Establishments and Characterization of Colonic Organoids for Application in Intra-Tumor Heterogeneity and Microbiome Studies = 대장 오가노이드 수립과 이를 활용한 종양내이질성(intra-tumor heterogeneity) 및 마이크로바이옴 연구

국문 초록 (Abstract)

오가노이드는 지난 10여 년간 전통적인 2차원 세포 배양의 한계를 극복하며, 생체 내 환경을 보다 정밀하게 모사할 수 있는 생리학적으로 유의미한 모델로 주목받아 왔다. 성체줄기세포로부터 유래된 정상 및 암 오가노이드를 기반으로, 단일 대장암 환자 내에서의 종양 내 이질성(intratumoral heterogeneity)을 탐구하였다.
환자에게 가장 적합한 항암제를 식별하기 위해, 고속 이미지 기반 고처리량 스크리닝(HTS, High-Throughput Screening) 기술을 활용하여 약물 반응성을 실시간으로 분석하였다. 이를 통해 단일 세포 유래 클론 간의 약물 감수성에서 나타나는 클론 수준의 이질성을 포착했을 뿐만 아니라, 최종 생존율이 유사하더라도 빠르고 완전한 세포사를 유도하는 화합물들을 효과적으로 식별할 수 있었다.
더불어, Bifidobacterium adolescentis 및 Bifidobacterium longum의 배양 상등액을 대장암 오가노이드에 처리함으로써, 장내 유익균이 가지는 잠재적인 항암 효과를 제시하였다.

다국어 초록 (Multilingual Abstract)

Organoids have emerged over the past decade as physiologically relevant models that overcome the limitations of traditional two-dimensional cell cultures, offering enhanced resemblance to in vivo systems. Successful differentiation of organoids has been achieved from various tissues using a range of methodologies. In this study, we established normal and tumor-derived colon organoids developed from adult stem cells. We investigated intratumoral heterogeneity within a single colorectal cancer patient and tumor-suppresive effects of commensal bacterium: Bifidobacterium adolescentis and bifidobacterium longum.
To identify the most suitable chemotherapeutic agents for the patient, we performed high-throughput screening (HTS)-based real-time image analysis to assess drug responsiveness. This approach enabled us to capture not only clonal heterogeneity in drug sensitivity among single-cell-derived clones but also to identify compounds that induced rapid and complete cell death, even when endpoint viability was similar across treatments.
Furthermore, we demonstrated the potential anticancer effects of beneficial gut microbes by treating colorectal cancer organoids with culture supernatants from Bifidobacterium adolescentis and Bifidobacterium longum, which showed efficacy via HTS-based analysis.

목차 (Table of Contents)

• Abstract i
• Contents iii-iv
• List of tables and figures v-x
• List of abbreviations xi-xii
• General Introduction 1
• Chapter 1 4
• Monoclonal Colon Cancer Organoids from Four Lesions in a Single Patient Reveal Tumor Heterogeneity and Differential Drug Responses
• Introduction 5
• Material and Methods 6
• Results 16
• Discussion 24
• Chapter 2 71
• Microbiome-Modulated Suppression of Colorectal Cancer: Comparative Efficacy of Beneficial Bacterial Metabolites
• Introduction 72
• Material and Methods 74
• Results 81
• Discussion 85
• General discussion 99
• Conclusion 101
• References 102
• Abstract in Korean 113