Granulocyte-colony stimulating factor gene therapy as a novel therapeutics for stroke in a mouse model

Journal of Biomedical Science

Michael Marshall¹  Kristen Medley²  Rui Tao³  Eugenia Jimenez³  Hongyaun Xu³  Jessica Shen³  Andrew Bent³  Jigar Modi⁴  Janet M. Menzie-Suderam⁵  Howard Prentice⁶  Jang-Yen Wu⁶  Paula Trujillo⁷ 

[1] AEURA Trust, 2525 Arapahoe Ave E4-138, 80302, Boulder, CO, USA;College of Medicine, New York University, 10003, New York, NY, USA;Department of Biomedical Sciences, Charles E. Schmidt College of Medicine, Florida Atlantic University, 33431, Boca Raton, FL, USA;Department of Biomedical Sciences, Charles E. Schmidt College of Medicine, Florida Atlantic University, 33431, Boca Raton, FL, USA;Complex Systems and Brain Sciences, Florida Atlantic University, Boca Raton, FL, USA;Department of Biomedical Sciences, Charles E. Schmidt College of Medicine, Florida Atlantic University, 33431, Boca Raton, FL, USA;Program in Integrative Biology, Florida Atlantic University, 33431, Boca Raton, FL, USA;Department of Biomedical Sciences, Charles E. Schmidt College of Medicine, Florida Atlantic University, 33431, Boca Raton, FL, USA;Program in Integrative Biology, Florida Atlantic University, 33431, Boca Raton, FL, USA;Complex Systems and Brain Sciences, Florida Atlantic University, Boca Raton, FL, USA;Program in Integrative Biology, Florida Atlantic University, 33431, Boca Raton, FL, USA;

关键词: Granulocyte-colony stimulating factor (g-CSF);    Gene therapy;    Endoplasmic reticulum stress apoptosis;    Mitochondrial dynamics;    Autophagy;    Transient global ischemia;   

DOI  :  10.1186/s12929-020-00692-5

来源: Springer

【 摘 要 】

BackgroundGlobal 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.MethodA 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.ResultsSeveral 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.ConclusionTaken 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.

【 授权许可】

CC BY