Protein Arginine Methylation in Chemotherapy Resistance

化疗耐药中的蛋白质精氨酸甲基化

• 批准号: 9242960
• 负责人: Wei Xu
• 金额: $ 16万
• 依托单位: UNIVERSITY OF WISCONSIN-MADISON
• 依托单位国家: 美国
• 财政年份: 2017
• 资助国家: 美国
• 起止时间: 2017-02-01 至 2022-01-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/9242960
• 关键词: Address; Adjuvant; Aging; Anthracyclines; Antineoplastic Agents; Arginine; BRAF gene; Biological Assay; Biological Markers; Breast Cancer Cell; Breast Cancer Patient; Breast Cancer cell line; Breast Cancer therapy; Cancer Patient; Categories; Cell Culture Techniques; Cell Line; Cells; ChIP-seq; Clinical; Clinical Management; Colon Carcinoma; Cytotoxic agent; DNA Damage; Databases; Development; Down-Regulation; Drug Combinations; Drug Controls; Drug resistance; Drug toxicity; Fluorouracil; Freezing; Gene Targeting; Genes; Genetic Transcription; Goals; Histone H3; Human; Implant; In Vitro; Individual; Knock-in; Knock-out; Link; Malignant Neoplasms; Malignant neoplasm of cervix uteri; Malignant neoplasm of lung; Mammary Neoplasms; Maps; Mass Spectrum Analysis; Mediating; Mediator of activation protein; Methylation; Methyltransferase; Mitosis; Modeling; Multi-Drug Resistance; Mutate; Mutation; Neoplasm Metastasis; Oncologist; Operative Surgical Procedures; Pathologic; Patients; Pharmaceutical Preparations; Prediction of Response to Therapy; Proteins; RNA Polymerase II; Recurrence; Reporting; Research; Resistance; Resistance development; Risk; Signal Transduction; Site; Specimen; Staining method; Stains; Testing; The Cancer Genome Atlas; Therapeutic Intervention; Tissue Microarray; Transforming Growth Factors; United States; Up-Regulation; Woman; Xenograft Model; Xenograft procedure; cancer diagnosis; cancer therapy; chemotherapy; clinically relevant; coactivator-associated arginine methyltransferase 1; cohort; drug development; drug sensitivity; exome sequencing; in vivo; inhibitor/antagonist; knock-down; lung Carcinoma; malignant breast neoplasm; melanoma; mortality; mouse model; mutant; new therapeutic target; novel; oncoprotein p21; outcome forecast; overexpression; personalized approach; polyclonal antibody; predicting response; prevent; receptor; resistance mechanism; response; taxane; triple-negative invasive breast carcinoma; tumor; tumor progression; tumor xenograft; zinc finger nuclease

PROJECT SUMMARY Chemotherapy continues to be an important component of therapy for breast cancer, given to reduce the risk of metastatic recurrence. Although response (sensitivity) or lack of response (resistance) to chemotherapy is associated with prognosis, there is currently no validated commercial assay available which specifically predicts response to chemotherapy. Personalized approaches to cancer therapy are needed to select a drug or combination of drugs to which a tumor is most sensitive, and to avoid the toxicity of drugs to which the tumor is or becomes resistant. Such biomarker will assist oncologists in the daily clinical management of aging patients who are fragile to multiple chemotherapies and triple negative breast cancer patients whose major treatment option is chemotherapy. Coactivator associated arginine methyltransferase 1 (CARM1) is a protein arginine (R) methyltransferase which can methylate histone H3 and a variety of non-histone substrates. We recently identified a mediator of RNA polymerase II transcription subunit 12 (MED12) as a novel substrate for CARM1. The proposed project will determine whether methylation of MED12 is a predictor for chemo-sensitivity in breast cancer. We have mapped the methylation sites of MED12 to R1862 and R1912. Coincidently, mutations on R1862 had been reported in lung and cervical cancers, and mutations on R1912 had been found in a melanoma patient who developed resistance to BRAF inhibitor. We found that overexpression of MED12 wild-type, but not MED12R1862K,R1912K mutant, increased sensitivity of HEK293 cells to 5-FU and anthracyclines. Interestingly, the MED12 methylation dependent mechanism is distinct from activation of TGF-βR signaling as reported for complete knockout of MED12 in lung and colon cancers. Further, we have identified targets regulated by methylated MED12 that may determine chemo-sensitivity. We hypothesize that methylation of MED12 by CARM1 represents an important mechanism conferring chemosensitivity. The proposed research will directly address (1) whether MED12 methylation predicts sensitivity of breast cancer cells to commonly used chemotherapies in cell culture and xenograft tumor models; (2) define the mechanism of methylated-MED12 controlled chemosensitivity; and (3) test the clinical relevance of the MED12 methylation in chemo- resistance using large cohorts of clinical specimens. The goal is to delineate the methylation dependent mechanism for chemotherapy resistance and uncover new targets for therapeutic intervention.

项目摘要 化疗仍然是乳腺癌治疗的一个重要组成部分，以减少乳腺癌的复发。 转移复发的风险。尽管对以下物质有反应（敏感性）或无反应（耐药性）， 化疗与预后相关，目前尚无有效的商业检测方法 它能特异性预测化疗的反应。癌症治疗的个性化方法是 需要选择肿瘤最敏感的药物或药物组合，并避免毒性 肿瘤已经或正在产生耐药性的药物。这种生物标志物将有助于肿瘤学家在日常工作中， 对多种化疗敏感的老年三阴性乳腺癌患者的临床管理 主要治疗选择为化疗的癌症患者。 辅激活因子相关精氨酸甲基转移酶1（CARM 1）是一种蛋白质精氨酸（R）甲基转移酶 其可以甲基化组蛋白H3和多种非组蛋白底物。我们最近找到了一个调解人 RNA聚合酶II转录亚基12（MED 12）作为CARM 1的新底物。拟议 该项目将确定MED 12甲基化是否是乳腺癌化疗敏感性的预测因子。 我们已经将MED 12的甲基化位点定位到R1862和R1912。巧合的是，R1862上的突变 在肺癌和宫颈癌中有报道，在黑色素瘤中发现了R1912的突变 对BRAF抑制剂产生耐药性的患者。我们发现MED 12野生型的过度表达， 而MED 12 R1862 K、R1912 K突变体则不能增加HEK 293细胞对5-FU和蒽环类药物的敏感性。 有趣的是，MED 12甲基化依赖性机制与TGF-βR信号传导的激活不同， 如报道的在肺癌和结肠癌中完全敲除MED 12。此外，我们还确定了目标， 由甲基化的MED 12调节，可以决定化学敏感性。我们假设， CARM 1的MED 12代表了赋予化学敏感性的重要机制。拟议 研究将直接解决（1）MED 12甲基化是否预测乳腺癌细胞对 在细胞培养和异种移植肿瘤模型中常用的化学疗法;（2）定义 甲基化的MED 12控制的化学敏感性;和（3）测试MED 12甲基化的临床相关性。 使用大量的临床样本来研究化疗耐药性。我们的目标是描绘甲基化 依赖性机制的化疗耐药性和发现新的治疗干预的目标。