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Research Articles: Therapeutics, Targets, and Development

Relevance of the Fanconi anemia pathway in the response of human cells to trabectedin

¹ Division of Hematopoiesis and Gene Therapy Program, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas and Centro de Investigación Biomédica en Red de Enfermedades Raras; Departments of ² Pharmacology, ³ Biochemistry and Molecular Biology, and ⁴ Organic Chemistry, Universidad de Alcalá, Alcalá de Henares; and ⁵ PharmaMar S.A., Madrid, Spain

Requests for reprints: Juan A. Bueren, Division of Hematopoiesis and Gene Therapy Program, Centro de Investigaciones Energéticas, Medioambientales y Tecnológicas, Avenida Complutense 22, 28040 Madrid, Spain. Phone: 34-91-3466518; Fax: 34-91-3466484. E-mail: juan.bueren{at}ciemat.es

Abstract

Trabectedin (Yondelis; ET-743) is a potent anticancer drug that binds to DNA by forming a covalent bond with a guanine in one strand and one or more hydrogen bonds with the opposite strand. Using a fluorescence-based melting assay, we show that one single trabectedin-DNA adduct increases the thermal stability of the double helix by >20°C. As deduced from the analysis of phosphorylated H2AX and Rad51 foci, we observed that clinically relevant doses of trabectedin induce the formation of DNA double-strand breaks in human cells and activate homologous recombination repair in a manner similar to that evoked by the DNA interstrand cross-linking agent mitomycin C (MMC). Because one important characteristic of this drug is its marked cytotoxicity on cells lacking a functional Fanconi anemia (FA) pathway, we compared the response of different subtypes of FA cells to MMC and trabectedin. Our data clearly show that human cells with mutations in FANCA, FANCC, FANCF, FANCG, or FANCD1 genes are highly sensitive to both MMC and trabectedin. However, in marked contrast to MMC, trabectedin does not induce any significant accumulation of FA cells in G₂-M. The critical relevance of FA proteins in the response of human cells to trabectedin reported herein, together with observations showing the role of the FA pathway in cancer suppression, strongly suggest that screening for mutations in FA genes may facilitate the identification of tumors displaying enhanced sensitivity to this novel anticancer drug. [Mol Cancer Ther 2008;7(5):1309–18]

Grant support: Comisión Interministerial de Ciencia y Tecnología SAF2005-00058 (J.A. Bueren) and SAF2006-12713-C02-02 (F.G.), Comunidad de Madrid S-BIO/0214/2006 (F. Gago), and Marcelino Botín Foundation to promote Translational Research on Biomedicine 2005-2009 (J.A. Bueren).

The costs of publication of this article were defrayed in part by the payment of page charges. This article must therefore be hereby marked advertisement in accordance with 18 U.S.C. Section 1734 solely to indicate this fact.

⁶ Supplementary data for this article are available at Molecular Cancer Therapeutics Online (http://mct.aacrjournals.org/).

Received 12/27/07; revised 2/21/08; accepted 2/29/08.