Characterization of putative pluripotency-regulating genes and defining the role of germ cell-specific genes during reprogramming

• Antragstellerinnen / Antragsteller: Dr. Jessica Nolte; Professor Ulrich Zechner, Ph.D.
• Fachliche Zuordnung: Zellbiologie ; Humangenetik
• Förderung: Förderung von 2008 bis 2016
• Projektkennung: Deutsche Forschungsgemeinschaft (DFG) - Projektnummer 66168673

• Erstellungsjahr: 2014

Zusammenfassung der Projektergebnisse

In the first funding period, we further substantiated the pluripotency of germ line-derived mouse maGSCs in comparison to ESCs and iPSCs as references using several approaches and different genetic backgrounds (mouse strains). We could show that there is no significant difference on the level of transcriptome, proteome, miRNAome, epigenome (imprinted gene methylation, promoter methylation of pluripotency marker genes, and genome-wide methylation, global and gene-specific histone modifications), telomerase activity and immune response. By analyzing the transcriptomes of maGSCs and ESCs under undifferentiated and differentiated conditions we were further able to identify novel putative pluripotency-associated genes. In the second funding period, we focused our work on more specific aspects of pluripotency by the characterization of two new putative pluripotency genes identified in the first funding period, namely Zfp819 and Lrrc34. For Zfp819, we could not only show that it is a novel pluripotency-related factor but also uncover its role in maintenance of genome integrity in mouse ESCs. For Lrrc34, we demonstrated that it is a novel ESC-specific nucleolar protein possibly involved in ribosome biogenesis and regulation of the pluripotency gene network of pluripotent stem cells. In a further subproject that was partly pursued with personnel, consumables and lab animals funded by other institutions we found that temporal expression of Dppa3 is critical for generation of fully-reprogrammed iPSCs and for maintaining the Dlk1-Dio3 imprinting during somatic cell reprogramming.

Projektbezogene Publikationen (Auswahl)

• (2008) Multipotent adult germline stem cells and embryonic stem cells have similar microRNA profiles. Mol Hum Reprod. 14, 521-529
  Zovoilis A., Nolte J., Drusenheimer N., Zechner U., Hada H., Guan K., Hasenfuss G., Nayernia K., Engel W.
  
• (2009) Comparative methylation profiles and telomerase biology of mouse multipotent adult germline stem cells and embryonic stem cells. Mol Hum Reprod. 15, 345-353
  Zechner U., Nolte J., Wolf M., Shirneshan K., Hajj N.E., Weise D., Kaltwasser B., Zovoilis A., Haaf T., Engel W.
  
• (2009) Members of the miR-290 cluster modulate in vitro differentiation of mouse embryonic stem cells. Differentiation 78, 69-78
  Zovoilis A., Smorag L., Pantazi A., Engel W.
  
• (2009) Multipotent adult germ-line stem cells, like other pluripotent stem cells, can be killed by cytotoxic T lymphocytes despite low expression of major histocompatibility complex class I molecules. Biol. Direct. 4, 31
  Dressel R., Guan K., Nolte J., Elsner L., Monecke S., Nayernia K., Hasenfuss G., Engel W.
  
• (2009) Multipotent adult germline stem cells and embryonic stem cells: comparative proteomic approach. J Proteome Res. 8, 5497-5510
  Dihazi H., Dihazi G.H., Nolte J., Meyer S., Jahn O., Müller G.A., Engel W.
  
• (2010) Embryonic stem cell-related miRNAs are involved in differentiation of pluripotent cells originating from the germ line. Mol Hum Reprod. 16, 793-803
  Zovoilis A., Pantazi A., Smorag L., Opitz L., Riester G.S., Wolf M., Zechner U., Holubowska A., Stewart C.L., Engel W.
  
• (2010) Pluripotent embryonic stem cells and multipotent adult germline stem cells reveal similar transcriptomes including pluripotency-related genes. Mol Hum Reprod. 16, 846-855
  Meyer S., Nolte J., Opitz L., Salinas-Riester G., Engel W.
  
• (2010). Pluripotent stem cells are highly susceptible targets for syngeneic, allogeneic, and xenogeneic natural killer cells. FASEB J. 24, 2164-2177
  Dressel R., Nolte J., Elsner L., Novota P., Guan K., Hasenfuss G., Jaenisch R., Engel W.
  
• (2011) Global and gene-specific histone modification profiles of mouse multipotent adult germline stem cells. Mol Hum Reprod. 17, 166-174
  Khromov T., Krishna Pantakani D.V., Nolte J., Wolf M., Dressel R., Engel W., Zechner U.
  
• (2011) Multipotent adult germline stem cells and embryonic stem cells functional proteomics revealed an important role of eukaryotic initiation factor 5A (Eif5a) in stem cell differentiation. J Proteome Res. 10, 1962-1973
  Dihazi H., Dihazi G.H., Jahn O., Meyer S., Nolte J., Asif A.R., Mueller G.A., Engel W.
  
• (2011) Stage-specific germ-cell marker genes are expressed in all mouse pluripotent cell types and emerge early during induced pluripotency. PLoS One. 6, e22413
  Xu X., Pantakani D.V., Lührig S., Tan X., Khromov T., Nolte J., Dressel R., Zechner U., Engel W.
  
• (2012) Apoptosis-related gene expression profiles of mouse ESCs and maGSCs: role of Fgf4 and Mnda in pluripotent cell responses to genotoxicity. PLoS One. 7, e48869
  Khromov T., Dressel R., Siamishi I., Nolte J., Opitz L., Engel W., Pantakani D.V.
  
• (2012) Gene Expression and Epigenetic Signatures of Germ Cell-Derived Pluripotent Stem Cells and Embryonic Stem Cells in: Stem Cells and Cancer Stem Cells, Volume 6, Therapeutic Applications in Disease and Injury, Ed,: M.A. Hayat, Springer Dordrecht
  Nolte J., Pantakani D.V., Dihazi H., Zechner U.
  
• 2012) Generation and Characterization of Yeast Two-Hybrid cDNA Libraries Derived From Two Distinct Mouse Pluripotent Cell Types. Mol Biotechnol.
  Zheng Y., Tan X., Pyczek J., Nolte J., Pantakani D.V., Engel W.
  
• (2013) Impact of the antiproliferative agent ciclopirox olamine treatment on stem cells proteome. World J Stem Cells. 5, 9-25
  Dihazi G.H., Bibi A., Jahn O., Nolte J., Mueller G.A., Engel W., Dihazi H.
  
• (2013) The novel BTB-kelch protein, KBTBD8, is located in the Golgi apparatus and translocates to the spindle apparatus during mitosis. Cell Div. 8, 3
  Lührig S., Kolb S., Mellies N., Nolte J.
  
• (2013) Zfp819, a novel KRAB-zinc finger protein, interacts with KAP1 and functions in genomic integrity maintenance of mouse embryonic stem cells. Stem Cell Res. 11, 1045-1059
  Tan T., Xu X., Zechner U., Nolte J., Engel W., Krishna Pantakani D.V.
  
• (2014) Lrrc34, a novel nucleolar protein, interacts with Npm1 and Ncl and has an impact on pluripotent stem cells. Stem Cells Dev.
  Lührig S., Siamishi I., Tesmer-Wolf M., Zechner U., Engel W., Nolte J.