Fully synthetic polymer vesicles for intracellular delivery of antibodies in live cells

There is an emerging need both in pharmacology and within the biomedical industry to develop new tools to target intracellular mechanisms. The efficient delivery of functionally active proteins within cells is potentially a powerful research strategy, especially through the use of antibodies. In this work, we report on a nanovector for the efficient encapsulation and delivery of antibodies into live cells with no significant loss of cell viability or any deleterious effect on cell metabolic activity. This delivery system is based on poly[2‐(methacryloyloxy)ethyl phosphorylcholine]‐block‐[2‐(diisopropylamino)ethyl methacrylate] (PMPC‐PDPA), a pH‐sensitive diblock copolymer that self‐assembles to form nanometer‐sized vesicles, also known as polymersomes, at physiological pH. Polymersomes can successfully deliver relatively high antibody payloads within different types of live cells. We demonstrate that these antibodies can target their respective epitope showing immunolabeling of γ‐tubulin, actin, Golgi protein, and the transcription factor NF‐κB in live cells. Finally, we demonstrate that intracellular delivery of antibodies can control specific subcellular events, as well as modulate cell activity and proinflammatory processes.—Canton, I., Massignani, M., Patikarnmonthon, N., Chierico, L., Robertson, J., Renshaw, S. A., Warren, N. J., Madsen, J. P., Armes, S P., Lewis, A. L., Battaglia, G. Fully synthetic polymer vesicles for intracellular delivery of antibodies in live cells. FASEB J. 27, 98–108 (2013). www.fasebj.org

[1]  A. Philimonenko,et al.  Nuclear γ‐tubulin associates with nucleoli and interacts with tumor suppressor protein C53 , 2012, Journal of Cellular Physiology.

[2]  E. Baba,et al.  [Development trends for therapeutic antibody]. , 2012, Nihon rinsho. Japanese journal of clinical medicine.

[3]  Kristoffer von Stedingk,et al.  Nuclear localization of γ-tubulin affects E2F transcriptional activity and S-phase progression , 2011, FASEB journal : official publication of the Federation of American Societies for Experimental Biology.

[4]  S. Armes,et al.  Controlling polymersome surface topology at the nanoscale by membrane confined polymer/polymer phase separation. , 2011, ACS nano.

[5]  Zhen Gu,et al.  Endoprotease-mediated intracellular protein delivery using nanocapsules. , 2011, ACS nano.

[6]  Tayyaba Hasan,et al.  Ki-67 as a molecular target for therapy in an in vitro three-dimensional model for ovarian cancer. , 2010, Cancer research.

[7]  S. MacNeil,et al.  Internalization and biodistribution of polymersomes into oral squamous cell carcinoma cells in vitro and in vivo. , 2010, Nanomedicine.

[8]  P. Sieving,et al.  Probing potassium channel function in vivo by intracellular delivery of antibodies in a rat model of retinal neurodegeneration , 2010, Proceedings of the National Academy of Sciences.

[9]  S. MacNeil,et al.  Enhanced Fluorescence Imaging of Live Cells by Effective Cytosolic Delivery of Probes , 2010, PloS one.

[10]  Zhen Gu,et al.  A novel intracellular protein delivery platform based on single-protein nanocapsules. , 2010, Nature nanotechnology.

[11]  S. Armes,et al.  Controlling cellular uptake by surface chemistry, size, and surface topology at the nanoscale. , 2009, Small.

[12]  M. Bettencourt-Dias,et al.  γ-Tubulin-containing abnormal centrioles are induced by insufficient Plk4 in human HCT116 colorectal cancer cells , 2009, Journal of Cell Science.

[13]  Giuseppe Battaglia,et al.  Polymersomes: nature inspired nanometer sized compartments , 2009 .

[14]  Janice M Reichert,et al.  Development trends for therapeutic antibody fragments , 2009, Nature Biotechnology.

[15]  A. Banga,et al.  In vitro transdermal delivery of therapeutic antibodies using maltose microneedles. , 2009, International journal of pharmaceutics.

[16]  S. MacNeil,et al.  Non-cytotoxic polymer vesicles for rapid and efficient intracellular delivery. , 2008, Faraday discussions.

[17]  A. Cattaneo,et al.  In vivo selection of intrabodies specifically targeting protein-protein interactions: a general platform for an "undruggable" class of disease targets. , 2008, Journal of biotechnology.

[18]  K. Miyata,et al.  Efficient delivery of antibody into living cells using a novel HVJ envelope vector system. , 2008, Journal of immunological methods.

[19]  S. Ghosh,et al.  Shared Principles in NF-κB Signaling , 2008, Cell.

[20]  S. MacNeil,et al.  Biomimetic pH Sensitive Polymersomes for Efficient DNA Encapsulation and Delivery , 2007 .

[21]  W. Denny,et al.  Genotoxicity of non-covalent interactions: DNA intercalators. , 2007, Mutation research.

[22]  Jiri Aubrecht,et al.  Toxicogenomics: overview and potential applications for the study of non-covalent DNA interacting chemicals. , 2007, Mutation research.

[23]  F. Grosse,et al.  Centrosomal localization of DNA damage checkpoint proteins , 2007, Journal of cellular biochemistry.

[24]  A. Merdes,et al.  γ-tubulin complexes and microtubule organization , 2007 .

[25]  A. Merdes,et al.  Gamma-tubulin complexes and microtubule organization. , 2007, Current opinion in cell biology.

[26]  N. Perkins,et al.  Integrating cell-signalling pathways with NF-κB and IKK function , 2007, Nature Reviews Molecular Cell Biology.

[27]  Sachdev S Sidhu,et al.  Synthetic therapeutic antibodies , 2006, Nature chemical biology.

[28]  S. Armes,et al.  Phosphorylcholine-based pH-responsive diblock copolymer micelles as drug delivery vehicles: light scattering, electron microscopy, and fluorescence experiments. , 2006, Biomacromolecules.

[29]  S. Armes,et al.  pH-sensitive vesicles based on a biocompatible zwitterionic diblock copolymer. , 2005, Journal of the American Chemical Society.

[30]  B. Monsarrat,et al.  DNA damage induce γ-tubulin–RAD51 nuclear complexes in mammalian cells , 2005, Oncogene.

[31]  A. Ryan,et al.  Bilayers and interdigitation in block copolymer vesicles. , 2005, Journal of the American Chemical Society.

[32]  B. Monsarrat,et al.  DNA damage induce gamma-tubulin-RAD51 nuclear complexes in mammalian cells. , 2005, Oncogene.

[33]  M. Feldmann,et al.  Heterogeneous requirement of IkappaB kinase 2 for inflammatory cytokine and matrix metalloproteinase production in rheumatoid arthritis: implications for therapy. , 2003, Arthritis and rheumatism.

[34]  G. Donnarumma,et al.  Bacterial components induce cytokine and intercellular adhesion molecules-1 and activate transcription factors in dermal fibroblasts. , 2003, Research in microbiology.

[35]  P. Khavari,et al.  Divergent gene regulation and growth effects by NF-κB in epithelial and mesenchymal cells of human skin , 2003, Oncogene.

[36]  K. Brand,et al.  Involvement of NF-κB signalling in skin physiology and disease , 2003 .

[37]  S. Armes,et al.  Synthesis of controlled-structure sulfate-based copolymers via atom transfer radical polymerisation and their use as crystal habit modifiers for BaSO4 , 2002 .

[38]  E. Kieff,et al.  Electroporation of antibodies, DNA, and other macromolecules into cells: a highly efficient method. , 2000, Journal of immunological methods.

[39]  D. Hammer,et al.  Polymersomes: tough vesicles made from diblock copolymers. , 1999, Science.

[40]  A. Cattaneo,et al.  The selection of intracellular antibodies. , 1998, Trends in biotechnology.

[41]  A. Baldwin,et al.  THE NF-κB AND IκB PROTEINS: New Discoveries and Insights , 1996 .

[42]  D. Curiel,et al.  Receptor-mediated gene delivery employing lectin-binding specificity. , 1994, Gene therapy.

[43]  M. Kay,et al.  Hepatic gene therapy: efficient gene delivery and expression in primary hepatocytes utilizing a conjugated adenovirus-DNA complex. , 1993, Proceedings of the National Academy of Sciences of the United States of America.

[44]  M. Cotten,et al.  Adenovirus enhancement of transferrin-polylysine-mediated gene delivery. , 1991, Proceedings of the National Academy of Sciences of the United States of America.