Silencing BMAL1 promotes M1/M2 polarization through the LDHA/lactate axis to promote GBM sensitivity to bevacizumab

Fan Wang; Wenjun Liao; Caiyan Li; Ling Zhu

doi:10.1016/j.intimp.2024.112187

Silencing BMAL1 promotes M1/M2 polarization through the LDHA/lactate axis to promote GBM sensitivity to bevacizumab

Int Immunopharmacol. 2024 May 10:134:112187. doi: 10.1016/j.intimp.2024.112187. Online ahead of print.

Authors

Fan Wang¹, Wenjun Liao¹, Caiyan Li¹, Ling Zhu²

Affiliations

¹ Department of Neurosurgery, Jingmen Central Hospital, No. 168 Xiangshan Avenue, Jingmen, 448000, Hubei province, China.
² Department of Neurosurgery, Jingmen Central Hospital, No. 168 Xiangshan Avenue, Jingmen, 448000, Hubei province, China. Electronic address: LingzhuVIP@yeah.net.

PMID: 38733825
DOI: 10.1016/j.intimp.2024.112187

Abstract

Objective: Glioblastoma (GBM) has poor clinical prognosis due to limited treatment options. In addition, the current treatment regimens for GBM may only slightly prolong patient survival. The aim of this study was to assess the role of BMAL1 in the immune microenvironment and drug resistance of GBM.

Methods: GBM cell lines with stable BMAL1 knockdown or LDHA overexpression were constructed, and functionally characterized by the CCK8, EdU incorporation, and transwell assays. In vivo GBM model was established in C57BL/6J mice. Flow cytometry, ELISA, immunofluorescence, and RT-qPCR were performed to detect macrophage polarization. Lactate production, pathological changes, and the expression of glycolytic proteins were analyzed by HE staining, immunohistochemistry, biochemical assays, and Western blotting.

Results: BMAL1 silencing inhibited the malignant characteristics, lactate production, and expression of glycolytic proteins in GBM cells, and these changes were abrogated by overexpression of LDHA or exogenous lactate supplementation. Furthermore, BMAL1 knockdown induced M1 polarization of macrophages, and inhibited M2 polarization and angiogenesis in GBM cells in conditioned media. Overexpression of LDHA or presence of exogenous lactate inhibited BMAL1-induced M1 polarization and angiogenesis. Finally, BMAL1 silencing and bevacizumab synergistically inhibited glycolysis, angiogenesis and M2 polarization, and promoted M1 polarization in vivo, thereby suppressing GBM growth.

Conclusion: BMAL1 silencing can sensitize GBM cells to bevacizumab by promoting M1/M2 polarization through the LDHA/lactate axis.

Keywords: BMAL1; Bevacizumab; Glioblastoma; LDHA/lactate; Macrophage polarization.