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New fusion gene detected in lung cancer

A detected and gene fusion technology, applied in fusion polypeptide, genetic engineering, detection of programmed cell death, etc., can solve problems such as differences in drug reactivity

Inactive Publication Date: 2016-06-08
NAT CANCER CENT +2
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

[0004] Advanced lung cancer is mainly treated with drugs, but since the response to drugs varies greatly from case to case, a method for predicting which drug will have a therapeutic effect is required

Method used

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  • New fusion gene detected in lung cancer
  • New fusion gene detected in lung cancer
  • New fusion gene detected in lung cancer

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0455] This example describes the identification of novel fusion transcripts in lung cancer tissue.

[0456] To identify novel fusion transcripts that could serve as therapeutic targets, whole-transcriptome sequencing of 114 lung adenocarcinomas and 3 noncancerous lung tissues was performed (RNA sequencing, Meyerson, M. et al., NatRev Genet, 2010, 11, 685-696 ).

[0457] Analyze the Paired-endRead obtained by RNA sequencing, and perform Sanger sequencing of reverse transcription (RT)-PCR products. The results are shown in Table 2 and figure 1 As shown, four novel fusion gene products were identified.

[0458] [Table 2]

[0459]

[0460] EZR-ERBB4 is produced by chromosomal transposition t(2;6), and is a fusion gene of the EZR gene present on chromosome 6q25 and the ERBB4 gene present on chromosome 2q34.

[0461] KIAA1468-RET is produced by chromosomal transposition t(10;18), and is a fusion gene of the KIAA1468 gene present on chromosome 18q21 and the RET gene present o...

Embodiment 2

[0466] This example describes the RT-PCR detection of the gene fusions discovered in Example 1.

[0467] For each fusion gene, PCR primers (forward primer and reverse primer, respectively) derived from the cDNA sequences of the 5' side and 3' side gene parts were prepared (Table 1). Using these primers, PCR amplification was performed using cDNA synthesized from cancer tissue-derived RNA as a template.

[0468] exist figure 2 Electropherogram showing PCR products. Amplification of a specific band was confirmed in a part of the samples, and the base sequence of the PCR product was further confirmed. As a result, it was confirmed that a part of the fusion gene was amplified.

[0469] From the above results, it was shown that the gene fusion found in Example 1 could be detected by checking whether the amplification of a specific band was obtained by RT-PCR and determining the base sequence of the PCR product.

Embodiment 3

[0471]This example shows that the gene fusion found in Example 1 is highly likely to be the responsible mutation for lung cancer.

[0472] In the five cases of lung cancer in which novel gene fusions were found in Example 1, whether there were EGFR point mutations-in-frame deletion mutations (EGFR point mutations, EGFRin-flamedeletion mutations), KRAS point mutations (KRAS point mutations), BRAF mutations, etc. Point mutation (BRAF pointmutation), HER2 in-frame insertion mutation (HER2in-flameinsertionmutation), EML4-ALK gene fusion (EML4-ALKfusion), KIF5B-RET gene fusion (KIF5B-RETfusion), CCDC6-RET gene fusion (CCDC6-RETfusion), CD74-ROS1 gene fusion (CD74-ROS1fusion), EZR-ROS1 gene fusion (EZR-ROS1fusion) and SLC34A2-ROS1 gene fusion (SLC34A2-ROS1fusion).

[0473] As a result, all of the above-mentioned 5 cases of lung cancer were negative for other known mutations, and the 4 novel gene fusions were mutually exclusive with other known mutations responsible for cancer.

[0...

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Abstract

A method for detecting gene fusion that is a mutation responsible for cancer (driver mutation), wherein the method includes a step for detecting, in an isolated sample derived from a subject having cancer, any fusion polynucleotide among an EZR-ERBB4 fusion polynucleotide, KIAA1468-RET fusion polynucleotide, TRIM24-BRAF fusion polynucleotide, CD74-NRG1 fusion polynucleotide, or SLC3A2-NRG1 fusion polynucleotide, or a polypeptide encoded thereby.

Description

technical field [0001] The present invention relates to a method for detecting gene fusion as a responsible mutation of cancer, identifying a substance that inhibits the expression and / or activity of a polypeptide encoded by the fusion polynucleotide produced by the gene fusion, and can bring therapeutic effects to cancer patients or patients with cancer Methods of testing subjects of cancer risk, etc. Background technique [0002] Cancer is the leading cause of death in Japan, and improvement of its treatment is required. In particular, lung cancer is the leading cause of cancer death not only in Japan but also in the whole world, causing more than 1 million deaths every year. Lung cancer is broadly divided into small cell lung cancer and non-small cell lung cancer, and non-small cell lung cancer is divided into three types: lung adenocarcinoma (LADC), squamous cell carcinoma of the lung, and large cell carcinoma. Among these lung cancers, lung adenocarcinoma accounts for...

Claims

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Application Information

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IPC IPC(8): C12Q1/68G01N33/574C12N15/09
CPCC12Q2600/106C12Q2600/156C12N15/113C12N15/62C12N2310/11C12N2310/141C12N9/12C12N9/93A61K31/517C07K14/47C07K14/4756C07K14/70596C07K2319/00C12N15/1135C12N2310/14C12Q1/6886A61P35/00C12Q2600/118C12Q2600/158G01N33/5011G01N2500/04G01N2500/10
Inventor 河野隆志茑幸治安田和基
Owner NAT CANCER CENT
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