Biomedical Research Funded in 2004

1) Jonathan Widdicombe, PhD, University of Davis, Davis, California, Principal Investigator

“Technical Support for Production of Primary Cultures of Human Tracheal Epithelium”

This grant is to provide funding to maintain a CORE facility for the supply of primary cultures

of airway epithelium. These researchers were the first to show that airway epithelial cells

could be grown in culture with retention of vectorial Cl secretion. They then used these cells

to provide the first demonstration of reduced Cl conductance of the apical membrane of

airway epithelium in CF. Additionally, cells grown using their technique have been widely

used by others.

Amount Funded: $30,000.00

2) Dennis Nielson, MD, PhD, University of California San Francisco, San Francisco, California, Principal Investigator and

Danieli Salinas, MD, Post Doctoral Fellow

“Novel Methods to Study the Link Between the Gene Defect and the Pathophysiology of CF”

The purpose of this study is to determine the link between the CF gene defect and clinical

disease. This is of critical importance in developing therapies to treat CF. New biophysical

measurement methods and model systems will be used to define the role of the ASL and

submucosal glands in CF. So far, researchers have found that the pH of submucosal

gland fluid from nasal biopsies is more acidic in CF than in non-CF patients. Further

research could help in understanding airway pH regulation, which could be key to

understanding airway defenses and chronic infection in CF. Additionally, researchers have

successfully demonstrated that submucosal gland dysfunction can be considered a primary

defect in CF. Other results indicate that the fluid secretion rate from submucosal glands

was reduced 2.7-fold and secreted fluid viscosity was elevated 2.2-fold in early CF. These

results provide evidence for submucosal gland dysfunction as an intrinsic defect in CF,

furthering earlier findings supporting the involvement of CFTR in gland fluid secretion in intact

airways, and of reduced fluid secretion and hyperviscosity in severely diseased CF airways.

Amount Funded: $33,333.34

3) Ron Kopito, PhD, Stanford University, Principal Investigator and

Wei Zhang, PhD, Post Doctoral Fellow

“Small Molecule Screening Approach to Search for Suppressors of Delta508”

The purpose of this study is to identify small molecules capable of enhancing the folding

and/or stability of misfolded Delta F508-CFTR. Another goal is to develop ultra-sensitive high

throughput homogeneous assay for detecting the cell surface expression of CFTR. As of now

researchers have generated a reporter that not only contains live cell surface displayed S-tags,

but also maintains Delta F508-CFTR folding and traffics defective properties. Also, the

proposed assay was demonstrated to be valid. Further research is required to finish up the

experiment, specifically generating a stable CHO cell line expressing Delta F508-4S-CFTR.

This stable cell line will then be tested on the optimized assay, and then they will set up a

homogenous cell-based assay for screening small molecules capable of enhancing folding,

cell surface delivery, stability and functional expression of Delta F508-CFTR.

Amount Funded: $40,000.00

4) Beate Illek, PhD, Children’s Hospital Oakland Research Institute, Oakland, California, Principal Investigator

“Vitamin C Uptake into Cystic Fibrosis Airways”

The purpose of this study is to introduce Vitamin C to CF airways in hopes of easing

airflow through CF lungs once again. CF lungs produce a thickened airway

mucus that inhibits transport of sputum, forming plugs that partially block airways. Recent

studies have shown that Vitamin C replenishes local Vitamin C deficits that contribute to

the inhibition of normal hydration of airway surface liquid/mucus. Another aspect has been

added to the study and that is to examine the properties of Vitamin C that act as an

antioxidant by scavenging free radicals. Researchers recently started to study the effects of

an oxidizing environment on CFTR Cl transport and found that an oxidizing environment

blocked CFTR Cl transport across human tracheal cultures. The goal of this project is to

determine the beneficial effects of Vitamin C and the combination of Vitamin C with other

antioxidants during oxidative stress of the airways.

Amount Funded: $20,000.00