title: Sonoporation delivery of monoclonal antibodies against human papillomavirus 16 E6 restores p53 expression in transformed cervical keratinocytes.
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.
authors.affiliation: Probe Development and Biomarker Exploration, Thunder Bay Regional Research Institute, Thunder Bay, Ontario, Canada.
authors.forename: Melissa
authors.initials: M
authors.lastname: Togtema
authors.forename: Samuel
authors.initials: S
authors.lastname: Pichardo
authors.forename: Robert
authors.initials: R
authors.lastname: Jackson
authors.forename: Paul F
authors.initials: PF
authors.lastname: Lambert
authors.forename: Laura
authors.initials: L
authors.lastname: Curiel
authors.forename: Ingeborg
authors.initials: I
authors.lastname: Zehbe
mesh_terms: D000911:Antibodies, Monoclonal / Q000008:administration & dosage / Q000276:immunology*; D045744:Cell Line, Tumor; D002471:Cell Transformation, Neoplastic; D002584:Cervix Uteri / Q000166:cytology; D016503:Drug Delivery Systems / Q000295:instrumentation*; D005240:Feasibility Studies; D005260:Female; D005786:Gene Expression Regulation; D052162:Human papillomavirus 16 / Q000502:physiology; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism / Q000473:pathology* / Q000821:virology; D045423:Microbubbles; D009856:Oncogene Proteins, Viral / Q000276:immunology*; D059748:Proteolysis; D012097:Repressor Proteins / Q000276:immunology*; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism*; D014465:Ultrasonics / Q000295:instrumentation*; D002583:Uterine Cervical Neoplasms / Q000473:pathology
chemical_list: D000911:Antibodies, Monoclonal; C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D012097:Repressor Proteins; D016159:Tumor Suppressor Protein p53