High-Throughput Screen Detects Calcium Signaling Dysfunction in Hutchinson-Gilford Progeria Syndrome
Hutchinson–Gilford progeria syndrome (HGPS) is a deadly childhood disorder, which is considered a very rare disease. It is caused by an autosomal dominant mutation on the LMNA gene, and it is characterized by accelerated aging. Human cell lines from HGPS patients and healthy parental controls were s...
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doaj-1fd3dfcb422647aea32f54460da5196b2021-07-23T13:45:26ZengMDPI AGInternational Journal of Molecular Sciences1661-65961422-00672021-07-01227327732710.3390/ijms22147327High-Throughput Screen Detects Calcium Signaling Dysfunction in Hutchinson-Gilford Progeria SyndromeJuan A. Fafián-Labora0Miriam Morente-López1Fco. Javier de Toro2María C. Arufe3Grupo de Terapia Celular y Medicina Regenerativa, Departamento de Fisioterapia, Ciencias Biomédicas y Medicina, Universdidade da Coruña, Agrupación Estratégica INIBIC-CICA, 15006 A Coruña, SpainGrupo de Terapia Celular y Medicina Regenerativa, Departamento de Fisioterapia, Ciencias Biomédicas y Medicina, Universdidade da Coruña, Agrupación Estratégica INIBIC-CICA, 15006 A Coruña, SpainGrupo de Terapia Celular y Medicina Regenerativa, Departamento de Fisioterapia, Ciencias Biomédicas y Medicina, Universdidade da Coruña, Agrupación Estratégica INIBIC-CICA, 15006 A Coruña, SpainGrupo de Terapia Celular y Medicina Regenerativa, Departamento de Fisioterapia, Ciencias Biomédicas y Medicina, Universdidade da Coruña, Agrupación Estratégica INIBIC-CICA, 15006 A Coruña, SpainHutchinson–Gilford progeria syndrome (HGPS) is a deadly childhood disorder, which is considered a very rare disease. It is caused by an autosomal dominant mutation on the LMNA gene, and it is characterized by accelerated aging. Human cell lines from HGPS patients and healthy parental controls were studied in parallel using next-generation sequencing (NGS) to unravel new non-previously altered molecular pathways. Nine hundred and eleven transcripts were differentially expressed when comparing healthy versus HGPS cell lines from a total of 21,872 transcripts; ITPR1, ITPR3, CACNA2D1, and CAMK2N1 stood out among them due to their links with calcium signaling, and these were validated by Western blot analysis. It was observed that the basal concentration of intracellular Ca<sup>2+</sup> was statistically higher in HGPS cell lines compared to healthy ones. The relationship between genes involved in Ca<sup>2+</sup> signaling and mitochondria-associated membranes (MAM) was demonstrated through cytosolic calcium handling by means of an automated fluorescent plate reading system (FlexStation 3, Molecular Devices), and apoptosis and mitochondrial ROS production were examined by means of flow cytometry analysis. Altogether, our data suggest that the Ca<sup>2+</sup> signaling pathway is altered in HGPS at least in part due to the overproduction of reactive oxygen species (ROS). Our results unravel a new therapeutic window for the treatment of this rare disease and open new strategies to study pathologies involving both accelerated and healthy aging.https://www.mdpi.com/1422-0067/22/14/7327HGPSCa<sup>2+</sup> signalingGRP75GRP78NACROS |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Juan A. Fafián-Labora Miriam Morente-López Fco. Javier de Toro María C. Arufe |
spellingShingle |
Juan A. Fafián-Labora Miriam Morente-López Fco. Javier de Toro María C. Arufe High-Throughput Screen Detects Calcium Signaling Dysfunction in Hutchinson-Gilford Progeria Syndrome International Journal of Molecular Sciences HGPS Ca<sup>2+</sup> signaling GRP75 GRP78 NAC ROS |
author_facet |
Juan A. Fafián-Labora Miriam Morente-López Fco. Javier de Toro María C. Arufe |
author_sort |
Juan A. Fafián-Labora |
title |
High-Throughput Screen Detects Calcium Signaling Dysfunction in Hutchinson-Gilford Progeria Syndrome |
title_short |
High-Throughput Screen Detects Calcium Signaling Dysfunction in Hutchinson-Gilford Progeria Syndrome |
title_full |
High-Throughput Screen Detects Calcium Signaling Dysfunction in Hutchinson-Gilford Progeria Syndrome |
title_fullStr |
High-Throughput Screen Detects Calcium Signaling Dysfunction in Hutchinson-Gilford Progeria Syndrome |
title_full_unstemmed |
High-Throughput Screen Detects Calcium Signaling Dysfunction in Hutchinson-Gilford Progeria Syndrome |
title_sort |
high-throughput screen detects calcium signaling dysfunction in hutchinson-gilford progeria syndrome |
publisher |
MDPI AG |
series |
International Journal of Molecular Sciences |
issn |
1661-6596 1422-0067 |
publishDate |
2021-07-01 |
description |
Hutchinson–Gilford progeria syndrome (HGPS) is a deadly childhood disorder, which is considered a very rare disease. It is caused by an autosomal dominant mutation on the LMNA gene, and it is characterized by accelerated aging. Human cell lines from HGPS patients and healthy parental controls were studied in parallel using next-generation sequencing (NGS) to unravel new non-previously altered molecular pathways. Nine hundred and eleven transcripts were differentially expressed when comparing healthy versus HGPS cell lines from a total of 21,872 transcripts; ITPR1, ITPR3, CACNA2D1, and CAMK2N1 stood out among them due to their links with calcium signaling, and these were validated by Western blot analysis. It was observed that the basal concentration of intracellular Ca<sup>2+</sup> was statistically higher in HGPS cell lines compared to healthy ones. The relationship between genes involved in Ca<sup>2+</sup> signaling and mitochondria-associated membranes (MAM) was demonstrated through cytosolic calcium handling by means of an automated fluorescent plate reading system (FlexStation 3, Molecular Devices), and apoptosis and mitochondrial ROS production were examined by means of flow cytometry analysis. Altogether, our data suggest that the Ca<sup>2+</sup> signaling pathway is altered in HGPS at least in part due to the overproduction of reactive oxygen species (ROS). Our results unravel a new therapeutic window for the treatment of this rare disease and open new strategies to study pathologies involving both accelerated and healthy aging. |
topic |
HGPS Ca<sup>2+</sup> signaling GRP75 GRP78 NAC ROS |
url |
https://www.mdpi.com/1422-0067/22/14/7327 |
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