Spirovation

The histology core provides expertise in all aspects of transmission and scanning electron microscopy. Access to a high pressure freezer and automated freeze substitution is available.

Novel Biomarkers for Pulmonary Drug Development

• Novel biomarkers for measuring the pharmacodynamic and therapeutic effects of inhaled therapeutic agents in sputum
  
• Measures of absolute mucin concentrations, mucin/mucus osmotic pressure/modulus, and measures of mucus adhesion in induced or spontaneously expectorated sputum
  
• The Sputum Core measures cell counts, cytokines, and growth factors in sputum and BAL
  
• Resources, including the Sputum Core, for development of novel biomarkers for pulmonary drug development

The Histology Core provides high quality light and electron microscopy services:

• Light microscopy services include: prosection, tissue fixation, processing, embedding, sectioning, staining and cover slipping of paraffin, frozen and soft plastic tissue sections.
  
• The histology core also provides expertise in all aspects of transmission and scanning electron microscopy. Access to a high pressure freezer and automated freeze substitution is available.
  
• A highlight of the core facility is our specialized procedure for histological evaluation of airway cells grown on porous membrane supports.

Isolation, storage, and analysis of high quality RNA.

PK, target engagement, initial clinical benefit/ risk

We offer numerous animal models of lung diseases, including the first “CF mouse” (UNCtm null).

The Mouse Models Core Laboratory operates under the supervision of Dr. Wanda O’Neal and is dedicated to in vivo studies involving transgenic and knock-out mice generated or procured through the Marsico Lung Institute.

Capabilities

• Generation and characterization of congenic βENaC -Tg lines (in C57BL6N, C3H/HeN, BALBc/J, FVB/NJ, 129/Sv genetic backgrounds).
  
• Transgenic lines that overexpress different combinations of ENaC subunits (a and/or β and/or g).
  
• Transgenic lines that overexpress low amount of βENaC transgene (low-overexpressor line).
  Crossings of βENaC-Tg mice with selected gene-deleted mice (TNFa, IL-4Ra, MyD88, TLR2, TLR4, Muc1, Muc 4, Muc16, Muc5ac, Muc5b, CFTRDF508, NLRP3).
  
• Administration of pharmacologic treatments aimed at preventing and/or blunting the insurgence of obstructing lung pathology.

Use recent advances in cell immortalization technology to create new cell lines and characterize their biochemical properties, morphology and electrophysiologic function

Obtain nasal tissues and tracheo-bronchiolar segments from normal, CF and disease control humans as sources of airway epithelial cells for culture, for ex vivo experiments, for RNA and proteins representative of in vivo conditions, and for in situ hybridization and immunohistochemistry

• Innovative animal models and in vitro models utilizing patient-derived materials
  
• Expertise in pathobiology of disease, enabling a pathway-based approach to target ID/ validation

• Innovative animal models and in vitro models utilizing patient-derived materials
  
• Expertise in pathobiology of disease, enabling a pathway-based approach to target ID/ validation

Ussing chamber ion current measurement in bronchial and GI epithelial cells

We have extensive capabilities to perform classical Ussing chamber studies on large numbers of tissues for screening purposes. Extensive experience has accrued in developing the appropriate quality control procedures to assure accurate and reproducible testing of compound activities in this assay

We offer an extensive array of technologies to measure ciliary function both in vitro and in vivo. Thus, we have available techniques for measurement of ciliary beat frequency in human and animal culture systems, measurements with high-speed videomicroscopy of ciliary beat form, ultrastructural measurement capacities to evaluate ciliary shaft structure, and the ability to perform proteomics of ciliary shaft proteins.

We have extensive experience with cell culture and freshly excised murine tracheal models to measure baseline and regulated mucin secretion. These models are coupled to a spectrum of mucin measurement technologies, including antibody/ELISA based, biochemical/UPLC - light scattering technologies, and techniques that couple both technologies.

In addition, we have extensive experience in measuring biophysical properties of mucus that are important for drug penetration through mucus, mucus flow, and mucus adhesion.

We have extensive experience in characterizing airway surface liquid physiology, in response to candidate therapeutic agents, under the “thin film” ASL technique. In “standard” versions of this technique, a combination of airway surface liquid (ASL) volume as measured by confocal techniques, indices of ion transport processes mediating ASL volume homeostasis from transepithelial microelectrode voltage measurements, coupled with micropipette liquid sampling for measurements of drug concentrations, can be made. Note, drugs can be added to the basolateral compartment of these preparations, the luminal compartment in perfluorocarbon, or indeed, more recently, by an interface to ultrasonic nebulizers that deliver aerosols to cultured airway surfaces at rates that mimic aerosol deposition in vivo.