Biomanufacturing of glioblastoma organoids exhibiting hierarchical and spatially organized tumor microenvironment via transdifferentiation
Biomanufacturing of glioblastoma organoids exhibiting hierarchical and spatially organized tumor microenvironment via transdifferentiation
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DOI：
10.1002/bit.28191
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发表时间：
2022-07
影响因子：
3.8
通讯作者：
Seungjo Park;Alexandra D. Avera;Yonghyun Kim
中科院分区：
文献类型：
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作者：
Seungjo Park;Alexandra D. Avera;Yonghyun Kim
胶质母细胞瘤（GBM）是最具侵袭性的脑肿瘤类型，起源于胶质母细胞瘤干细胞（GSCs）。在大脑中，GSCs由位于血管周围生态位和缺氧生态位的肿瘤微环境（TME）支持。GBM TME具有高度异质性，并表现出复杂的细胞间相互作用。在干细胞富集培养基中培养的三维肿瘤球常被用作体外模型。GBM肿瘤球保留了一些转录和翻译的GSC特征，但往往不能再现肿瘤间的异质性。在这里，我们在小型生物反应器中利用患者来源的异种移植GBM细胞系开发了一种简单的、无基质的、体内样的GBM类器官（GBOs）。剪切应力优化后，可在4-5周内生产直径超过1mm的高重复性gbo。与传统的肿瘤球培养相比，GBOs表现出高干性和强细胞间相互作用。它们显示出CD133阳性细胞所在的缺氧诱导因子1α阳性核心的空间梯度和NOTCH及其配体的空间异质性表达。我们还观察到，通过GBM转分化为内皮细胞、周细胞和星形胶质细胞，形成了一个自我建立的、分层组织的、异质性的TME。总的来说，我们证明了生物制造均匀大小的gbo的能力，这些gbo概括了体内GBM TME的特征，可以作为改进的GBM体外模型。
Glioblastoma (GBM) is the most aggressive type of brain tumor that originates from glioblastoma stem cells (GSCs). In the brain, GSCs are supported by a tumor microenvironment (TME) residing in the perivascular niche and the hypoxic niche. The GBM TME is highly heterogenous and exhibits complex cell‐to‐cell interactions. Three‐dimensional tumorspheres cultured in stem cell‐enriching media is often used as an in vitro model. The GBM tumorspheres retain some of the transcriptional and translational GSC features but often fails to recapitulate intertumor heterogeneity. Here, we developed a simple, matrix‐free, and in vivo‐like GBM organoids (GBOs) using patient‐derived xenograft GBM lines in small‐scale bioreactors. Shear stress was optimized to produce highly reproducible GBOs over 1 mm diameter within 4–5 weeks. GBOs exhibited high stemness and strong cell‐to‐cell interactions compared to conventional tumorsphere cultures. They displayed spatial gradients of hypoxia‐inducible factor 1α positive hypoxic cores where CD133‐positive cells resided and spatially heterogeneous expression of NOTCH and its ligands. We also observed a self‐established, hierarchically organized, and heterogeneous TME by GBM transdifferentiation into endothelial cells, pericytes, and astrocytes. Collectively, we demonstrate the ability to biomanufacture uniformly sized GBOs that recapitulate in vivo GBM TME features that can serve as an improved GBM in vitro model.