Metabolic Drug Response Phenotyping in Colorectal Cancer Organoids by LC-QTOF-MS

Neef, Sylvia K.; Janssen, Nicole; Winter, Stefan; Wallisch, Svenja K.; Hofmann, Ute; Dahlke, Marc H.; Schwab, Matthias; Mürdter, Thomas E.; Haag, Mathias

doi:10.3390/metabo10120494

Cited by 23 publications

(14 citation statements)

References 65 publications

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“…Metabolites were extracted using 80% organic solvent as described. 37 , 38 In brief, pellets were taken up in 150 μl ice cold methanol:acetonitrile:water (2:2:1, v/v/v) and lysed by three cycles of flash‐freezing, thawing and ultrasonic treatment. Afterwards, lysates were frozen for 1 h at ‐20°C, and debris was removed by centrifugation.…”

Section: Methodsmentioning

confidence: 99%

Nicotinamide‐N‐methyltransferase is a promising metabolic drug target for primary and metastatic clear cell renal cell carcinoma

Reustle

Menig

Leuthold

et al. 2022

Clinical & Translational Med

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Background The metabolic enzyme nicotinamide‐N‐methyltransferase (NNMT) is highly expressed in various cancer entities, suggesting tumour‐promoting functions. We systematically investigated NNMT expression and its metabolic interactions in clear cell renal cell carcinoma (ccRCC), a prominent RCC subtype with metabolic alterations, to elucidate its role as a drug target. Methods NNMT expression was assessed in primary ccRCC (n = 134), non‐tumour tissue and ccRCC‐derived metastases (n = 145) by microarray analysis and/or immunohistochemistry. Findings were validated in The Cancer Genome Atlas (kidney renal clear cell carcinoma [KIRC], n = 452) and by single‐cell analysis. Expression was correlated with clinicopathological data and survival. Metabolic alterations in NNMT‐depleted cells were assessed by nontargeted/targeted metabolomics and extracellular flux analysis. The NNMT inhibitor (NNMTi) alone and in combination with the inhibitor 2‐deoxy‐D‐glucose for glycolysis and BPTES (bis‐2‐(5‐phenylacetamido‐1,3,4‐thiadiazol‐2‐yl)ethyl‐sulfide) for glutamine metabolism was investigated in RCC cell lines (786‐O, A498) and in two 2D ccRCC‐derived primary cultures and three 3D ccRCC air–liquid interface models. Results NNMT protein was overexpressed in primary ccRCC (p = 1.32 × 10–16) and ccRCC‐derived metastases (p = 3.92 × 10–20), irrespective of metastatic location, versus non‐tumour tissue. Single‐cell data showed predominant NNMT expression in ccRCC and not in the tumour microenvironment. High NNMT expression in primary ccRCC correlated with worse survival in independent cohorts (primary RCC—hazard ratio [HR] = 4.3, 95% confidence interval [CI]: 1.5–12.4; KIRC—HR = 3.3, 95% CI: 2.0–5.4). NNMT depletion leads to intracellular glutamine accumulation, with negative effects on mitochondrial function and cell survival, while not affecting glycolysis or glutathione metabolism. At the gene level, NNMT‐depleted cells upregulate glycolysis, oxidative phosphorylation and apoptosis pathways. NNMTi alone or in combination with 2‐deoxy‐D‐glucose and BPTES resulted in inhibition of cell viability in ccRCC cell lines and primary tumour and metastasis‐derived models. In two out of three patient‐derived ccRCC air–liquid interface models, NNMTi treatment induced cytotoxicity. Conclusions Since efficient glutamine utilisation, which is essential for ccRCC tumours, depends on NNMT, small‐molecule NNMT inhibitors provide a novel therapeutic strategy for ccRCC and act as sensitizers for combination therapies.

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Section: Methodsmentioning

confidence: 99%

Nicotinamide‐N‐methyltransferase is a promising metabolic drug target for primary and metastatic clear cell renal cell carcinoma

Reustle

Menig

Leuthold

et al. 2022

Clinical & Translational Med

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“…Patient-derived colorectal cancer organoids were generated and cultured as described earlier [ 21 , 22 ]. In brief, the tissue was cut into small pieces and dissociated at 37 °C.…”

Section: Methodsmentioning

confidence: 99%

A human ex vivo coculture model to investigate peritoneal metastasis and innovative treatment options

Mönch

Koch

Maaß

et al. 2021

Pleura and Peritoneum

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Objectives Peritoneal metastasis (PM) is commonly observed in patients with colorectal cancer (CRC). The outcome of these patients is poor, with an average survival of only six months without therapy, which requires a better understanding of PM biology and new treatment strategies. Methods We established and characterized a human ex vivo peritoneal model to investigate the mechanisms of peritoneal seeding and possible treatment options. For this, CRC cell lines and patient-derived tumor organoids were cultured together with human peritoneum to investigate the invasion of malignant cells and the effects of local chemotherapy. Results Fresh human peritoneum was cultured for up to three weeks in a stainless steel ring system, allowing for survival of all peritoneal structures. Peritoneal cell survival was documented by light microscopy and immunohistochemical staining. Further, immunohistological characterization of the tissue revealed CD3-positive T-lymphocytes and vimentin-positive fibroblasts within the peritoneum. In addition, extracellular matrix components (collagens, matrix metalloproteinases) were localized within the tissue. Coculture with CRC cell lines and patient-derived CRC organoids revealed that cancer cells grew on the peritoneum and migrated into the tissue. Coculture with CRC cells confirmed that hyperthermal treatment at 41 °C for 90 min significantly enhanced the intracellular entry of doxorubicin. Moreover, treatment with mitomycin C under hyperthermic conditions significantly reduced the amount of cancer cells within the peritoneum. Conclusions This human ex vivo peritoneal model provides a stringent and clinically relevant platform for the investigation of PM and for further elucidation of possible treatment options.

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“…The ability to expand organoid lines to relatively high passage numbers has enabled their use for high-throughput multi-omics technologies, producing transcriptomics, proteomics, metabolomics, and lipidomics data sets that can be explored to better unravel the various interactions taking place in a tissue. 69 , 173 , 174 , 175 , 176 …”

Section: What To Consider When Adapting Organoid-based Models To Unex...mentioning

confidence: 99%

Everything You Always Wanted to Know About Organoid-Based Models (and Never Dared to Ask)

Hautefort

Poletti

Papp

et al. 2022

Cellular and Molecular Gastroenterology and Hepatology

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Metabolic Drug Response Phenotyping in Colorectal Cancer Organoids by LC-QTOF-MS

Cited by 23 publications

References 65 publications

Nicotinamide‐N‐methyltransferase is a promising metabolic drug target for primary and metastatic clear cell renal cell carcinoma

Nicotinamide‐N‐methyltransferase is a promising metabolic drug target for primary and metastatic clear cell renal cell carcinoma

A human ex vivo coculture model to investigate peritoneal metastasis and innovative treatment options

Everything You Always Wanted to Know About Organoid-Based Models (and Never Dared to Ask)

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