Glutamine deficiency promotes stemness and chemoresistance in tumor cells through DRP1-induced mitochondrial fragmentation

Abstract

Glutamine is essential for maintaining the TCA cycle in cancer cells yet they undergo glutamine starvation in the core of tumors. Cancer stem cells (CSCs), responsible for tumor recurrence are often found in the nutrient limiting cores. Our study uncovers the molecular basis and cellular links between glutamine deprivation and stemness in the cancer cells. We showed that glutamine is dispensable for the survival of ovarian and colon cancer cells while it is required for their proliferation. Glutamine starvation leads to the metabolic reprogramming in tumor cells with enhanced glycolysis and unaltered oxidative phosphorylation. Production of reactive oxygen species (ROS) in glutamine limiting condition induces MAPK-ERK1/2 signaling pathway to phosphorylate dynamin-related protein-1(DRP1) at Ser616. Moreover, p-DRP1 promotes mitochondrial fragmentation and enhances numbers of CD44 and CD117/CD45 positive CSCs. Besides the established features of cancer stem cells, glutamine deprivation induces perinuclear localization of fragmented mitochondria and reduction in proliferation rate which are usually observed in CSCs. Treatment with glutaminase inhibitor (L-DON) mimics the effects of glutamine starvation without altering cell survival in in vitro as well as in in vivo model. Interestingly, the combinatorial treatment of L-DON with DRP1 inhibitor (MDiVi-1) reduces the stem cell population in tumor tissue in mouse model. Collectively our data suggest that glutamine deficiency in the core of tumors can increase the cancer stem cell population and the combination therapy with MDiVi-1 and L-DON is a useful approach to reduce CSCs population in tumor.

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Availability of data

Data are available on request from the corresponding author.

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Acknowledgements

We thankfully acknowledge Mr. Sounak Bhattacharya (confocal microscopy), Mr. Tanmoy Dalui, & Mrs. Debalina Chakraborty (flow cytometry), Dr. E. Padmanaban (NMR spectroscopy), and Mr. T. Muruganandan (AFM) of Central Instrumentation Facility of IICB. Prof. Pijush K. Das (CSIR-IICB) and Dr. Partha Chakrabarti (CSIR-IICB) are gratefully acknowledged for their valuable suggestions in preparing the manuscript. We are also thankful to Prof. Susanta Roychoudhury and Dr. Damayanti Das Ghosh (both from Saroj Gupta Cancer Centre & Research Institute, Kolkata, India) for providing human patient tissue samples and also for their valuable suggestions. Technical assistance of Mr. Prabir Kumar Dey is acknowledged. Other members of SSR laboratory are acknowledged for their co-operation.

Funding

This work was supported by grants from Science and Engineering Research Board (SERB) project GAP-360 (EMR/2016/002578) and the Council of Scientific and Industrial Research (CSIR) in house projects.

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Contributions

PP, SG and SSR conceptualized the study; PP and SG investigated, performed analysis and validation of the data and wrote the original draft. SSR provided the financial support, supervised the work, reviewed, and approved the final manuscript.

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Correspondence to Sib Sankar Roy.

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The authors declare that they have no conflict of interest.

Ethical approval for animal studies

All animal experiments were approved by the institutional animal ethics committee (IAEC), CSIR-Indian Institute of Chemical Biology, India, (Registration no. 147/GO/ReBi/S/99/CPSCEA) following the guidelines of Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA), Govt. of India.

Ethical approval for human studies

The human patients' tissue samples were collected from Saroj Gupta Cancer Centre and Research Institute (SGCCRI), Kolkata, India with proper human ethics clearance from the Institutional Ethics Committee (approval number -IEC SGCCRI REF NO.- 16/2/2018/Non-Reg/SSR/3).

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Prasad, P., Ghosh, S. & Roy, S.S. Glutamine deficiency promotes stemness and chemoresistance in tumor cells through DRP1-induced mitochondrial fragmentation. Cell. Mol. Life Sci. (2021). https://doi.org/10.1007/s00018-021-03818-6

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Keywords

  • Glutaminase
  • Glutamine metabolism
  • ROS
  • Tumor growth
  • Mitochondrial fission