Date of Award

January 2014

Document Type

Open Access 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.

Comments

This is an Open Access Thesis.

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