[논문]Development of a 3D Tissue Culture–Based High-Content Screening Platform That Uses Phenotypic Profiling to Discriminate Selective Inhibitors of Receptor Tyrosine Kinases

Booij, Tijmen H.; Klop, Maarten J. D.; Yan, Kuan; Szá; ntai-Kis, Csaba; Szokol, Balint; Orfi, Laszlo; van de Water, Bob; Keri, Gyorgy; Price, Leo S.

doi:10.1177/1087057116657269

Development of a 3D Tissue Culture–Based High-Content Screening Platform That Uses Phenotypic Profiling to Discriminate Selective Inhibitors of Receptor Tyrosine Kinases 원문보기

Journal of biomolecular screening : the official journal of the Society for Biomolecular Screening, v.21 no.9, 2016년, pp.912 - 922

Booij, Tijmen H. (Division of Toxicology, Leiden Academic Centre for Drug Research, Leiden University, Leiden, The Netherlands) , Klop, Maarten J. D. (OcellO B.V., Leiden, The Netherlands) , Yan, Kuan (OcellO B.V., Leiden, The Netherlands) , Szántai-Kis, Csaba (Vichem Chemie Research Ltd., Budapest, Hungary) , Szokol, Balint (Vichem Chemie Research Ltd., Budapest, Hungary) , Orfi, Laszlo (Vichem Chemie Research Ltd., Budapest, Hungary) , van de Water, Bob (Division of Toxicology, Leiden Academic Centre for Drug Research, Leiden University, Leiden, The Netherlands) , Keri, Gyorgy (Vichem Chemie Research Ltd., Budapest, Hungary) , Price, Leo S. (Division of Toxicology, Leiden Academic Centre for Drug Research, Leiden University, Leiden, The Netherlands)

초록
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초록

3D 조직 배양은 2D 세포 배양에 비해 화합물의 스크리닝을 위한 더 많은 생리학적 관련성을 지닌 컨텍스트를 제공한다. 또한 3D 히드로겔에서 배양된 세포는 표현형 프로파일의 범위를 증가시키는 복잡한 표현형을 나타낸다.

여기서 우리는 표준 384정 분석 플레이트의 3D에서 배양된 침습성 전립선 암세포를 사용하여 잠재적 치료 소분자 및 항체 바이오로직스의 활동을 연구하는 하이콘텐츠 스크리닝 플랫폼을 설명한다. 이미지 분석 도구는 3D 화상 데이터를 처리하여 800개 이상의 표현형 파라미터를 측정하기 위해 개발되었다. 조직 형태학에서 화합물의 효과를 평가하기 위해 다중 파라미터 분석이 사용되었다.

우리는 3D 배양 전립선 암세포에서 c-Met와 EGF 수용체(EGFR) 티로신 인산화효소의 억제제 활동 및 선택성을 측정하기 위해 이 스크리닝 플랫폼을 적용했다. c-Met와 EGFR 활동은 각각의 리간드 HGF와 EGF에 의해 유도된 표현형 프로필을 기반으로 정량화되었다. 스크리닝 방법은 80개의 c-Met와 EGFR 추정 억제제의 신규 컬렉션에 적용되었다. 화합물은 c-Met, EGFR의 선택적 억제 또는 모든 대상의 이중특이성 억제를 나타내는 표현형 프로필을 유도하는 것으로 확인되었다.

결론적으로, 우리는 표현형 프로파일링을 사용하여 생리학적으로 관련된 3D 세포 배양 세팅에서 선택적 및 비선택적(오프타깃) 억제제를 구별하는 완벽하게 확장 가능한 하이콘텐츠 스크리닝 플랫폼을 설명한다.

초록

Abstract ▼ AI-Helper

3D tissue cultures provide a more physiologically relevant context for the screening of compounds, compared with 2D cell cultures. Cells cultured in 3D hydrogels also show complex phenotypes, increasing the scope for phenotypic profiling. Here we describe a high-content screening platform that uses invasive human prostate cancer cells cultured in 3D in standard 384-well assay plates to study the activity of potential therapeutic small molecules and antibody biologics. Image analysis tools were developed to process 3D image data to measure over 800 phenotypic parameters. Multiparametric analysis was used to evaluate the effect of compounds on tissue morphology. We applied this screening platform to measure the activity and selectivity of inhibitors of the c-Met and epidermal growth factor (EGF) receptor (EGFR) tyrosine kinases in 3D cultured prostate carcinoma cells. c-Met and EGFR activity was quantified based on the phenotypic profiles induced by their respective ligands, hepatocyte growth factor and EGF. The screening method was applied to a novel collection of 80 putative inhibitors of c-Met and EGFR. Compounds were identified that induced phenotypic profiles indicative of selective inhibition of c-Met, EGFR, or bispecific inhibition of both targets. In conclusion, we describe a fully scalable high-content screening platform that uses phenotypic profiling to discriminate selective and nonselective (off-target) inhibitors in a physiologically relevant 3D cell culture setting.

주제어

참고문헌 (30)

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표제어: PCR

동의어: Packet Collision Rate

용어 설명 출처 목록 (6)

용어 설명: PCR은 세균 특이성이 있는 primer를 이용하여 적은 수의 세균이 있을지라도 쉽게 검출할 수 있는 유용한 방법이며, 이를 이용하여 구강 내 치면세균막이나 타액에서 직접 세균을 검출할 수 있게 되었다[8].

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