Keratinocyte Growth Factor Gene Electroporation into Skeletal Muscle as a Novel Gene Therapeutic Approach for Elastase-Induced Pulmonary Emphysema in Mice

Abstract

Pulmonary emphysema is a progressive disease with airspace destruction and an effective therapy is needed. Keratinocyte growth factor (KGF) promotes pulmonary epithelial proliferation and has the potential to induce lung regeneration. The aim of this study was to determine the possibility of using KGF gene therapy for treatment of a mouse emphysema model induced by porcine pancreatic elastase (PPE). Eight-week-old BALB/c male mice treated with intra-tracheal PPE administration were transfected with 80 μg of a recombinant human KGF (rhKGF)-expressing FLAG-CMV14 plasmid (pKGF-FLAG gene), or with the pFLAG gene expressing plasmid as a control, into the quadriceps muscle by electroporation. In the lung, the expression of proliferating cell nuclear antigen (PCNA) was augmented, and surfactant protein A (SP-A) and KGF receptor (KGFR) were co-expressed in PCNA-positive cells. Moreover, endogenous KGF and KGFR gene expression increased significantly by pKGF-FLAG gene transfection. Arterial blood gas analysis revealed that the PaO2 level was not significantly reduced on day 14 after PPE instillation with pKGF-FLAG gene transfection compared to that of normal mice. These results indicated that KGF gene therapy with electroporation stimulated lung epithelial proliferation and protected depression of pulmonary function in a mouse emphysema model, suggesting a possible method of treating pulmonary emphysema.

Therapeutic and pathological roles of fibroblast growth factors in pulmonary diseases

“…Oropharyngeal aspiration of FGF7 after elastase instillation cures mice from lung emphysema by inducing TGFb1 expression in AEC2 (Yildirim et al, ). Fgf7 gene delivery into skeletal muscle is also protective against elastase‐induced lung emphysema in mice (Tobinaga et al, ). Attenuating FGFR2b ligand activity blocks compensatory lung growth following pneumonectomy (Chen et al, ).…”

Changes in DNA Methylation of Oocytes and Granulosa Cells Assessed by HELMET during Folliculogenesis in Mouse Ovary

“…Immunohistochemical detection of 5-methylcytosine was performed on the paraffin sections of mouse ovaries, as described in detail previously [ 11 , 18 , 20 ]. After they were dewaxed, the sections were immersed in 10 mM citrate buffer , pH 6.0, autoclaved at 120°C for 15 min, and then pre-incubated with 500 μg/ml normal goat IgG dissolved in 1% BSA in PBS, pH 7.2, for 1 hr.…”

Electroporation: characteristics and therapeutic application of the method (a review)

“…Для элек-НАУЧНЫЕ ОБЗОРЫ тропорации использовался ген фактора роста кератиноци-тов (KGF). Внутримышечная электропорация гена KGF со-провождалась стимуляцией пролиферации легочного эпи-телия и протекцией депрессии функции легких у мышей с моделированной эмфиземой [54]. Метод электропораци-онной терапии в экспериментальной и клинической меди-цине используется и по другим направлениям [51,[55][56][57][58].…”