초록 ▼

연구목표
<최종목표>
● 최신의 암 유전체 분석 기법인 NGS를 이용하여 폐암의 암 유전체 연구를 수행, 새로운 유전자 변형 등을 발굴하여 폐암의 진단 및 치료에 기여를 할 수 있는 원천 지식을 제공하고 암 유전체 연구를 위한 기반 확보
<당해연도목표>
● 비흡연 폐선암 환자에서 EGFR-TKI 내성에 관여하는 새로운 유전체 변형을 발굴하여 EGFR-TKI 일차적 내성기전 제시
● 비흡연 폐선암의 새로운 driver oncogene 발굴
● NGS 분석을 통한 소세소폐암의 발생기전 분석
연구내

연구목표
<최종목표>
● 최신의 암 유전체 분석 기법인 NGS를 이용하여 폐암의 암 유전체 연구를 수행, 새로운 유전자 변형 등을 발굴하여 폐암의 진단 및 치료에 기여를 할 수 있는 원천 지식을 제공하고 암 유전체 연구를 위한 기반 확보
<당해연도목표>
● 비흡연 폐선암 환자에서 EGFR-TKI 내성에 관여하는 새로운 유전체 변형을 발굴하여 EGFR-TKI 일차적 내성기전 제시
● 비흡연 폐선암의 새로운 driver oncogene 발굴
● NGS 분석을 통한 소세소폐암의 발생기전 분석
연구내용 및 방법
● EGFR-TKI 일차내성 관련 기전 분석 및 폐선암 새로운 driver oncogene 발굴
1) 수술 후 재발하여 EGFR-TKI 치료를 받은 비흡연 폐선암 환자 37명의 암 조직으로부터 DNA 분리함.
2) IonTorrnet PGM을 이용하여 targeted sequencing 실시함.
3) IonTorrent 분석법으로 내성 기전 발굴에 실패한 7명 환자에서 Whole exome sequencing과 RNA sequencing 실시함.
● Whole genome sequencing을 통한 소세포폐암 발생 기전 분석
1) 수술적 적제술을 시행한 stage I SCLC (pT2N0) 1례 종양 및 정상조직에서 DNA 분리
2) whole genome sequencing using 90 bp paired-end reads & validation variations by Sanger sequencing & SNP genotyping
연구개발에 따른 기대성과
<정량적 성과>
<정성적 성과>

Abstract ▼

Clinical application of targeted next generation sequencing for predicting responsiveness to epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) in never-smoking lung adenocarcinoma
Background: To investigate the clinical utility of targeted next-generation sequencing (NGS) for

Clinical application of targeted next generation sequencing for predicting responsiveness to epidermal growth factor receptor-tyrosine kinase inhibitor (EGFR-TKI) in never-smoking lung adenocarcinoma
Background: To investigate the clinical utility of targeted next-generation sequencing (NGS) for predicting the responsiveness to epidermal growth factor receptor (EGFR)-tyrosine kinase inhibitor (TKI) therapy, we compared the efficacy with conventional sequencing in never-smokers with lung adenocarcinoma (NSLAs).
Methods: We obtained DNA from 48 NSLAs who received gefitinib or erlotinib for their recurrent disease after surgery. Sanger sequencing and peptide nucleic acid clamp polymerase chain reaction (PCR) were used to analyze EGFR, KRAS, BRAF, and PIK3CA mutations. We analyzed ALK, RET, and ROS1 rearrangements by fluorescent in situ hybridization or reverse transcriptase-PCR and quantitative real-time PCR. After molecular screening, Ion Torrent NGS was performed in 31 cases harboring only EGFR exon 19 deletions (19DEL), an L858R mutation, or none of the above mutations.
Results: The 31 samples were divided into four groups: 1) responders to EGFR-TKIs with only 19DEL or L858R (n=15); 2) primary resistance to EGFR-TKI with only 19DEL or L858R (n=4); 3) primary resistance to EGFR-TKI without any mutations (n=8); 4) responders to EGFR-TKI without any mutations (n=4). With NGS, all conventionally detected mutations were confirmed except for one L858R in group 2. Additional uncovered predictive mutations with NGS included one PIK3CA E542K in group 2, two KRAS (G12V and G12D), one PIK3CA E542K, one concomitant PIK3CA and EGFR L858R in group 3, and one EGFR 19DEL in group 4.
Conclusions: Targeted NGS provided a more accurate and clinically useful molecular classification of NSLAs. It may improve the efficacy of EGFR-TKI therapy in lung cancer.
Identification of novel oncogenes in a stage I small cell lung cancer through whole-genome sequencing
Background: Small cell lung cancer (SCLC) is rapidly progressive and most cases are presented with advanced stage. It hampers access to early stage cancer samples, which are essential to understand the early development of this disease. We performed whole-genome sequencing (WGS) of a stage I SCLC to analyze the genomic feature of early tumorigenesis of SCLC.
Methods: We obtained normal and tumor genomic DNAs from a patient with stage I SCLC underwent curative resection and analyzed the normal and cancer genomes by WGS using 90 bp paired-end reads and validated the cancer variations by Sanger sequencing and single-nucleotide polymorphism (SNP) genotyping.
Results: WGS revealed a lot of SNVs, small insertion/deletions, and chromosomal abnormality.
Chromosomes 4p, 5q, 13q, 15q, 17p, and 22q contained many block deletions. Especially, copy loss was observed in tumor suppressor genes RB1 and TP53, and copy gain in oncogene hTERT. Recurrent somatic mutations were found in two genes, TP53 and CREBBP. Novel non-synonymous (ns) single nucleotide variations (SNV) were found in C6ORF103 and SLC5A4 genes. Sequencing of SLC5A4 gene in 23 independent limited disease SCLC samples revealed another ns SNV in SLC5A4 gene, indicating ns SNVs in SLC5A4 gene are recurrent in early –S stage SCLC. This cancer genome had mutated genes associated with Notch and WNT signaling pathways.
Conclusion: WGS analysis in this study provides the genomic landscape of early-stage SCLC, which may provide new insight into the early development of SCLC.

목차 Contents

• 기관고유연구사업 최종보고서 ... 1
• 작 성 요 령 ... 2
• 목 차 ... 3
• 요 약 문 ... 4
• Project Summary ... 6
• 1. 연구의 최종목표 ... 8
• 2. 연구의 내용 및 결과 ... 8
• 3. 연구결과 고찰 및 결론 ... 37
• 4. 연구성과 및 목표달성도 ... 44
• (1) 연구성과 ... 44
• (2) 목표달성도 ... 47
• 5. 연구결과의 활용계획 ... 49
• (1) 연구종료 2년후 예상 연구성과 ... 49
• (2) 연구성과의 활용계획 ... 49
• 6. 참고문헌 ... 50
• 7. 첨부서류 ... 55
• 끝페이지 ... 55