脑器官/所有共培养模型揭示了与BCP的CNS参与至关重要的AP-1途径

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:7.10000

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

摘要

中枢神经系统（CNS）的参与仍然是治疗儿童B细胞前体急性淋巴细胞白血病（BCP-ALL）的临床障碍。 CNS白血病的疾病机制主要使用二维细胞培养和小鼠模型研究。鉴于人与鼠中枢神经系统之间的细胞身份和结构的变化，必须寻求互补模型来研究CNS白血病。在这里，我们提出了一种结合人脑器官和BCP-ALL细胞的首个三维共培养模型。我们注意到，BCP所有细胞系和患者衍生的异种移植（PDX）细胞的植入明显高于非所有细胞。为了验证器官共培养和体内鼠模型之间的可翻译性，我们确认针对CNS白血病相关的途径，例如CD79A/IGα或C-X-C基矩阵趋化因子受体4层细胞衍生的因子1降低了BCP-ALL细胞的入侵。与未感染的馏分相比，摄取器官的白血病细胞的RNA测序和功能验证表明，激活蛋白1（AP-1）转录因子复合构件在涉及器官的细胞中的显着上调。此外，我们检测到PDX中AP-1途径基因的显着富集所有从中枢神经系统中回收的细胞，与接受TCF3 :: PBX1+ PDX细胞进行移植的小鼠的脾脏爆炸相比，证实了AP-1信号在CNS疾病中的作用。因此，我们发现与CNS阴性病例相比，最初被诊断为CNS阳性BCP的患者的AP-1基因Jun Proto-Cogene的水平明显更高，而在100名患有BCP-ALL的患者中，与CNS阴性病例相比，与CNS-Relapse vs non-CNS-RELAPSE病例相比。我们的结果表明，CNS器官是一种新型模型，以研究中枢神经系统的参与并确定AP-1途径是BCP all的关键驱动力的关键驱动力。

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.