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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.

癌细胞对化疗产生耐药性是由于特定蛋白表达的内在或获得性变化。我们已经研究了天然产物化疗药物的耐药性，如阿霉素、长春花碱和紫杉醇，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抗药性。对于天然产物药物，这种交叉耐药性通常是由于被称为P-糖蛋白(P-gp)的能量依赖的药物外排系统(ABC转运体)的表达，P-糖蛋白(P-gp)是MDR1或ABCB1基因的产物，或者ABC转运体家族的其他成员，包括ABCG2和ABCB5。在Suresh Ambukar小组的合作下，我们研究了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-糖蛋白跨膜区的一组氨基酸残基，这些残基可以改变某些罗丹明化合物从细胞外到细胞内的运输方向。这一过程依赖于浓度和ATP，并对P-糖蛋白如何决定转运的方向性提供了重要的见解。Ambukar实验室正在进行进一步的研究，以澄清特定残留物的作用和逆转运输方向的机制。与比利时内穆斯大学的Jean-Pierre Gillet合作，我们进一步探索了ABCB5在癌症耐药性、黑色素瘤恶性表型(经常发生突变)中的作用，以及作为异源二聚体与其他ABC B型转运蛋白的合作伙伴。在表达ABCB5的有色细胞中，有来自不同启动子的全长和部分转录本。部分转录本编码一个半转运蛋白，它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。铁下垂是一种由致命性脂质过氧化引起的非凋亡性细胞死亡形式。小分子铁下垂诱导剂(FINs)已经被报道，但关于它们与三磷酸腺苷结合盒(ABC)转运体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，这可能提供关于口服生物利用度和脑渗透的信息，并预测药物-药物相互作用。P-gp的过表达使人对FIN56和erastin的衍生物咪唑酮、erastin和哌嗪erastin产生抗性。CRISPR介导的ABCB1基因敲除也逆转了P-gp介导的对咪唑酮、erastin和哌嗪erastin的耐药性。10微摩尔的FINS ML-162、GPX抑制剂26a和PACMA31能使表达P-gp的MDR-19细胞内罗丹明123的荧光增强10倍以上。在表达ABCG2的R-5细胞中，GPX抑制剂26a能使细胞内紫红素-18的荧光增加4倍以上。我们得出结论，P-gp的表达可能会降低这些FINS在表达转运蛋白的癌症中的疗效，并可能阻止对脑等避难所的访问。一些FINS抑制P-gp和ABCG2的能力暗示了潜在的药物-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）

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

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

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.

Identification of a Cryptic Bacterial Promoter in Mouse (mdr1a) P-Glycoprotein cDNA.

小鼠 (mdr1a) P-糖蛋白 cDNA 中隐秘细菌启动子的鉴定。

DOI：
10.1371/journal.pone.0136396
发表时间：
2015
期刊：
PloS one
影响因子：
3.7
作者：
Pluchino,KristenM;Esposito,Dominic;Moen,JannaK;Hall,MatthewD;Madigan,JamesP;Shukla,Suneet;Procter,LaurenV;Wall,VanessaE;Schneider,ThomasD;Pringle,Ian;Ambudkar,SureshV;Gill,DeborahR;Hyde,StevenC;Gottesman,MichaelM
通讯作者：
Gottesman,MichaelM

Targeting the Achilles heel of multidrug-resistant cancer by exploiting the fitness cost of resistance.

DOI：
10.1021/cr4006236
发表时间：
2014-06-11
期刊：
CHEMICAL REVIEWS
影响因子：
62.1
作者：
Szakacs, Gergely;Hall, Matthew D.;Gottesman, Michael M.;Boumendjel, Ahcene;Kachadourian, Remy;Day, Brian J.;Baubichon-Cortay, Helene;Di Pietro, Attilio
通讯作者：
Di Pietro, Attilio