Different Trafficking Phenotypes of Niemann-Pick C1 Gene Mutations Correlate with Various Alterations in Lipid Storage, Membrane Composition and Miglustat Amenability
Niemann-Pick Type C (NPC) is an autosomal recessive lysosomal storage disease leading to progressive neurodegeneration. Mutations in the NPC1 gene, which accounts for 95% of the cases, lead to a defect in intra-lysosomal trafficking of cholesterol and an accumulation of storage material including ch...
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| مؤلفون آخرون: | , , , , , |
| التنسيق: | article |
| منشور في: |
2020
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| الوصول للمادة أونلاين: | http://hdl.handle.net/10725/11979 https://doi.org/10.3390/ijms21062101 http://libraries.lau.edu.lb/research/laur/terms-of-use/articles.php https://www.mdpi.com/1422-0067/21/6/2101 |
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| _version_ | 1864513489663426560 |
|---|---|
| author | Brogden, Graham |
| author2 | Shammas, Hadeel Walters, Friederike Maalouf, Katia Das, Anibh M. Naim, Hassan Y. Rizk, Sandra |
| author2_role | author author author author author author |
| author_facet | Brogden, Graham Shammas, Hadeel Walters, Friederike Maalouf, Katia Das, Anibh M. Naim, Hassan Y. Rizk, Sandra |
| author_role | author |
| dc.creator.none.fl_str_mv | Brogden, Graham Shammas, Hadeel Walters, Friederike Maalouf, Katia Das, Anibh M. Naim, Hassan Y. Rizk, Sandra |
| dc.date.none.fl_str_mv | 2020-07-15T10:37:37Z 2020-07-15T10:37:37Z 2020 2020-07-15 |
| dc.identifier.none.fl_str_mv | 1661-6596 http://hdl.handle.net/10725/11979 https://doi.org/10.3390/ijms21062101 Brogden, G., Shammas, H., Walters, F., Maalouf, K., Das, A. M., Naim, H. Y., & Rizk, S. (2020). Different Trafficking Phenotypes of Niemann-Pick C1 Gene Mutations Correlate with Various Alterations in Lipid Storage, Membrane Composition and Miglustat Amenability. International Journal of Molecular Sciences, 21(6), 1-13. http://libraries.lau.edu.lb/research/laur/terms-of-use/articles.php https://www.mdpi.com/1422-0067/21/6/2101 |
| dc.language.none.fl_str_mv | en |
| dc.relation.none.fl_str_mv | International Journal of Molecular Sciences |
| dc.rights.*.fl_str_mv | info:eu-repo/semantics/openAccess |
| dc.title.none.fl_str_mv | Different Trafficking Phenotypes of Niemann-Pick C1 Gene Mutations Correlate with Various Alterations in Lipid Storage, Membrane Composition and Miglustat Amenability |
| dc.type.none.fl_str_mv | Article info:eu-repo/semantics/publishedVersion info:eu-repo/semantics/article |
| description | Niemann-Pick Type C (NPC) is an autosomal recessive lysosomal storage disease leading to progressive neurodegeneration. Mutations in the NPC1 gene, which accounts for 95% of the cases, lead to a defect in intra-lysosomal trafficking of cholesterol and an accumulation of storage material including cholesterol and sphingolipids in the endo-lysosomal system. Symptoms are progressive neurological and visceral deterioration, with variable onset and severity of the disease. This study investigates the influence of two different NPC1 mutations on the biochemical phenotype in fibroblasts isolated from NPC patients in comparison to healthy, wild type (WT) cells. Skin derived fibroblasts were cultured from one patient compound-heterozygous for D874V/D948Y mutations, which presented wild-type like intracellular trafficking of NPC1, and a second patient compound- heterozygous for I1061T/P887L mutations, which exhibited a more severe biochemical phenotype as revealed in the delayed trafficking of NPC1. Biochemical analysis using HPLC and TLC indicated that lipid accumulations were mutation-dependent and correlated with the trafficking pattern of NPC1: higher levels of cholesterol and glycolipids were associated with the mutations that exhibited delayed intracellular trafficking, as compared to their WT-like trafficked or wild type (WT) counterparts. Furthermore, variations in membrane structure was confirmed in these cell lines based on alteration in lipid rafts composition as revealed by the shift in flotillin-2 (FLOT2) distribution, a typical lipid rafts marker, which again showed marked alterations only in the NPC1 mutant showing major trafficking delay. Finally, treatment with N-butyldeoxynojirimycin (NB-DNJ, Miglustat) led to a reduction of stored lipids in cells from both patients to various extents, however, no normalisation in lipid raft structure was achieved. The data presented in this study help in understanding the varying biochemical phenotypes observed in patients harbouring different mutations, which explain why the effectiveness of NB-DNJ treatment is patient specific. |
| eu_rights_str_mv | openAccess |
| format | article |
| id | LAURepo_4a0e741cfcd1025416bc6ba5ff0aff48 |
| identifier_str_mv | 1661-6596 Brogden, G., Shammas, H., Walters, F., Maalouf, K., Das, A. M., Naim, H. Y., & Rizk, S. (2020). Different Trafficking Phenotypes of Niemann-Pick C1 Gene Mutations Correlate with Various Alterations in Lipid Storage, Membrane Composition and Miglustat Amenability. International Journal of Molecular Sciences, 21(6), 1-13. |
| language_invalid_str_mv | en |
| network_acronym_str | LAURepo |
| network_name_str | Lebanese American University repository |
| oai_identifier_str | oai:laur.lau.edu.lb:10725/11979 |
| publishDate | 2020 |
| repository.mail.fl_str_mv | |
| repository.name.fl_str_mv | |
| repository_id_str | |
| spelling | Different Trafficking Phenotypes of Niemann-Pick C1 Gene Mutations Correlate with Various Alterations in Lipid Storage, Membrane Composition and Miglustat AmenabilityBrogden, GrahamShammas, HadeelWalters, FriederikeMaalouf, KatiaDas, Anibh M.Naim, Hassan Y.Rizk, SandraNiemann-Pick Type C (NPC) is an autosomal recessive lysosomal storage disease leading to progressive neurodegeneration. Mutations in the NPC1 gene, which accounts for 95% of the cases, lead to a defect in intra-lysosomal trafficking of cholesterol and an accumulation of storage material including cholesterol and sphingolipids in the endo-lysosomal system. Symptoms are progressive neurological and visceral deterioration, with variable onset and severity of the disease. This study investigates the influence of two different NPC1 mutations on the biochemical phenotype in fibroblasts isolated from NPC patients in comparison to healthy, wild type (WT) cells. Skin derived fibroblasts were cultured from one patient compound-heterozygous for D874V/D948Y mutations, which presented wild-type like intracellular trafficking of NPC1, and a second patient compound- heterozygous for I1061T/P887L mutations, which exhibited a more severe biochemical phenotype as revealed in the delayed trafficking of NPC1. Biochemical analysis using HPLC and TLC indicated that lipid accumulations were mutation-dependent and correlated with the trafficking pattern of NPC1: higher levels of cholesterol and glycolipids were associated with the mutations that exhibited delayed intracellular trafficking, as compared to their WT-like trafficked or wild type (WT) counterparts. Furthermore, variations in membrane structure was confirmed in these cell lines based on alteration in lipid rafts composition as revealed by the shift in flotillin-2 (FLOT2) distribution, a typical lipid rafts marker, which again showed marked alterations only in the NPC1 mutant showing major trafficking delay. Finally, treatment with N-butyldeoxynojirimycin (NB-DNJ, Miglustat) led to a reduction of stored lipids in cells from both patients to various extents, however, no normalisation in lipid raft structure was achieved. The data presented in this study help in understanding the varying biochemical phenotypes observed in patients harbouring different mutations, which explain why the effectiveness of NB-DNJ treatment is patient specific.PublishedN/A2020-07-15T10:37:37Z2020-07-15T10:37:37Z20202020-07-15Articleinfo:eu-repo/semantics/publishedVersioninfo:eu-repo/semantics/article1661-6596http://hdl.handle.net/10725/11979https://doi.org/10.3390/ijms21062101Brogden, G., Shammas, H., Walters, F., Maalouf, K., Das, A. M., Naim, H. Y., & Rizk, S. (2020). Different Trafficking Phenotypes of Niemann-Pick C1 Gene Mutations Correlate with Various Alterations in Lipid Storage, Membrane Composition and Miglustat Amenability. International Journal of Molecular Sciences, 21(6), 1-13.http://libraries.lau.edu.lb/research/laur/terms-of-use/articles.phphttps://www.mdpi.com/1422-0067/21/6/2101enInternational Journal of Molecular Sciencesinfo:eu-repo/semantics/openAccessoai:laur.lau.edu.lb:10725/119792021-03-19T10:47:40Z |
| spellingShingle | Different Trafficking Phenotypes of Niemann-Pick C1 Gene Mutations Correlate with Various Alterations in Lipid Storage, Membrane Composition and Miglustat Amenability Brogden, Graham |
| status_str | publishedVersion |
| title | Different Trafficking Phenotypes of Niemann-Pick C1 Gene Mutations Correlate with Various Alterations in Lipid Storage, Membrane Composition and Miglustat Amenability |
| title_full | Different Trafficking Phenotypes of Niemann-Pick C1 Gene Mutations Correlate with Various Alterations in Lipid Storage, Membrane Composition and Miglustat Amenability |
| title_fullStr | Different Trafficking Phenotypes of Niemann-Pick C1 Gene Mutations Correlate with Various Alterations in Lipid Storage, Membrane Composition and Miglustat Amenability |
| title_full_unstemmed | Different Trafficking Phenotypes of Niemann-Pick C1 Gene Mutations Correlate with Various Alterations in Lipid Storage, Membrane Composition and Miglustat Amenability |
| title_short | Different Trafficking Phenotypes of Niemann-Pick C1 Gene Mutations Correlate with Various Alterations in Lipid Storage, Membrane Composition and Miglustat Amenability |
| title_sort | Different Trafficking Phenotypes of Niemann-Pick C1 Gene Mutations Correlate with Various Alterations in Lipid Storage, Membrane Composition and Miglustat Amenability |
| url | http://hdl.handle.net/10725/11979 https://doi.org/10.3390/ijms21062101 http://libraries.lau.edu.lb/research/laur/terms-of-use/articles.php https://www.mdpi.com/1422-0067/21/6/2101 |