Comprehensive proteomic analysis of exosomes and biomarker development with patient derived organoids from ESCC patients

2022 Fiscal Year Final Research Report

• Project/Area Number: 18K16307
• Research Category: Grant-in-Aid for Early-Career Scientists
• Allocation Type: Multi-year Fund
• Review Section: Basic Section 55020:Digestive surgery-related
• Research Institution: Osaka University
• Principal Investigator: HARA TAKEO 大阪大学, 大学院医学系研究科, 招へい教員 (00781775)
• Project Period (FY): 2021-11-01 – 2023-03-31
• Keywords: Organoid / Exosome / Biomarker / ESCC

Outline of Final Research Achievements

We collected the supernatant from patient derived organoid, and attempted to isolate exosome, but we could not get enough amount of exosome. So, we tried to improve the organoid formation rate with adjusting the component of organoid media. We investigated which component is essential for esophageal organoid, and decided to remove some components, including p38 MAPK inhibitor from organoid media. After improvement of organoid media, we found the organoid formation rate was improved more than 70%. Organoids were harvested and assessed the similarities with original cancer by IHC, PCR-RT, Western blotting, and we confirmed the organoids heritage the characteristics of original cancer. Next, we assessed the proliferation of organoid after seeding, and confirmed the peak of proliferation is Day 7 and the best term to collect the supernatant is Day7-11.

Free Research Field

Cancer biology

Academic Significance and Societal Importance of the Research Achievements

オルガノイド培養では患者由来腫瘍細胞をディシュ上で培養し、直接アッセイに利用可能であり、将来的に個別化医療への応用も期待される。様々な癌腫でオルガノイド培養の報告があるが、食道扁平上皮癌(ESCC)に対する報告はほとんどなく、培養効率も不十分でさらなる改良が望まれる。
今回我々はオルガノイド培養からESCC由来のExosomeの分離を試みたが、培養効率が不十分で分離困難であった。しかしオルガノイド培地内の成長因子の調整で培養効率の改善が可能となった。本研究で改良されたオルガノイド培養は、Exosomeの分離のみならず、感受性試験やオルガノイドバンクといった様々な研究に応用可能と考えられる。