Identifying Dihydropyrimidine Dehydrogenase as a Novel Regulator of Hepatic Steatosis

Pyrimidine catabolism is implicated in hepatic steatosis. Dihydropyrimidine Dehydrogenase (DPYD) is an enzyme responsible for uracil as well as thymine catabolism, and human genetic variability in this enzyme has been described in relation to clinically observed toxicity following 5-Fluorouracil (5-FU) administration. We have demonstrated that pharmacologic inhibition of DPYD is protective in a human in vitro model of diet-induced steatosis. A gain-of-function mutation in DPYD through CRISPR-Cas9 engineering leads to an increased lipid burden associated with altered mitochondrial functionality in a hepatocarcionma cell line. These studies uncovered a novel role DPYD plays in regulating the metabolic phenotype of hepatocytes and therefore identifies DPYD as a key modulator of hepatic steatosis.

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Identifying Dihydropyrimidine Dehydrogenase as a Novel Regulator of Hepatic Steatosis

10.21203/rs.3.rs-971571/v1 ◽

2021 ◽

Author(s):

Kelly E. Sullivan ◽

Sheetal Kumar ◽

Xin Liu ◽

Ye Zhang ◽

Emily Koning ◽

...

Keyword(s):

Genetic Variability ◽

Cell Line ◽

Hepatic Steatosis ◽

Lipid Accumulation ◽

Metabolic Regulation ◽

Dihydropyrimidine Dehydrogenase ◽

Gain Of Function ◽

Pyrimidine Catabolism ◽

Pharmacologic Inhibition

Abstract Pyrimidine catabolism is implicated in hepatic steatosis. Dihydropyrimidine Dehydrogenase (DPYD) is an enzyme responsible for uracil and thymine catabolism, and DPYD human genetic variability affects clinically observed toxicity following 5-Fluorouracil (5-FU) administration. In an in vitro model of diet-induced steatosis, the pharmacologic inhibition of DPYD resulted in protection from lipid accumulation. Additionally, a gain-of-function mutation of DPYD, created through clustered regularly interspaced short palindromic repeats associated protein 9 (CRISPR-Cas9) engineering, led to an increased lipid burden, which was associated with altered mitochondrial functionality in a hepatocarcionma cell line. The studies presented herein describe a novel role for DPYD in hepatocyte metabolic regulation as a modulator of hepatic steatosis.

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Lipid accumulation in hepatocytes induces activation and pro-fibrotic gene expression in hepatic stellate cells.–A new in vitro model to study fibrogenesis in hepatic steatosis

Zeitschrift für Gastroenterologie ◽

10.1055/s-2008-1037459 ◽

2008 ◽

Vol 46 (01) ◽

Author(s):

H Wobser ◽

C Dorn ◽

T Weiss ◽

T Amann ◽

C Bollheimer ◽

...

Keyword(s):

Gene Expression ◽

Hepatic Steatosis ◽

Hepatic Stellate Cells ◽

Lipid Accumulation ◽

In Vitro Model ◽

Stellate Cells ◽

Vitro Model

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The Human Leukemic HL-60 Cell Line: An in Vitro Model for Cell Death Endpoint Identification

Alternatives to Laboratory Animals ◽

10.1177/026119299602400419 ◽

1996 ◽

Vol 24 (4) ◽

pp. 581-587

Author(s):

Cristiana Zanetti ◽

Arrnalaura Stammati ◽

Orazio Sapora ◽

Flavia Zucco

Keyword(s):

Cell Death ◽

Cell Line ◽

In Vitro Model ◽

Leukemia Cell ◽

Promyelocytic Leukemia ◽

Leukemia Cell Line ◽

Cell Type ◽

Vitro Model ◽

Promyelocytic Leukemia Cell Line

The aim of this study was to investigate the endpoints related to cell death, either necrosis or apoptosis, induced by four chemicals in the promyelocytic leukemia cell line, HL-60. Cell morphology, DNA fragmentation, cytofluorimetric analysis and oxygen consumption were used to classify the type of cell death observed. In our analysis, we found that not all the selected parameters reproduced the differences observed in the cell death caused by the four chemicals tested. As cell death is a very complex phenomenon, several factors should be taken into account (cell type, exposure time and chemical concentration), if chemicals are to be classified according to differences in the mechanisms more directly involved in cell death.

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