Alterations of mTOR signaling impact metabolic stress resistance in colorectal carcinomas with BRAF and KRAS mutations

Fritsche‐Guenther, Raphaela; Zasada, Christin; Mastrobuoni, Guido; Royla, Nadine; Rainer, Roman; Roßner, Florian; Pietzke, Matthias; Klipp, Edda; Sers, Christine; Kempa, Stefan

doi:10.1038/s41598-018-27394-1

Cited by 27 publications

(24 citation statements)

References 56 publications

(59 reference statements)

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“…Finally, two groups have recently demonstrated that KRAS G12v and BRAF V600E mutations affects metabolic pathways differently in CRC [109, 110]. A proteomics and metabolomics approach by Fritsche-Guenther and colleagues showed that glycolytic flux was upregulated in Caco-2- KRAS G12v cells but decreased in Caco-2- BRAF V600E cells as compared to Caco-2 controls at physiologic glucose levels [110].…”

Section: Rtk-ras Signalingmentioning

confidence: 99%

Colorectal Cancer and Metabolism

Brown

Short

Williams

2018

Curr Colorectal Cancer Rep

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Purpose of Review-Metabolic reprogramming is essential for the rapid proliferation of cancer cells and is thus recognized as a hallmark of cancer. In this review, we will discuss the etiologies and effects of metabolic reprogramming in colorectal cancer. Recent Findings-Changes in cellular metabolism may precede the acquisition of driver mutations ultimately leading to colonocyte transformation. Oncogenic mutations and loss of tumor suppressor genes further reprogram CRC cells to upregulate glycolysis, glutaminolysis, onecarbon metabolism, and fatty acid synthesis. These metabolic changes are not uniform throughout tumors, as subpopulations of tumor cells may rely on different pathways to adapt to nutrient availability in the local tumor microenvironment. Finally, metabolic cross-communication between stromal cells, immune cells, and the gut microbiota enable CRC growth, invasion, and metastasis. Summary-Altered cellular metabolism occurs in CRC at multiple levels, including in the cells that make up the bulk of CRC tumors, cancer stem cells, the tumor microenvironment, and hostmicrobiome interactions. This knowledge may inform the development of improved screening and therapeutics for CRC.

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Section: Rtk-ras Signalingmentioning

confidence: 99%

Colorectal Cancer and Metabolism

Brown

Short

Williams

2018

Curr Colorectal Cancer Rep

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“…A previous protocol used in our lab for adherent cells growing under 10% FCS was a modified Bligh-Dyer extraction to separate lipid and polar metabolites [23,24]. The protocol includes the use of 50% MeOH for cell quenching followed by an extraction using a 1:0.4:1, v/v/v MeOH:CHCl 3 :H 2 O ratio.…”

Section: Preliminary Experiments On Phase Separationmentioning

confidence: 99%

Modified Protocol of Harvesting, Extraction, and Normalization Approaches for Gas Chromatography Mass Spectrometry-Based Metabolomics Analysis of Adherent Cells Grown Under High Fetal Calf Serum Conditions

et al. 2019

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A gas chromatography mass spectrometry (GC-MS) metabolomics protocol was modified for quenching, harvesting, and extraction of metabolites from adherent cells grown under high (20%) fetal calf serum conditions. The reproducibility of using either 50% or 80% methanol for quenching of cells was compared for sample harvest. To investigate the efficiency and reproducibility of intracellular metabolite extraction, different volumes and ratios of chloroform were tested. Additionally, we compared the use of total protein amount versus cell mass as normalization parameters. We demonstrate that the method involving 50% methanol as quenching buffer followed by an extraction step using an equal ratio of methanol:chloroform:water (1:1:1, v/v/v) followed by the collection of 6 mL polar phase for GC-MS measurement was superior to the other methods tested. Especially for large sample sets, its comparative ease of measurement leads us to recommend normalization to protein amount for the investigation of intracellular metabolites of adherent human cells grown under high (or standard) fetal calf serum conditions. To avoid bias, care should be taken beforehand to ensure that the ratio of total protein to cell number are consistent among the groups tested. For this reason, it may not be suitable where culture conditions or cell types have very different protein outputs (e.g., hypoxia vs. normoxia). The full modified protocol is available in the Supplementary Materials.

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“…It is already known that the glycolysis and glutaminolysis of cancer cells can be blocked by using glycolytic inhibitors like 3-Bromopyruvic acid (BrPy) [ 7 , 23 ]. Therefore, we assessed the viability of the OS cell lines treated with BrPy.…”

Section: Resultsmentioning

confidence: 99%

“…The extracts were centrifuged for 10 min at 18,213× g , and aliquots of 25 µL were transferred into glass vials for GC-MS measurements. An identification mixture for reliable compound identification was prepared and derivatized in the same way, and an alkane mixture for a reliable retention index calculation was included [ 7 , 23 , 41 ]. The metabolite analysis was performed on a Pegasus 4D GCxGC TOFMS-System (LECO Corporation, St. Joseph, MN, USA) complemented with an auto-sampler (Gerstel MPS DualHead with CAS4 injector, Mühlheim an der Ruhr, Germany).…”

Section: Methodsmentioning

confidence: 99%

“…Thus, metabolites may serve as biomarkers for cancer diagnosis and/or prognosis. To date, metabolomic signatures have already been published for several epithelial cancers, such as colon, liver, lung, pancreatic, ovarian and breast [ 6 , 7 ]. Previous research in OS is more modest and has focused on understanding the pathogenesis and development of OS.…”

Section: Introductionmentioning

confidence: 99%

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Progression-Dependent Altered Metabolism in Osteosarcoma Resulting in Different Nutrient Source Dependencies

Fritsche‐Guenther

Gloaguen

Kirchner

et al. 2020

Cancers

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Osteosarcoma (OS) is a primary malignant bone tumor and OS metastases are mostly found in the lung. The limited understanding of the biology of metastatic processes in OS limits the ability for effective treatment. Alterations to the metabolome and its transformation during metastasis aids the understanding of the mechanism and provides information on treatment and prognosis. The current study intended to identify metabolic alterations during OS progression by using a targeted gas chromatography mass spectrometry approach. Using a female OS cell line model, malignant and metastatic cells increased their energy metabolism compared to benign OS cells. The metastatic cell line showed a faster metabolic flux compared to the malignant cell line, leading to reduced metabolite pools. However, inhibiting both glycolysis and glutaminolysis resulted in a reduced proliferation. In contrast, malignant but non-metastatic OS cells showed a resistance to glycolytic inhibition but a strong dependency on glutamine as an energy source. Our in vivo metabolic approach hinted at a potential sex-dependent metabolic alteration in OS patients with lung metastases (LM), although this will require validation with larger sample sizes. In line with the in vitro results, we found that female LM patients showed a decreased central carbon metabolism compared to metastases from male patients.

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Alterations of mTOR signaling impact metabolic stress resistance in colorectal carcinomas with BRAF and KRAS mutations

Cited by 27 publications

References 56 publications

Colorectal Cancer and Metabolism

Colorectal Cancer and Metabolism

Modified Protocol of Harvesting, Extraction, and Normalization Approaches for Gas Chromatography Mass Spectrometry-Based Metabolomics Analysis of Adherent Cells Grown Under High Fetal Calf Serum Conditions

Progression-Dependent Altered Metabolism in Osteosarcoma Resulting in Different Nutrient Source Dependencies

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