Cellular and Molecular Biomarkers Indicate Premature Aging in Pseudoxanthoma Elasticum Patients
Janina Tiemann1, Thomas Wagner1, Olivier M. Vanakker2, Matthias van Gils2, José-Luis Bueno Cabrera3, Bettina Ibold1, Isabel Faust1, Cornelius Knabbe1, Doris Hendig1,*
1Institut für Laboratoriums- und Transfusionsmedizin, Herz- und Diabeteszentrum Nordrhein-Westfalen, Universitätsklinik der Ruhr-Universität Bochum, Bad Oeynhausen, Germany 2Center for Medical Genetics, Ghent University Hospital, Ghent, Belgium 3Haematology Department, Hospital Universitario Puerta de Hierro-Majadahonda, Majadahonda, Spain
The molecular processes of aging are very heterogenic and not fully understood. Studies on rare progeria syndromes, which display an accelerated progression of physiological aging, can help to get a better understanding. Pseudoxanthoma elasticum (PXE) caused by mutations in the ATP-binding cassette sub-family C member 6 (ABCC6) gene shares some molecular characteristics with such premature aging diseases. Thus, this is the first study trying to broaden the knowledge of aging processes in PXE patients. In this study, we investigated aging associated biomarkers in primary human dermal fibroblasts and sera from PXE patients compared to healthy controls. Determination of serum concentrations of the aging biomarkers eotaxin-1 (CCL11), growth differentiation factor 11 (GDF11) and insulin-like growth factor 1 (IGF1) showed no significant differences between PXE patients and healthy controls. Insulin-like growth factor binding protein 3 (IGFBP3) showed a significant increase in serum concentrations of PXE patients older than 45 years compared to the appropriate control group. Tissue specific gene expression of GDF11 and IGFBP3 were significantly decreased in fibroblasts from PXE patients compared to normal human dermal fibroblasts (NHDF). IGFBP3 protein concentration in supernatants of fibroblasts from PXE patients were decreased compared to NHDF but did not reach statistical significance due to potential gender specific variations. The minor changes in concentration of circulating aging biomarkers in sera of PXE patients and the significant aberrant tissue specific expression seen for selected factors in PXE fibroblasts, suggests a link between ABCC6 deficiency and accelerated aging processes in affected peripheral tissues of PXE patients.
Table 1 Characterization of human dermal fibroblasts from PXE patients and healthy controls.
Annealing temperature (°C)
GAPDH glyceraldehyde-3-phosphate dehydrogenase
GDF11growth differentiation factor 11
IGFBP3insulin-like growth factor binding protein 3
Table 2 Primer sequences used for quantitative real-time PCR.
Figure 1. CCL11 protein concentration in sera from PXE patients (grey) and healthy controls (white). Data are shown as Box-Plot with median, 25th and 75th percentile and Tukey whiskers (± 1.5 times interquartile range). Control/PXE: ns p>0.05. Cohorts <45 years (n=23) vs. cohorts >45 years (n=22): ##/++ p≤0.01.
Figure 2. Systemic concentration and local mRNA expression of GDF11. (A) GDF11 protein concentration in sera from PXE patients (grey) and healthy controls (white). Data are shown as Box-Plot with median, 25th and 75th percentile and Tukey whiskers (± 1.5 times interquartile range). (B) Relative GDF11 mRNA-expression of PXE fibroblasts (grey) and NHDF (white). Data are shown as mean ± SEM. Control/PXE: *** p ≤0.001. Cohorts <45 years (n=23) vs. cohorts >45 years (n=22): ns p>0.05.
Figure 3. Systemic IGF1 and IGFBP3 protein concentration in sera from PXE patients (grey) and healthy controls (white). (A) IGF1 serum protein concentrations of PXE patients (grey) and healthy controls (white) (B) IGFBP3 serum protein concentrations of PXE patients (grey) and healthy controls (white) (C) molar IGF1/IGFBP3 ratio of serum protein concentrations of PXE patients (grey) and healthy controls (white). Data are shown as Box-Plot with median, 25th and 75th percentile and Tukey whiskers (± 1.5 times interquartile range). Control/PXE: * p≤0.05; ns p>0.05. Cohorts <45 years (n=23) vs. cohorts >45 years (n=22): ns p>0.05.
Figure 4. Local IGFBP3 mRNA expression and protein concentration. (A) Relative IGFBP3 mRNA expression of PXE fibroblasts (grey) and NHDF (white). (B) IGFBP3 protein concentration in supernatant of PXE fibroblasts (grey) and NHDF (white). Data are shown as mean ± SEM. Control/PXE: * p≤0.05; ns p>0.05.
Figure 5. Male and female specific IGFBP3 mRNA expression and protein concentration. (A) Relative IGFBP3 mRNA expression of female PXE fibroblasts (grey) and NHDF (white). (B) IGFBP3 protein concentration in supernatant of female PXE fibroblasts (grey) and NHDF (white). (C) Relative IGFBP3 mRNA expression of male PXE fibroblasts (grey) and NHDF (white). (D) IGFBP3 protein concentration in supernatant of male PXE fibroblasts (grey) and NHDF (white). Data are shown as mean ± SEM. Control/PXE: ** p≤0.01; ns p>0.05.
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