Genetic Modification of Airway Progenitors after Lentiviral Gene Delivery to the Amniotic Fluid of Murine Fetuses
| Autor: | Carolyn Lutzko, Chang Mu Hong, Nancy Smiley, Xingchao Wang, Kim Chi Bui, Ping Xia, Dinithi Senadheera, Suparna Mishra |
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| Rok vydání: | 2011 |
| Předmět: |
Pulmonary and Respiratory Medicine
Amniotic fluid Transgene Green Fluorescent Proteins Clinical Biochemistry Fluorescent Antibody Technique Gene delivery Biology Green fluorescent protein Mice Fetus Pregnancy Transduction Genetic Animals Whole Body Imaging Luciferase Transgenes Luciferases Lung Molecular Biology Cellular Senescence Wound Healing Stem Cells Lentivirus Gene Transfer Techniques Epithelial Cells Surfactant protein C Articles Cell Biology respiratory system Amniotic Fluid Survival Analysis Molecular biology Trachea Phosphoglycerate Kinase Organ Specificity Female Stem cell Cell aging Subcellular Fractions |
| Zdroj: | American Journal of Respiratory Cell and Molecular Biology. 44:562-570 |
| ISSN: | 1535-4989 1044-1549 |
| DOI: | 10.1165/rcmb.2009-0235oc |
| Popis: | Lentiviral vectors with the firefly luciferase or enhanced green fluorescent protein (EGFP) transgenes were delivered to the amniotic fluid of murine fetuses at Embryonic Day (E) 14.5 or E16.5. Whole-body imaging of luciferase recipients after birth demonstrated transgene expression in the peritoneal and thoracic regions. Organ imaging showed luciferase expression in lung, skin, stomach, and/or intestine. Histological immunofluorescence analysis of EGFP recipients demonstrated that small clusters (≤ three cells) of EGFP-positive epithelial cells were present in the large and small airways of recipients at up to 7 months (n = 11). There was no difference in the frequency of transgene expression in mice injected at E14.5 or E16.5 in respiratory or nonrespiratory organs. Analysis of the bronchoalveolar duct junctions on tissue sections of recipient mice identified multiple EGFP-positive epithelial cells. Cells coexpressing EGFP, Clara cell 10-kd protein, and surfactant protein C (SPC) were also found in lungs, consistent with the transduction of bronchoalveolar stem cells. Next, naphthalene lung injury in both luciferase and EGFP recipients was performed to determine whether transduced cells could contribute to tissue repair. In luciferase recipients, the whole-body luciferase signal increased 2- to 20-fold at 2 weeks after naphthalene treatment. Remarkably, immunohistological analysis of the lungs of EGFP recipients after lung injury repair demonstrated repopulation of airways with long stretches of EGFP-positive epithelial cells (n = 4). Collectively, these data demonstrate that lentiviral gene delivery to the amniotic fluid of murine fetuses genetically modifies long-lived epithelial progenitors capable of contributing to lung injury repair. |
| Databáze: | OpenAIRE |
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