In the absence of apoptosis, myeloid cells arrest when deprived of growth factor, but remain viable by consuming extracellular glucose

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Abstract

Withdrawal of the growth factor interleukin-3 (IL-3) from IL-3-dependent myeloid cells causes them to undergo Bax/Bak1-dependent apoptosis, whereas factor-deprived Bax−/−Bak1−/− cells remain viable, but arrest and shrink. It was reported that withdrawal of IL-3 from Bax−/−Bak1−/− cells caused decreased expression of the glucose transporter Glut1, leading to reduced glucose uptake, so that arrested cells required Atg5-dependent autophagy for long-term survival. In other cell types, a decrease in Glut1 is mediated by the thioredoxin-interacting protein (Txnip), which is induced in IL-3-dependent myeloid cells when growth factor is removed. We mutated Atg5 and Txnip by CRISPR/Cas9 and found that Atg5-dependent autophagy was not necessary for the long-term viability of cycling or arrested Bax−/−Bak1−/− cells, and that Txnip was not required for the decrease in Glut1 expression in response to IL-3 withdrawal. Surprisingly, Atg5-deficient Bax/Bak1 double mutant cells survived for several weeks in medium supplemented with 10% fetal bovine serum (FBS), without high concentrations of added glucose or glutamine. When serum was withdrawn, the provision of an equivalent amount of glucose present in 10% FBS (~0.5 mM) was sufficient to support cell survival for more than a week, in the presence or absence of IL-3. Thus, Bax−/−Bak1−/− myeloid cells deprived of growth factor consume extracellular glucose to maintain long-term viability, without a requirement for Atg5-dependent autophagy.

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Acknowledgements

We thank Gabriela Brumatti, Rebecca Feltham, James Vince, Simon Monard, David Huang, and Andre Samson for discussion, technical advice, and reagents. Funding for this project was provided by the Australian National Health and Medical Research Council (NHMRC) Program Grant #1113133 and NHMRC fellowship 1020136 to DLV; and the Leukemia and Lymphoma Society SCOR grant #7001-13. Work in the authors’ laboratory is made possible by operational infrastructure grants through the Australian Government Independent Research Institutes Infrastructure Support (IRISS) and the Victorian State Government OIS.

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Correspondence to Hoanh Tran.

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