Novel Therapeutic Strategy for Stroke in Rats by Bone Marrow Stromal Cells and ex vivo HGF Gene Transfer with HSV-1 Vector

Abstract

Occlusive cerebrovascular disease leads to brain ischemia that causes neurological deficits. Here we introduce a new strategy combining mesenchymal stromal cells (MSCs) and ex vivo hepatocyte growth factor (HGF) gene transferring with a multimutated herpes simplex virus type-1 vector in a rat transient middle cerebral artery occlusion (MCAO) model. Gene-transferred MSCs were intracerebrally transplanted into the rats' ischemic brains at 2 h (superacute) or 24 h (acute) after MCAO. Behavioral tests showed significant improvement of neurological deficits in the HGF-transferred MSCs (MSC-HGF)-treated group compared with the phosphatebuffered saline (PBS)-treated and MSCs-only-treated group. The significant difference of infarction areas on day 3 was detected only between the MSC-HGF group and the PBS group with the superacute treatment, but was detected among each group on day 14 with both transplantations. After the superacute transplantation, we detected abundant expression of HGF protein in the ischemic brain of the MSC-HGF group compared with others on day 1 after treatment, and it was maintained for at least 2 weeks. Furthermore, we determined that the increased expression of HGF was derived from the transferred HGF gene in gene-modified MSCs. The percentage of apoptosis-positive cells in the ischemic boundary zone (IBZ) was significantly decreased, while that of remaining neurons in the cortex of the IBZ was significantly increased in the MSC-HGF group compared with others. The present study shows that combined therapy is more therapeutically efficient than MSC cell therapy alone, and it may extend the therapeutic time window from superacute to acute phase.

Bone marrow-derived mesenchymal stromal cells for the repair of central nervous system injury

“…60 Ex vivo gene transfer has also been attempted with hepatocyte growth factor (HGF) which resulted in decreased apoptosis, increased neuron survival in the ischemic boundary zone and functional improvement. 61 …”

Hepatocyte growth factor incorporated into herpes simplex virus vector accelerates facial nerve regeneration after crush injury

“…Thereafter, HGF was measured with rat HGF EIA (Institute of Immunology, Tokyo, Japan), as previously reported. 19,46 This kit is known to react with mouse HGF because rat HGF has about 90.9% amino-acid sequence homology with mouse HGF. 47 Expression of the reporter protein around the inoculated nerve and in the facial nucleus Mice were transcardially perfused with 4% paraformaldehyde under deep anesthesia on day 3, 7 or 14.…”

“…These vectors coded for LacZ (HSV-LacZ) or rat HGF (HSV-HGF), as described previously. 46 The former vector contained the CMV enhancer followed by the LacZ sequence, and the latter contained the full-length rat HGF complementary DNA tagged with the KT3 (large T antigen of simian virus 40) epitope. The two viruses were prepared with the same titers (1Â10 8 pfu ml À1 ).…”

Hypoxia-Inducible Factor 1-α-AA-Modified Bone Marrow Stem Cells Protect PC12 Cells from Hypoxia-Induced Apoptosis, Partially Through VEGF/PI3K/Akt/FoxO1 Pathway

“…Many studies on stroke using BMSCs as treatment suggest genetic modification of BMSCs to improve therapeutic potential. Indeed, overexpression of trophic factors such as brain-derived neurotrophic factor (BDNF), fibroblast growth factor-2, and hematopoietic growth factor in gene-modified BMSCs has been demonstrated to improve neurologic outcome in animal models of ischemic stroke [7][8][9] Hypoxia-inducible factor-1 (HIF-1) is an oxygen-sensitive transcription factor [10]. HIF-1 is a heterodimeric protein that consists of a functional HIF-1a subunit and a constitutively expressed HIF-1b subunit.…”