Granulocyte-colony stimulating factor gene therapy as a novel therapeutics for stroke in a mouse model
Open Access
- 30 October 2020
- journal article
- research article
- Published by Springer Science and Business Media LLC in Journal of Biomedical Science
- Vol. 27 (1), 1-22
- https://doi.org/10.1186/s12929-020-00692-5
Abstract
Global ischemia is the resulting effect of a cardiopulmonary arrest (CPA). Presently there is no effective treatment to address neurological deficits in patients who survived a CPA. Granulocyte-colony stimulating factor is a growth factor (G-CSF) with a plethora of beneficial effects, including neuroprotection. Clinical application of human G-CSF (hG-CSF) is limited due to its plasma half-life of 4 h. Therefore, novel approaches need to be investigated that would (1) enable prolonged manifestation of hG-CSF and (2) demonstrate G-CSF efficacy from studying the underlying protective mechanisms of hG-CSF. In our previous work, we used the self-complementary adeno-associated virus (stereotype2: scAAV2) as a vector to transfect the hG-CSF gene into the global ischemic brain of a mouse. As an extension of that work, we now seek to elucidate the protective mechanisms of hG-CSF gene therapy against endoplasmic reticulum induced stress, mitochondrial dynamics and autophagy in global ischemia. A single drop of either AAV-CMV-hG-CSF or AAV-CMV-GFP was dropped into the conjunctival sac of the Swiss Webster mouse’s left eye, 30–60 min after bilateral common artery occlusion (BCAO). The efficacy of the expressed hG-CSF gene product was analyzed by monitoring the expression levels of endoplasmic reticulum stress (ER), mitochondrial dynamics and autophagic proteins over 4- and 7-days post-BCAO in vulnerable brain regions including the striatum, overlying cortex (frontal brain regions) and the hippocampus (middle brain regions). Statistical analysis was performed using mostly One-Way Analysis of variance (ANOVA), except for behavioral analysis, which used Repeated Measures Two-Way ANOVA, post hoc analysis was performed using the Tukey test. Several biomarkers that facilitated cellular death, including CHOP and GRP78 (ER stress) DRP1 (mitochondrial dynamics) and Beclin 1, p62 and LC3-ll (autophagy) were significantly downregulated by hG-CSF gene transfer. hG-CSF gene therapy also significantly upregulated antiapoptotic Bcl2 while downregulating pro-apoptotic Bax. The beneficial effects of hG-CSF gene therapy resulted in an overall improvement in functional behavior. Taken together, this study has substantiated the approach of sustaining the protein expression of hG-CSF by eye drop administration of the hG-CSF gene. In addition, the study has validated the efficacy of using hG-CSF gene therapy against endoplasmic reticulum induced stress, mitochondrial dynamics and autophagy in global ischemia.This publication has 83 references indexed in Scilit:
- The Unfolded Protein Response and Cell Fate ControlMolecular Cell, 2018
- GABA-containing liposomes: neuroscience applications and translational perspectives for targeting neurological diseasesNanomedicine: Nanotechnology, Biology and Medicine, 2017
- Mechanisms of Action of Bcl-2 Family ProteinsCold Spring Harbor Perspectives in Biology, 2013
- Impaired Autophagosome Clearance Contributes to Cardiomyocyte Death in Ischemia/Reperfusion InjuryJournal of the American College of Cardiology, 2012
- Endogenous brain protection by granulocyte-colony stimulating factor after ischemic strokeExperimental Neurology, 2009
- ER Stress Triggers Apoptosis by Activating BH3-Only Protein BimCell, 2007
- The autophagic response to nutrient deprivation in the hl‐1 cardiac myocyte is modulated by Bcl‐2 and sarco/endoplasmic reticulum calcium storesThe FEBS Journal, 2007
- G-CSF Reduces Infarct Volume and Improves Functional Outcome after Transient Focal Cerebral Ischemia in MiceJournal of Cerebral Blood Flow & Metabolism, 2005
- LC3, GABARAP and GATE16 localize to autophagosomal membrane depending on form-II formationJournal of Cell Science, 2004
- The development of a new mouse model of global ischemia: focus on the relationships between ischemia duration, anesthesia, cerebral vasculature, and neuronal injury following global ischemia in miceBrain Research, 1998