Using induced pluripotent stem cells (iPSC) to evaluate new CFTR-directed and microRNA-based therapies for cystic fibrosis (CF)
Prof C Greene
Dr K Hurley
Monday, December 02, 2019
Competition Funded PhD Project (Students Worldwide)
The recent development of drugs that directly modulate the CFTR protein in patients with CF has delivered a paradigm shift in how CF is treated using personalised medicine. However, a significant number of patients with CF do not have the better understood and more common CFTR mutations and therefore cannot access these lifesaving medications because CFTR modulator medications are only licensed and available to patients with specific mutations (e.g. Ivacaftor for G551D).
Several CF preclinical models exist but none are lung-specific and reflect an individual patient’s genetic background. Here we propose to engineer a novel in vitro human system to enable the derivation of lungspecific organoids, called bronchospheres, from induced pluripotent stem cells (iPSC) isolated from the peripheral blood of patients with such CFTR mutations. Using forskolin induced swelling of these bronchospheres we will measure lung-specific CFTR function in a high throughput fashion to screen approved and experimental CF therapies. The experimental therapies that will be tested are proprietary inhibitors of microRNA that downregulate CFTR expression. In addition to the high throughput methods we will also assess CFTR function using MQAE and YFP fluorescence-based chloride ion conductance assays in air-liquid interface (ALI) cultures, and immunoprecipitation and western blotting for mature CFTR.
Aims: • To establish iPSC-derived lung-specific models of CF from patients with rare or unknown CFTR mutations. A placement in Boston University, School of Medicine will be provided to learn the methodology.
• To interrogate these models to study CFTR protein, translation, trafficking and function.
• To test the efficiency of existing and novel CFTR-targetting drugs for patients with rare CFTR mutations.
This PhD project will be co-supervised by a scientist and a clinician-scientist. It is designed to facilitate the acquisition of in-depth cutting-edge and state-of-the-art cell and molecular biology skills in a translational and personalised medicine setting.