Targeted delivery of siRNA using transferrin-coupled lipoplexes specifically sensitizes CD71 high expressing malignant cells to antibody-mediated complement attack
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The overexpression of membrane-bound complement regulatory proteins (mCRP; CD46, CD55, CD59) preventing opsonization and complement-dependent cytotoxicity (CDC) is considered a major barrier for successful antibody-based cancer immunotherapy. To avoid a potential deleterious effect of mCRP neutralization on normal tissue cells, complement regulation has to be selectively targeted to the malignant cells. In this study, anti-mCRP small interfering RNAs (siRNAs) were encapsulated in transferrin-coupled lipoplexes for the specific delivery to transferrin receptor CD71high expressing BT474, DU145, and SW480 as well as corresponding CD71-knockdown (CD71low) tumor cells. Targeted delivery with transferrin-siRNA-lipoplexes became possible by charge neutralization and resulted in efficient silencing of all three mCRPs up to 90 %, which is dependent on their CD71 expression. The mCRP knockdown led to a significant increase of CDC on CD71high tumor cells by 68 % in BT474, 58 % in DU145, and 40 % in SW480 cells but only slightly increased on CD71low cells. Downregulation of CD46 and CD55 significantly increased C3 opsonization only on CD71high tumor cells. Our results demonstrate for the first time that by specific delivery of anti-mCRP siRNA through transferrin receptor, complement regulation can be selectively neutralized, allowing specific antibody-mediated killing of tumor cells without affecting healthy bystander cells, which appears to be a suited strategy to improve antibody-based cancer immunotherapy.
KeywordsComplement-dependent cytotoxicity Immunotherapy Membrane complement regulatory proteins siRNA Transferrin receptor Tumor cell targeting
The study was supported by the “BMBF BIODISC (0315503)” and the “Exzellenzinitiative, Innovationfonds Frontier Programme, University of Heidelberg,” and by the “Stiftung für Krebs–und Scharlachforschung Mannheim”.
Conflict of interest
The authors disclose no potential conflicts of interest.
- 13.Czuczman MS, Olejniczak S, Gowda A, Kotowski A, Binder A, Kaur H, Knight J, Starostik P, Deans J, Hernandez-Ilizaliturri FJ (2008) Acquirement of rituximab resistance in lymphoma cell lines is associated with both global CD20 gene and protein down-regulation regulated at the pretranscriptional and posttranscriptional levels. Clin Cancer Res Off J Am Assoc Cancer Res 14:1561–1570CrossRefGoogle Scholar
- 14.Ge X, Wu L, Hu W, Fernandes S, Wang C, Li X, Brown JR, Qin X (2011) rILYd4, a human CD59 inhibitor, enhances complement-dependent cytotoxicity of ofatumumab against rituximab-resistant B-cell lymphoma cells and chronic lymphocytic leukemia. Clin Cancer Res Off J Am Assoc Cancer Res 17:6702–6711CrossRefGoogle Scholar
- 15.Golay J, Lazzari M, Facchinetti V, Bernasconi S, Borleri G, Barbui T, Rambaldi A, Introna M (2001) CD20 levels determine the in vitro susceptibility to rituximab and complement of B-cell chronic lymphocytic leukemia: further regulation by CD55 and CD59. Blood 98:3383–3389CrossRefPubMedGoogle Scholar
- 23.Filleur S, Courtin A, Ait-Si-Ali S, Guglielmi J, Merle C, Harel-Bellan A, Clezardin P, Cabon F (2003) SiRNA-mediated inhibition of vascular endothelial growth factor severely limits tumor resistance to antiangiogenic thrombospondin-1 and slows tumor vascularization and growth. Cancer Res 63:3919–3922PubMedGoogle Scholar
- 24.Tolentino MJ, Brucker AJ, Fosnot J, Ying GS, Wu IH, Malik G, Wan S, Reich SJ (2004) Intravitreal injection of vascular endothelial growth factor small interfering RNA inhibits growth and leakage in a nonhuman primate, laser-induced model of choroidal neovascularization. Retina 24:132–138CrossRefPubMedGoogle Scholar
- 30.Liu Y, Tao J, Li Y, Yang J, Yu Y, Wang M, Xu X, Huang C, Huang W, Dong J, Li L, Liu J, Shen G, Tu Y (2009) Targeting hypoxia-inducible factor-1alpha with Tf-PEI-shRNA complex via transferrin receptor-mediated endocytosis inhibits melanoma growth. Molec Ther J Am Soc Gene Ther 17:269–277CrossRefGoogle Scholar
- 36.Horl S, Banki Z, Huber G, Ejaz A, Mullauer B, Willenbacher E, Steurer M, Stoiber H (2013) Complement factor H-derived short consensus repeat 18-20 enhanced complement-dependent cytotoxicity of ofatumumab on chronic lymphocytic leukemia cells. Haematologica 98:1939–1947PubMedCentralCrossRefPubMedGoogle Scholar
- 44.Smilevska T, Stamatopoulos K, Samara M, Belessi C, Tsompanakou A, Paterakis G, Stavroyianni N, Athanasiadou I, Chiotoglou I, Hadzidimitriou A, Athanasiadou A, Douka V, Saloum R, Laoutaris N, Anagnostopoulos A, Fassas A, Stathakis N, Kollia P (2006) Transferrin receptor-1 and 2 expression in chronic lymphocytic leukemia. Leuk Res 30:183–189CrossRefPubMedGoogle Scholar
- 45.Qing Y, Shuo W, Zhihua W, Huifen Z, Ping L, Lijiang L, Xiaorong Z, Liming C, Daiwen X, Yu H, Wei X, Min F, Zuohua F, Guanxin S (2006) The in vitro antitumor effect and in vivo tumor-specificity distribution of human-mouse chimeric antibody against transferrin receptor. Cancer Immunol Immunother CII 55:1111–1121CrossRefPubMedGoogle Scholar
- 47.Brooks D, Taylor C, Dos Santos B, Linden H, Houghton A, Hecht TT, Kornfeld S, Taetle R (1995) Phase Ia trial of murine immunoglobulin A antitransferrin receptor antibody 42/6. Clin Cancer Res Off J Am Assoc Cancer Res 1:1259–1265Google Scholar