Date of Award
Medical Doctor (MD)
Objective. To determine if poractant alfa (Curosurf®) could serve as a viable vehicle for silencing RNA (siRNA) delivery. Specifically, we hypothesize that use of Curosurf® as an enhanced delivery vehicle for C/EBP homologous protein (CHOP) and Angiopoietin 2 (Ang2) siRNA would augment gene silencing, preventing cell death and bronchopulmonary dysplasia (BPD) in the mouse lung.
Design. Transfection efficiency of Curosurf® was compared to naked siRNA and Lipofectamine 2000®. MLE-12 cells were transfected with CHOP and Ang2 siRNA using Curosurf®, then exposed to hyperoxic conditions. Cultures were evaluated for respective protein expression and markers of cell death. Curosurf® enhanced siRNA therapy was tested in a hyperoxia-induced mouse model of BPD. Protein expression and lung morphology were assessed and compared to naked siRNA.
Results. Curosurf® significantly improved cell transfection rates when compared to naked siRNA alone. Cell cultures treated with Curosurf® enhanced CHOP and Ang2 siRNA delivery had significantly lower levels of expression of their respective protein products. Cleaved caspase 3, a marker of cell death, was also decreased in cells transfected using Curosurf®. Curosurf® enhanced delivery of Ang2 siRNA had a modest, but significant, improvement in mouse lung morphology in a hyperoxia-induced model of BPD.
Conclusions. Curosurf® enhances silencing RNA delivery of C/EBP homologous protein and Angiopoietin 2, resulting in an improvement in mouse lung phenotype in a hyperoxia-induced model of BPD.
Ramirez, John Gregory, "Surfactant Enhanced Delivery Of C/ebp Homologous Protein And Angiopoietin 2 Silencing Rna To Prevent Bronchopulmonary Dysplasia In Mouse Lung" (2014). Yale Medicine Thesis Digital Library. 1917.