In utero gene editing for monogenic lung disease.
Deepthi AlapatiWilliam J ZachariasHeather A HartmanAvery C RossidisJohn D StratigisNicholas J AhnBarbara E CoonsSu ZhouHiaying LiKshitiz SinghJeremy KatzenYaniv TomerAlexandra C ChadwickKiran MusunuruMichael F BeersEdward E MorriseyWilliam H PeranteauPublished in: Science translational medicine (2020)
Monogenic lung diseases that are caused by mutations in surfactant genes of the pulmonary epithelium are marked by perinatal lethal respiratory failure or chronic diffuse parenchymal lung disease with few therapeutic options. Using a CRISPR fluorescent reporter system, we demonstrate that precisely timed in utero intra-amniotic delivery of CRISPR-Cas9 gene editing reagents during fetal development results in targeted and specific gene editing in fetal lungs. Pulmonary epithelial cells are predominantly targeted in this approach, with alveolar type 1, alveolar type 2, and airway secretory cells exhibiting high and persistent gene editing. We then used this in utero technique to evaluate a therapeutic approach to reduce the severity of the lethal interstitial lung disease observed in a mouse model of the human SFTPCI73T mutation. Embryonic expression of SftpcI73T alleles is characterized by severe diffuse parenchymal lung damage and rapid demise of mutant mice at birth. After in utero CRISPR-Cas9-mediated inactivation of the mutant SftpcI73T gene, fetuses and postnatal mice showed improved lung morphology and increased survival. These proof-of-concept studies demonstrate that in utero gene editing is a promising approach for treatment and rescue of monogenic lung diseases that are lethal at birth.
Keyphrases
- crispr cas
- genome editing
- interstitial lung disease
- genome wide
- mouse model
- respiratory failure
- systemic sclerosis
- pulmonary hypertension
- gestational age
- wild type
- endothelial cells
- induced apoptosis
- poor prognosis
- rheumatoid arthritis
- preterm infants
- extracorporeal membrane oxygenation
- dna methylation
- low grade
- oxidative stress
- cancer therapy
- idiopathic pulmonary fibrosis
- mechanical ventilation
- pregnant women
- type diabetes
- cell death
- cell proliferation
- cell cycle arrest
- metabolic syndrome
- long non coding rna
- skeletal muscle
- living cells
- sensitive detection
- pregnancy outcomes
- induced pluripotent stem cells
- african american
- pluripotent stem cells
- single molecule
- transcription factor