title: An inducible, isogenic cancer cell line system for targeting the state of mismatch repair deficiency.
abstract: The DNA mismatch repair system (MMR) maintains genome stability through recognition and repair of single-base mismatches and small insertion-deletion loops. Inactivation of the MMR pathway causes microsatellite instability and the accumulation of genomic mutations that can cause or contribute to cancer. In fact, 10-20% of certain solid and hematologic cancers are MMR-deficient. MMR-deficient cancers do not respond to some standard of care chemotherapeutics because of presumed increased tolerance of DNA damage, highlighting the need for novel therapeutic drugs. Toward this goal, we generated isogenic cancer cell lines for direct comparison of MMR-proficient and MMR-deficient cells. We engineered NCI-H23 lung adenocarcinoma cells to contain a doxycycline-inducible shRNA designed to suppress the expression of the mismatch repair gene MLH1, and compared single cell subclones that were uninduced (MLH1-proficient) versus induced for the MLH1 shRNA (MLH1-deficient). Here we present the characterization of these MMR-inducible cell lines and validate a novel class of rhodium metalloinsertor compounds that differentially inhibit the proliferation of MMR-deficient cancer cells.
authors.affiliation: Oncology Research, Amgen Inc., South San Francisco, California, United States of America.
authors.forename: Julie M
authors.initials: JM
authors.lastname: Bailis
authors.forename: Marcia L
authors.initials: ML
authors.lastname: Gordon
authors.forename: Jesse L
authors.initials: JL
authors.lastname: Gurgel
authors.forename: Alexis C
authors.initials: AC
authors.lastname: Komor
authors.forename: Jacqueline K
authors.initials: JK
authors.lastname: Barton
authors.forename: Ilan R
authors.initials: IR
authors.lastname: Kirsch
mesh_terms: D048868:Adaptor Proteins, Signal Transducing / Q000172:deficiency / Q000235:genetics; D000970:Antineoplastic Agents / Q000737:chemistry / Q000494:pharmacology; D045744:Cell Line, Tumor; D002999:Clone Cells / Q000473:pathology; D004249:DNA Damage; D053843:DNA Mismatch Repair / Q000187:drug effects / Q000235:genetics*; D015536:Down-Regulation / Q000187:drug effects / Q000235:genetics; D055785:Gene Knockdown Techniques; D006801:Humans; D053842:Microsatellite Instability / Q000187:drug effects; D000070957:MutL Protein Homolog 1; D009687:Nuclear Proteins / Q000172:deficiency / Q000235:genetics; D009942:Organometallic Compounds / Q000737:chemistry / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000235:genetics; D012238:Rhodium / Q000737:chemistry
chemical_list: D048868:Adaptor Proteins, Signal Transducing; D000970:Antineoplastic Agents; C501486:MLH1 protein, human; D009687:Nuclear Proteins; D009942:Organometallic Compounds; D034741:RNA, Small Interfering; D012238:Rhodium; D000070957:MutL Protein Homolog 1