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Intracellular Expression of an Antibody Fragment-neutralizing p21 Ras Promotes Tumor Regression¹

Laboratoire de Biotechnologie des Anticorps, Institut Curie, 75005 Paris [O. C., J-L., T.]; Gene Medicine Department, Rhône-Poulenc Rorer Central Research [M. K., I. D., M-C. M., F. S., B. T.], and Gencell, Centre de Recherche de Vitry Alfortville [A. V-O.], 94403 Vitry sur Seine; and Unité 255, Institut National de la Santé et de la Recherche Médicale, Institut Curie, 75005 Paris [W. H. F., J-L. T.], France

Mutated ras genes are found in a large number of human tumors and, therefore, constitute one of the primary targets for cancer treatment. Micro-injection of the neutralizing anti-Ras monoclonal antibody Y13-259 was previously reported to induce transient phenotypic reversion of ras-transformed rodent fibroblasts in vitro. We have prepared a single-chain Fv fragment (scFv) derived from Y13-259, and here, we show that intracellular expression of the scFv led to the specific inhibition of the Ras signaling pathway in Xenopus laevis oocytes and NIH3T3 fibroblasts. Moreover, neutralizing Ras with the scFv specifically promoted apoptosis in vitro in human cancer cells but not in untransformed cells. As a step toward cancer gene therapy, we finally demonstrated that intratumor transduction of HCT116 colon carcinoma cells with the anti-Ras scFv using an adenoviral vector elicited sustained tumor regression in nude mice.

¹ This work was supported by the French Ministries of Industry and Research (Bio-Avenir Program) and by the Institut Curie.

² To whom requests for reprints should be addressed, at Gene Medicine Department and Gencell, Centre de Recherche de Vitry Alfortville, 13 quai J. Guesde, 94403 Vitry sur Seine, France. Phone: 331 55 71 80 83; Fax: 331 55 71 37 96; E-mail: bruno.tocque@rhone-poulenc.com.

Received 9/ 2/97. Accepted 1/15/98.

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