title: SLFN11 Is a Transcriptional Target of EWS-FLI1 and a Determinant of Drug Response in Ewing Sarcoma.
abstract: OBJECTIVE
  SLFN11 was identified as a critical determinant of response to DNA-targeted therapies by analyzing gene expression and drug sensitivity of NCI-60 and CCLE datasets. However, how SLFN11 is regulated in cancer cells remained unknown. Ewing sarcoma, which is characterized by the chimeric transcription factor EWS-FLI1, has notably high SLFN11 expression, leading us to investigate whether EWS-FLI1 drives SLFN11 expression and the role of SLFN11 in the drug response of Ewing sarcoma cells.
  
  
  METHODS
  Binding sites of EWS-FLI1 on the SLFN11 promoter were analyzed by chromatin immunoprecipitation sequencing and promoter-luciferase reporter analyses. The relationship between SLFN11 and EWS-FLI1 were further examined in EWS-FLI1-knockdown or -overexpressing cells and in clinical tumor samples.
  
  
  RESULTS
  EWS-FLI1 binds near the transcription start site of SLFN11 promoter and acts as a positive regulator of SLFN11 expression in Ewing sarcoma cells. EWS-FLI1-mediated SLFN11 expression is responsible for high sensitivity of Ewing sarcoma to camptothecin and combinations of PARP inhibitors with temozolomide. Importantly, Ewing sarcoma patients with higher SLFN11 expression showed better tumor-free survival rate. The correlated expression between SLFN11 and FLI1 extends to leukemia, pediatric, colon, breast, and prostate cancers. In addition, expression of other ETS members correlates with SLFN11 in NCI-60 and CCLE datasets, and molecular experiments demonstrate that ETS1 acts as a positive regulator for SLFN11 expression in breast cancer cells.
  
  
  CONCLUSIONS
  Our results imply the emerging relevance of SLFN11 as an ETS transcription factor response gene and for therapeutic response to topoisomerase I inhibitors and temozolomide-PARP inhibitor combinations in ETS-activated cancers.
authors.affiliation: Laboratory of Molecular Pharmacology, Developmental Therapeutics Branch, NCI, NIH, Bethesda, Maryland.
authors.forename: Sai-Wen
authors.initials: SW
authors.lastname: Tang
authors.affiliation: Genetics Branch, NCI, NIH, Bethesda, Maryland.
authors.forename: Sven
authors.initials: S
authors.lastname: Bilke
authors.affiliation: Genetics Branch, NCI, NIH, Bethesda, Maryland.
authors.forename: Liang
authors.initials: L
authors.lastname: Cao
authors.affiliation: Laboratory of Molecular Pharmacology, Developmental Therapeutics Branch, NCI, NIH, Bethesda, Maryland.
authors.forename: Junko
authors.initials: J
authors.lastname: Murai
authors.affiliation: Laboratory of Molecular Pharmacology, Developmental Therapeutics Branch, NCI, NIH, Bethesda, Maryland. CETROGEN, PPGFARM, UFMS, Campo Grande, Brazil.
authors.forename: Fabricio G
authors.initials: FG
authors.lastname: Sousa
authors.affiliation: Laboratory of Molecular Pharmacology, Developmental Therapeutics Branch, NCI, NIH, Bethesda, Maryland.
authors.forename: Mihoko
authors.initials: M
authors.lastname: Yamade
authors.affiliation: Laboratory of Molecular Pharmacology, Developmental Therapeutics Branch, NCI, NIH, Bethesda, Maryland.
authors.forename: Vinodh
authors.initials: V
authors.lastname: Rajapakse
authors.affiliation: Laboratory of Molecular Pharmacology, Developmental Therapeutics Branch, NCI, NIH, Bethesda, Maryland.
authors.forename: Sudhir
authors.initials: S
authors.lastname: Varma
authors.affiliation: Pediatric Oncology Branch, Center for Cancer Research, NCI, NIH, Bethesda, Maryland.
authors.forename: Lee J
authors.initials: LJ
authors.lastname: Helman
authors.affiliation: Genetics Branch, NCI, NIH, Bethesda, Maryland.
authors.forename: Javed
authors.initials: J
authors.lastname: Khan
authors.affiliation: Genetics Branch, NCI, NIH, Bethesda, Maryland.
authors.forename: Paul S
authors.initials: PS
authors.lastname: Meltzer
authors.affiliation: Laboratory of Molecular Pharmacology, Developmental Therapeutics Branch, NCI, NIH, Bethesda, Maryland. pommier@nih.gov.
authors.forename: Yves
authors.initials: Y
authors.lastname: Pommier
mesh_terms: D020816:Amino Acid Motifs; D000970:Antineoplastic Agents / Q000494:pharmacology*; D002166:Camptothecin / Q000737:chemistry; D045744:Cell Line, Tumor; D047369:Chromatin Immunoprecipitation; D004249:DNA Damage; D003606:Dacarbazine / Q000031:analogs & derivatives / Q000737:chemistry; D020869:Gene Expression Profiling; D015972:Gene Expression Regulation, Neoplastic; D057809:HEK293 Cells; D006801:Humans; D009687:Nuclear Proteins / Q000235:genetics* / Q000378:metabolism*; D020411:Oligonucleotide Array Sequence Analysis; D015514:Oncogene Proteins, Fusion / Q000378:metabolism*; D011401:Promoter Regions, Genetic; D011485:Protein Binding; D051789:Proto-Oncogene Protein c-fli-1 / Q000378:metabolism*; D034802:RNA-Binding Protein EWS / Q000378:metabolism*; D060888:Real-Time Polymerase Chain Reaction; D012512:Sarcoma, Ewing / Q000378:metabolism* / Q000473:pathology; D017422:Sequence Analysis, DNA; D000077204:Temozolomide
chemical_list: D000970:Antineoplastic Agents; C106031:EWS-FLI fusion protein; D009687:Nuclear Proteins; D015514:Oncogene Proteins, Fusion; D051789:Proto-Oncogene Protein c-fli-1; D034802:RNA-Binding Protein EWS; C576668:SLFN11 protein, human; D003606:Dacarbazine; D002166:Camptothecin; D000077204:Temozolomide