Advances in Cancer Drug Targets

Volume: 3

Indexed in: EBSCO, Ulrich's Periodicals Directory

Advances in Cancer Drug Targets is an e-book series that brings together recent expert reviews published on the subject with a focus on strategies for synthesizing and isolating organic compounds and ...
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Inhibition of Membrane Complement Inhibitor Expression (CD46, CD55, CD59) by siRNA Sensitizes Tumor Cells to Complement Attack

Pp. 50-77 (28)

Srinivas Mamidi, Simon Höne, Nicolas Geis, Stefanie Zell, Renate Rutz, Wenhan Li, Thomas Giese, Stefan Schultz and Michael Kirschfink

Abstract

The efficacy of cancer-immunotherapy with complement-activating monoclonal antibodies is limited by over-expression of one or more membrane-bound complement regulatory proteins (mCRPs: CD46, CD55, CD59) on the surface of neoplastic cells.

In this study we designed small interfering RNAs (siRNAs) for posttranscriptional gene knock down of CD46, CD55 and CD59 aiming to sensitize tumor cells to complement attack and thereby exploiting complement for tumor cell destruction. Tumor cell lines of different origin, such as Du145 (prostate), BT474 (breast) and K562 (erythroleukemia) were selected for the study. FACS-analysis demonstrated that siRNA anti-CD46 (301) reduced CD46 protein expression up to 80%, siRNA anti-CD55 (255) diminished CD55 protein expression up to 49%, and CD59 protein expression was inhibited up to 82% by siRNA anti-CD59 (1339). Time course experiments revealed a long-lasting silencing effect with >50% complement regulator inhibition up to day 13. Upon mCRP knock down, complement-dependent cytotoxicity (CDC) was augmented by 20-30% for CD46, by up to 24% for CD55 and by up to 55% for CD59. The combined inhibition of all three inhibitors further enhanced CDC by up to 66%. Dependent on the cell line, CD46 and CD55 downregulation increased significantly C3 opsonization, which is known to support cell-mediated defense mechanisms. mCRP blocking antibodies were only partly able to further augment the tumor cells’ susceptibility to complement lysis.

Thus, siRNA-induced inhibition of complement regulator expression clearly sensitizes malignant cells to complement attack and, if specifically targeted to the tumor, appears suited as adjuvant to improve antibody-based cancer immunotherapy.

Keywords:

Complement resistance, Membrane complement regulatory proteins, siRNA, Tumor immunology.

Affiliation:

Institute of Immunology, University of Heidelberg, Germany, Im Neuenheimer Feld 305, 69120 Heidelberg, Germany.