Forward genetic screens in mice uncover mediators and suppressors of metastatic reactivation.
We have developed a screening platform for the isolation of genetic entities involved in metastatic reactivation. Retroviral libraries of cDNAs from fully metastatic breast-cancer cells or pooled microRNAs were transduced into breast-cancer cells that become dormant upon infiltrating the lung. Upon inoculation in the tail vein of mice, the cells that had acquired the ability to undergo reactivation generated metastatic lesions. Integrated retroviral vectors were recovered from these lesions, sequenced, and subjected to a second round of validation. By using this strategy, we isolated canonical genes and microRNAs that mediate metastatic reactivation in the lung. To identify genes that oppose reactivation, we screened an expression library encoding shRNAs, and we identified target genes that encode potential enforcers of dormancy. Our screening strategy enables the identification and rapid biological validation of single genetic entities that are necessary to maintain dormancy or to induce reactivation. This technology should facilitate the elucidation of the molecular underpinnings of these processes.
Research Center for Translational Medicine, East Hospital, School of Life Sciences and Technology, Tongji University, Shanghai 200092, China; and Cell Biology Program and Metastasis Research Center and f-giancotti@ski.mskcc.org gaoh@tongji.edu.cn.
Hua
H
Gao
Cell Biology Program and Metastasis Research Center and.
Goutam
G
Chakraborty
Cell Biology Program and Metastasis Research Center and.
Ai Ping
AP
Lee-Lim
Cancer Biology and Genetics Program, Sloan Kettering Institute for Cancer Research, Memorial Sloan Kettering Cancer Center, New York, NY 10065.
Konstantinos J
KJ
Mavrakis
Cancer Biology and Genetics Program, Sloan Kettering Institute for Cancer Research, Memorial Sloan Kettering Cancer Center, New York, NY 10065.
Hans-Guido
HG
Wendel
Cell Biology Program and Metastasis Research Center and f-giancotti@ski.mskcc.org gaoh@tongji.edu.cn.
Filippo G
FG
Giancotti
D000818:Animals; D002277:Carcinoma / Q000556:secondary; D045744:Cell Line, Tumor; D018076:DNA, Complementary / Q000235:genetics; D004273:DNA, Neoplasm / Q000235:genetics; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D015723:Gene Library; D052138:Genes, Neoplasm; D005822:Genetic Vectors / Q000235:genetics / Q000302:isolation & purification; D057166:High-Throughput Screening Assays / Q000379:methods*; D008175:Lung Neoplasms / Q000556:secondary; D008325:Mammary Neoplasms, Experimental / Q000473:pathology*; D051379:Mice; D008807:Mice, Inbred BALB C; D035683:MicroRNAs / Q000235:genetics / Q000302:isolation & purification; D009362:Neoplasm Metastasis / Q000235:genetics* / Q000473:pathology; D009363:Neoplasm Proteins / Q000235:genetics / Q000502:physiology; D009368:Neoplasm Transplantation; D011533:Proviruses / Q000235:genetics / Q000302:isolation & purification; D012334:RNA, Neoplasm / Q000235:genetics / Q000302:isolation & purification; D034741:RNA, Small Interfering / Q000235:genetics; D012190:Retroviridae / Q000235:genetics / Q000302:isolation & purification; D014161:Transduction, Genetic; D016662:Virus Integration
D018076:DNA, Complementary; D004273:DNA, Neoplasm; D035683:MicroRNAs; D009363:Neoplasm Proteins; D012334:RNA, Neoplasm; D034741:RNA, Small Interfering
cDNA library screen; forward genetic screens; metastatic reactivation; microRNA library screen; shRNA library screen