title: Generation of induced Pluripotent Stem Cells as disease modelling of NLSDM.
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.
authors.affiliation: 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.
authors.forename: D
authors.initials: D
authors.lastname: Tavian
authors.affiliation: 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.
authors.forename: S
authors.initials: S
authors.lastname: Missaglia
authors.affiliation: Laboratory of Human Genetics, E.O. Ospedali Galliera, Via Volta 6, Genoa 16128, Italy.
authors.forename: M
authors.initials: M
authors.lastname: Castagnetta
authors.affiliation: 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.
authors.forename: D
authors.initials: D
authors.lastname: Degiorgio
authors.affiliation: UOC Neurologia, San Filippo Neri Hospital, via Martinotti 20, Rome 00135, Italy.
authors.forename: E M
authors.initials: EM
authors.lastname: Pennisi
authors.affiliation: Department of Nutrition, University of North Carolina, Chapel Hill, NC 27599, USA.
authors.forename: R A
authors.initials: RA
authors.lastname: Coleman
authors.affiliation: Cellular Fate Reprogramming Unit, Department of Molecular and Translational Medicine, University of Brescia, Brescia 25123, Italy.
authors.forename: P
authors.initials: P
authors.lastname: Dell'Era
authors.affiliation: Cellular Fate Reprogramming Unit, Department of Molecular and Translational Medicine, University of Brescia, Brescia 25123, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Mora
authors.affiliation: IRCCS Fondazione Ospedale S. Camillo, Venice, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Angelini
authors.affiliation: Laboratory of Human Genetics, E.O. Ospedali Galliera, Via Volta 6, Genoa 16128, Italy.
authors.forename: D A
authors.initials: DA
authors.lastname: Coviello
mesh_terms: D018929:Cell Culture Techniques; D002454:Cell Differentiation; D005347:Fibroblasts / Q000166:cytology* / Q000473:pathology; D006801:Humans; D008049:Lipase / Q000235:genetics*; D008052:Lipid Metabolism, Inborn Errors / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D008066:Lipolysis; D008954:Models, Biological; D009135:Muscular Diseases / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D009154:Mutation; D039904:Pluripotent Stem Cells / Q000166:cytology* / Q000378:metabolism / Q000473:pathology; D014280:Triglycerides / Q000378:metabolism; D000217:Acyltransferases
chemical_list: D008049:Lipase; D014280:Triglycerides; C494794:PNPLA2 protein, human; D000217:Acyltransferases
keywords: Lipid droplets; Lipid metabolism; Myopathy; NLSDM; PNPLA2; iPSCs