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PLoS One. 2017 Nov 17;12(11):e0188256. doi: 10.1371/journal.pone.0188256. eCollection 2017.

Age of heart disease presentation and dysmorphic nuclei in patients with LMNA mutations.

Author information

1
Departments of Biomedical Engineering, University of California, Irvine, CA, United States of America.
2
The Edwards Lifesciences Center for Advanced Cardiovascular Technology, University of California, Irvine, CA, United States of America.
3
Pediatrics-Genetics & Genomics Division-School of Medicine, University of California, Irvine, CA, United States of America.
4
Biological Chemistry-School of Medicine, University of California, Irvine, CA, United States of America.
5
Chemical Engineering and Materials Science, University of California, Irvine, CA, United States of America.

Abstract

Nuclear shape defects are a distinguishing characteristic in laminopathies, cancers, and other pathologies. Correlating these defects to the symptoms, mechanisms, and progression of disease requires unbiased, quantitative, and high-throughput means of quantifying nuclear morphology. To accomplish this, we developed a method of automatically segmenting fluorescently stained nuclei in 2D microscopy images and then classifying them as normal or dysmorphic based on three geometric features of the nucleus using a package of Matlab codes. As a test case, cultured skin-fibroblast nuclei of individuals possessing LMNA splice-site mutation (c.357-2A>G), LMNA nonsense mutation (c.736 C>T, pQ246X) in exon 4, LMNA missense mutation (c.1003C>T, pR335W) in exon 6, Hutchinson-Gilford Progeria Syndrome, and no LMNA mutations were analyzed. For each cell type, the percentage of dysmorphic nuclei, and other morphological features such as average nuclear area and average eccentricity were obtained. Compared to blind observers, our procedure implemented in Matlab codes possessed similar accuracy to manual counting of dysmorphic nuclei while being significantly more consistent. The automatic quantification of nuclear defects revealed a correlation between in vitro results and age of patients for initial symptom onset. Our results demonstrate the method's utility in experimental studies of diseases affecting nuclear shape through automated, unbiased, and accurate identification of dysmorphic nuclei.

PMID:
29149195
PMCID:
PMC5693421
DOI:
10.1371/journal.pone.0188256
[Indexed for MEDLINE]
Free PMC Article

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