Identification of target genes for chimeric transcription factors in malignant mesenchymal tumors

恶性间质肿瘤嵌合转录因子靶基因的鉴定

• 批准号: 12470041
• 负责人: NAKAMURA Takuro
• 金额: $ 10.18万
• 依托单位: Japanese Foundation for Cancer Research
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 2000
• 资助国家: 日本
• 起止时间: 2000 至 2002
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/en/grant/KAKENHI-PROJECT-12470041/
• 关键词: chimeric transcription factor; target gene; Ewing sarcoma; clear cell sarcoma; transcriptional regulation; EWS -FLI1; EWS -ATF1; DIGR method; 標的遣伝子; 転写因子

A number of chimeric transcription factors due to tumor-specific chromosomal translocation have been reported in human sarcomas. In order to understand fundamental function of the chimeric transcription factors we have isolated target genes for EWS-ATF1 and EWS-FLI1 in clear cell sarcoma and Ewing sarcoma, respectively, by using a DNA-protein crosslinking/immunopurification/GFP reporter assay (DIGR). Expression of candidate target genes was examined in original tumor cells, and difference of expression induction by chimeric and wild type transcription factors was compared. We have identified POSH, ATM, ARNT2, GPP34, NKX6.1 and NYD-SP28 as target genes for EWS-ATF1 in clear cell sarcoma. EWS-ATF1 repressed POSH expression while it upregulated the other targets. Endogenous POSH expression was suppressed in the clear cell sarcoma cell line, and induction of POSH brought apoptotic cell death to the same cell, suggesting that repressed expression of POSH in clear cell sarcoma may be relevant to the normal signaling pathway in apoptosis. Moreover, BCAR3, RPS18 and p29 have been identified as EWS-FLI1 targets in Ewing sarcoma, indicating that the DIGR method could be applied to broad range of transcription factors. Recently, we have identified a novel chimera in Ewing Sarcoma with t(4;19) translocation. The chimera consists of the HMG box gene CIC and the double homeobox gene DUX4, and it exemplifies the EWS-ETS independent molecular pathway in Ewing sarcoma.

许多嵌合转录因子由于肿瘤特异性染色体易位已被报道在人类肉瘤。为了了解嵌合转录因子的基本功能，我们通过使用DNA-蛋白质交联/免疫纯化/GFP报告基因测定（DIGR）分别分离了透明细胞肉瘤和尤文肉瘤中EWS-ATF 1和EWS-FLI 1的靶基因。在原始肿瘤细胞中检测候选靶基因的表达，并比较嵌合和野生型转录因子诱导表达的差异。我们已经确定POSH、ATM、ARNT 2、GPP 34、NKX6.1和NYD-SP28为透明细胞肉瘤中EWS-ATF 1的靶基因。EWS-ATF 1抑制POSH表达，同时上调其他靶点。透明细胞肉瘤细胞系中内源性POSH表达被抑制，POSH的诱导导致同一细胞的凋亡性细胞死亡，这表明透明细胞肉瘤中POSH的抑制表达可能与凋亡中的正常信号通路有关。此外，BCAR 3、RPS 18和p29已被鉴定为尤文肉瘤中的EWS-FLI 1靶标，表明DIGR方法可应用于广泛的转录因子。最近，我们发现了一种新的嵌合体尤文肉瘤与t（4;19）易位。该嵌合体由HMG盒基因CIC和双同源盒基因DUX 4组成，证实了尤文肉瘤中EWS-ETS独立的分子通路。

项目成果

Saiki Y, Yamazaki Y, Yoshida M, Katoh O, Nakamura T.: "Human EVI9, a homologue of the mouse myeloid leukemia gene, is expressed in the hematopoietic progenitors and down-regulated during myeloid differentiation of HL60 cells."Genomics. 70. 387-391 (2000)

Saiki Y、Yamazaki Y、Yoshida M、Katoh O、Nakamura T.：“人类 EVI9 是小鼠骨髓性白血病基因的同源物，在造血祖细胞中表达，并在 HL60 细胞的骨髓分化过程中下调。”基因组学。

Fujino T: "Single-iranslocation and double-chimeric transcripts : detection of NUP98-HOXA9 in myeloid leukemlas with HOXA11 or HOXA13 breaks of the chromosomal translocation t(7 ; 11)(p15 ; p15)"Blood. 99・(4). 1428-1433 (2002)

Fujino T：“单异位和双嵌合转录本：在染色体易位 t(7; 11)(p15; p15) 发生 HOXA11 或 HOXA13 断裂的骨髓性白血病中检测 NUP98-HOXA9”血液 99・(4)。 1428-1433 (2002)

Liu P, Keller JR, Ortiz M, Tessarollo L, Rachel RA, Nakamura T. Jenkins NA, Copeland NG.: "Bel11a is essential for normal lymphoid development."Nat. Immunol.. in press..

Liu P、Keller JR、Ortiz M、Tessarollo L、Rachel RA、Nakamura T. Jenkins NA、Copeland NG.：“Bel11a 对于正常淋巴发育至关重要。”Nat。

Suzuki A: "t(7 ; 11)(p15 ; p15) chronic myeloid leukaemia developed into blastic transformation showing a novel NUP98/HOXA11 fusion"Br J Haematol. 116・1. 170-172 (2002)

Suzuki A：“t(7; 11)(p15; p15) 慢性粒细胞白血病发展为原始细胞转化，显示出新型 NUP98/HOXA11 融合”Br J Haematol. 116·1 (2002)。

Suzuki A: "t(7 ; 11)(p15 ; p15) chronic myeloid leukaemia developed into blastic transformation showing a novel NUP98/HOXA11 fusion."Br.J.Haematol.. 116・(1). 170-172 (2002)

Suzuki A：“t(7; 11)(p15; p15) 慢性粒细胞白血病发展为母细胞转化，显示出新型 NUP98/HOXA11 融合。”Br.J.Haematol.. 116・(1) (2002)。