Gene delivery by the hSP-B promoter to lung alveolar type II epithelial cells in LAL-knockout mice through bone marrow mesenchymal stem cells

Abstract

Tissue damage and inflammation promote bone marrow stem cells (BMSCs) to differentiate into a variety of cell types in residing tissues. BMSCs can stably maintain their plasticity and are an ideal cell population for delivery of therapeutic genes to non-hematopoietic tissues. Using lacZ as a reporter gene, we demonstrated that the lung-specific human surfactant protein B (hSP-B) 1.5-kb promoter is able to deliver the lacZ gene into the lung of lysosomal acid lipase (LAL) gene-knockout (lalÀ/À) mice by b-galactosidase staining, flow cytometry and double immunofluorescence staining. Around 10-18% alveolar type II epithelial cells (AT II cells) exhibited positive lacZ gene expression after 8 weeks of BMSC injection in recipient lalÀ/À mice. The wild-type mice exhibited no expression after the same treatment. BMSCs from hSP-B 1.5-kb lacZ transgenic mice entered and repopulated in lalÀ/À bone marrow. The study supports a concept that pulmonary inflammation caused by LAL deficiency can trigger BMSC residing in lalÀ/À bone marrow, migrating into the lung and converting into residential AT II cells. The hSP-B 1.5 kb promoter is an ideal tool to deliver therapeutic genes into AT II cells through BMSCs to cure pulmonary inflammation-triggered diseases.

Overexpression of Dominant Negative Peroxisome Proliferator-Activated Receptor-γ (PPARγ) in Alveolar Type II Epithelial Cells Causes Inflammation and T-Cell Suppression in the Lung

“…ϪDOX, doxycycline-untreated bitransgenic mice; ϩDOX, doxycycline-treated bitransgenic mice previous study, LAL deficiency caused inflammation-induced BMSC conversion into AT II epithelial cells. 17 Because PPAR␥ serves as a downstream effector of LAL, it is conceivable that BMSC-to-AT II cell conversion is regulated by PPAR␥. To test this assumption, the purified adherent BMSCs from hSP-B 1.5-kb lacZ transgenic mice were injected into 2-month doxycycline-treated or untreated CCSPrtTA/(tetO) 7 -CMV-dnPPAR␥ recipient bitransgenic mice.…”

“…17 BMSCs were isolated from femur and tibia of hSP-B 1.5kb lacZ transgenic mice, 2 to 3 months old, 18 and then were cultured in Mesencult MSC basal medium (StemCell Technologies, Vancouver, BC, Canada) at 37°C with 5% CO 2 . After 1 day, the unattached cells were discarded and, for attached BMSCs, the medium was completely replaced.…”

“…We have previously reported that inactivation of the endogenous PPAR␥ function by blocking ligand synthesis promoted BMSCs to convert into AT II epithelial cells in lal Ϫ/Ϫ mice as part of pathogenesis. 17 Based on the observations of double-immunofluorescence staining and FACS analysis in the present study, BMSCs from hSP-B 1.5-kb lacZ donor mice showed the ability to convert into AT II epithelial cells during pulmonary inflammation and injury in the doxycycline-treated CCSP-rtTA/ (tetO) 7 -CMV-dnPPAR␥ bitransgenic lung ( Figure 8A). Conversion did not occur in doxycycline-untreated bitransgenic mice.…”

Critical Roles of Lysosomal Acid Lipase in T Cell Development and Function

“…Bone marrow cells were flushed from femurs and resuspended in FACS buffer as previously described. 10 Peripheral blood mononuclear cells were obtained after red blood cell lysis. Lung single cell suspension was prepared as previously described.…”

Gene Therapy and Stem Cell Therapy to Improve the Outcome of Human Islet Transplantation

“…We are the first group to report the combination strategy of Adv-based gene therapy and BMSC-based cell therapy to improve the outcome of islet transplantation. Our data indicated that BMSCs were efficient gene delivery vehicles to improve the outcome of islet transplantation, as they did to treat other diseases [138,139]. Adv-hHGF-hIL-1Ra-transduced primary BMSCs secreted elevated levels of HGF and IL-1Ra and retained their protective capability for the long-term survival of islet grafts.…”