Cargando…
Pathophysiological In Vitro Profile of Neuronal Differentiated Cells Derived from Niemann-Pick Disease Type C2 Patient-Specific iPSCs Carrying the NPC2 Mutations c.58G>T/c.140G>T
Niemann-Pick type C2 (NP-C2) disease is a rare hereditary disease caused by mutations in the NPC2 gene. NPC2 is a small, soluble protein consisting of 151 amino acids, primarily expressed in late endosomes and lysosomes (LE/LY). Together with NPC1, a transmembrane protein found in these organelles,...
Autores principales: | , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2021
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8069078/ https://www.ncbi.nlm.nih.gov/pubmed/33924575 http://dx.doi.org/10.3390/ijms22084009 |
_version_ | 1783683152917561344 |
---|---|
author | Liedtke, Maik Völkner, Christin Jürs, Alexandra V. Peter, Franziska Rabenstein, Michael Hermann, Andreas Frech, Moritz J. |
author_facet | Liedtke, Maik Völkner, Christin Jürs, Alexandra V. Peter, Franziska Rabenstein, Michael Hermann, Andreas Frech, Moritz J. |
author_sort | Liedtke, Maik |
collection | PubMed |
description | Niemann-Pick type C2 (NP-C2) disease is a rare hereditary disease caused by mutations in the NPC2 gene. NPC2 is a small, soluble protein consisting of 151 amino acids, primarily expressed in late endosomes and lysosomes (LE/LY). Together with NPC1, a transmembrane protein found in these organelles, NPC2 accomplishes the exclusion of cholesterol; thus, both proteins are essential to maintain cellular cholesterol homeostasis. Consequently, mutations in the NPC2 or NPC1 gene result in pathophysiological accumulation of cholesterol and sphingolipids in LE/LY. The vast majority of Niemann-Pick type C disease patients, 95%, suffer from a mutation of NPC1, and only 5% display a mutation of NPC2. The biochemical phenotype of NP-C1 and NP-C2 appears to be indistinguishable, and both diseases share several commonalities in the clinical manifestation. Studies of the pathological mechanisms underlying NP-C2 are mostly based on NP-C2 animal models and NP-C2 patient-derived fibroblasts. Recently, we established induced pluripotent stem cells (iPSCs), derived from a donor carrying the NPC2 mutations c.58G>T/c.140G>T. Here, we present a profile of pathophysiological in vitro features, shared by NP-C1 and NP-C2, of neural differentiated cells obtained from the patient specific iPSCs. Profiling comprised a determination of the NPC2 protein level, detection of cholesterol accumulation by filipin staining, analysis of oxidative stress, and determination of autophagy. As expected, the NPC2-deficient cells displayed a significantly reduced amount of NPC2 protein, and, accordingly, we observed a significantly increased amount of cholesterol. Most notably, NPC2-deficient cells displayed only a slight increase of reactive oxygen species (ROS), suggesting that they do not suffer from oxidative stress and express catalase at a high level. As a site note, comparable NPC1-deficient cells suffer from a lack of catalase and display an increased level of ROS. In summary, this cell line provides a valuable tool to gain deeper understanding, not only of the pathogenic mechanism of NP-C2, but also of NP-C1. |
format | Online Article Text |
id | pubmed-8069078 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2021 |
publisher | MDPI |
record_format | MEDLINE/PubMed |
spelling | pubmed-80690782021-04-26 Pathophysiological In Vitro Profile of Neuronal Differentiated Cells Derived from Niemann-Pick Disease Type C2 Patient-Specific iPSCs Carrying the NPC2 Mutations c.58G>T/c.140G>T Liedtke, Maik Völkner, Christin Jürs, Alexandra V. Peter, Franziska Rabenstein, Michael Hermann, Andreas Frech, Moritz J. Int J Mol Sci Article Niemann-Pick type C2 (NP-C2) disease is a rare hereditary disease caused by mutations in the NPC2 gene. NPC2 is a small, soluble protein consisting of 151 amino acids, primarily expressed in late endosomes and lysosomes (LE/LY). Together with NPC1, a transmembrane protein found in these organelles, NPC2 accomplishes the exclusion of cholesterol; thus, both proteins are essential to maintain cellular cholesterol homeostasis. Consequently, mutations in the NPC2 or NPC1 gene result in pathophysiological accumulation of cholesterol and sphingolipids in LE/LY. The vast majority of Niemann-Pick type C disease patients, 95%, suffer from a mutation of NPC1, and only 5% display a mutation of NPC2. The biochemical phenotype of NP-C1 and NP-C2 appears to be indistinguishable, and both diseases share several commonalities in the clinical manifestation. Studies of the pathological mechanisms underlying NP-C2 are mostly based on NP-C2 animal models and NP-C2 patient-derived fibroblasts. Recently, we established induced pluripotent stem cells (iPSCs), derived from a donor carrying the NPC2 mutations c.58G>T/c.140G>T. Here, we present a profile of pathophysiological in vitro features, shared by NP-C1 and NP-C2, of neural differentiated cells obtained from the patient specific iPSCs. Profiling comprised a determination of the NPC2 protein level, detection of cholesterol accumulation by filipin staining, analysis of oxidative stress, and determination of autophagy. As expected, the NPC2-deficient cells displayed a significantly reduced amount of NPC2 protein, and, accordingly, we observed a significantly increased amount of cholesterol. Most notably, NPC2-deficient cells displayed only a slight increase of reactive oxygen species (ROS), suggesting that they do not suffer from oxidative stress and express catalase at a high level. As a site note, comparable NPC1-deficient cells suffer from a lack of catalase and display an increased level of ROS. In summary, this cell line provides a valuable tool to gain deeper understanding, not only of the pathogenic mechanism of NP-C2, but also of NP-C1. MDPI 2021-04-13 /pmc/articles/PMC8069078/ /pubmed/33924575 http://dx.doi.org/10.3390/ijms22084009 Text en © 2021 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/). |
spellingShingle | Article Liedtke, Maik Völkner, Christin Jürs, Alexandra V. Peter, Franziska Rabenstein, Michael Hermann, Andreas Frech, Moritz J. Pathophysiological In Vitro Profile of Neuronal Differentiated Cells Derived from Niemann-Pick Disease Type C2 Patient-Specific iPSCs Carrying the NPC2 Mutations c.58G>T/c.140G>T |
title | Pathophysiological In Vitro Profile of Neuronal Differentiated Cells Derived from Niemann-Pick Disease Type C2 Patient-Specific iPSCs Carrying the NPC2 Mutations c.58G>T/c.140G>T |
title_full | Pathophysiological In Vitro Profile of Neuronal Differentiated Cells Derived from Niemann-Pick Disease Type C2 Patient-Specific iPSCs Carrying the NPC2 Mutations c.58G>T/c.140G>T |
title_fullStr | Pathophysiological In Vitro Profile of Neuronal Differentiated Cells Derived from Niemann-Pick Disease Type C2 Patient-Specific iPSCs Carrying the NPC2 Mutations c.58G>T/c.140G>T |
title_full_unstemmed | Pathophysiological In Vitro Profile of Neuronal Differentiated Cells Derived from Niemann-Pick Disease Type C2 Patient-Specific iPSCs Carrying the NPC2 Mutations c.58G>T/c.140G>T |
title_short | Pathophysiological In Vitro Profile of Neuronal Differentiated Cells Derived from Niemann-Pick Disease Type C2 Patient-Specific iPSCs Carrying the NPC2 Mutations c.58G>T/c.140G>T |
title_sort | pathophysiological in vitro profile of neuronal differentiated cells derived from niemann-pick disease type c2 patient-specific ipscs carrying the npc2 mutations c.58g>t/c.140g>t |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8069078/ https://www.ncbi.nlm.nih.gov/pubmed/33924575 http://dx.doi.org/10.3390/ijms22084009 |
work_keys_str_mv | AT liedtkemaik pathophysiologicalinvitroprofileofneuronaldifferentiatedcellsderivedfromniemannpickdiseasetypec2patientspecificipscscarryingthenpc2mutationsc58gtc140gt AT volknerchristin pathophysiologicalinvitroprofileofneuronaldifferentiatedcellsderivedfromniemannpickdiseasetypec2patientspecificipscscarryingthenpc2mutationsc58gtc140gt AT jursalexandrav pathophysiologicalinvitroprofileofneuronaldifferentiatedcellsderivedfromniemannpickdiseasetypec2patientspecificipscscarryingthenpc2mutationsc58gtc140gt AT peterfranziska pathophysiologicalinvitroprofileofneuronaldifferentiatedcellsderivedfromniemannpickdiseasetypec2patientspecificipscscarryingthenpc2mutationsc58gtc140gt AT rabensteinmichael pathophysiologicalinvitroprofileofneuronaldifferentiatedcellsderivedfromniemannpickdiseasetypec2patientspecificipscscarryingthenpc2mutationsc58gtc140gt AT hermannandreas pathophysiologicalinvitroprofileofneuronaldifferentiatedcellsderivedfromniemannpickdiseasetypec2patientspecificipscscarryingthenpc2mutationsc58gtc140gt AT frechmoritzj pathophysiologicalinvitroprofileofneuronaldifferentiatedcellsderivedfromniemannpickdiseasetypec2patientspecificipscscarryingthenpc2mutationsc58gtc140gt |