Silencing of PNPLA6, the Neuropathy Target Esterase (NTE) codifying gene, alters neurodifferentiation of human embryonal carcinoma stem cells (NT2)

Neuropathy Target Esterase (NTE) is a protein involved in the development of a polyneuropathy caused by exposure to certain organophosphorus compounds. In vivo and in vitro studies have also associated NTE with embryonic development since NTE null mice embryos are non-viable, and silencing the NTE-codifying gene (Pnpla6) in mouse embryonic stem cells strongly alters the differentiation of vascular and nervous systems. In this paper, human embryonal carcinoma stem cells (hNT2) are used as an in vitro neurodifferentiation model to determine whether PNPLA6 silencing is able to alter the differentiation process. In control cultures, PNPLA6 mRNA levels increased in parallel with other neuroectodermal markers during neurodifferentiation. PNPLA6 silencing with specific interference RNA reached a 97% decrease in gene expression 3 days after transfection and with a maximum reduction in NTE enzymatic activity (50%), observed on day 4. Silencing PNPLA6 showed an 80% decrease in quantifiable neuronal cells after 13 days in vitro (DIV) compared to controls and absence of different neuronal markers after 66 DIV. Microarray data analysis of the PNPLA6-silenced cells showed alterations in several developmental processes, mainly neurogenesis and epithelium tube morphogenesis. PNPLA6 silencing also led to a reduction in electrical activity and an altered neuronal phenotype. This work is the first proof supporting the hypothesis that NTE plays a role in human early neurodevelopment using a human cell differentiation model.

PAMIES David;  PRICE Anna;  FABBRI Marco;  GRIBALDO Laura;  SCELFO Bibiana;  HARRIS Georgina;  COLLOTTA Angelo;  VILANOVA Eugenio;  SOGORB Miguel A.; 

2015-01-13

PERGAMON-ELSEVIER SCIENCE LTD

JRC91488

0306-4522,   

http://www.sciencedirect.com/science/article/pii/S0306452214007088,    https://publications.jrc.ec.europa.eu/repository/handle/JRC91488,   

10.1016/j.neuroscience.2014.08.031,