Functional and Multi-Omics Effects of an Optimized CRISPR-Mediated FURIN Depletion in U937 Monocytes.
Ruiming ChuaLijin WangRoshni R SingarajaSujoy GhoshPublished in: Cells (2024)
The pro-protein convertase FURIN (PCSK3) is implicated in a wide range of normal and pathological biological processes such as infectious diseases, cancer and cardiovascular diseases. Previously, we performed a systemic inhibition of FURIN in a mouse model of atherosclerosis and demonstrated significant plaque reduction and alterations in macrophage function. To understand the cellular mechanisms affected by FURIN inhibition in myeloid cells, we optimized a CRISPR-mediated gene deletion protocol for successfully deriving hemizygous (HZ) and nullizygous (NZ) FURIN knockout clones in U937 monocytic cells using lipotransfection-based procedures and a dual guide RNA delivery strategy. We observed differences in monocyte and macrophage functions involving phagocytosis, lipid accumulation, cell migration, inflammatory gene expression, cytokine release patterns, secreted proteomics (cytokines) and whole-genome transcriptomics between wild-type, HZ and NZ FURIN clones. These studies provide a mechanistic basis on the possible roles of myeloid cell FURIN in cardiovascular disorders.
Keyphrases
- gene expression
- induced apoptosis
- dendritic cells
- cell migration
- single cell
- genome wide
- cardiovascular disease
- wild type
- infectious diseases
- mouse model
- cell cycle arrest
- dna methylation
- crispr cas
- oxidative stress
- adipose tissue
- acute myeloid leukemia
- randomized controlled trial
- type diabetes
- squamous cell carcinoma
- endothelial cells
- endoplasmic reticulum stress
- signaling pathway
- coronary artery disease
- metabolic syndrome
- young adults
- peripheral blood
- cell death
- immune response
- small molecule
- papillary thyroid
- cell proliferation
- cell therapy
- binding protein
- mesenchymal stem cells
- cardiovascular risk factors
- genome wide identification