This proposal is a NHLBI-RFA (R43) to develop and validate a novel human cell-based diagnostic tool for predicting individual cystic fibrosis (CF) patient responses to drugs that improve the function of mutated forms of the cystic fibrosis transmembrane conductance regulator (CFTR) protein. Large- scale in vitro expansion of human nasal epithelial cells is now feasible. The primary cells, when differentiated as air/liquid interface cultures (ALI), recapitulate a patient-specific CFTR phenotype and provide a sensitive and reproducible platform for testing CFTR-directed therapeutics. Transepithelial ion transport is a direct measure of CFTR function and in vitro demonstration of improved function of mutated CFTR has been shown to predict clinical benefit in specific patients. Nasal epithelial cells are collected from CF subjects, cultured, and cryopreserved to generate a bank of cells of known CFTR genotype. Cells are expanded, seeded onto filter supports for ALI culture (up to ~5,000 cultures/subject), differentiated, tested for CFTR expression and activity. The immediate goals of this project are to use the primary HNE cell cultures to: 1) validate protocols for expansion, differentiation, and testing, 2) demonstrate mutation-specific drug efficacy of known CFTR-directed drugs, 3) establish reproducibility of inter-patient CFTR-drug response variability, and 4) determine if in vitro responses predict in vivo clinical benefit. The long term goal is to show that this in vitro human cell model predicts individual patient responses to CFTR-directed therapeutics and to develop a diagnostic platform to guide optimization of personalized therapies for all CF patients.
Public Health Relevance Statement: Project Narrative The goal of the work described in this proposal is to develop and validate a novel human cell-based diagnostic tool for predicting individual patient responses to drugs that improve the function of mutated forms of the cystic fibrosis transmembrane conductance regulator (CFTR) protein, which is defective in cystic fibrosis (CF) patients. We will collect human nasal epithelial cells from living-donors using a simple, cost-effective, minimally-invasive nasal brushing procedure. The cells are expanded in the laboratory and electrophysiological methods are used to measure CFTR ion transport activity. Previous studies have shown that drug-induced increases ion transport activity in airway epithelial cells predict clinical benefit. This work will establish the protocols for collecting, growing, and testing the cells and will generate data to validate the use of this model for predicting responses of individual CF patients to CFTR-directed drugs. Ultimately the goal is to be able to take advantage of new CFTR drugs as they come to market and deliver personalized therapies for all CF patients.
Project Terms: abstracting; Air; base; Biological Models; Biological Sciences; cell bank; Cell Culture Techniques; Cell model; Cells; Clinical; Clinical Data; clinical efficacy; cohort; Collection; cost effective; Coupled; Cryopreservation; Cultured Cells; Cystic Fibrosis; cystic fibrosis patients; Cystic Fibrosis Transmembrane Conductance Regulator; Data; Data Set; Diagnostic; drug development; drug efficacy; Electrophysiology (science); Epithelial; Epithelial Cells; Evaluation; Genotype; Goals; Human; improved functioning; In Vitro; in vivo; Individual; individual patient; innovation; Ion Transport; Laboratories; Liquid substance; Living Donors; Lung; Marketing; Measures; Methods; minimally invasive; Modeling; mutant; Mutate; Mutation; Mutation Spectra; National Heart, Lung, and Blood Institute; Nose; novel; novel drug combination; novel therapeutics; Patient Care; Patients; Pediatric Hospitals; personalized medicine; Pharmaceutical Preparations; Phenotype; pre-clinical; predicting response; Procedures; Proteins; Protocols documentation; Pulmonary Cystic Fibrosis; Reproducibility; response; Sampling; success; System; Testing; Therapeutic; tool; Translations; Validation; Variant; VX-770; VX-809; Work
