지원연구분야 ( 코드 ) G0205 과제번호 1210500 창의일반과제프로그램공개가능여부과제성격 ( 기초, 응용, 개발 ) 응용실용화대상여부실용화비공개 ( 공개, 비공개 ) ( 국문 ) 비소세포폐암환자에서 matrix-assisted laser desorption 연구과제명 과제책임자 세부과제 ionization-time of flight mass spectrometry (MALDI-TOF MS) 를이용한 제피티닙내성관련유전자의빈도분석과임상학적의의검증 ( 영문 ) A MALDI-TOF MS study of EGFR T790M mutation gene resistant to gefitinib in non-small cell lung cancer patients 구분 1 2 3 소속폐암연구과직위주임연구원 성명이영주전공의학 세부과제명 세부과제책임자 성명 소속 ( 직위 ) 전공 총연구기간 2012 년 1 원 ~ 2013 년 12 월 ( 총 2 년 ) 참여연구원수 ( 단위 : 명, MY) 4 명 연구기간및 연구비 ( 단위 : 천원 ) 구분연구기간계국립암센터 기업부담금소계현금현물 계 2012.01.01~2013.12.31 100 110 0 0 0 제 1 차 2012.01.01~2012.12.31 60 60 0 0 0 제 2 차 2013.01.01~2013.12.31 50 50 0 0 0 제 3 차 ~ 참여기업명칭전화 FAX 기관고유연구사업관리규칙에따라본연구개발사업을성실히수행하였으며아래와같이최종보고서 를제출합니다. 2013 년 10 월 31 일 과제책임자이영주 ( 서명 )
< 최종목표 > - EGFR tyrosine kinase 억제제인제피티닙에대한대표적인획득내성기전인 EGFR T790M 유전자돌연변이의약물노출이전존재유무와그빈도를고민감도염기서열분석방법중에하나인 MALDI-TOF MS를통하여알아보고이유전자돌연변이를갖는환자군의병태생리적특징을규명하고검증함으로써비소세포폐암환자의제피티닙치료효과를향상시킬수있는방안을제시한다. 연구목표 (200 자이내 ) 연구내용및방법 (500 자이내 ) < 당해연도목표 > 1. 후향적이행성연구를위한적절한비소세포폐암환자대상자선정 2. 환자종양조직을이용하여 EGFR T790M 유전자돌연변이에대한 MALDI-TOF MS 시행 3. 제피티닙치료효과및예후와 MALDI-TOF MS 결과분석 4. EGFR 돌연변이양성비소세포폐암세포주을이용하여제피티닙내성세포주확립 5. EGFR T790M 유전자돌연변이제피티닙내성세포주와민감성세포주의 cell mixture 실험을통하여 EGFR T790M 유전자돌연변이의영향을정량적으로측정 연구내용 - EGFR 돌연변이비소세포폐암환자는 EGFR tyrosine kinase 억제제인제피티닙에 70%~ 80% 의높은반응률을보이지만약 8~9개월이후에는획득내성을보이고있으며내성발생시점또한환자마다개인차이가있어서약 30% 는 3개월미만의짧은반응기간을보임. - EGFR 돌연변이폐암환자에서각기다른제피티닙효과를보이는원인에대하여 EGFR T790M 유전자돌연변이가제피티닙사용이전부터적은수로존재하고있다가제피티닙사용이후에선택적으로증폭되어내성을보이게할수있다는주장이있으나이를뒷받침할수있는검증된임상적데이터는없음. - 기존의 direct sequencing 방법을이용한돌연변이검사는종양조직내에소량으로존재할수있는 EGFR T790M 유전자를발견하기에는민감도가낮을수있어이를극복할수있는다른 sequencing 방법들이시도되고있지만아직까지일관성있는결과를보이지못함. - 따라서, 종양조직내에존재하는이질적인유전자돌연변이들의식별이쉽고적은 DNA 양으로도측정가능한 MALDI-TOF MS를이용하여 EGFR T790M 유전자돌연변이의약물노출이전존재유무와그빈도를알아보고자함. 연구방법 1. 후향적이행성연구를위한적절한비소세포폐암환자대상자선정
1) 2005년 1월이전진단 2) 조직학적혹은세포학적으로확인된비소세포폐암 4기 3) 선암종혹은비흡연자 4) 제피티닙혹은타세바를진행된병기에서사용 5) 치료전종양조직검체 ( 조직검사혹은수술적제거로채집, 파라핀블록으로보관 ) 6) Direct sequensing method으로 the kinase domain of EGFR gene에서 exons19, exon 21L858r mutation 존재 2. 환자조직검체를이용하여 EGFR 유전자에대하여염기서열분석시행 1) genotyping by MALDI-TOF MS for EGFR T790M 3. 제피티닙치료효과및예후와 MALDI-TOF MS 결과분석 1) Active mutation (EGFR 19del, EGFR L858R) 를가진환자군에서 resistant mutation (EGFR T790M) 을동시에가진환자들의빈도를측정 2) MALDI-TOF MS로확인된 T790M 양성군과음성군사이의 EGFR-TKI 반응률과반응기간비교 3) MALDI-TOF MS로확인된 T790M 양성군에서 1차치료제로 EGFR-TKI를사용한군과 2차이상의치료제로 EGFR-TKI를사용한군간의반응률과반응기간비교 4) T790M 양성종양의병리생태적특징규명 ( 흡연, 종양의병기, 크기혹은분화도등과의관련성 ) 5) 종양내의 EGFR T790M 분포양에따른 EGFR-TKI 반응기간에대한효과 4. EGFR 돌연변이양성비소세포폐암세포주을이용하여제피티닙내성세포주확립 5. Cell mixture 실험에서 MALDI-TOF MS 민감도확인및 EGFR T790M의영향정량분석 < 정량적성과 1) > 연구개발에따른기대성과 < 정성적성과 > 구분 달성치 / 목표치 1) 달성도 (%) SCI 논문편수 0/1 0% IF 합 0/4 0% 기타성과 - 임상적발견을실험실내에서재현하고그원리, 기전을밝혀내어다시임상모델 에적용하는모범적인성공사례제시 - 신진의과학자의연구역량고취 색인어 국문 영문 비소세포폐암 EGFR 유전자돌연변이 제피티닙 약물내성 non-small cell lung cancer EGFR gene mutation gefitinib drug resistance
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