Patient-Derived Organotypic Epithelial Rafts Model Phenotypes in Juvenile-Onset Recurrent Respiratory Papillomatosis
Mary C. Bedard,
Marion G. Brusadelli,
Adrean Carlile,
Sonya Ruiz-Torres,
Hannah Lodin,
Denis Lee,
Matthew Kofron,
Paul F. Lambert,
Adam Lane,
Najim Ameziane,
El Mustapha Bahassi,
Kathryn A. Wikenheiser-Brokamp,
Alessandro de Alarcon,
David F. Smith,
Susanne I. Wells
Affiliations
Mary C. Bedard
Division of Oncology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH 45229, USA
Marion G. Brusadelli
Medpace, Cincinnati, OH 45227, USA
Adrean Carlile
Division of Oncology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH 45229, USA
Sonya Ruiz-Torres
Division of Oncology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH 45229, USA
Hannah Lodin
Mayo Clinic Alix School of Medicine, Mayo Clinic, Phoenix, AZ 85259, USA
Denis Lee
McArdle Laboratory for Cancer Research, University of Wisconsin School of Medicine and Public Health, Madison, WI 53705, USA
Matthew Kofron
Division of Developmental Biology, Cincinnati Children’s Hospital Research Foundation, Cincinnati, OH 45229, USA
Paul F. Lambert
McArdle Laboratory for Cancer Research, University of Wisconsin School of Medicine and Public Health, Madison, WI 53705, USA
Adam Lane
Department of Pediatrics, University of Cincinnati College of Medicine, Cincinnati, OH 45267, USA
Najim Ameziane
Centogene, Am Strande 7, 18055 Rostock, Germany
El Mustapha Bahassi
Medpace, Cincinnati, OH 45227, USA
Kathryn A. Wikenheiser-Brokamp
Division of Pathology & Laboratory Medicine and The Perinatal Institute Division of Pulmonary Biology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH 45229, USA
Alessandro de Alarcon
Division of Pediatric Otolaryngology–Head and Neck Surgery, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH 45229, USA
David F. Smith
Division of Pediatric Otolaryngology–Head and Neck Surgery, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH 45229, USA
Susanne I. Wells
Division of Oncology, Cincinnati Children’s Hospital Medical Center, Cincinnati, OH 45229, USA
Juvenile-onset recurrent respiratory papillomatosis (JoRRP) is driven by human papillomavirus (HPV) low-risk strains and is associated with significant morbidity. While previous studies of 2D cultures have shed light on disease pathogenesis and demonstrated the utility of personalized medicine approaches, monolayer cultures lack the 3D tissue architecture and physiology of stratified, sequentially differentiated mucosal epithelium important in RRP disease pathogenesis. Herein we describe the establishment of JoRRP-derived primary cell populations that retain HPV genomes and viral gene expression in culture. These were directly compared to cells from matched adjacent non-diseased tissue, given the known RRP patient-to-patient variability. JoRRP papilloma versus control cells displayed decreased growth at subconfluency, with a switch to increased growth after reaching confluency, suggesting relative resistance to cell-cell contact and/or differentiation. The same papilloma cells grown as 3D organotypic rafts harbored hyperproliferation as compared to controls, with increased numbers of proliferating basal cells and inappropriately replicating suprabasal cells, mimicking phenotypes in the patient biopsies from which they were derived. These complementary model systems provide novel opportunities to elucidate disease mechanisms at distinct stages in JoRRP progression and to identify diagnostic, prognostic and therapeutic factors to personalize patient management and treatment.
