This project will develop a cell and gene therapy for cystic fibrosis (CF). CF, the most common inherited disease in the Caucasian population with > 30,000 affected individuals in the US. The efficacy of CF therapies has been limited and does not address the underlying genetic cause or disease associated tissue damage. Mutation corrected, patient-derived CF-induced pluripotent stem (iPS) cells have the potential to repair tissue affected by disease processes with cells/tissue that function normally. The studies test the hypotheses that: 1) CF cells can be converted into iPS cells by Sendai virus reprogramming and maintain long-term DNA integrity, 2) CF mutations can be corrected in iPS cells by small DNA fragments (SDFs) in conjunction with sequence specific transcription activator-like effector nucleases (TALENs) or helper-dependent adenovirus (HDAV) vectors and maintain DNA integrity, and 3) corrected iPS cells can differentiate into chromosomally stable airway epithelial and inflammatory cell progenitors that can engraft and repair disease damaged tissues. The proposal will: (yr 1) reprogram CF patient iPS cells with a Sendai virus vector containing Oct4, Sox2, Klf4, and/or L-Myc transgenes, (yrs 1-2) repair the disease causing mutation by gene editing using SDFs with TALENs or by HDAV) homologous recombination, and (yrs 2-3) direct the differentiation of corrected cells into human lung airway epithelium and inflammatory cell progenitors associated with CF pathology.
There are > 2500 individuals with CF in California and of the ~ 500,000 new births in California every year, >100 babies are born with CF. Patients with CF require lifelong care that involves expensive therapies to treat the manifestations that result from opportunistic infections and multi-organ dysfunction. While there has been progress in the treatment of CF and the mean survival of someone with CF has increased to 38 yrs, there is no cure for CF, the underlying pathology, and the extensive organ and tissue damage. The only effective, albeit short-term, amelioration of the airway disease pathology has been allogenic lung transplantation. Aside from the human cost and suffering caused by CF, the economic impact to California is substantial. The cost of treating someone with CF is between $10,000 to $25,000/yr/patient, with an estimated lifetime cost of $300,000 to $450.000 per patient. One approach to prevent the disease has been through early detection and prenatal diagnosis, where the usual outcome is selective abortion of the affected fetus. This project offers an alternative approach for possibility curing both newborn and adult patients with their own cells that have been reprogrammed, corrected and converted to cells that will regenerate tissues affected by CF pathology. The benefit to California would be to eliminate emotional and physical suffering in patients and mitigate the economic impact that the life-long treatment has on the medical system.