脑类器官/ALL共培养模型揭示AP-1通路与BCP-ALL中枢神经系统（CNS）累及的关键关联

A brain organoid/ALL coculture model reveals the AP-1 pathway as critically associated with CNS involvement of BCP-ALL

Blood Advances

影响因子:7.1

分区:医学1区 Top / 血液学2区

发表日期:2024 Oct 08

作者: Philip Gebing, Stefanos Loizou, Sebastian Hänsch, Julian Schliehe-Diecks, Lea Spory, Pawel Stachura, Vera H Jepsen, Melina Vogt, Aleksandra A Pandyra, Herui Wang, Zhengping Zhuang, Johannes Zimmermann, Martin Schrappe, Gunnar Cario, Ameera Alsadeq, Denis M Schewe, Arndt Borkhardt, Lennart Lenk, Ute Fischer, Sanil Bhatia

DOI: 10.1182/bloodadvances.2023011145

摘要

中枢神经系统（CNS）累及仍是儿童B细胞前体急性淋巴细胞白血病（BCP-ALL）治疗中的一大临床难题。CNS白血病的发病机制主要通过二维细胞培养和小鼠模型进行研究。鉴于人类与鼠类CNS在细胞身份和结构上的差异，亟需寻找补充模型以研究CNS白血病。在此，我们首次提出结合人脑类器官与BCP-ALL细胞的三维共培养模型。我们观察到BCP-ALL细胞系和患者来源的异种移植（PDX）细胞在脑类器官中的移植效率显著高于非ALL细胞。为了验证类器官共培养模型与体内小鼠模型的可转化性，我们确认靶向CNS白血病相关通路（如CD79a/Igα或C-X-C基序趋化因子受体4-基质细胞源因子1）能减少BCP-ALL细胞侵入类器官的能力。RNA测序和功能验证显示，类器官侵入细胞中，激活蛋白1（AP-1）转录因子复合物的成员显著上调。进一步分析发现，从中枢神经系统（CNS）中回收的PDX ALL细胞中，AP-1通路基因显著富集，相比之下，输注到小鼠的脾脏血细胞中未观察到此富集，验证了AP-1信号在CNS疾病中的作用。在一项涵盖100例BCP-ALL患者的队列中，初诊为CNS阳性患者中AP-1基因jun原癌基因表达明显高于CNS阴性病例，以及CNS复发与非CNS复发病例中也表现出更高的水平。我们的研究表明，脑类器官是一种研究CNS累及的新模型，且AP-1通路是BCP-ALL CNS疾病的重要驱动因子。依据知识共享署名-非商业性使用-禁止演绎 4.0国际许可协议（CC BY-NC-ND 4.0），仅允许非商业和非衍生性使用，需注明出处。

Abstract

Central nervous system (CNS) involvement remains a clinical hurdle in treating childhood B-cell precursor acute lymphoblastic leukemia (BCP-ALL). The disease mechanisms of CNS leukemia are primarily investigated using 2-dimensional cell culture and mouse models. Given the variations in cellular identity and architecture between the human and murine CNS, it becomes imperative to seek complementary models to study CNS leukemia. Here, we present a first-of-its-kind 3-dimensional coculture model combining human brain organoids and BCP-ALL cells. We noticed significantly higher engraftment of BCP-ALL cell lines and patient-derived xenograft (PDX) cells in cerebral organoids than non-ALL cells. To validate translatability between organoid coculture and in vivo murine models, we confirmed that targeting CNS leukemia-relevant pathways such as CD79a/Igα or C-X-C motif chemokine receptor 4-stromal cell-derived factor 1 reduced the invasion of BCP-ALL cells into organoids. RNA sequencing and functional validations of organoid-invading leukemia cells compared with the noninvaded fraction revealed significant upregulation of activator protein 1 (AP-1) transcription factor-complex members in organoid-invading cells. Moreover, we detected a significant enrichment of AP-1 pathway genes in PDX ALL cells recovered from the CNS compared with spleen blasts of mice that had received transplantation with TCF3::PBX1+ PDX cells, substantiating the role of AP-1 signaling in CNS disease. Accordingly, we found significantly higher levels of the AP-1 gene, jun proto-oncogene, in patients initially diagnosed as CNS-positive BCP-ALL compared with CNS-negative cases as well as CNS-relapse vs non-CNS-relapse cases in a cohort of 100 patients with BCP-ALL. Our results suggest CNS organoids as a novel model to investigate CNS involvement and identify the AP-1 pathway as a critical driver of CNS disease in BCP-ALL.Licensed under Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International (CC BY-NC-ND 4.0), permitting only noncommercial, nonderivative use with attribution.