Development of Glandular Models from Human Nasal Progenitor Cells

Abstract

Hyperplasia/hypertrophy of submucosal glands contributes to mucus overproduction in chronic diseases of the upper and lower respiratory tracts, especially in adult and pediatric chronic rhinosinusitis. Mechanisms that lead to glandular hyperplasia/ hypertrophy are markedly understudied, reflecting a lack of in vitro model systems wherein airway epithelial progenitor cells differentiate into glandular cells. In this study, we developed and compared several in vitro three-dimensional systems using human nasal epithelial basal cells (HNEBCs) cultured by different methods on two types of extracellular matrices. We demonstrate that HNEBCs cultured on Matrigel (Corning, Tewksbury, MA) form glandular acini-like structures, whereas HNEBCs embedded in a collagen type I matrix form a network of tubules. Fibroblast-conditioned medium increases tubule formation in collagen type I. In contrast, HNEBCs cocultured with fibroblasts self-aggregate into organotypic structures with tubules and acini. These observations provide morphological evidence that HNEBCs are pluripotent and retain the capacity to differentiate into structures resembling specific structural components of submucosal glands depending on the extracellular matrices and culture conditions. The resultant models should prove useful in targeting cross-talk between epithelial cells and fibroblasts to decipher molecular mechanisms and specific signals responsible for the development of glandular hyperplasia/hypertrophy, which in turn may lead to new therapeutic strategies for chronic rhinosinusitis and other inflammatory respiratory diseases characterized by glandular hyperplasia/hypertrophy.Keywords: human nasal epithelial cells; sinonasal basal cells; glandular hyperplasia/hypertrophy; extracellular matrix; chronic rhinosinusitis Clinical RelevanceThe resultant three-dimensional models should be useful for studying the development of sinonasal submucosal glands and submucosal glandular hyperplasia/hypertrophy in chronic rhinosinusitis and for deciphering the molecular mechanisms and specific signals responsible for sinonasal basal cell activation. This in turn would enable studies on how to reverse glandular hyperplasia, thereby decreasing mucin overproduction and morbidity in patients with chronic rhinosinusitis. This model may also be applicable to diseases of the lower respiratory tract that exhibit glandular hyperplasia.

Airway Gland Structure and Function

“…Growth of dispersed isolated gland cells in collagen gels produces tubular structures very similar to native glands (298), and the same result can be obtained with surface epithelial cells or purified basal cells from human bronchi (753,754). These preparations are less suitable than planar sheets for many studies but have special advantages for studying phenomena such as the gland hyperplasia and hypertrophy characteristic of many airway diseases.…”

Detection of CFTR function and modulation in primary human nasal cell spheroids

“…Acquisition of GI tissue remains invasive, requiring endoscopy or suction biopsy for rectal specimens, with minor safety risks [10,22]. Limited existing work in 3-dimensional culture of nasal tissue is not clearly adaptable to monitoring CFTR function [23–25]. We hypothesized that primary human nasal epithelial cells (NECs) obtained non-surgically could be grown in 3-dimensional culture to yield a patient-derived, swelling-based model of CFTR function.…”

Toward inclusive therapy with CFTR modulators: Progress and challenges

“…126 Despite these advantages, controversy remains in their application as a surrogate for the lung, as CFTR and other expression profiles may be different from the lower airway, sustenance in culture is more limited, and propensity to differentiate fully is more problematic than lung-derived cells; some of the discrepancies may be mitigated by improved culturing techniques, and there is a concerted effort to optimize culturing technique. To combat the small number of cells obtained from nasal epithelial biopsy, conditional reprogramming 92,[127][128][129] or induced pluripotent stem cells [129][130][131][132][133] represent attractive approaches. The development of three-dimensional organoid models similar to intestinal organoids may also improve ability to discern useful information from small sample sizes.…”