Increase of Neuronal Differentiation Yield by Preventing Cell Death

Autor: Almeida, Ana S, Soares, Nuno L, Vieira, Melissa, Gramsbergen, Jan Bert, Vieira, Helena L A
Přispěvatelé: NOVA Medical School|Faculdade de Ciências Médicas (NMS|FCM), Centro de Estudos de Doenças Crónicas (CEDOC), Instituto de Tecnologia Química e Biológica António Xavier (ITQB)
Jazyk: angličtina
Rok vydání: 2016
Předmět:
Zdroj: Repositório Científico de Acesso Aberto de Portugal
Repositório Científico de Acesso Aberto de Portugal (RCAAP)
instacron:RCAAP
Popis: This work was supported by: Grant 1 "Translational and mechanistic approaches on carbon monoxide as a modulator of neuroinflammation and neuroprotection in glial cells", ANR/NEU-NMC/0022/2012 (http://www.fct.pt); Fellowship 1 - salary for HLAV, "Translational and mechanistic approaches on carbon monoxide/heme-oxygenase for cytoprotection following cerebral ischemia: integrative role of apoptosis, autophagy, metabolism and neurogenesis", IF/00185/2012 (http://www.fct.pt); Fellowship 2 - salary for ASA, "Carbon Monoxide modulation of neuronal differentiation - novel cell therapy strategies for neurological disorders", SFRH/BD/78440/2011 (http://www.fct.pt); and COST Action BM1005, European Network on Gasotransmitters (http://gasotransmitters.eu). Cerebral ischemia and neurodegenerative diseases lead to impairment or death of neurons in the central nervous system. Stem cell based therapies are promising strategies currently under investigation. Carbon monoxide (CO) is an endogenous product of heme degradation by heme oxygenase (HO) activity. Administration of CO at low concentrations produces several beneficial effects in distinct tissues, namely anti-apoptotic and anti-inflammatory. Herein the CO role on modulation of neuronal differentiation was assessed. Three different models with increasing complexity were used: human neuroblastoma SH-S5Y5 cell line, human teratocarcinoma NT2 cell line and organotypic hippocampal slice cultures (OHSC). Cell lines were differentiated into post-mitotic neurons by treatment with retinoic acid (RA) supplemented with CO-releasing molecule A1 (CORM-A1). CORM-A1 positively modulated neuronal differentiation, since it increased final neuronal production and enhanced the expression of specific neuronal genes: Nestin, Tuj1 and MAP2. Furthermore, during neuronal differentiation process, there was an increase in proliferative cell number (ki67 mRNA expressing cells) and a decrease in cell death (lower propidium iodide (PI) uptake, limitation of caspase-3 activation and higher Bcl-2 expressing cells). CO supplementation did not increase the expression of RA receptors. In the case of SH-S5Y5 model, small amounts of reactive oxygen species (ROS) generation emerges as important signaling molecules during CO-promoted neuronal differentiation. CO's improvement of neuronal differentiation yield was validated using OHSC as ex vivo model. CORM-A1 treatment of OHSC promoted higher levels of cells expressing the neuronal marker Tuj1. Still, CORM-A1 increased cell proliferation assessed by ki67 expression and also prevented cell death, which was followed by increased Bcl-2 expression, decreased levels of active caspase-3 and PI uptake. Likewise, ROS signaling emerged as key factors in CO's increasing number of differentiated neurons in OHSC. In conclusion, CO's increasing number of differentiated neurons is a novel biological role disclosed herein. CO improves neuronal yield due to its capacity to reduce cell death, promoting an increase in proliferative population. However, one cannot disregard a direct CO's effect on specific cellular processes of neuronal differentiation. Further studies are needed to evaluate how CO can potentially modulate cell mechanisms involved in neuronal differentiation. In summary, CO appears as a promising therapeutic molecule to stimulate endogenous neurogenesis or to improve in vitro neuronal production for cell therapy strategies. publishersversion published
Databáze: OpenAIRE
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