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1. Intestinal Bacteroides modulates inflammation, systemic cytokines, and microbial ecology via propionate in a mouse model of cysfibrosis

   https://doi.org/10.1128/mbio.03144-23  

   Price, Courtney E; Valls, Rebecca A; Ramsey, Alexis R; Loeven, Nicole A; Jones, Jane T; Barrack, Kaitlyn E; Schwartzman, Joseph D; Royce, Darlene B; Cramer, Robert A; Madan, Juliette C; et al (January 2024, mBio)

   Harwood, Caroline S (Ed.)

   Persons with cystic fibrosis (CF), starting in early life, show intestinal microbiome dysbiosis characterized in part by a decreasedrelative abundance of the genus Bacteroides. Bacteroides is a major producer of the intestinal short chain fatty acid propionate. Wdemonstrate here that cystic fibrosis transmembrane conductance regulator-defective (CFTR−/−) Caco-2 intestinal epithelial cellsresponsive to the anti-inflammatory effects of propionate. Furthermore, Bacteroides isolates inhibit the IL-1β-induced inflammatorresponse of CFTR−/− Caco-2 intestinal epithelial cells and do so in a propionate-dependent manner. The introduction of Bacteroisupplemented stool from infants with cystic fibrosis into the gut of CftrF508del mice results in higher propionate in the stool as wethe reduction in several systemic pro-inflammatory cytokines. Bacteroides supplementation also reduced the fecal relativeabundance of Escherichia coli, indicating a potential interaction between these two microbes, consistent with previous clinicalstudies. For a Bacteroides propionate mutant in the mouse model, pro-inflammatory cytokine KC is higher in the airway and serucompared with the wild-type (WT) strain, with no significant difference in the absolute abundance of these two strains. Takentogether, our data indicate the potential multiple roles of Bacteroides-derived propionate in the modulation of systemic and airwayinflammation and mediating the intestinal ecology of infants and children with CF. The roles of Bacteroides and the propionate itproduces may help explain the observed gut-lung axis in CF and could guide the development of probiotics to mitigate systemic aairway inflammation for persons with CF. 

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2. Development of the intestinal microbiome in cystic fibrosis in early life

   https://doi.org/10.1128/msphere.00046-23  

   Price, Courtney E.; Hampton, Thomas H.; Valls, Rebecca A.; Barrack, Kaitlyn E.; O’Toole, George A.; Madan, Juliette C.; Coker, Modupe O. (August 2023, mSphere)

   Young, Vincent B. (Ed.)

   Cystic fibrosis is a heritable disease that disrupts ion transport at mucosal surfaces, causing a buildup of mucus and dysregulation of microbial communities in both the lungs and the intestines. Persons with CF are known to have dysbiotic gut microbial communities, but the development of these communities over time beginning at birth has not been thoroughly studied. Here, we describe an observation study following the development of the gut microbiome of cwCF throughout the first 4 years of life, during the critical window of both gut microbiome and immune development. Our findings indicate the possibility of the gut microbiota as a reservoir of airway pathogens and a surprisingly early indication of a microbiota associated with inflammatory bowel disease. 

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3. Experimental and statistical methods for microrheological characterization of heterogeneity in human respiratory mucus mimics of health and disease progression

   https://doi.org/10.1122/8.0000789  

   Caughman, Neall; Papanikolas, Micah; Markovetz, Matthew; Freeman, Ronit; Hill, David B; Forest, M Gregory; Lysy, Martin (November 2024, Journal of Rheology)

   Human respiratory mucus (HRM) is extremely soft, compelling passive microrheology for linear viscoelastic characterization. We focus this study on the use of passive microrheology to characterize HRM heterogeneity, a phenomenon in normal HRM that becomes extreme during cystic fibrosis (CF) disease. Specifically, a fraction of the mucin polymers comprising HRM phase-separate into insoluble structures, called flakes, dispersed in mucin-depleted solution. We first reconstitute HRM samples to the MUC5B:MUC5AC mucin ratios consistent with normal and CF clinical samples, which we show recapitulate progressive flake formation and heterogeneity. We then employ passive particle tracking with 200 nm and 1 μm diameter beads in each reconstituted sample. To robustly analyze the tracking data, we introduce statistical denoising methods for low signal-to-noise tracking data within flakes, tested and verified using model-generated synthetic data. These statistical methods provide a fractional Brownian motion classifier of all successfully denoised, tracked beads in flakes and the dilute solution. From the ensemble of classifier data, per bead diameter and mucus sample, we then employ clustering methods to learn and infer multiple levels of heterogeneity: (i) tracked bead data within vs. outside flakes and (ii) within-flake data buried within or distinguishable from the experimental noise floor. Simulated data consistent with experimental data (within and outside flakes) are used to explore form(s) of the generalized Stokes–Einstein relation (GSER) that recover the dynamic moduli of homogeneous and heterogeneous truth sets of purely flakelike, dilute solution, and mixture samples. The appropriate form of GSER is applied to experimental data to show (i) flakes are heterogeneous with gel and sol domains; (ii) dilute solutions are heterogeneous with only sol domains; and (iii) flake and dilute solution properties vary with probe diameter. 

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4. Molecular dynamics simulations to explore the structure and rheological properties of normal and hyperconcentrated airway mucus

   https://doi.org/10.1111/sapm.12433  

   Ford, Andrew G.; Cao, Xue‐Zheng; Papanikolas, Micah J.; Kato, Takafumi; Boucher, Richard C.; Markovetz, Matthew R.; Hill, David B.; Freeman, Ronit; Forest, Mark Gregory (August 2021, Studies in Applied Mathematics)

   Abstract We develop the first molecular dynamics model of airway mucus based on the detailed physical properties and chemical structure of the predominant gel‐forming mucin MUC5B. Our airway mucus model leverages the LAMMPS open‐source code [https://lammps.sandia.gov], based on the statistical physics of polymers, from single molecules to networks. On top of the LAMMPS platform, the chemical structure of MUC5B is used to superimpose proximity‐based, noncovalent, transient interactions within and between the specific domains of MUC5B polymers. We explore feasible ranges of hydrophobic and electrostatic interaction strengths between MUC5B domains with 9 nm spatial and 1 ns temporal resolution. Our goal here is to propose and test a mechanistic hypothesis for a striking clinical observation with respect to airway mucus: a 10‐fold increase in nonswellable, dense structures called flakes during progression of cystic fibrosis disease. Among the myriad possible effects that might promote self‐organization of MUC5B networks into flake structures, we hypothesize and confirm that the clinically confirmed increase in mucin concentration, from 1.5 to 5 mg/ml, alone is sufficient to drive the structure changes observed with scanning electron microscopy images from experimental samples. We postprocess the LAMMPS simulated data sets at 1.5 and 5 mg/ml, both to image the structure transition and compare with scanning electron micrographs and to show that the 3.33‐fold increase in concentration induces closer proximity of interacting electrostatic and hydrophobic domains, thereby amplifying the proximity‐based strength of the interactions. 

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5. Roles of mucus adhesion and cohesion in cough clearance

   https://doi.org/10.1073/pnas.1811787115  

   Button, Brian; Goodell, Henry P.; Atieh, Eyad; Chen, Yu-Cheng; Williams, Robert; Shenoy, Siddharth; Lackey, Elijah; Shenkute, Nathan T.; Cai, Li-Heng; Dennis, Robert G.; et al (November 2018, Proceedings of the National Academy of Sciences)

   Clearance of intrapulmonary mucus by the high-velocity airflow generated by cough is the major rescue clearance mechanism in subjects with mucoobstructive diseases and failed cilial-dependent mucus clearance, e.g., subjects with cystic fibrosis (CF) or chronic obstructive pulmonary disease (COPD). Previous studies have investigated the mechanical forces generated at airway surfaces by cough but have not considered the effects of mucus biophysical properties on cough efficacy. Theoretically, mucus can be cleared by cough from the lung by an adhesive failure, i.e., breaking mucus-cell surface adhesive bonds and/or by cohesive failure, i.e., directly fracturing mucus. Utilizing peel-testing technologies, mucus-epithelial surface adhesive and mucus cohesive strengths were measured. Because both mucus concentration and pH have been reported to alter mucus biophysical properties in disease, the effects of mucus concentration and pH on adhesion and cohesion were compared. Both adhesive and cohesive strengths depended on mucus concentration, but neither on physiologically relevant changes in pH nor bicarbonate concentration. Mucus from bronchial epithelial cultures and patient sputum samples exhibited similar adhesive and cohesive properties. Notably, the magnitudes of both adhesive and cohesive strength exhibited similar velocity and concentration dependencies, suggesting that viscous dissipation of energy within mucus during cough determines the efficiency of cough clearance of diseased, hyperconcentrated, mucus. Calculations of airflow-induced shear forces on airway mucus related to mucus concentration predicted substantially reduced cough clearance in small versus large airways. Studies designed to improve cough clearance in subjects with mucoobstructive diseases identified reductions of mucus concentration and viscous dissipation as key therapeutic strategies. 

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