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Pharmacological inhibition of serine synthesis enhances temozolomide efficacy by decreasing O6-methylguanine DNA methyltransferase (MGMT) expression and reactive oxygen species (ROS)-mediated DNA damage in glioblastoma

Abstract

Glioblastoma (GBM) is the most malignant primary tumor in the central nervous system of adults. Temozolomide (TMZ), an alkylating agent, is the first-line chemotherapeutic agent for GBM patients. However, its efficacy is often limited by innate or acquired chemoresistance. Cancer cells can rewire their metabolic programming to support rapid growth and sustain cell survival against chemotherapies. An example is the de novo serine synthesis pathway (SSP), one of the main branches from glycolysis that is highly activated in multiple cancers in promoting cancer progression and inducing chemotherapy resistance. However, the roles of SSP in TMZ therapy for GBM patients remain unexplored. In this study, we employed NCT503, a highly selective inhibitor of phosphoglycerate dehydrogenase (PHGDH, the first rate-limiting enzyme of SSP), to study whether inhibition of SSP may enhance TMZ efficacy in MGMT-positive GBMs. 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT), flowcytometry and colony formation assays demonstrated that NCT503 worked synergistically with TMZ in suppressing GBM cell growth and inducing apoptosis in T98G and U118 cells in vitro. U118 and patient-derived GBM subcutaneous xenograft models showed that combined NCT503 and TMZ treatment inhibited GBM growth and promoted apoptosis more significantly than would each treatment alone in vivo. Mechanistically, we found that NCT503 treatment decreased MGMT expression possibly by modulating the Wnt/β-catenin pathway. Moreover, intracellular levels of reactive oxygen species were elevated especially when NCT503 and TMZ treatments were combined, and the synergistic effects could be partially negated by NAC, a classic scavenger of reactive oxygen species. Taken together, these results suggest that NCT503 may be a promising agent for augmenting TMZ efficacy in the treatment of GBM, especially in TMZ-resistant GBMs with high expression of MGMT.

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Fig. 1: NCT503 augments TMZ-mediated chemotherapy in MGMT-positive GBM cells.
Fig. 2: NCT503 enhances TMZ efficacy in vivo.
Fig. 3: NCT503 inhibits MGMT expression in MGMT-positive GBM cells and xenografts.
Fig. 4: NCT503 inhibits Wnt/β-catenin pathway activity.
Fig. 5: NCT503 augments TMZ efficacy partially by ROS-mediated DNA damage in MGMT-positive GBM cells.

Data availability

The datasets used during the current study are available from the corresponding author on reasonable request.

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Acknowledgements

We thank Dr. Michael E. Pacold (Grossman School of Medicine, New York University) and Prof. David M. Sabatini (Massachusetts Institute of Technology) for their kind sharing of the detailed protocol for in vivo administration of NCT503.

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All authors made substantial contributions to this study. Lei Jin was responsible for conceptualization, study design, investigation, acquisition, analysis and interpretation of data and original manuscript drafting. Karrie Mei-Yee Kiang and Stephen Yin Cheng were responsible for conceptualization and review of the manuscript. Gilberto Ka-Kit Leung was responsible for supervision, conceptualization and revision of manuscript and gave final approval of submission and publication.

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Correspondence to Gilberto Ka-Kit Leung.

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The research committee of the University of Hong Kong has reviewed and approved the study according to the principles expressed in the Declaration of Helsinki.

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Jin, L., Kiang, K.MY., Cheng, S.Y. et al. Pharmacological inhibition of serine synthesis enhances temozolomide efficacy by decreasing O6-methylguanine DNA methyltransferase (MGMT) expression and reactive oxygen species (ROS)-mediated DNA damage in glioblastoma. Lab Invest 102, 194–203 (2022). https://doi.org/10.1038/s41374-021-00666-7

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