MetSearch: A screening platform for the discovery of genes implicated in metastatic dormancy and reactivation

ABSTRACT: We have developed a screening platform for the isolation of genetic entities involved in metastatic reactivation. Retroviral libraries encoding cDNAs from highly 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 identified 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. We conducte miRNA microarray analysis (Agilent Technologies) as a complementary technique to examine miRNA expression profiles in our tumor progression series model, comparing the non-metastatic cells lines (67NR, 168FARN, 4TO7) with that of the metastatic cell line (4T1). By studying these profiles, we can identify sets of miRNAs that regulate the different steps of metastasis, namely intravasation, survival in the circulatory system and at distant sites, and extravasation and metastases formation, as represented by the different cell lines.

ORGANISM(S): Mus musculus

SUBMITTER: Jeffrey Zhao 

PROVIDER: E-GEOD-43483 | biostudies-arrayexpress |

REPOSITORIES: biostudies-arrayexpress