Determination of the Pathological Features of NPC1 Variants in a Cellular Complementation Test
Niemann-Pick Type C (NP-C) is a rare disorder of lipid metabolism caused by mutations within the <i>NPC1</i> and <i>NPC2</i> genes. NP-C is a neurovisceral disease leading to a heterogeneous, multisystemic spectrum of symptoms in those affected. Until now, there is no investi...
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| Format: | Article |
| Language: | English |
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MDPI AG
2019-10-01
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| Series: | International Journal of Molecular Sciences |
| Subjects: | |
| Online Access: | https://www.mdpi.com/1422-0067/20/20/5185 |
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doaj-9ccf745efeff4f5e878797a15cc61680 |
|---|---|
| record_format |
Article |
| collection |
DOAJ |
| language |
English |
| format |
Article |
| sources |
DOAJ |
| author |
Xiao Feng Claudia Cozma Supansa Pantoom Christina Hund Katharina Iwanov Janine Petters Christin Völkner Claudia Bauer Florian Vogel Peter Bauer Frank U. Weiss Markus M. Lerch Anne-Marie Knospe Andreas Hermann Moritz J. Frech Jiankai Luo Arndt Rolfs Jan Lukas |
| spellingShingle |
Xiao Feng Claudia Cozma Supansa Pantoom Christina Hund Katharina Iwanov Janine Petters Christin Völkner Claudia Bauer Florian Vogel Peter Bauer Frank U. Weiss Markus M. Lerch Anne-Marie Knospe Andreas Hermann Moritz J. Frech Jiankai Luo Arndt Rolfs Jan Lukas Determination of the Pathological Features of NPC1 Variants in a Cellular Complementation Test International Journal of Molecular Sciences variant of unknown significance lipid metabolism pharmacological chaperone liver disease |
| author_facet |
Xiao Feng Claudia Cozma Supansa Pantoom Christina Hund Katharina Iwanov Janine Petters Christin Völkner Claudia Bauer Florian Vogel Peter Bauer Frank U. Weiss Markus M. Lerch Anne-Marie Knospe Andreas Hermann Moritz J. Frech Jiankai Luo Arndt Rolfs Jan Lukas |
| author_sort |
Xiao Feng |
| title |
Determination of the Pathological Features of NPC1 Variants in a Cellular Complementation Test |
| title_short |
Determination of the Pathological Features of NPC1 Variants in a Cellular Complementation Test |
| title_full |
Determination of the Pathological Features of NPC1 Variants in a Cellular Complementation Test |
| title_fullStr |
Determination of the Pathological Features of NPC1 Variants in a Cellular Complementation Test |
| title_full_unstemmed |
Determination of the Pathological Features of NPC1 Variants in a Cellular Complementation Test |
| title_sort |
determination of the pathological features of npc1 variants in a cellular complementation test |
| publisher |
MDPI AG |
| series |
International Journal of Molecular Sciences |
| issn |
1422-0067 |
| publishDate |
2019-10-01 |
| description |
Niemann-Pick Type C (NP-C) is a rare disorder of lipid metabolism caused by mutations within the <i>NPC1</i> and <i>NPC2</i> genes. NP-C is a neurovisceral disease leading to a heterogeneous, multisystemic spectrum of symptoms in those affected. Until now, there is no investigative tool to demonstrate the significance of single variants within the <i>NPC</i> genes. Hence, the aim of the study was to establish a test that allows for an objective assessment of the pathological potential of <i>NPC1</i> gene variants. Chinese hamster ovary cells defective in the <i>NPC1</i> gene accumulate cholesterol in lysosomal storage organelles. The cells were transfected with NPC1-GFP plasmid vectors carrying distinct sequence variants. Filipin staining was used to test for complementation of the phenotype. The known variant p.Ile1061Thr showed a significantly impaired cholesterol clearance after 12 and 24 h compared to the wild type. Among the investigated variants, p.Ser954Leu and p.Glu1273Lys showed decelerated cholesterol clearance as well. The remaining variants p.Gln60His, p.Val494Met, and p.Ile787Val showed a cholesterol clearance indistinguishable from wild type. Further, p.Ile1061Thr acquired an enhanced clearance ability upon 25-hydroxycholesterol treatment. We conclude that the variants that caused an abnormal clearance phenotype are highly likely to be of clinical relevance. Moreover, we present a system that can be utilized to screen for new drugs. |
| topic |
variant of unknown significance lipid metabolism pharmacological chaperone liver disease |
| url |
https://www.mdpi.com/1422-0067/20/20/5185 |
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AT xiaofeng determinationofthepathologicalfeaturesofnpc1variantsinacellularcomplementationtest AT claudiacozma determinationofthepathologicalfeaturesofnpc1variantsinacellularcomplementationtest AT supansapantoom determinationofthepathologicalfeaturesofnpc1variantsinacellularcomplementationtest AT christinahund determinationofthepathologicalfeaturesofnpc1variantsinacellularcomplementationtest AT katharinaiwanov determinationofthepathologicalfeaturesofnpc1variantsinacellularcomplementationtest AT janinepetters determinationofthepathologicalfeaturesofnpc1variantsinacellularcomplementationtest AT christinvolkner determinationofthepathologicalfeaturesofnpc1variantsinacellularcomplementationtest AT claudiabauer determinationofthepathologicalfeaturesofnpc1variantsinacellularcomplementationtest AT florianvogel determinationofthepathologicalfeaturesofnpc1variantsinacellularcomplementationtest AT peterbauer determinationofthepathologicalfeaturesofnpc1variantsinacellularcomplementationtest AT frankuweiss determinationofthepathologicalfeaturesofnpc1variantsinacellularcomplementationtest AT markusmlerch determinationofthepathologicalfeaturesofnpc1variantsinacellularcomplementationtest AT annemarieknospe determinationofthepathologicalfeaturesofnpc1variantsinacellularcomplementationtest AT andreashermann determinationofthepathologicalfeaturesofnpc1variantsinacellularcomplementationtest AT moritzjfrech determinationofthepathologicalfeaturesofnpc1variantsinacellularcomplementationtest AT jiankailuo determinationofthepathologicalfeaturesofnpc1variantsinacellularcomplementationtest AT arndtrolfs determinationofthepathologicalfeaturesofnpc1variantsinacellularcomplementationtest AT janlukas determinationofthepathologicalfeaturesofnpc1variantsinacellularcomplementationtest |
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doaj-9ccf745efeff4f5e878797a15cc616802020-11-25T01:56:46ZengMDPI AGInternational Journal of Molecular Sciences1422-00672019-10-012020518510.3390/ijms20205185ijms20205185Determination of the Pathological Features of NPC1 Variants in a Cellular Complementation TestXiao Feng0Claudia Cozma1Supansa Pantoom2Christina Hund3Katharina Iwanov4Janine Petters5Christin Völkner6Claudia Bauer7Florian Vogel8Peter Bauer9Frank U. Weiss10Markus M. Lerch11Anne-Marie Knospe12Andreas Hermann13Moritz J. Frech14Jiankai Luo15Arndt Rolfs16Jan Lukas17Translational Neurodegeneration Section „Albrecht-Kossel“, Department of Neurology, University Medical Center Rostock, University of Rostock, 18147 Rostock, GermanyCentogene AG, 18055 Rostock, GermanyTranslational Neurodegeneration Section „Albrecht-Kossel“, Department of Neurology, University Medical Center Rostock, University of Rostock, 18147 Rostock, GermanyTranslational Neurodegeneration Section „Albrecht-Kossel“, Department of Neurology, University Medical Center Rostock, University of Rostock, 18147 Rostock, GermanyTranslational Neurodegeneration Section „Albrecht-Kossel“, Department of Neurology, University Medical Center Rostock, University of Rostock, 18147 Rostock, GermanyTranslational Neurodegeneration Section „Albrecht-Kossel“, Department of Neurology, University Medical Center Rostock, University of Rostock, 18147 Rostock, GermanyTranslational Neurodegeneration Section „Albrecht-Kossel“, Department of Neurology, University Medical Center Rostock, University of Rostock, 18147 Rostock, GermanyCentogene AG, 18055 Rostock, GermanyCentogene AG, 18055 Rostock, GermanyCentogene AG, 18055 Rostock, GermanyDepartment of Medicine A, University Medicine Greifswald, 17489 Greifswald, GermanyDepartment of Medicine A, University Medicine Greifswald, 17489 Greifswald, GermanyTranslational Neurodegeneration Section „Albrecht-Kossel“, Department of Neurology, University Medical Center Rostock, University of Rostock, 18147 Rostock, GermanyTranslational Neurodegeneration Section „Albrecht-Kossel“, Department of Neurology, University Medical Center Rostock, University of Rostock, 18147 Rostock, GermanyTranslational Neurodegeneration Section „Albrecht-Kossel“, Department of Neurology, University Medical Center Rostock, University of Rostock, 18147 Rostock, GermanyTranslational Neurodegeneration Section „Albrecht-Kossel“, Department of Neurology, University Medical Center Rostock, University of Rostock, 18147 Rostock, GermanyCentogene AG, 18055 Rostock, GermanyTranslational Neurodegeneration Section „Albrecht-Kossel“, Department of Neurology, University Medical Center Rostock, University of Rostock, 18147 Rostock, GermanyNiemann-Pick Type C (NP-C) is a rare disorder of lipid metabolism caused by mutations within the <i>NPC1</i> and <i>NPC2</i> genes. NP-C is a neurovisceral disease leading to a heterogeneous, multisystemic spectrum of symptoms in those affected. Until now, there is no investigative tool to demonstrate the significance of single variants within the <i>NPC</i> genes. Hence, the aim of the study was to establish a test that allows for an objective assessment of the pathological potential of <i>NPC1</i> gene variants. Chinese hamster ovary cells defective in the <i>NPC1</i> gene accumulate cholesterol in lysosomal storage organelles. The cells were transfected with NPC1-GFP plasmid vectors carrying distinct sequence variants. Filipin staining was used to test for complementation of the phenotype. The known variant p.Ile1061Thr showed a significantly impaired cholesterol clearance after 12 and 24 h compared to the wild type. Among the investigated variants, p.Ser954Leu and p.Glu1273Lys showed decelerated cholesterol clearance as well. The remaining variants p.Gln60His, p.Val494Met, and p.Ile787Val showed a cholesterol clearance indistinguishable from wild type. Further, p.Ile1061Thr acquired an enhanced clearance ability upon 25-hydroxycholesterol treatment. We conclude that the variants that caused an abnormal clearance phenotype are highly likely to be of clinical relevance. Moreover, we present a system that can be utilized to screen for new drugs.https://www.mdpi.com/1422-0067/20/20/5185variant of unknown significancelipid metabolismpharmacological chaperoneliver disease |