Date of Award

January 2014

Document Type

Thesis

Degree Name

Medical Doctor (MD)

Department

Medicine

First Advisor

Vineet Bhandari

Subject Area(s)

Medicine

Abstract

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.

Share

COinS