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Mol Genet Metab. 2017 May;121(1):28-34. doi: 10.1016/j.ymgme.2017.03.009. Epub 2017 Apr 3.

Generation of induced Pluripotent Stem Cells as disease modelling of NLSDM.

Author information

1
Laboratory of Cellular Biochemistry and Molecular Biology, CRIBENS, Catholic University of the Sacred Heart, pz Buonarroti 30, Milan 20145, Italy; Psychology Department, Catholic University of the Sacred Heart, Largo Gemelli 1, Milan 20123, Italy. Electronic address: daniela.tavian@unicatt.it.
2
Laboratory of Cellular Biochemistry and Molecular Biology, CRIBENS, Catholic University of the Sacred Heart, pz Buonarroti 30, Milan 20145, Italy; Psychology Department, Catholic University of the Sacred Heart, Largo Gemelli 1, Milan 20123, Italy.
3
Laboratory of Human Genetics, E.O. Ospedali Galliera, Via Volta 6, Genoa 16128, Italy.
4
Laboratory of Human Genetics, E.O. Ospedali Galliera, Via Volta 6, Genoa 16128, Italy; Stem Cell Laboratory, Department of Experimental Medicine, University of Genoa, c/o Advanced Biotechnology Center, L.go R. Benzi, 10, Genoa 16132, Italy.
5
UOC Neurologia, San Filippo Neri Hospital, via Martinotti 20, Rome 00135, Italy.
6
Department of Nutrition, University of North Carolina, Chapel Hill, NC 27599, USA.
7
Cellular Fate Reprogramming Unit, Department of Molecular and Translational Medicine, University of Brescia, Brescia 25123, Italy.
8
IRCCS Fondazione Ospedale S. Camillo, Venice, Italy.

Abstract

Neutral Lipid Storage Disease with Myopathy (NLSDM) is a rare defect of triacylglycerol metabolism, characterized by the abnormal storage of neutral lipid in organelles known as lipid droplets (LDs). The main clinical features are progressive myopathy and cardiomyopathy. The onset of NLSDM is caused by autosomal recessive mutations in the PNPLA2 gene, which encodes adipose triglyceride lipase (ATGL). Despite its name, this enzyme is present in a wide variety of cell types and catalyzes the first step in triacylglycerol lipolysis and the release of fatty acids. Here, we report the derivation of NLSDM-induced pluripotent stem cells (NLSDM-iPSCs) from fibroblasts of two patients carrying different PNPLA2 mutations. The first patient was homozygous for the c.541delAC, while the second was homozygous for the c.662G>C mutation in the PNPLA2 gene. We verified that the two types of NLSDM-iPSCs possessed properties of embryonic-like stem cells and could differentiate into the three germ layers in vitro. Immunofluorescence analysis revealed that iPSCs had an abnormal accumulation of triglycerides in LDs, the hallmark of NLSDM. Furthermore, NLSDM-iPSCs were deficient in long chain fatty acid lipolysis, when subjected to a pulse chase experiment with oleic acid. Collectively, these results demonstrate that NLSDM-iPSCs are a promising in vitro model to investigate disease mechanisms and screen drug compounds for NLSDM, a rare disease with few therapeutic options.

KEYWORDS:

Lipid droplets; Lipid metabolism; Myopathy; NLSDM; PNPLA2; iPSCs

PMID:
28391974
PMCID:
PMC5434246
DOI:
10.1016/j.ymgme.2017.03.009
[Indexed for MEDLINE]
Free PMC Article

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