲低X淘汰赛。这 将检查小鼠交叉是否对乳腺癌的敏感性和涉及的机制进行检查 正常和肿瘤腺的发展。 1

项目成果

Fhit loss-associated initiation and progression of neoplasia in vitro.

• DOI: 10.1111/cas.13032
• 发表时间: 2016-11
• 期刊: Cancer science
• 影响因子: 5.7
• 作者: Karras JR;Schrock MS;Batar B;Zhang J;La Perle K;Druck T;Huebner K
• 通讯作者: Huebner K

FHIT loss-induced DNA damage creates optimal APOBEC substrates: Insights into APOBEC-mediated mutagenesis.

• DOI: 10.18632/oncotarget.2636
• 发表时间: 2015-02-20
• 期刊: Oncotarget
• 作者: Waters CE;Saldivar JC;Amin ZA;Schrock MS;Huebner K
• 通讯作者: Huebner K

WWOX: a fragile tumor suppressor.

• DOI: 10.1177/1535370214561590
• 发表时间: 2015-03
• 期刊: Experimental biology and medicine (Maywood, N.J.)
• 作者: Schrock MS;Huebner K
• 通讯作者: Huebner K

Fhit down-regulation is an early event in pancreatic carcinogenesis.

• DOI: 10.1007/s00428-017-2105-3
• 发表时间: 2017-06
• 期刊: Virchows Archiv : an international journal of pathology
• 作者: Fassan M;Rusev B;Corbo V;Gasparini P;Luchini C;Vicentini C;Mafficini A;Paiella S;Salvia R;Cataldo I;Scarpa A;Huebner K
• 通讯作者: Huebner K

Fragile histidine triad protein, WW domain-containing oxidoreductase protein Wwox, and activator protein 2gamma expression levels correlate with basal phenotype in breast cancer.

• DOI: 10.1002/cncr.24103
• 发表时间: 2009-02-15
• 期刊: CANCER
• 影响因子: 6.2
• 作者: Guler, Gulnur;Huebner, Kay;Himmetoglu, Cigdem;Jimenez, Rafael E.;Costinean, Stefan;Volinia, Stefano;Pilarski, Robert T.;Hayran, Mutlu;Shapiro, Charles L.
• 通讯作者: Shapiro, Charles L.