CRISPR-Cas9 screen identifies oxidative phosphorylation as essential for cancer cell survival at low extracellular pH

Michl, Johanna; Wang, Yunyi; Monterisi, Stefania; Błaszczak, Wiktoria; Beveridge, Ryan; Bridges, Esther; Koth, Jana; Bodmer, Walter F.; Swietach, Pawel

doi:10.1016/j.celrep.2022.110493

Cited by 25 publications

(43 citation statements)

References 34 publications

(46 reference statements)

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“…A large part of the confusion on mitochondrial dysfunction in cancer comes from the incorrect assumption that oxygen consumption observed in cancer cells is linked to ATP synthesis through OxPhos ( 14 , 28 , 29 , 40 , 50 – 53 ). Many cancers, including GBM, can survive in hypoxia (0.1% oxygen) or in a solution of potassium cyanide, a Complex IV inhibitor, findings that would exclude normal OxPhos as a source of ATP synthesis ( 54 – 57 ).…”

Section: Fermentation Metabolism Is Responsible For Gbm Growthmentioning

confidence: 99%

“…In light of these structural and functional abnormalities, it would not be possible for GBM mitochondria to synthesize much if any ATP through OxPhos based on the foundational principle in evolutionary biology that structure determines function (14,26,27). The numerous reports suggesting that OxPhos is either normal or not seriously impaired in GBM cells is inconsistent with this foundational principle (28)(29)(30)(31)(32)(33)(34)(35)(36)(37)(38)(39)(40). It is important to recognize that oxygen consumption is not a reliable marker for OxPhos function in cancer cells (see below).…”

Section: Introductionmentioning

confidence: 99%

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Metabolic management of microenvironment acidity in glioblastoma

et al. 2022

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Glioblastoma (GBM), similar to most cancers, is dependent on fermentation metabolism for the synthesis of biomass and energy (ATP) regardless of the cellular or genetic heterogeneity seen within the tumor. The transition from respiration to fermentation arises from the documented defects in the number, the structure, and the function of mitochondria and mitochondrial-associated membranes in GBM tissue. Glucose and glutamine are the major fermentable fuels that drive GBM growth. The major waste products of GBM cell fermentation (lactic acid, glutamic acid, and succinic acid) will acidify the microenvironment and are largely responsible for drug resistance, enhanced invasion, immunosuppression, and metastasis. Besides surgical debulking, therapies used for GBM management (radiation, chemotherapy, and steroids) enhance microenvironment acidification and, although often providing a time-limited disease control, will thus favor tumor recurrence and complications. The simultaneous restriction of glucose and glutamine, while elevating non-fermentable, anti-inflammatory ketone bodies, can help restore the pH balance of the microenvironment while, at the same time, providing a non-toxic therapeutic strategy for killing most of the neoplastic cells.

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Section: Fermentation Metabolism Is Responsible For Gbm Growthmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Metabolic management of microenvironment acidity in glioblastoma

et al. 2022

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“…In order to sustain glycolytic metabolism, accumulating lactate is dissipated to neighbouring cells via gap junctions formed by connexin 43 or exported by the H + /lactate cotransporter (e.g. monocarboxylate transporter, MCT), fuelling a vicious circle of microenvironmental acidification (Cardone et al., 2005; Michl et al., 2022). And very recently MCTs have been linked to ECM remodelling in pancreatic cancer (Ufuk et al., 2022).…”

Section: Introductionmentioning

confidence: 99%

When healing turns into killing – the pathophysiology of pancreatic and hepatic fibrosis

Ferdek

Krzysztofik

Stopa

et al. 2022

The Journal of Physiology

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“…DLD1 cells were selected for these experiments based on their high glycolytic rate. A recent whole-genome CRISPR-Cas9 screen identified ALDOA , a gene coding for the glycolytic enzyme aldolase A, as essential for CRC cell survival under physiological pH (29). Ablation of ALDOA using virally transduced gRNAs reduced expression ( Figure 6A ) and glycolytic rate, measured in terms of medium acidification ( Figure 6B ) and lactate production ( Figure 6C ).…”

Section: Resultsmentioning

confidence: 99%

“…The third test for metabolite exchange used cells with genetically inactivated mitochondrial respiration on the basis that this organellar process influences the levels of diffusible substances in cytoplasm, including ATP. SW1222 cells were selected on the basis of their high respiratory rate ( Figure 7B ), which could be genetically inactivated by knockout of NDUFS1 , a gene coding for a component of complex I ( Figure 7A ) (29). Oxidative phosphorylation was blocked in NDUFS1 KO cells, and a compensatory increase in glycolytic rate was observed ( Figure 7B ).…”

Section: Resultsmentioning

confidence: 99%

Solute exchange through gap junctions lessens the adverse effects of inactivating mutations in metabolite-handling genes

Monterisi¹,

Michl²,

Hill³

et al. 2022

Preprint

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Experimental inactivation of certain genes involved in metabolism attenuates cancer cell growth in vitro. However, loss-of-function mutations in metabolic pathways are not negatively selected in human cancers, indicating that these genes are not essential in vivo. We hypothesize that spontaneous mutations affecting metabolic pathways do not necessarily result in a functional defect because affected cells may be rescued by exchanging metabolites with neighboring wild-type cells via gap junctions. Using fluorescent substances to probe inter-cellular diffusion, we show that colorectal cancer (CRC) cells are coupled by gap junctions assembled from connexins, particularly the constitutively expressed Cx26. In co-cultures of wild-type cells with cells that had inactivated components of pH regulation (SLC9A1), glycolysis (ALDOA), or mitochondrial metabolism (NDUFS1), we show that diffusive coupling was able to rescue the functional defect associated with the inactivation of metabolite-handling genes. Function rescue was dependent on Cx26 channels and reduced phenotypic heterogeneity among cells. Since the phenotypic landscape did not map onto genotype, an individual cell should not be considered as the unit under selection, at least in the case of metabolite-handling processes. Our findings can explain why certain loss-of-function mutations in genes, previously ascribed as being 'essential', do not influence the growth of human cancers.

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CRISPR-Cas9 screen identifies oxidative phosphorylation as essential for cancer cell survival at low extracellular pH

Cited by 25 publications

References 34 publications

Metabolic management of microenvironment acidity in glioblastoma

Metabolic management of microenvironment acidity in glioblastoma

When healing turns into killing – the pathophysiology of pancreatic and hepatic fibrosis

Solute exchange through gap junctions lessens the adverse effects of inactivating mutations in metabolite-handling genes

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