ZNF521 Has an Inhibitory Effect on the Adipogenic Differentiation of Human Adipose-Derived Mesenchymal Stem Cells

Abstract

Mesenchymal stem cells (MSCs) are multipotent progenitors present in the bone marrow stroma and in subcutaneous abdominal fat, an abundant and easily accessible source of MSCs with the ability to differentiate along multiple lineage pathways. The stem cell-associated transcription co-factor Zinc Finger Protein 521 (ZNF521/zfp521) has been implicated in the control of the homeostasis of hematopoietic, neural and osteo-adipogenic progenitors. Here we document through the analysis of a panel of human adipose-derived stem cells (hADSCs), that ZNF521 strongly inhibits the generation of mature adipocytes. Enforced overexpression of ZNF521 in these cells resulted in a significant delay and reduction in adipocyte differentiation upon exposure to inducers of adipogenesis. Of particular relevance, ZNF521 was able to inhibit the expression of ZNF423, recently identified as an essential commitment factor necessary for the generation of pre-adipocytes. Conversely, silencing of ZNF521 was found to significantly enhance the adipogenic differentiation of hADSCs. Inhibition of adipogenesis by ZNF521 was at least in part due to inhibition of EBF1. Taken together, these results confirm a role for ZNF521 as a key negative regulator of adipocyte differentiation of hADSCs.

An Approach towards a GMP Compliant In-Vitro Expansion of Human Adipose Stem Cells for Autologous Therapies

“…In recent years, several important genes have been discovered that are crucial for the maintenance of stemness and for the differentiation of hASCs [ 34 , 45 , 46 ]. Further information about the different genes, including their relationships to each other and a short description, can be found in “ Supplementary Materials ” or in the literature [ 47 , 48 , 49 , 50 , 51 , 52 , 53 , 54 , 55 , 56 , 57 , 58 , 59 , 60 , 61 , 62 , 63 , 64 , 65 , 66 , 67 , 68 , 69 , 70 , 71 , 72 , 73 , 74 , 75 , 76 , 77 , 78 , 79 , 80 , 81 , 82 , 83 ]. The selected genes can be used as markers in RT-qPCR tests to assess and compare the differentiation status of hASCs expanded in static 2D (see Section 3.2 ) and/or dynamic 3D conditions (see Section 3.3 ).…”

Chemoresistance in H-Ferritin Silenced Cells: The Role of NF-κB

“…To this end we used a homemade pRc/CMV-HA-IκBα plasmid and its scrambled control kindly provided by Ileana Quinto (Magna Graecia University of Catanzaro, Italy). To study the role of FHC amounts in controlling NF-κB activation instead we transfected K562 shFHC cells with the expression vector containing the full length of human FHC cDNA (pc3FHC) for 48 h. In brief, 1 × 10 6 cells were suspended in 100 μL of pre-warmed Nucleofector Solution Kit V (Amaza, Basel, Switzerland), and mixed with 3 µg of the expression vectors or with 3 µg of scrambled controls as previously described by Chiarella et al [ 40 ]. Then, DNA-cell suspensions were transferred to Amaxa certified cuvettes and electroporated using the Nucleofector program T-016.…”

Nasal Polyposis: Insights in Epithelial-Mesenchymal Transition and Differentiation of Polyp Mesenchymal Stem Cells

“…When the PO-MSCs are grown in an adipogenic induction medium for 21 days, some cells showed a tendency to form spherical accumulations of multiple intra-cellular lipid-filled droplets [ 50 , 62 ], which can be detected by Oil Red O staining. These PO-MSCs express an increased gene expression of the PPARγ, a key player in controlling the transcriptional pathway of adipogenesis, as well as the target gene FABP4 [ 57 ] and the transcription factor ZNF423 as found in adipocytes derived from mesenchymal stem cells [ 69 , 70 ]. In addition, recent evidence suggests that the fine balance between transcription zinc finger proteins, such as ZNF521/ZNF423, is relevant for maintenance of stemness in mesenchymal stem and progenitor cells [ 69 , 71 , 72 ].…”