Histone demethylase JARID1C inactivation triggers genomic instability in sporadic renal cancer
Beatrice Rondinelli, … , Davide Cittaro, Giovanni Tonon
Beatrice Rondinelli, … , Davide Cittaro, Giovanni Tonon
Published November 9, 2015
Citation Information: J Clin Invest. 2015;125(12):4625-4637. https://doi.org/10.1172/JCI81040.
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Research Article Genetics Oncology

Histone demethylase JARID1C inactivation triggers genomic instability in sporadic renal cancer

Abstract

Mutations in genes encoding chromatin-remodeling proteins are often identified in a variety of cancers. For example, the histone demethylase JARID1C is frequently inactivated in patients with clear cell renal cell carcinoma (ccRCC); however, it is largely unknown how JARID1C dysfunction promotes cancer. Here, we determined that JARID1C binds broadly to chromatin domains characterized by the trimethylation of lysine 9 (H3K9me3), which is a histone mark enriched in heterochromatin. Moreover, we found that JARID1C localizes on heterochromatin, is required for heterochromatin replication, and forms a complex with established players of heterochromatin assembly, including SUV39H1 and HP1α, as well as with proteins not previously associated with heterochromatin assembly, such as the cullin 4 (CUL4) complex adaptor protein DDB1. Transcription on heterochromatin is tightly suppressed to safeguard the genome, and in ccRCC cells, JARID1C inactivation led to the unrestrained expression of heterochromatic noncoding RNAs (ncRNAs) that in turn triggered genomic instability. Moreover, ccRCC patients harboring JARID1C mutations exhibited aberrant ncRNA expression and increased genomic rearrangements compared with ccRCC patients with tumors endowed with other genetic lesions. Together, these data suggest that inactivation of JARID1C in renal cancer leads to heterochromatin disruption, genomic rearrangement, and aggressive ccRCCs. Moreover, our results shed light on a mechanism that underlies genomic instability in sporadic cancers.

Authors

Beatrice Rondinelli, Dalia Rosano, Elena Antonini, Michela Frenquelli, Laura Montanini, DaChuan Huang, Simona Segalla, Kosuke Yoshihara, Samir B. Amin, Dejan Lazarevic, Bin Tean The, Roel G.W. Verhaak, P. Andrew Futreal, Luciano Di Croce, Lynda Chin, Davide Cittaro, Giovanni Tonon

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Figure 3

Loss of heterochromatic histone marks at pericentric and centromeric repeats of JARID1C-silenced cells.

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Loss of heterochromatic histone marks at pericentric and centromeric rep...
(A) Western blot analysis of total cell lysates of CTRsh- and J1CshA-transfected NIH-3T3 cells. (B) Chromatin of control and silenced NIH-3T3 cells was immunoprecipitated with anti-H3K4me3– and anti-H3K9me3–specific Abs and analyzed for enrichment by qPCR with specific minor and major primers, as listed in the Supplemental Methods. Isotypic IgGs were used as controls. Results are expressed as the percentage of input. Error bars represent the SEM of 3 independent experiments. *P < 0.05, Student’s t test. (C) Confocal immunofluorescence images of NIH-3T3 cells stained with anti-H3K9me3–specific Ab and DAPI. Representative cells with heterochromatic H3K9me3 dots (positive) and without dots (negative) are shown. The table scores the mean ± SEM of 2 independent experiments (100 cells per condition analyzed; P < 0.01, Fisher’s exact test). (D) Rescue experiments were performed on NIH-3T3 cells, as reported in the Supplemental Methods. After 72 hours, chromatin was immunoprecipitated with an anti-H3K4me3–specific Ab. Enrichment was analyzed by qPCR with minor and major primers. Results are expressed as the percentage of input. Error bars represent the SEM of 3 independent replicates. Orange and yellow bars represent IgGs detected with minor and major qPCRs, respectively. *P < 0.05, Student’s t test. Mut, mutant.