Dube Collin J, Zhang Ying, Saha Shekhar, Lai Michelle, Gibert Myron K, Escalante Miguel, Hudson Kadie, Wong Doris, Marcinkiewicz Pawel, Yener Ulas, Sun Yunan, Xu Esther, Sorot Aditya, Mulcahy Elizabeth, Kefas Benjamin, Hanif Farina, Guessous Fadilla, Vernon Ashley, Patel Manoj K, Schiff David, Zong Hui, Purow Benjamin, Holland Eric, Sonkusare Swapnil, Sontheimer Harald, Abounader Roger
Department of Microbiology, Immunology & Cancer Biology, University of Virginia, Charlottesville, VA 22908, USA.
Department of Neuroscience, University of Virginia, Charlottesville, VA 22908, USA.
bioRxiv. 2024 Aug 23:2024.08.22.609229. doi: 10.1101/2024.08.22.609229.
Glioblastoma (GBM) is the most common primary malignant brain tumor. The aim of this study was to elucidate the role of microenvironment and intrinsic T-type calcium channels (Cav3) in regulating tumor growth and progression.
We grafted syngeneic GBM cells into Cav3.2 knockout mice to assess the role of microenvironment T-Type calcium channels on GBM tumor growth. We performed single-cell RNA-seq (scRNA-seq) of tumors from WT and Cav3.2 KO mice to elucidate the regulation of tumors by the microenvironment. We used neurons from WT and Cav3.2 KO mice in co-culture with GBM stem cells (GSC) to assess the effects of Cav3.2 on neuron/GSC synaptic connections and tumor cell growth.
Cav3.2 KO in the microenvironment led to significant reduction of GBM growth and prolongation of animal survival. scRNA-seq showed that microenvironment Cav3.2 regulates neuronal and glial biological processes. Microenvironment Cav3.2 downregulated numerous genes associated with regulating the OPC cell state in GBM tumors such as SOX10 and Olig2. Neuronal Cav3.2 promoted neuron/GSC synaptic connections and GSC growth. Treatment of GSCs with the Cav3 blocker mibefradil downregulated genes associated with neuronal processes. The Cav3 blocker drug mibefradil synergized with temozolomide (TMZ) and radiation to reduce in vivo tumor growth and prolong animal survival.
Together these data reveal a role for microenvironment Cav3 in promoting GBM tumor progression through regulating neuronal and glial processes particularly associated with the OPC-cell state. Targeting both intrinsic and microenvironment Cav3 with the inhibitor mibefradil significantly enhanced the anti-GBM effects of TMZ and radiation.
胶质母细胞瘤（GBM）是最常见的原发性恶性脑肿瘤。本研究的目的是阐明微环境和内在T型钙通道（Cav3）在调节肿瘤生长和进展中的作用。
我们将同基因GBM细胞移植到Cav3.2基因敲除小鼠体内，以评估微环境T型钙通道对GBM肿瘤生长的作用。我们对野生型（WT）和Cav3.2基因敲除小鼠的肿瘤进行单细胞RNA测序（scRNA-seq），以阐明微环境对肿瘤的调节作用。我们将野生型和Cav3.2基因敲除小鼠的神经元与GBM干细胞（GSC）共培养，以评估Cav3.2对神经元/GSC突触连接和肿瘤细胞生长的影响。
微环境中的Cav3.2基因敲除导致GBM生长显著减少，动物存活时间延长。scRNA-seq显示微环境Cav3.2调节神经元和胶质细胞的生物学过程。微环境Cav3.2下调了GBM肿瘤中许多与调节少突胶质前体细胞（OPC）状态相关的基因，如SOX10和Olig2。神经元Cav3.2促进神经元/GSC突触连接和GSC生长。用Cav3阻滞剂米贝拉地尔处理GSC可下调与神经元过程相关的基因。Cav3阻滞剂药物米贝拉地尔与替莫唑胺（TMZ）和放疗协同作用，可减少体内肿瘤生长，延长动物存活时间。
这些数据共同揭示了微环境Cav3通过调节神经元和胶质细胞过程，特别是与OPC细胞状态相关的过程，在促进GBM肿瘤进展中的作用。用抑制剂米贝拉地尔靶向内在和微环境Cav3可显著增强TMZ和放疗的抗GBM效果。
