Sonoporation delivery of monoclonal antibodies against human papillomavirus 16 E6 restores p53 expression in transformed cervical keratinocytes

PLoS One. 2012;7(11):e50730. doi: 10.1371/journal.pone.0050730. Epub 2012 Nov 30.

Abstract

High-risk types of human papillomavirus (HPV), such as HPV16, have been found in nearly all cases of cervical cancer. Therapies targeted at blocking the HPV16 E6 protein and its deleterious effects on the tumour suppressor pathways of the cell can reverse the malignant phenotype of affected keratinocytes while sparing uninfected cells. Through a strong interdisciplinary collaboration between engineering and biology, a novel, non-invasive intracellular delivery method for the HPV16 E6 antibody, F127-6G6, was developed. The method employs high intensity focused ultrasound (HIFU) in combination with microbubbles, in a process known as sonoporation. In this proof of principle study, it was first demonstrated that sonoporation antibody delivery into the HPV16 positive cervical carcinoma derived cell lines CaSki and SiHa was possible, using chemical transfection as a baseline for comparison. Delivery of the E6 antibody using sonoporation significantly restored p53 expression in these cells, indicating the antibody is able to enter the cells and remains active. This delivery method is targeted, non-cytotoxic, and non-invasive, making it more easily translatable for in vivo experiments than other transfection methods.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Antibodies, Monoclonal / administration & dosage
  • Antibodies, Monoclonal / immunology*
  • Cell Line, Tumor
  • Cell Transformation, Neoplastic
  • Cervix Uteri / cytology
  • Drug Delivery Systems / instrumentation*
  • Feasibility Studies
  • Female
  • Gene Expression Regulation*
  • Human papillomavirus 16 / physiology
  • Humans
  • Keratinocytes / metabolism
  • Keratinocytes / pathology*
  • Keratinocytes / virology
  • Microbubbles
  • Oncogene Proteins, Viral / immunology*
  • Proteolysis
  • Repressor Proteins / immunology*
  • Tumor Suppressor Protein p53 / metabolism*
  • Ultrasonics / instrumentation*
  • Uterine Cervical Neoplasms / pathology

Substances

  • Antibodies, Monoclonal
  • E6 protein, Human papillomavirus type 16
  • Oncogene Proteins, Viral
  • Repressor Proteins
  • Tumor Suppressor Protein p53

Grant support

This work was mainly funded by a Natural Sciences and Engineering Research Council of Canada (NSERC) grant to IZ (#355858-2008) and partially funded by Philips Medical Healthcare. E6 antibodies were kindly provided by Arbor Vita Corporation. The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.