Urine-sample-derived human induced pluripotent stem cells as a model to study PCSK9-mediated autosomal dominant hypercholesterolemia
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 card...
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doaj-25a1d3f503b14c22b8fd68999092740d2020-11-24T21:46:25ZengThe Company of BiologistsDisease Models & Mechanisms1754-84111754-84032016-01-0191819010.1242/dmm.022277022277Urine-sample-derived human induced pluripotent stem cells as a model to study PCSK9-mediated autosomal dominant hypercholesterolemiaKarim Si-Tayeb0Salam Idriss1Benoite Champon2Amandine Caillaud3Matthieu Pichelin4Lucie Arnaud5Patricia Lemarchand6Cédric Le May7Kazem Zibara8Bertrand Cariou9 INSERM, UMR1087, L’institut du thorax, Nantes F-44000, France INSERM, UMR1087, L’institut du thorax, Nantes F-44000, France INSERM, UMR1087, L’institut du thorax, Nantes F-44000, France INSERM, UMR1087, L’institut du thorax, Nantes F-44000, France INSERM, UMR1087, L’institut du thorax, Nantes F-44000, France INSERM, UMR1087, L’institut du thorax, Nantes F-44000, France INSERM, UMR1087, L’institut du thorax, Nantes F-44000, France INSERM, UMR1087, L’institut du thorax, Nantes F-44000, France ER045 - Laboratory of Stem Cells, PRASE, DSST, Beirut, Lebanon INSERM, UMR1087, L’institut du thorax, Nantes F-44000, France 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.http://dmm.biologists.org/content/9/1/81Urine-derived somatic cellsHuman induced pluripotent stem cellsHepatocyte differentiationAutosomal dominant hypercholesterolemiaPCSK9 |
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DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Karim Si-Tayeb Salam Idriss Benoite Champon Amandine Caillaud Matthieu Pichelin Lucie Arnaud Patricia Lemarchand Cédric Le May Kazem Zibara Bertrand Cariou |
spellingShingle |
Karim Si-Tayeb Salam Idriss Benoite Champon Amandine Caillaud Matthieu Pichelin Lucie Arnaud Patricia Lemarchand Cédric Le May Kazem Zibara Bertrand Cariou Urine-sample-derived human induced pluripotent stem cells as a model to study PCSK9-mediated autosomal dominant hypercholesterolemia Disease Models & Mechanisms Urine-derived somatic cells Human induced pluripotent stem cells Hepatocyte differentiation Autosomal dominant hypercholesterolemia PCSK9 |
author_facet |
Karim Si-Tayeb Salam Idriss Benoite Champon Amandine Caillaud Matthieu Pichelin Lucie Arnaud Patricia Lemarchand Cédric Le May Kazem Zibara Bertrand Cariou |
author_sort |
Karim Si-Tayeb |
title |
Urine-sample-derived human induced pluripotent stem cells as a model to study PCSK9-mediated autosomal dominant hypercholesterolemia |
title_short |
Urine-sample-derived human induced pluripotent stem cells as a model to study PCSK9-mediated autosomal dominant hypercholesterolemia |
title_full |
Urine-sample-derived human induced pluripotent stem cells as a model to study PCSK9-mediated autosomal dominant hypercholesterolemia |
title_fullStr |
Urine-sample-derived human induced pluripotent stem cells as a model to study PCSK9-mediated autosomal dominant hypercholesterolemia |
title_full_unstemmed |
Urine-sample-derived human induced pluripotent stem cells as a model to study PCSK9-mediated autosomal dominant hypercholesterolemia |
title_sort |
urine-sample-derived human induced pluripotent stem cells as a model to study pcsk9-mediated autosomal dominant hypercholesterolemia |
publisher |
The Company of Biologists |
series |
Disease Models & Mechanisms |
issn |
1754-8411 1754-8403 |
publishDate |
2016-01-01 |
description |
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. |
topic |
Urine-derived somatic cells Human induced pluripotent stem cells Hepatocyte differentiation Autosomal dominant hypercholesterolemia PCSK9 |
url |
http://dmm.biologists.org/content/9/1/81 |
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