Identification of an agent selectively targeting DPC4 (deleted in pancreatic cancer locus 4)-deficient pancreatic cancer cells.
One of the most common types of genetic alterations in cancer is the loss-of-function mutations in tumor-suppressor genes. Such mutations are usually very specific to cancer cells and present attractive and unique opportunities for therapeutic interventions. However, for various reasons, antitumor agents that target loss-of-function mutations have not been readily identified. In this report, using the deleted in pancreatic cancer locus 4 (DPC4) gene in pancreatic cancer as an example, we show the feasibility of a novel screening strategy, which we have named Pharmacological Synthetic Lethal Screening, for the identification of agents that selectively target cancer cells with loss-of-function mutations. We created DPC4 isogenic cell lines through the restoration of wild-type DPC4 in a pancreatic cancer cell line, BxPC-3, whose DPC4 gene was homozygously deleted. The isogenic cell lines were then used in the Pharmacological Synthetic Lethal Screening to evaluate compound libraries for antiproliferative activity and selectivity against DPC4 deficiency. After screening 19,590 compounds, we identified one lead compound, UA62001, which showed 4.6-fold selectivity against DPC4 deficiency in the DPC4 isogenic cell lines. UA62001 selectivity was also seen in another set of DPC4 isogenic cell lines generated by small interfering RNA knockdown. In addition, UA62001 was evaluated in commonly used pancreatic cancer cell lines. A fairly good correlation between DPC4 deficiency and UA62001 sensitivity was observed. Cell cycle analysis indicates that UA62001 arrests cells in S and G(2)-M phases. The results of microarray gene expression profiling and quantitative real-time reverse transcription-PCR suggest that cyclin B/CDC2 and minichromosome maintenance complexes might be the downstream cellular targets of UA62001.
Wang
Hong
H
Pancreatic Cancer Research Laboratory, Clinical Translational Research Division, Translational Genomics Research Institute, Phoenix, AZ 85004, USA. hwang@tgen.org
Han
Haiyong
H
Von Hoff
Daniel D
DD
D000230:Adenocarcinoma / Q000235:genetics / Q000473:pathology*; D000880:Anthraquinones / Q000494:pharmacology*; D000970:Antineoplastic Agents / Q000494:pharmacology*; D016203:CDC2 Protein Kinase / Q000378:metabolism; D045744:Cell Line, Tumor / Q000187:drug effects; D019926:Cyclin B / Q000378:metabolism; D004354:Drug Screening Assays, Antitumor / Q000379:methods*; D005240:Feasibility Studies; D005347:Fibroblasts / Q000187:drug effects; D016195:G2 Phase / Q000187:drug effects; D020869:Gene Expression Profiling; D006801:Humans; D007191:Indazoles / Q000494:pharmacology*; D009363:Neoplasm Proteins / Q000172:deficiency* / Q000235:genetics; D020411:Oligonucleotide Array Sequence Analysis; D010190:Pancreatic Neoplasms / Q000235:genetics / Q000473:pathology*; D034622:RNA Interference; D034741:RNA, Small Interfering / Q000494:pharmacology; D011993:Recombinant Fusion Proteins / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016196:S Phase / Q000187:drug effects; D051901:Smad4 Protein / Q000172:deficiency* / Q000235:genetics
C524064:2-phenyl-1,2-dihydronaphtho(2,3-g)indazole-3,6,11-trione; D000880:Anthraquinones; D000970:Antineoplastic Agents; D019926:Cyclin B; D007191:Indazoles; D009363:Neoplasm Proteins; D034741:RNA, Small Interfering; D011993:Recombinant Fusion Proteins; C497796:SMAD4 protein, human; D051901:Smad4 Protein; D016203:CDC2 Protein Kinase