ATR inhibition controls aggressive prostate tumors deficient in Y-linked histone demethylase KDM5D
Kazumasa Komura, … , Christopher J. Sweeney, Philip W. Kantoff
Kazumasa Komura, … , Christopher J. Sweeney, Philip W. Kantoff
Published June 4, 2018
Citation Information: J Clin Invest. 2018;128(7):2979-2995. https://doi.org/10.1172/JCI96769.
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Research Article Genetics Oncology

ATR inhibition controls aggressive prostate tumors deficient in Y-linked histone demethylase KDM5D

Abstract

Epigenetic modifications control cancer development and clonal evolution in various cancer types. Here, we show that loss of the male-specific histone demethylase lysine-specific demethylase 5D (KDM5D) encoded on the Y chromosome epigenetically modifies histone methylation marks and alters gene expression, resulting in aggressive prostate cancer. Fluorescent in situ hybridization demonstrated that segmental or total deletion of the Y chromosome in prostate cancer cells is one of the causes of decreased KDM5D mRNA expression. The result of ChIP-sequencing analysis revealed that KDM5D preferably binds to promoter regions with coenrichment of the motifs of crucial transcription factors that regulate the cell cycle. Loss of KDM5D expression with dysregulated H3K4me3 transcriptional marks was associated with acceleration of the cell cycle and mitotic entry, leading to increased DNA-replication stress. Analysis of multiple clinical data sets reproducibly showed that loss of expression of KDM5D confers a poorer prognosis. Notably, we also found stress-induced DNA damage on the serine/threonine protein kinase ATR with loss of KDM5D. In KDM5D-deficient cells, blocking ATR activity with an ATR inhibitor enhanced DNA damage, which led to subsequent apoptosis. These data start to elucidate the biological characteristics resulting from loss of KDM5D and also provide clues for a potential novel therapeutic approach for this subset of aggressive prostate cancer.

Authors

Kazumasa Komura, Yuki Yoshikawa, Teppei Shimamura, Goutam Chakraborty, Travis A. Gerke, Kunihiko Hinohara, Kalyani Chadalavada, Seong Ho Jeong, Joshua Armenia, Shin-Yi Du, Ying Z. Mazzu, Kohei Taniguchi, Naokazu Ibuki, Clifford A. Meyer, Gouri J. Nanjangud, Teruo Inamoto, Gwo-Shu Mary Lee, Lorelei A. Mucci, Haruhito Azuma, Christopher J. Sweeney, Philip W. Kantoff

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Figure 2

Aggressive phenotype in loss of KDM5D.

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Aggressive phenotype in loss of KDM5D. (A) Immunoblotting in indicated c...
(A) Immunoblotting in indicated cell lines. Nuclear fractions were collected in indicated cells and subjected to immunoblotting with the indicated antibodies. pLenti-C, control; pLenti-K, KDM5D. (B) Cell growth of indicated cell lines in CSS medium. LNCaP sh-control, KDM5D#1, and KDM5D#3 were cultured in 6-well plates with 0.15 μg/ml of doxycycline, and medium was changed every 3 days. 104R2-control and KDM5D overexpression cells were cultured in 6-well plates, and medium was changed every 3 days. (C) Soft agar colony-formation assay in LNCaP sh-control, KDM5D#1, and KDM5D#3. Representative images after 21 days are shown. (D) Number of colonies in C were counted in 5 random fields in 21 days, and results are shown as mean ± SD. *P < 0.05, 1-way ANOVA with post hoc Tukey’s HSD test. (E) Immunoblotting in indicated cell lines. Nuclear fractions were collected in indicated cells and subjected to immunoblotting with the indicated antibodies. (F) Representative images of soft colony formation assay in indicated cell lines. (G) Schematic representation of orthotopic xenograft mouse model. After the orthotopic inoculation, cells were allowed to form the tumor in 2 weeks, followed by surgical castration, and then the luciferase activity was measured every 2 weeks (n = 5 in each group). (H) Representative images of the quantitative luminescence measurement for each group of 4 in an orthotopic xenograft model. (I) Representative images of the tumor orthotopically inoculated for 8 weeks in each group of 4. (J) Quantitative evaluation of the developed tumor in orthotopic xenograft mice. Total flux (photons/s) in the region of interest (ROI) was recorded every 2 weeks. *P < 0.05, 1-way ANOVA with post hoc Tukey’s HSD test.