Molecular characterization of gene regulatory networks in primary human tracheal and bronchial epithelial cells

Background: Robust methods to culture primary airway epithelial cells were developed several decades ago and these cells provide the model of choice to investigate many diseases of the human lung. However, the molecular signature of cells from different regions of the airway epithelium has not been well characterized. Methods: We utilize DNase-seq and RNA-seq to examine the molecular signatures of primary cells derived from human tracheal and bronchial tissues, as well as healthy and diseased (cystic fibrosis (CF)) donor lung tissue. Results: Our data reveal an airway cell signature that is divergent from other epithelial cell types and from common airway epithelial cell lines. The differences between tracheal and bronchial cells are clearly evident as are common regulatory features. Only minor variation is seen between bronchial cells from healthy or CF donors. Conclusions: These data are a valuable resource for functional genomics analysis of airway epithelial tissues in human disease

A BAC Transgene Expressing Human CFTR under Control of Its Regulatory Elements Rescues Cftr Knockout Mice

Small-molecule modulators of cystic fibrosis transmembrane conductance regulator (CFTR) biology show promise in the treatment of cystic fibrosis (CF). A Cftr knockout (Cftr KO) mouse expressing mutants of human CFTR would advance in vivo testing of new modulators. A bacterial artificial chromosome (BAC) carrying the complete hCFTR gene including regulatory elements within 40.1 kb of DNA 5′ and 25 kb of DNA 3′ to the gene was used to generate founder mice expressing hCFTR. Whole genome sequencing indicated a single integration site on mouse chromosome 8 (8qB2) with ~6 gene copies. hCFTR+ offspring were bred to murine Cftr KO mice, producing hCFTR+/mCftr− (H+/m−) mice, which had normal survival, growth and goblet cell function as compared to wild-type (WT) mice. Expression studies showed hCFTR protein and transcripts in tissues typically expressing mCftr. Functionally, nasal potential difference and large intestinal short-circuit (Isc) responses to cAMP stimulation were similar in magnitude to WT mice, whereas small intestinal cAMP ΔIsc responses were reduced. A BAC transgenic mouse with functional hCFTR under control of its regulatory elements has been developed to enable the generation of mouse models of hCFTR mutations by gene editing for in vivo testing of new CF therapies. © 2019, The Author(s)

A Numerical Study of the Hydrodynamic Stable Concentration Boundary Layers in a Membrane System Under Microgravitational Conditions

On the basis of the classic formula of the concentration Rayleigh number and the Kedem–Katchalsky equation for diffusive membrane transport, we derived the equations of sixteenth order which show the dependence of the thicknesses of the concentration boundary layers on the difference of the solution concentrations, the concentration Rayleigh number, the solute permeability coefficient of the membrane and the diffusion coefficients in the solution, the kinematic viscosity of the solution, the density of solutions, the temperature and gravitational acceleration. The obtained equation has numerical solutions in the first, third and fourth quadrant of a co-ordinate system. However, only two solutions from the first quadrant of the co-ordinate system have physical meaning. Confining ourselves to the set of solutions with physical meaning only, the thicknesses of concentration boundary layers for different parameters occurring in the obtained equation were calculated numerically