Hutchinson–Gilford progeria syndrome (HGPS) is a rare and fatal human premature ageing disease1, 2, 3, 4, 5, characterized by premature arteriosclerosis and degeneration of vascular smooth muscle cells (SMCs)6, 7, 8. HGPS is caused by a single point mutation in the lamin A (LMNA) gene, resulting in the generation of progerin, a truncated splicing mutant of lamin A. Accumulation of progerin leads to various ageing-associated nuclear defects including disorganization of nuclear lamina and loss of heterochromatin9, 10, 11, 12. Here we report the generation of induced pluripotent stem cells (iPSCs) from fibroblasts obtained from patients with HGPS. HGPS-iPSCs show absence of progerin, and more importantly, lack the nuclear envelope and epigenetic alterations normally associated with premature ageing. Upon differentiation of HGPS-iPSCs, progerin and its ageing-associated phenotypic consequences are restored. Specifically, directed differentiation of HGPS-iPSCs to SMCs leads to the appearance of premature senescence phenotypes associated with vascular ageing. Additionally, our studies identify DNA-dependent protein kinase catalytic subunit (DNAPKcs, also known as PRKDC) as a downstream target of progerin. The absence of nuclear DNAPK holoenzyme correlates with premature as well as physiological ageing. Because progerin also accumulates during physiological ageing6, 12, 13, our results provide an in vitro iPSC-based model to study the pathogenesis of human premature and physiological vascular ageing.
At a glance
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- Supplementary Figures (11.4M)
This file contains Supplementary Figures 1-18 with legends.
- Supplementary Table 1 (30K)
This table contains the DNA methylation parameters.
- Supplementary Table 4 (58K)
This table contains the peptides identified by MudPIT for the indicated candidate progerin-associated partners.
- Supplementary Table 5 (101K)
This table contains the primers used in this study.
- Supplementary Table 2 (356K)
This table contains the genes within 10kb of a differentially methylated region (DMR) found between BJ and HGPS fibroblasts.
- Supplementary Table 3 (77K)
This table contains the genes within 10kb of a differentially methylated region (DMR) found between BJ-iPSCs and HGPS-iPSCs.
- Supplementary Movie 1 (6.3M)
In this movie we see the contracting area derived from BJ-iPSC colony following directed differentiation into cardiac tissue. Movies were taken at day 12 of embryoid body development.
- Supplementary Movie 2 (9.7M)
In this movie we see HGPS-iPSCs develop into contractile cardiac tissue with pacemaker activity in vitro. Movies were taken at day 12 of embryoid body development.