Patient-Derived Organotypic Epithelial Rafts Model Phenotypes in Juvenile-Onset Recurrent Respiratory Papillomatosis.

Mary C Bedard,David F Smith,Najim Ameziane,Marion G Brusadelli,Alessandro De Alarcon,Sonya Ruiz-Torres,Adam Lane,Adrean Carlile,Denis Lee,El Mustapha Bahassi,Paul F Lambert,Matthew Kofron,Susanne I Wells,Kathryn A Wikenheiser-Brokamp,Hannah Lodin

doi:10.3390/v13010068

Abstract

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.

Highlights

Recurrent respiratory papillomatosis (RRP) is a hyperproliferative condition of regenerative epithelial papillomas that occur along the respiratory tract, often requiring repeated surgical removal [1]
Our studies demonstrate that 2D cultures of primary papilloma versus non-diseased cells are optimally complemented by 3D
Primary cellcell culture, wewe hypothesized that models els routinely used in studies of high-risk might broaden the range of tools availroutinely used in studies of high-risk human papillomavirus (HPV) [11] might broaden the range of tools available for able for mechanistic studies, screening, therapeutic andmedicine personalized medicine apmechanistic studies, therapeutic andscreening, personalized approaches for Juvenile-onset recurrent respiratory papillomatosis (JoRRP)

Summary

Introduction

Recurrent respiratory papillomatosis (RRP) is a hyperproliferative condition of regenerative epithelial papillomas that occur along the respiratory tract, often requiring repeated surgical removal [1]. Clinical presentation of RRP is divided into two distinct subgroups, adult-onset (AoRRP) and juvenile-onset (JoRRP) disease [3]. JoRRP is etiologically associated with low-risk strains of human papillomavirus (HPV). Acquired by vertical transmission at birth [4,5]; most patients with JoRRP are positive for HPV6 or HPV11, with greater prevalence for HPV6 [6]. AoRRP has a higher frequency of HPV-negative papillomas [6] and may involve extended viral latency [7]. JoRRP is more common and more aggressive than AoRRP [8], with JoRRP patients undergoing 4–5 surgeries on average in the year following diagnosis [9]. JoRPP is the most common benign neoplasm of the larynx in children [3]

Methods

Results

Conclusion