Interruption of progerin–lamin A/C binding ameliorates Hutchinson-Gilford progeria syndrome phenotype
Su-Jin Lee, … , Gyu Yong Song, Bum-Joon Park
Su-Jin Lee, … , Gyu Yong Song, Bum-Joon Park
Published September 12, 2016
Citation Information: J Clin Invest. 2016;126(10):3879-3893. https://doi.org/10.1172/JCI84164.
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Research Article Aging

Interruption of progerin–lamin A/C binding ameliorates Hutchinson-Gilford progeria syndrome phenotype

Abstract

Hutchinson-Gilford progeria syndrome (HGPS) is a rare autosomal dominant genetic disease that is caused by a silent mutation of the LMNA gene encoding lamins A and C (lamin A/C). The G608G mutation generates a more accessible splicing donor site than does WT and produces an alternatively spliced product of LMNA called progerin, which is also expressed in normal aged cells. In this study, we determined that progerin binds directly to lamin A/C and induces profound nuclear aberrations. Given this observation, we performed a random screening of a chemical library and identified 3 compounds (JH1, JH4, and JH13) that efficiently block progerin–lamin A/C binding. These 3 chemicals, particularly JH4, alleviated nuclear deformation and reversed senescence markers characteristic of HGPS cells, including growth arrest and senescence-associated β-gal (SA–β-gal) activity. We then used microarray-based analysis to demonstrate that JH4 is able to rescue defects of cell-cycle progression in both HGPS and aged cells. Furthermore, administration of JH4 to LmnaG609G/G609G-mutant mice, which phenocopy human HGPS, resulted in a marked improvement of several progeria phenotypes and an extended lifespan. Together, these findings indicate that specific inhibitors with the ability to block pathological progerin–lamin A/C binding may represent a promising strategy for improving lifespan and health in both HGPS and normal aging.

Authors

Su-Jin Lee, Youn-Sang Jung, Min-Ho Yoon, So-mi Kang, Ah-Young Oh, Jee-Hyun Lee, So-Young Jun, Tae-Gyun Woo, Ho-Young Chun, Sang Kyum Kim, Kyu Jin Chung, Ho-Young Lee, Kyeong Lee, Guanghai Jin, Min-Kyun Na, Nam Chul Ha, Clea Bárcena, José M.P. Freije, Carlos López-Otín, Gyu Yong Song, Bum-Joon Park

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Figure 2

Ameliorating effect of JH chemicals on nuclear deformation.

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Ameliorating effect of JH chemicals on nuclear deformation. (A) Specific...
(A) Specific effect of JH chemicals on the binding of lamin A–progerin but not on that of lamin A–lamin A. GST pull-down assay using lysates from HEK293 cells transfected with GFP-LMNA after incubation with GST-LMNA or GST-progerin under treatment with the indicated chemicals. (B) Dissociation of lamin A and progerin by JH4 in human cells. Dissociation of progerin from lamin A/C was confirmed by IP in HEK293 cells. For this, GFP-progerin–transfected HEK293 cells were incubated with the indicated chemicals (5 μM) for 24 hours. A reduction of p16INK4A was also detected in whole-cell lysate. Actin was used as a loading control. SUP, supernatent. (C) JH4 blocked the colocalization of lamin A and progerin. For this, HEK293 cells were cotransfected with GFP-LMNA and nontagged progerin (NT progerin) for 24 hours and incubated with JH4 for an additional 24 hours. JH4 could abolish the colocalization of lamin A and progerin (red) in the nuclear membrane or in nuclear speckles. DAPI was used for DNA staining. Scale bar: 10 μm. (D) JH4 blocks the interaction of lamin A and progerin in HGPS cells. Treatment with 5 μM JH4, but not FTI-277, for 24 hours blocked the interaction of lamin A and progerin in HGPS cells. Cell lysates were immunoprecipitated with a lamin A–specific Ab (H-102, sc-20680) and immunoblotted with a progerin-specific Ab. Normal cells (9 yr; N9) were used as a negative control for progerin. (E) Effect of JH chemicals on nuclear deformation of HGPS cells. Three kinds of HGPS cells were incubated with the indicated chemicals for 24 hours and fixed for lamin A/C immunofluorescence analysis (green). The same effect was observed in other HGPS cells (Supplemental Figure 4F). Scale bars: 10 μm. (F) Quantification of nuclear abnormalities in HGPS cells. JH chemicals, in particular JH4, ameliorated nuclear deformation of HGPS cells. P values were determined by Student’s t test. PPT, protein pellet.