The Effect of SETD3 on Beta-Catenin and Canonical Wnt Signaling Pathway Activity in Mouse Embryonic Stem Cells

2023-1-16
Alganatay, Ceren
Show full item record
Creative Commons License
This work is licensed under a Creative Commons Attribution-NonCommercial-NoDerivatives 4.0 International License.
282
views
61
downloads
Mouse embryonic stem cells (mESCs) are self-renewing, pluripotent cells that can differentiate into endoderm, mesoderm and neuroectoderm. As differentiation requires deactivation of pluripotency factors and activation of germ layer specific genes, many epigenetic factors play an essential role. A previously performed shRNA screen showed that SETD3 which is a SET-domain containing epigenetic factor, is an essential factor for mesendoderm differentiation of mESCs. Our preliminary data demonstrated that when mESCs were grown in standard serum-containing mESC medium prior to directed differentiation, wild type mESCs can successfully differentiate into mesendoderm, while SETD3 knockout (setd3∆) mESCs cannot. According to our preliminary data, we aimed to identify the effect of SETD3 on the levels and localization of β-catenin as well as on the canonical Wnt signaling pathway activity in mESCs. Proximity ligation assay revealed that SETD3 and β-catenin are interacting in mESCs, further supporting the role of canonical Wnt signaling pathway. Even though immunocytochemistry assay performed wild type and setd3∆ mESCs grown in the same medium did not show significant localization differences, western blot experiments performed with nuclear fractions showed that setd3∆ mESCs grown in standard serum-containing mESC medium had lower levels of β-catenin than wild type mESCs. Additionally, luciferase assay results demonstrated that, regardless of the growth medium, absence of SETD3 results with significantly lower β-catenin transcriptional activity. Observed lower nuclear βcatenin levels and lower canonical Wnt signaling pathway activity in the absence of SETD3 might explain the mesendoderm differentiation defect of setd3∆ mESCs grown in standard serum-containing mESC medium.
SETD3, Mouse Embryonic Stem Cells, Canonical Wnt Signaling Pathway, β-catenin
https://hdl.handle.net/11511/102077
Graduate School of Natural and Applied Sciences, Thesis

Suggestions

SETD3-dependent gene expression changes during endoderm differentiation of mouse embryonic stem cells
Balbaşı, Emre; Terzi Çizmecioğlu, Nihal; Department of Biology (2022-2-09)
Mouse embryonic stem cells (mESCs) are pluripotent cells that have self renewal capability. They can differentiate into all three primary germ layers: mesoderm, endoderm, and ectoderm during embryonic development. The embryonic development is controlled via spatiotemporal regulation of gene expression changes. The collaborative effects of Wnt, Nodal, BMP signaling pathways help form the primitive streak, and the subsequent definitive endoderm layer in the gastrulating embryo. Deactivation of core pluripoten...
Characterization and identification of human mesenchymal stem cells at molecular level
Aksoy, Ceren; Severcan, Feride; Çetinkaya, Duygu Uçkan; Department of Biotechnology (2012)
Bone marrow mesenchymal stem cells (BM-MSCs) are pluripotent cells that can differentiate into a variety of non-hematopoietic tissues. They also maintain healthy heamatopoiesis by providing supportive cellular microenvironment into BM. In this thesis, MSCs were characterized in terms of their morphological, immunophenotypical and differentiation properties. Then, they were examined by attenuated total reflection-Fourier transform infrared (ATR-FTIR) spectroscopy together with hierarchical clustering, and FT...
Evaluating Oxygen Tensions Related to Bone Marrow and Matrix for MSC Differentiation in 2D and 3D Biomimetic Lamellar Scaffolds
Sayin, Esen; Baran, Erkan Turker; Elsheikh, Ahmed; Mudera, Vivek; Cheema, Umber; Hasırcı, Vasıf Nejat (2021-04-01)
The physiological O-2 microenvironment of mesenchymal stem cells (MSCs) and osteoblasts and the dimensionality of a substrate are known to be important in regulating cell phenotype and function. By providing the physiologically normoxic environments of bone marrow (5%) and matrix (12%), we assessed their potential to maintain stemness, induce osteogenic differentiation, and enhance the material properties in the micropatterned collagen/silk fibroin scaffolds that were produced in 2D or 3D. Expression of ost...
Directed Differentiation of Mouse Embryonic Stem Cells to Mesoderm, Endoderm, and Neuroectoderm Lineages.
Balbasi, Emre; Sezginmert, Dersu; Alganatay, Ceren; Terzi Cizmecioglu, Nihal (2021-10-06)
The self-renewal and pluripotency features of mouse embryonic stem cells (mESCs) make them a great tool to study early mammalian development. Various signaling pathways that shape early mammalian development can be mimicked for in vitro mESC differentiation toward primitive lineages first and more specialized cell types later. Since the precise nature of the molecular mechanisms and the crosstalk between these signaling pathways is yet to be fully understood, there is a high level of variability in the effi...
Influence of STRO-1 selection on osteogenic potential of human tooth germ derived mesenchymal stem cells
Ercal, Pinar; Pekozer, Gorke G.; Gumru, Osman Z.; KÖSE, GAMZE; Ramazanoglu, Mustafa (Elsevier BV, 2017-10-01)
Mesenchymal stem cells derived from the human tooth germ (hTGSCs) are a heterogeneous cell population that can differentiate into osteogenic, neurogenic, and adipogenic lineages. The aim of this study was to compare the osteogenic differentiation capacity of STRO-1 positive (STRO-1 +) hTGSCs and unsorted heterogeneous hTGSCs and to establish if STRO-1 + cells are more committed to osteogenic differentiation. HTGSCs were isolated from impacted third molar tooth germ tissues of adolescents, and a subpopulatio...
Citation Formats
C. Alganatay, “The Effect of SETD3 on Beta-Catenin and Canonical Wnt Signaling Pathway Activity in Mouse Embryonic Stem Cells,” M.S. - Master of Science, Middle East Technical University, 2023.
METU IIS - Integrated Information Service open@metu.edu.tr