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Intratumoral gene delivery of anti-cathepsin L single-chain variable fragment by lentiviral vector inhibits tumor progression induced by human melanoma cells

Cancer Gene Therapy volume 15, pages 591–604 (2008)Cite this article

Abstract

We previously demonstrated that the switch from non- to highly tumorigenic phenotype of human melanoma cells is directly related to procathepsin L secretion, which increased cell resistance to complement-mediated cell lysis. Involvement of procathepsin L secretion in tumor growth was clearly demonstrated by three different strategies: (1) inhibition of secreted procathepsin L activity; (2) increase of procathepsin L secretion; and (3) inhibition of procathepsin L secretion. This latter strategy was triggered by intracellular expression of anti-human cathepsin L single-chain variable fragment (ScFv). These previous experiments were performed by processing melanoma cells before their injection into nude mice. We herein designed a new lentiviral vector in which this anti-cathepsin L ScFv was cloned. This lentiviral vector was optimized to allow the highest intracellular expression of anti-cathepsin L ScFv in transduced melanoma cells. In these transduced cells, procathepsin L secretion was strongly inhibited. In addition, injection of this anti-cathepsin L ScFv lentiviral vector into tumors already induced in nude mice inhibited tumor growth and associated angiogenesis. This is the first report to demonstrate that targeting procathepsin L secretion with anti-cathepsin L ScFv lentiviral construct constitutes a new gene therapy in the challenge to inhibit the growth of tumors induced by human melanoma cells.

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Abbreviations

AcGFP:

Aequorea coerulescens green fluorescent protein

cPPT:

central polypurine tract

anti-CTSL:

anti-cathepsin L

ScFv:

single-chain variable fragment

moAb:

monoclonal antibody

s.c.:

subcutaneously

MOI:

multiplicity of infection

MVD:

microvessel density

sp:

signal peptide

5′-UTR:

5′-untranslated region

WPRE:

woodchuck hepatitis virus post-transcriptional regulatory element

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Acknowledgements

We thank Pr Menashe Bar-Eli (MD Anderson Cancer Center, Houston, TX) for helpful discussions. The experiments on mice were performed by Cellvax Inc. (France) with the technical assistance of Dr Delphine Allard and Dr Ming Wei, in the Center for Exploration and Experimental Functional Research (CERFE) of Evry (France). This work was supported by INSERM, the Institut National contre le Cancer (INCa), Association de Recherche contre le Cancer (ARC), Ligue National contre le Cancer (Comité de Paris) and Ministère de l'Education Nationale et de la Recherche Scientifique (MENRS).

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  1. INSERM U.672 (former U.354), Immunochemistry of Cell Regulations and Viral Interactions, Bâtiment G8, Evry Cedex, France

    R Frade, N Rousselet & D Jean

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Correspondence to R Frade.

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Frade, R., Rousselet, N. & Jean, D. Intratumoral gene delivery of anti-cathepsin L single-chain variable fragment by lentiviral vector inhibits tumor progression induced by human melanoma cells. Cancer Gene Ther 15, 591–604 (2008). https://doi.org/10.1038/cgt.2008.51

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