Brain tumoroids: Treatment prediction and drug development for brain tumors with fast, reproducible, and easy-to-use personalized models

Aurélie Soubéran, Carine Jiguet-Jiglaire, SolineToutain, Philippe Morando, Nathalie Baeza-Kallee , Romain Appay, Céline Boucard, Thomas Graillon, Mikael Meyer, Kaissar Farah, Dominique Figarella-Branger, EmelineTabouret#, , and AurélieTchoghandjian#

Neuro-oncology 27(2), 415–429, 2025

https://doi.org/10.1093/neuonc/noae184

Abstract (원문 요약)

Background: The generation of patient avatars is critically needed in neuro-oncology for treatment prediction and preclinical therapeutic development. Our objective was to develop a fast, reproducible, low-cost, and easy-to-use method of tumoroids generation and analysis, efficient for all types of brain tumors, primary and metastatic.

Methods: Tumoroids were generated from 89 patients: 81 primary tumors including 77 gliomas, and 8 brain metastases. Tumoroids morphology and cellular and molecular characteristics were compared with the ones of the parental tumor by using histology, methylome profiling, pTERT mutations, and multiplexed spatial immunofluorescences. Their cellular stability over time was validated by flow cytometry. Therapeutic sensitivity was evaluated and predictive factors of tumoroid generation were analyzed.

Results:  All the tumoroids analyzed had similar histological (n = 21) and molecular features (n = 7) to the parental tumor. The median generation time was 5 days. The success rate was 65%: it was higher for high-grade gliomas and brain metastases versus IDH mutated low-grade gliomas. For high-grade gliomas, neither other clinical, neuroimaging, histological nor molecular factors were predictive of tumoroid generation success. The cellular organization inside tumoroids analyzed by MACSima revealed territories dedicated to specific cell subtypes. Finally, we showed the correlation between tumoroid and patient treatment responses to radio-chemotherapy and their ability to respond to immunotherapy thanks to a dedicated and reproducible 3D analysis workflow.

Conclusions:  Patient-derived tumoroid model that we developed offers a robust, user-friendly, low-cost, and reproducible preclinical model valuable for therapeutic development of all types of primary or metastatic brain tumors, allowing their integration into forthcoming early-phase clinical trials.

국문 초록 요약

배경: 신경종양학에서 치료 반응 예측과 전임상 치료제 개발을 위해 환자 아바타(patient avatar)의 구축은 필수적이다. 본 연구의 목적은 원발 및 전이성 뇌종양을 포함한 모든 유형의 뇌종양에서 적용 가능한, 빠르고 재현 가능하며 저비용·간편한 tumoroid 생성 및 분석 방법을 개발하는 것이었다.

방법: 총 89명의 환자로부터 tumoroid를 제작하였다(원발성 81명, 이 중 교종 77명, 뇌 전이 8명). 제작된 tumoroid의 형태, 세포학적·분자학적 특징을 모종양과 비교하였다(조직학, 메틸화 프로파일링, pTERT 변이, 다중공간 면역형광 분석 활용). 시간 경과에 따른 세포 안정성은 유세포분석(flow cytometry)으로 검증하였다. 또한 약물 민감도를 평가하고 tumoroid 생성 성공을 예측하는 인자를 분석하였다.

결과: 분석된 tumoroid는 모두 원발 종양과 유사한 조직학적(n=21) 및 분자학적(n=7) 특징을 보였다. 평균 생성 시간은 5일이었으며, 성공률은 65%였다. 고등급 교종과 뇌 전이종양에서 성공률이 높았으며, IDH 변이 저등급 교종에서는 낮았다. 고등급 교종의 경우, 다른 임상·영상·조직·분자 인자는 생성 성공률과 관련이 없었다. MACSima 분석을 통해 tumoroid 내부의 세포 아형별 공간적 구획이 확인되었다. 또한, tumoroid의 방사선-화학요법 반응과 실제 환자의 치료 반응 간 상관관계, 면역치료 반응 가능성을 전용 3D 분석 워크플로우를 통해 입증하였다.

결론: 본 연구에서 개발된 환자 유래 tumoroid 모델은 원발 및 전이성 뇌종양 모두에서 적용 가능한, 견고하고 사용자 친화적이며 저비용·재현 가능한 전임상 모델로서, 향후 초기 임상시험(early-phase clinical trials)에 통합되어 치료제 개발에 기여할 수 있다.

 1. 연구 배경

• 신경종양학에서는 환자 아바타(patient avatar) 모델이 필요함 → 치료 반응 예측, 신약 개발 가속화를 위함.

• 기존 세포주/마우스 모델은 종양 미세환경과 환자 특이적 이질성을 충분히 반영하지 못함.

• 이를 보완하기 위해 tumoroid(환자 종양 유래 3D 모델) 개발이 필수적.

2. 연구 방법

• 환자 수집: 89명 환자의 종양 샘플 (81개 원발성, 8개 뇌 전이종양).

• Tumoroid 생성: 수술 후 24시간 이내 조직을 소량 절편으로 분리, 흔들어주는 조건(shaking)에서 배양.

• 분석:

  • 형태학적/조직학적 비교 (H&E, IHC).

  • 분자 수준 비교 (메틸화 패턴, pTERT 변이, CNV 분석).

  • 다중공간 면역형광 (MICS, MACSima).

  • 흐름세포분석으로 시간에 따른 면역세포, stem-like cell 변화 평가.

  • 약물 반응 (TMZ, 방사선, SMAC mimetic, IL2, BLZ945).

3. 주요 결과

-Tumoroid 성공률

o    전체 성공률: 65%

o    고등급 교모세포종(GBM), 뇌 전이종양 → 성공률 높음

o    IDH 변이 저등급 교종 → 성공률 낮음

o    예측인자: 종양 등급 & IDH 변이 여부

o    EGFR 증폭은 tumoroid 성공과 연관성 시사.

-재현성 및 특성 유지

o    원발 종양과 조직학적, 분자적 특징이 매우 유사.

o    CNV, MGMT 메틸화, pTERT 변이 등 원래 종양과 일치.

o    면역세포, 신경세포, 성상세포 등 이질적 세포 군집이 tumoroid 내에서 구획화됨.

-치료 반응 반영

o    환자 임상 반응과 tumoroid 반응이 일치

§  예: STUPP protocol (TMZ + RT)에 반응한 환자의 tumoroid도 반응.

§  비반응 환자 tumoroid는 약물 반응 없음.

o    신규 치료 실험 가능성

§  SMAC mimetic + TMZ 병용 → 세포사멸 증가, 증식 억제.

§  IL-2, BLZ945 등 면역치료 후보도 평가 가능.

-실용성

o    Tumoroid 생성까지 평균 5일

o    2주 이내에 치료 반응 평가 가능 → 조기 임상시험에 통합 가능성.

4. 결론

• 본 연구는 빠르고, 저비용, 재현 가능한 tumoroid 제작법을 제시.

• Tumoroid는 원래 종양의 분자·세포 이질성, 치료 반응을 잘 반영.

• 맞춤형 치료(prediction) 및 신약 개발(preclinical test)에 활용 가능.

• 향후 초기 임상시험(early-phase clinical trials)에 통합될 수 있는 잠재력을 입증.