title: Urine-sample-derived human induced pluripotent stem cells as a model to study PCSK9-mediated autosomal dominant hypercholesterolemia.
abstract: Proprotein convertase subtilisin kexin type 9 (PCSK9) is a critical modulator of cholesterol homeostasis. Whereas PCSK9 gain-of-function (GOF) mutations are associated with autosomal dominant hypercholesterolemia (ADH) and premature atherosclerosis, PCSK9 loss-of-function (LOF) mutations have a cardio-protective effect and in some cases can lead to familial hypobetalipoproteinemia (FHBL). However, limitations of the currently available cellular models preclude deciphering the consequences of PCSK9 mutation further. We aimed to validate urine-sample-derived human induced pluripotent stem cells (UhiPSCs) as an appropriate tool to model PCSK9-mediated ADH and FHBL. To achieve our goal, urine-sample-derived somatic cells were reprogrammed into hiPSCs by using episomal vectors. UhiPSC were efficiently differentiated into hepatocyte-like cells (HLCs). Compared to control cells, cells originally derived from an individual with ADH (HLC-S127R) secreted less PCSK9 in the media (-38.5%; P=0.038) and had a 71% decrease (P<0.001) of low-density lipoprotein (LDL) uptake, whereas cells originally derived from an individual with FHBL (HLC-R104C/V114A) displayed a strong decrease in PCSK9 secretion (-89.7%; P<0.001) and had a 106% increase (P=0.0104) of LDL uptake. Pravastatin treatment significantly enhanced LDL receptor (LDLR) and PCSK9 mRNA gene expression, as well as PCSK9 secretion and LDL uptake in both control and S127R HLCs. Pravastatin treatment of multiple clones led to an average increase of LDL uptake of 2.19 ± 0.77-fold in HLC-S127R compared to 1.38 ± 0.49 fold in control HLCs (P<0.01), in line with the good response to statin treatment of individuals carrying the S127R mutation (mean LDL cholesterol reduction=60.4%, n=5). In conclusion, urine samples provide an attractive and convenient source of somatic cells for reprogramming and hepatocyte differentiation, but also a powerful tool to further decipher PCSK9 mutations and function.
authors.affiliation: INSERM, UMR1087, L'institut du thorax, Nantes F-44000, France CNRS, UMR 6291, Nantes F-44000, France Université de Nantes, Nantes F-44000, France karim.si-tayeb@univ-nantes.fr.
authors.forename: Karim
authors.initials: K
authors.lastname: Si-Tayeb
authors.affiliation: INSERM, UMR1087, L'institut du thorax, Nantes F-44000, France CNRS, UMR 6291, Nantes F-44000, France Université de Nantes, Nantes F-44000, France ER045 - Laboratory of Stem Cells, PRASE, DSST, Beirut, Lebanon Biology Department, Faculty of Sciences-I, Lebanese University, Beirut 6573/14, Lebanon.
authors.forename: Salam
authors.initials: S
authors.lastname: Idriss
authors.affiliation: INSERM, UMR1087, L'institut du thorax, Nantes F-44000, France CNRS, UMR 6291, Nantes F-44000, France Université de Nantes, Nantes F-44000, France.
authors.forename: Benoite
authors.initials: B
authors.lastname: Champon
authors.affiliation: INSERM, UMR1087, L'institut du thorax, Nantes F-44000, France CNRS, UMR 6291, Nantes F-44000, France Université de Nantes, Nantes F-44000, France.
authors.forename: Amandine
authors.initials: A
authors.lastname: Caillaud
authors.affiliation: INSERM, UMR1087, L'institut du thorax, Nantes F-44000, France CNRS, UMR 6291, Nantes F-44000, France Université de Nantes, Nantes F-44000, France CHU Nantes, L'institut du thorax, CIC Endocrinology-Nutrition, Nantes F-44000, France.
authors.forename: Matthieu
authors.initials: M
authors.lastname: Pichelin
authors.affiliation: INSERM, UMR1087, L'institut du thorax, Nantes F-44000, France CNRS, UMR 6291, Nantes F-44000, France Université de Nantes, Nantes F-44000, France.
authors.forename: Lucie
authors.initials: L
authors.lastname: Arnaud
authors.affiliation: INSERM, UMR1087, L'institut du thorax, Nantes F-44000, France CNRS, UMR 6291, Nantes F-44000, France Université de Nantes, Nantes F-44000, France.
authors.forename: Patricia
authors.initials: P
authors.lastname: Lemarchand
authors.affiliation: INSERM, UMR1087, L'institut du thorax, Nantes F-44000, France CNRS, UMR 6291, Nantes F-44000, France Université de Nantes, Nantes F-44000, France.
authors.forename: Cédric
authors.initials: C
authors.lastname: Le May
authors.affiliation: ER045 - Laboratory of Stem Cells, PRASE, DSST, Beirut, Lebanon Biology Department, Faculty of Sciences-I, Lebanese University, Beirut 6573/14, Lebanon.
authors.forename: Kazem
authors.initials: K
authors.lastname: Zibara
authors.affiliation: INSERM, UMR1087, L'institut du thorax, Nantes F-44000, France CNRS, UMR 6291, Nantes F-44000, France Université de Nantes, Nantes F-44000, France CHU Nantes, L'institut du thorax, CIC Endocrinology-Nutrition, Nantes F-44000, France.
authors.forename: Bertrand
authors.initials: B
authors.lastname: Cariou
mesh_terms: D000818:Animals; D002454:Cell Differentiation; D049109:Cell Proliferation; D008078:Cholesterol, LDL / Q000378:metabolism; D005260:Female; D005347:Fibroblasts / Q000378:metabolism; D020869:Gene Expression Profiling; D022781:Hepatocytes / Q000166:cytology; D006801:Humans; D006938:Hyperlipoproteinemia Type II / Q000235:genetics* / Q000652:urine*; D057026:Induced Pluripotent Stem Cells / Q000166:cytology*; D007621:Karyotyping; D008077:Lipoproteins, LDL / Q000378:metabolism; D008297:Male; D051379:Mice; D009154:Mutation; D017035:Pravastatin / Q000627:therapeutic use; D000071449:Proprotein Convertase 9; D043484:Proprotein Convertases / Q000235:genetics*; D012333:RNA, Messenger / Q000378:metabolism; D012697:Serine Endopeptidases / Q000235:genetics*; D014556:Urine / Q000737:chemistry*
chemical_list: D008078:Cholesterol, LDL; D008077:Lipoproteins, LDL; D012333:RNA, Messenger; C472125:PCSK9 protein, human; D000071449:Proprotein Convertase 9; D043484:Proprotein Convertases; D012697:Serine Endopeptidases; D017035:Pravastatin
keywords: Autosomal dominant hypercholesterolemia; Hepatocyte differentiation; Human induced pluripotent stem cells; PCSK9; Urine-derived somatic cells