Genetic Analysis of the Multidrug Resistance Phenotype in Tumor Cells

肿瘤细胞多药耐药表型的遗传分析

基本信息

批准号：
10925952
负责人：
Michael Gottesman
金额：
$ 28.4万
依托单位：
DIVISION OF BASIC SCIENCES - NCI
依托单位国家：
美国
项目类别：
财政年份：
资助国家：
美国
起止时间：
至
项目状态：
未结题

来源：
https://reporter.nih.gov/project-details/10925952
关键词：
ABCB1 gene ABCB6 gene ABCC1 gene ABCG2 gene ATP-Binding Cassette Transporters Amino Acids Belgium Binding Biological Availability Blood - brain barrier anatomy Brain Cause of Death Cell Death Cells Clustered Regularly Interspaced Short Palindromic Repeats Collaborations Cytotoxic agent Dimerization Doxorubicin Drug Efflux Drug Interactions Failure Family Fluorescence Goals Histone Deacetylase Inhibitor Imidazole Ketones Knock-out Laboratories Length Lipid Peroxidation Malignant Neoplasms Mediating Molecular Multi-Drug Resistance Mutate Natural Product Drug Natural Products Oral Paclitaxel Penetration Pharmaceutical Preparations Phenotype Physiological Piperazines Process Proteins Renal Cell Carcinoma Reporting Resistance Rhodamine Rhodamine 123 Role Site System Transcript Transmembrane Domain Universities Vinca Alkaloids Work cancer cell cancer drug resistance chemotherapeutic agent chemotherapy erastin genetic analysis human tissue inhibitor insight kinase inhibitor malignant phenotype melanoma member multi drug transporter multidrug resistant cancer neoplastic cell overexpression prevent promoter resistance mechanism small molecule tool uptake

项目摘要

Resistance to chemotherapy occurs in cancer cells because of intrinsic or acquired changes in expression of specific proteins. We have studied resistance to natural product chemotherapeutic agents such as doxorubicin, Vinca alkaloids, and taxol and more recently, histone deacetylase inhibitors and targeted kinase inhibitors. In most cases, cells become simultaneously resistant to multiple drugs because of reductions in intracellular drug concentrations. For the natural product drugs, this cross-resistance is frequently due to expression of an energy-dependent drug efflux system (ABC transporter) known as P-glycoprotein (P-gp), the product of the MDR1 or ABCB1 gene, or to other members of the ABC transporter family, including ABCG2 and ABCB5. In collaboration with the group of Suresh Ambudkar, we have examined the basis of directional transport of compounds out of cells by P-glycoprotein. These studies have revealed a set of amino acid residues in the transmembrane regions of P-glycoprotein which can be altered to change the direction of transport of certain rhodamine compounds from out of the cell to into the cell. This process is concentration- and ATP-dependent, and gives important insight into how directionality of transport is determined in P-glycoprotein. Further studies are underway in the Ambudkar laboratory to clarify the role of specific residues and the mechanism by which the direction of transport is reversed. In collaboration with Jean-Pierre Gillet (University of Nemours, Belgium), we have further explored the role of ABCB5 in cancer drug resistance, in the malignant phenotype in melanoma (where it is frequently mutated), and as a partner as a heterodimer with other ABC B-type transporters. In pigmented cells that express ABCB5 there are both full-length and partial transcripts generated from different promoters. The partial transcripts encode a half-transporter which appears to be able to dimerize with other half transporters (notably ABCB6 and ABCB9). The localization and potential function of these heterodimeric transporters is under study. Ferroptosis is a non-apoptotic form of cell death caused by lethal lipid peroxidation. Several small molecule ferroptosis inducers (FINs) have been reported, yet little information is available regarding their interaction with the ATP-binding cassette (ABC) transporters P-glycoprotein (P-gp, ABCB1) and ABCG2. We thus sought to characterize the interactions of FINS with P-gp and ABCG2 which may provide information regarding oral bioavailability and brain penetration and predict drug-drug interactions. P-gp overexpression conferred resistance to FIN56 and the erastin derivatives imidazole ketone erastin and piperazine erastin. P-gp-mediated resistance to imidazole ketone erastin and piperazine erastin was also reversed in UO-31 renal cancer cells by CRISPR-mediated knockout of ABCB1. The FINs ML-162, GPX inhibitor 26a, and PACMA31 at 10 micromolar were able to increase intracellular rhodamine 123 fluorescence over 10-fold in P-gp-expressing MDR-19 cells. GPX inhibitor 26a was able to increase intracellular purpurin-18 fluorescence over 4-fold in ABCG2-expressing R-5 cells. We conclude that expression of P-gp may reduce the efficacy of these FINs in cancers that express the transporter and may prevent access to sanctuary sites such as the brain. The ability of some FINs to inhibit P-gp and ABCG2 suggests potential drug-drug interactions.

由于特定蛋白质表达的内在或后天变化，癌细胞对化疗产生耐药性。我们研究了对天然产物化疗药物的耐药性，如阿霉素、长春花生物碱和紫杉醇，以及最近的组蛋白脱乙酰酶抑制剂和靶向激酶抑制剂。在大多数情况下，由于细胞内药物浓度的降低，细胞同时对多种药物产生耐药性。对于天然产物药物，这种交叉耐药性通常是由于称为 P-糖蛋白 (P-gp) 的能量依赖性药物外排系统（ABC 转运蛋白）的表达、MDR1 或 ABCB1 基因的产物或其他原因所致。 ABC 转运蛋白家族的成员，包括 ABCG2 和 ABCB5。我们与 Suresh Ambudkar 团队合作，研究了 P-糖蛋白将化合物定向转运出细胞的基础。这些研究揭示了 P-糖蛋白跨膜区域中的一组氨基酸残基，可以改变这些残基来改变某些罗丹明化合物从细胞外转运到细胞内的方向。该过程依赖于浓度和 ATP，并且为了解如何确定 P-糖蛋白中的转运方向性提供了重要的见解。 Ambudkar 实验室正在进行进一步研究，以阐明特定残基的作用以及逆转转运方向的机制。我们与 Jean-Pierre Gillet（比利时内穆尔大学）合作，进一步探讨了 ABCB5 在癌症耐药性、黑色素瘤恶性表型（经常突变）中的作用，以及作为异二聚体的合作伙伴其他 ABC B 型运输机。在表达 ABCB5 的色素细胞中，不同启动子产生全长转录本和部分转录本。部分转录物编码一个半转运蛋白，该半转运蛋白似乎能够与其他半转运蛋白（特别是 ABCB6 和 ABCB9）二聚化。这些异二聚体转运蛋白的定位和潜在功能正在研究中。铁死亡是由致命的脂质过氧化引起的一种非凋亡形式的细胞死亡。已报道了几种小分子铁死亡诱导剂 (FIN)，但有关它们与 ATP 结合盒 (ABC) 转运蛋白 P-糖蛋白 (P-gp、ABCB1) 和 ABCG2 相互作用的信息很少。因此，我们试图表征 FINS 与 P-gp 和 ABCG2 的相互作用，这可能提供有关口服生物利用度和脑渗透的信息并预测药物间相互作用。 P-gp 过表达赋予对 FIN56 和erastin 衍生物咪唑酮erastin 和哌嗪erastin 的抗性。在 UO-31 肾癌细胞中，通过 CRISPR 介导的 ABCB1 敲除，P-gp 介导的对咪唑酮erastin 和哌嗪erastin 的耐药性也被逆转。在表达 P-gp 的 MDR-19 细胞中，10 微摩尔浓度的 FIN ML-162、GPX 抑制剂 26a 和 PACMA31 能够将细胞内罗丹明 123 荧光增强 10 倍以上。 GPX 抑制剂 26a 能够将表达 ABCG2 的 R-5 细胞中的细胞内 purpurin-18 荧光增加 4 倍以上。我们得出的结论是，P-gp 的表达可能会降低表达转运蛋白的癌症中这些 FIN 的功效，并可能阻止进入大脑等避难所部位。一些 FIN 抑制 P-gp 和 ABCG2 的能力表明潜在的药物相互作用。

项目成果

期刊论文数量（47）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

Pseudovirions as vehicles for the delivery of siRNA.

DOI：
10.1007/s11095-009-0012-2
发表时间：
2010-03
期刊：
PHARMACEUTICAL RESEARCH
影响因子：
3.7
作者：
Lund, Paul E.;Hunt, Ryan C.;Gottesman, Michael M.;Kimchi-Sarfaty, Chava
通讯作者：
Kimchi-Sarfaty, Chava

Identification of compounds selectively killing multidrug-resistant cancer cells.

DOI：
10.1158/0008-5472.can-09-2422
发表时间：
2009-11-01
期刊：
Cancer research
影响因子：
11.2
作者：
Türk D;Hall MD;Chu BF;Ludwig JA;Fales HM;Gottesman MM;Szakács G
通讯作者：
Szakács G

Drug resistance: still a daunting challenge to the successful treatment of AML.

DOI：
10.1016/j.drup.2012.02.001
发表时间：
2012-02
期刊：
DRUG RESISTANCE UPDATES
影响因子：
24.3
作者：
Shaffer, Brian C.;Gillet, Jean-Pierre;Patel, Chirayu;Baer, Maria R.;Bates, Susan E.;Gottesman, Michael M.
通讯作者：
Gottesman, Michael M.

Synthesis and structure-activity evaluation of isatin-β-thiosemicarbazones with improved selective activity toward multidrug-resistant cells expressing P-glycoprotein.

DOI：
10.1021/jm2006047
发表时间：
2011-08-25
期刊：
JOURNAL OF MEDICINAL CHEMISTRY
影响因子：
7.3
作者：
Hall, Matthew D.;Brimacombe, Kyle R.;Varonka, Matthew S.;Pluchino, Kristen M.;Monda, Julie K.;Li, Jiayang;Walsh, Martin J.;Boxer, Matthew B.;Warren, Timothy H.;Fales, Henry M.;Gottesman, Michael M.
通讯作者：
Gottesman, Michael M.

Revisiting the role of ABC transporters in multidrug-resistant cancer.