Silencing the long non-coding RNA antisense to ZEB2 facilitates reprogramming of aged fibroblasts and safeguards stem cell pluripotency

  • FCT Investigator, Instituto de Medicina Molecular (IMM), Faculdade de Medicina da Universidade de Lisboa
      Bruno Bernardes de Jesus is a FCT Investigator at the unit of Dr. Carmo-Fonseca, Instituto de Medicina Molecular (IMM), Faculdade de Medicina da Universidade de Lisboa, Lisboa, since 2015. From 2013 to 2015, he was a Post-Doctoral fellow in the unit of Dr. Carmo-Fonseca, Cell Biology Division at the Instituto de Medicina Molecular (IMM), Faculdade de Medicina da Universidade de Lisboa and, before that, from 2009 to 2013, Bruno was a Post-Doctoral fellow under the supervision of Dr. Maria A. Blasco, head of the Telomeres and Telomerase Laboratory, and Director of the Spanish National Cancer Research Centre (CNIO), Madrid, Spain. Bruno got his PhD in Molecular and Cellular Biology (University Louis Pasteur) at the Functional Genomic Department, IGBMC, Strasbourg, France, during 2008 and under supervision of Prof. Jean-Marc Egly. Bruno has a BSc in Biochemistry from the Faculty of Sciences and Technology, University of Coimbra, Portugal (1998 to 2003). Bruno Bernardes de Jesus principal research interests are Aging, stem cell biology, telomerase, cancer and lncRNAs. Bruno was the author of several scientific papers and patents on those research areas. Additionally, he collaborated in several MSc and PhD programs.


    Ageing imposes a barrier to somatic cell reprogramming through mechanisms that are poorly understood. Here we studied the age-associated decline in reprogramming efficiency of fibroblasts derived from transgenic mice carrying doxycycline-inducible Oct4, Sox2, Klf4, and c-Myc. We found that fibroblasts from old mice express higher levels of Zeb2, a transcription factor that activates epithelial-to-mesenchymal transition (EMT). As reprogramming requires suppression of pro-EMT signals, we hypothesized that Zeb2 overexpression contributes to the inefficient reprogramming of old fibroblasts. Synthesis of Zeb2 protein is controlled by a natural antisense transcript named Zeb2-NAT, and we show that silencing Zeb2-NAT suffices to enhance reprogramming of old fibroblasts. Transfection of adult fibroblasts with specific LNA Gapmers induced a robust downregulation of Zeb2-NAT transcripts and Zeb2 protein, and subsequent induction with doxycycline resulted in efficient reprogramming into pluripotent cells with capacity to spontaneously form differentiated tumors comprising the three germ layers. We further observed that Zeb2-NAT expression is precociously activated by differentiation stimuli in embryonic stem (ES) cells. Knocking-down Zeb2-NAT maintained ES cells challenged with commitment signals in the ground state of self-renewal and pluripotency. In conclusion, our study identifies a long non-coding RNA that is overlapping and antisense to the Zeb2 locus as a novel target for rejuvenation strategies.

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