ABCB5 P-Glycoprotein in Cancer Multidrug Resistance

ABCB5 P-糖蛋白在癌症多药耐药性中的作用

• 批准号: 7763930
• 负责人: Markus H. Frank
• 金额: $ 29.13万
• 依托单位: BOSTON CHILDREN'S HOSPITAL
• 依托单位国家: 美国
• 财政年份: 2006
• 资助国家: 美国
• 起止时间: 2006-04-14 至 2012-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7763930
• 关键词: ABCB1 gene; ATP-Binding Cassette Transporters; Antineoplastic Agents; Binding; Biological Assay; Calcium ion; Cancer Etiology; Cancer cell line; Cell Line; Cell fusion; Cell physiology; Cells; Chemosensitivity Assay; Chemotherapy-Oncologic Procedure; Clinical; Clinical Oncology; Developmental Therapeutics Program; Differentiation and Growth; Doxorubicin; Drug Efflux; Drug Transport; Drug resistance; Effectiveness; Experimental Animal Model; Family member; Gene Proteins; Genes; Growth; Human; Immunohistochemistry; In Vitro; Knock-out; Knockout Mice; Laboratories; Lesion; Malignant Neoplasms; Mediating; Melanoma Cell; Membrane; Membrane Potentials; Multi-Drug Resistance; Mus; National Cancer Institute; Neoplasm Metastasis; P-Glycoprotein; P-Glycoproteins; Pharmaceutical Preparations; Phenotype; Physiological; Population; Research Personnel; Resistance; Resistance profile; Reverse Transcriptase Polymerase Chain Reaction; Rhodamine; Rhodamines; Role; Skin; Small Interfering RNA; Staining method; Stains; Stem cells; Therapeutic; Tissue Microarray; Tissues; Tumor Stem Cells; Vesicle; Xenograft Model; base; cancer cell; cancer therapy; in vivo; melanoma; neoplastic cell; new therapeutic target; novel; programs; protein expression; self-renewal; success; tumor; tumor eradication; tumor progression; tumor xenograft; uptake

Multidrug resistance (MDR) mediated by MDR1 (ABCB1) P-glycoprotein and related ATP-binding cassette (ABC) transporters is an impediment to successful cancer therapy. ABCB5 P-glycoprotein is a novel ABC transporter, which mediates drug efflux in cancer cells and regulates cell fusion and resultant differentiation of progenitor cells in physiological tissues. It is hypothesized that ABCB5 mediates dual functions in cancer, conferring MDR via its function as a drug efflux transporter, and regulating tumor renewal as a determinant of cell fusion involving tumor stem cells. This study aims to (1) identify systematically ABCB5 drug efflux substrates and examine the role of ABCB5 in the chemoresistance to such compounds, (2) determine whether ABCBS-expressing cancer cells function as tumor stem cells, and (3) investigate the role of ABCB5 in cancer MDR and tumor formation in vivo and examine whether ABCB5 can be specifically targeted for tumor eradication. First, ABCB5 gene and protein expression will be assayed across the NCI-60 cancer cell lines with resistance profiles for >100,000 compounds and drug resistance correlations will be established by. COMPARE analysis. Candidate ABCB5 transport substrates will be validated experimentally using competitive drug efflux analysis in ABCB5 gene-transfected cell lines, and chemosensitivity assays performed in ABCBS-blocked human cancer cell lines and murine ABCB5 -/- knockout cells. Second, the capacity of ABCBS-positive tumor cells vis-a-vis ABCBS-negative tumor bulk populations for self-renewal and the mechanistic contribution of ABCBS-positive progenitor cells to tumor culture growth and differentiation will be examined in vitro using tumor clonogenicity and cell fusion assays. Third, the in vivo role of ABCB5 in chemoresistance to identified drug substrates and the in vivo relevance of ABCBS-positive tumor cells for tumor formation and growth will be studied in human to mouse tumor xenograft models. The results will define the role of ABCB5 in cancer MDR and will establish whether chemoresistant ABCBS- positive cells function as tumor stem cells in expressing cancers. Thus, the findings will identify whether ABCB5 represents a novel therapeutic target in clinical oncology.

MDR1 (ABCB1) P-糖蛋白和相关 ATP 结合盒介导的多药耐药性 (MDR) (ABC) 转运蛋白是成功癌症治疗的障碍。 ABCB5 P-糖蛋白是一种新型 ABC 转运蛋白，介导癌细胞中的药物流出并调节细胞融合和由此产生的分化 生理组织中的祖细胞。据推测 ABCB5 在癌症中介导双重功能， 通过其作为药物外流转运蛋白的功能赋予 MDR，并作为决定因素调节肿瘤更新 涉及肿瘤干细胞的细胞融合。本研究旨在 (1) 系统地识别 ABCB5 药物流出 底物并检查 ABCB5 在此类化合物的化学抗性中的作用，(2) 确定 表达ABCBS的癌细胞是否具有肿瘤干细胞的功能，以及（3）研究ABCB5的作用 癌症 MDR 和体内肿瘤形成，并检查 ABCB5 是否可以特异性靶向 肿瘤根除。首先，将在 NCI-60 癌细胞中检测 ABCB5 基因和蛋白质表达 将建立具有> 100,000种化合物的耐药性谱线和耐药性相关性的线。 比较分析。候选 ABCB5 传输底物将使用以下方法进行实验验证 ABCB5 基因转染细胞系中的竞争性药物流出分析和化学敏感性测定 在 ABCBS 阻断的人类癌细胞系和小鼠 ABCB5 -/- 敲除细胞中进行。其次， ABCBS 阳性肿瘤细胞相对于 ABCBS 阴性肿瘤群体的自我更新能力 以及 ABCBS 阳性祖细胞对肿瘤培养物生长和生长的机制贡献 将使用肿瘤克隆形成和细胞融合测定在体外检查分化。三、体内 ABCB5 在对已识别药物底物的化学耐药性中的作用以及 ABCBS 阳性的体内相关性 将在人对小鼠肿瘤异种移植模型中研究肿瘤细胞对肿瘤形成和生长的影响。这 结果将确定 ABCB5 在癌症 MDR 中的作用，并将确定化疗耐药性 ABCBS- 阳性细胞在表达癌症中充当肿瘤干细胞。因此，研究结果将确定是否 ABCB5代表了临床肿瘤学中的一个新的治疗靶点。