title: High-throughput in vivo genotoxicity testing: an automated readout system for the somatic mutation and recombination test (SMART).
abstract: Genotoxicity testing is an important component of toxicity assessment. As illustrated by the European registration, evaluation, authorization, and restriction of chemicals (REACH) directive, it concerns all the chemicals used in industry. The commonly used in vivo mammalian tests appear to be ill adapted to tackle the large compound sets involved, due to throughput, cost, and ethical issues. The somatic mutation and recombination test (SMART) represents a more scalable alternative, since it uses Drosophila, which develops faster and requires less infrastructure. Despite these advantages, the manual scoring of the hairs on Drosophila wings required for the SMART limits its usage. To overcome this limitation, we have developed an automated SMART readout. It consists of automated imaging, followed by an image analysis pipeline that measures individual wing genotoxicity scores. Finally, we have developed a wing score-based dose-dependency approach that can provide genotoxicity profiles. We have validated our method using 6 compounds, obtaining profiles almost identical to those obtained from manual measures, even for low-genotoxicity compounds such as urethane. The automated SMART, with its faster and more reliable readout, fulfills the need for a high-throughput in vivo test. The flexible imaging strategy we describe and the analysis tools we provide should facilitate the optimization and dissemination of our methods.
authors.affiliation: Image Mining Group, Institut Pasteur Korea, Sampyeong-dong 696, Bundang-gu, Seongnam-si, Gyeonggi-do, Korea.
authors.forename: Benoit
authors.initials: B
authors.lastname: Lombardot
authors.affiliation: Drug Biology Group, Institut Pasteur Korea, Sampyeong-dong 696, Bundang-gu, Seongnam-si, Gyeonggi-do, Korea.
authors.forename: Chun-Taek
authors.initials: CT
authors.lastname: Oh
authors.affiliation: Image Mining Group, Institut Pasteur Korea, Sampyeong-dong 696, Bundang-gu, Seongnam-si, Gyeonggi-do, Korea.
authors.forename: Jihoon
authors.initials: J
authors.lastname: Kwak
authors.affiliation: Image Mining Group, Institut Pasteur Korea, Sampyeong-dong 696, Bundang-gu, Seongnam-si, Gyeonggi-do, Korea.
authors.forename: Auguste
authors.initials: A
authors.lastname: Genovesio
authors.affiliation: Department of Mathematical Sciences, Seoul National University, 1 Gwanak-ro, Gwanak-gu, Seoul 151-747, Korea.
authors.forename: Myungjoo
authors.initials: M
authors.lastname: Kang
authors.affiliation: Image Mining Group, Institut Pasteur Korea, Sampyeong-dong 696, Bundang-gu, Seongnam-si, Gyeonggi-do, Korea.
authors.forename: Michael Adsett Edberg
authors.initials: MA
authors.lastname: Hansen
authors.affiliation: Drug Biology Group, Institut Pasteur Korea, Sampyeong-dong 696, Bundang-gu, Seongnam-si, Gyeonggi-do, Korea.
authors.forename: Sung-Jun
authors.initials: SJ
authors.lastname: Han
mesh_terms: D000818:Animals; D004330:Drosophila / Q000235:genetics* / Q000502:physiology; D006197:Hair / Q000378:metabolism; D007091:Image Processing, Computer-Assisted; D009152:Mutagenicity Tests / Q000379:methods*; D009154:Mutation; D011995:Recombination, Genetic; D014921:Wings, Animal / Q000378:metabolism
references.citation: J Cell Biol. 1993 Oct;123(1):209-21
references.pmid: 8408199
references.citation: IARC Sci Publ. 1999;(146):427-70
references.pmid: 10353398
references.citation: Br J Biomed Sci. 2009;66(3):160-5
references.pmid: 19839229
references.citation: J Microsc. 2004 Jul;215(Pt 1):67-76
references.pmid: 15230877
references.citation: Food Chem Toxicol. 1996 Jan;34(1):1-14
references.pmid: 8603789
references.citation: Cytometry. 1997 Aug 1;28(4):289-97
references.pmid: 9266748
references.citation: Environ Mol Mutagen. 2000;35(3):234-52
references.pmid: 10737958
references.citation: Nat Rev Drug Discov. 2004 Aug;3(8):711-5
references.pmid: 15286737
references.citation: IEEE Trans Image Process. 1998;7(4):555-70
references.pmid: 18276273
references.citation: Food Chem Toxicol. 2008 Jan;46(1):393-401
references.pmid: 17910989
references.citation: Science. 2005 Jan 21;307(5708):423-6
references.pmid: 15662015
references.citation: Int J Surg Pathol. 2006 Oct;14(4):285-305
references.pmid: 17041192
references.citation: Environ Mutagen. 1984;6(2):153-88
references.pmid: 6423380
references.citation: Cell. 2001 Apr 6;105(1):81-91
references.pmid: 11301004
references.citation: Inf Process Med Imaging. 2003 Jul;18:136-47
references.pmid: 15344453
references.citation: Toxicol Mech Methods. 2005;15(3):155-80
references.pmid: 20021080
references.citation: Mutat Res. 1995 Apr;334(2):247-58
references.pmid: 7885379
references.citation: Cytometry. 1999 Feb 1;35(2):134-44
references.pmid: 10554169
references.citation: Mutat Res. 2004 May 18;549(1-2):5-27
references.pmid: 15120960