title: Molecularly evolved thymidylate synthase inhibits 5-fluorodeoxyuridine toxicity in human hematopoietic cells.
abstract: Thymidylate synthase (TS) inhibitors, such as 5-fluorouracil (5-FU) and 5-fluorodeoxyuridine (5-FUdR), are amongst the most frequently used chemotherapeutic drugs available, although their efficacy is often limited by myelotoxicity. An emerging strategy for overcoming bone marrow toxicity involves ex vivo genetic transfer of drug resistance to autologous hematopoietic progenitor cells, followed by reimplantation of the transfected cells before chemotherapy. Here we establish that expression of mutant TS genes, selected from millions of engineered variants, renders human hematopoietic cells resistant to 5-FUdR, and identify the most efficacious variant for gene therapeutic rescue of drug-induced myelosuppression.
authors.affiliation: Molecular Diagnostics Program, Public Health Sciences Division, Fred Hutchinson Cancer Research Center, Seattle, WA 98109, USA. jbielas@fhcrc.org
authors.forename: Jason H
authors.initials: JH
authors.lastname: Bielas
authors.forename: Michael W
authors.initials: MW
authors.lastname: Schmitt
authors.forename: Amalia
authors.initials: A
authors.lastname: Icreverzi
authors.forename: Nolan G
authors.initials: NG
authors.lastname: Ericson
authors.forename: Lawrence A
authors.initials: LA
authors.lastname: Loeb
mesh_terms: D019943:Amino Acid Substitution; D000964:Antimetabolites, Antineoplastic / Q000009:adverse effects* / Q000627:therapeutic use; D004351:Drug Resistance / Q000235:genetics*; D005467:Floxuridine / Q000009:adverse effects* / Q000627:therapeutic use; D015870:Gene Expression; D015316:Genetic Therapy; D006412:Hematopoietic Stem Cells / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D011487:Protein Conformation; D013489:Suppression, Genetic; D013940:Thymidylate Synthase / Q000737:chemistry / Q000235:genetics*; D014161:Transduction, Genetic
chemical_list: D000964:Antimetabolites, Antineoplastic; D005467:Floxuridine; D013940:Thymidylate Synthase
references.citation: Br J Cancer. 2003 Sep 1;89(5):816-20
references.pmid: 12942110
references.citation: Annu Rev Biochem. 1995;64:721-62
references.pmid: 7574499
references.citation: Nat Rev Cancer. 2001 Nov;1(2):99-108
references.pmid: 11905809
references.citation: Ann Oncol. 2004 Jul;15(7):1025-32
references.pmid: 15205195
references.citation: Lancet Oncol. 2002 Mar;3(3):154-8
references.pmid: 11902501
references.citation: Clin Cancer Res. 2000 Apr;6(4):1322-7
references.pmid: 10778957
references.citation: J Biol Chem. 2002 Sep 27;277(39):36304-11
references.pmid: 12147691
references.citation: J Biol Chem. 2002 Nov 8;277(45):43224-32
references.pmid: 12202489
references.citation: Cancer Res. 2001 Jan 15;61(2):666-72
references.pmid: 11212266
references.citation: Nat Med. 1998 Oct;4(10):1136-43
references.pmid: 9771746
references.citation: Nat Rev Cancer. 2004 Apr;4(4):296-307
references.pmid: 15057289
references.citation: Clin Cancer Res. 2003 Oct 1;9(12):4363-7
references.pmid: 14555507
references.citation: Nat Biotechnol. 1999 Feb;17(2):143-7
references.pmid: 10052349
references.citation: Biochemistry. 2001 Feb 20;40(7):1897-902
references.pmid: 11329255
references.citation: Cancer Gene Ther. 2005 Nov;12(11):849-63
references.pmid: 16037821
references.citation: J Biol Chem. 1998 May 8;273(19):11611-8
references.pmid: 9565579