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Updated: Jun 29, 2025
Macrophage Differentiation and Polarization into an M2-Like Phenotype using a Human Monocyte-Like THP-1 Leukemia Cell Line
Published on: August 2, 2021
Mevalonate kinase-deficient THP-1 cells show a disease-characteristic pro-inflammatory phenotype
Frouwkje A Politiek1,2, Marjolein Turkenburg1, Rob Ofman1
1Laboratory Genetic Metabolic Diseases, Department of Laboratory Medicine, Amsterdam University Medical Centers, Location Academic Medical Center, Amsterdam, Netherlands.
Objective:
Bi-allelic pathogenic variants in the MVK gene, which encodes mevalonate kinase (MK), an essential enzyme in isoprenoid biosynthesis, cause the autoinflammatory metabolic disorder mevalonate kinase deficiency (MKD). We generated and characterized MK-deficient monocytic THP-1 cells to identify molecular and cellular mechanisms that contribute to the pro-inflammatory phenotype of MKD.
Methods:
Using CRISPR/Cas9 genome editing, we generated THP-1 cells with different MK deficiencies mimicking the severe (MKD-MA) and mild end (MKD-HIDS) of the MKD disease spectrum. Following confirmation of previously established disease-specific biochemical hallmarks, we studied the consequences of the different MK deficiencies on LPS-stimulated cytokine release, glycolysis versus oxidative phosphorylation rates, cellular chemotaxis and protein kinase activity.
Results:
Similar to MKD patients' cells, MK deficiency in the THP-1 cells caused a pro-inflammatory phenotype with a severity correlating with the residual MK protein levels. In the MKD-MA THP-1 cells, MK protein levels were barely detectable, which affected protein prenylation and was accompanied by a profound pro-inflammatory phenotype. This included a markedly increased LPS-stimulated release of pro-inflammatory cytokines and a metabolic switch from oxidative phosphorylation towards glycolysis. We also observed increased activity of protein kinases that are involved in cell migration and proliferation, and in innate and adaptive immune responses. The MKD-HIDS THP-1 cells had approximately 20% residual MK activity and showed a milder phenotype, which manifested mainly upon LPS stimulation or exposure to elevated temperatures.
Conclusion:
MK-deficient THP-1 cells show the biochemical and pro-inflammatory phenotype of MKD and are a good model to study underlying disease mechanisms and therapeutic options of this autoinflammatory disorder.
Insights
Mevalonate kinase deficiency (MKD) causes inflammation due to impaired isoprenoid synthesis. MK-deficient cells mimic MKD, revealing a pro-inflammatory state linked to metabolic shifts and altered kinase activity, aiding disease research.
Area of Science:
- Biochemistry
- Immunology
- Cell Biology
Background:
- Mevalonate kinase deficiency (MKD) is an autoinflammatory disorder caused by pathogenic variants in the MVK gene.
- MK deficiency disrupts isoprenoid biosynthesis, impacting cellular functions.
- Understanding the molecular mechanisms of MKD's pro-inflammatory phenotype is crucial for therapeutic development.
Purpose of the Study:
- To generate and characterize MK-deficient THP-1 cells as a model for MKD.
- To investigate the molecular and cellular consequences of MK deficiency on inflammatory responses.
- To correlate MK deficiency levels with disease severity and cellular phenotypes.
Main Methods:
- CRISPR/Cas9 genome editing was used to create THP-1 cells with varying MK deficiencies (MKD-MA and MKD-HIDS).
- Biochemical hallmarks of MKD were confirmed.
- Consequences of MK deficiency were assessed, including cytokine release, metabolic rates (glycolysis vs. oxidative phosphorylation), chemotaxis, and protein kinase activity.
Main Results:
- MK-deficient THP-1 cells exhibited a pro-inflammatory phenotype mirroring MKD patients' cells, with severity correlating to residual MK protein levels.
- Severe MK deficiency (MKD-MA) led to undetectable MK protein, affected protein prenylation, increased pro-inflammatory cytokine release, and a metabolic shift towards glycolysis.
- Mild MK deficiency (MKD-HIDS) showed a less pronounced phenotype, primarily upon LPS stimulation or heat stress, with approximately 20% residual MK activity.
Conclusions:
- MK-deficient THP-1 cells accurately model the biochemical and pro-inflammatory characteristics of MKD.
- These cells provide a valuable platform for studying MKD pathogenesis.
- The established cell model can be utilized to explore potential therapeutic strategies for MKD.
