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Published on: September 1, 2019
Transcriptional properties of human NANOG1 and NANOG2 in acute leukemic cells
Irina Eberle1, Birgit Pless, Miriam Braun
1Institute of Pharmaceutical Biology/DCAL/ZAFES/CEF, Goethe-University, Biocenter, Max-von-Laue-Str. 9, 60438 Frankfurt/Main, Germany.
Abstract:
Transcripts of NANOG and OCT4 have been recently identified in human t(4;11) leukemia and in a model system expressing both t(4;11) fusion proteins. Moreover, downstream target genes of NANOG/OCT4/SOX2 were shown to be transcriptionally activated. However, the NANOG1 gene belongs to a gene family, including a gene tandem duplication (named NANOG2 or NANOGP1) and several pseudogenes (NANOGP2-P11). Thus, it was unclear which of the NANOG family members were transcribed in t(4;11) leukemia cells. 5'-RACE experiments revealed novel 5'-exons of NANOG1 and NANOG2, which could give rise to the expression of two different NANOG1 and three different NANOG2 protein variants. Moreover, a novel PCR-based method was established that allows distinguishing between transcripts deriving from NANOG1, NANOG2 and all other NANOG pseudogenes (P2-P11). By applying this method, we were able to demonstrate that human hematopoietic stem cells and different leukemic cells transcribe NANOG2. Furthermore, we functionally tested NANOG1 and NANOG2 protein variants by recombinant expression in 293 cells. These studies revealed that NANOG1 and NANOG2 protein variants are functionally equivalent and activate a regulatory circuit that activates specific stem cell genes. Therefore, we pose the hypothesis that the transcriptional activation of NANOG2 represents a 'gain-of-stem cell function' in acute leukemia.
Insights
Researchers identified NANOG2 gene transcripts in leukemia cells, distinct from NANOG1. Both NANOG1 and NANOG2 variants function similarly, activating stem cell genes, suggesting NANOG2 contributes to leukemia
Area of Science:
- Molecular Biology
- Cancer Research
- Stem Cell Biology
Background:
- NANOG and OCT4 transcripts were found in t(4;11) leukemia.
- Downstream targets of NANOG/OCT4/SOX2 were transcriptionally activated.
- The NANOG gene family includes NANOG1, NANOG2 (NANOGP1), and pseudogenes (NANOGP2-P11).
Purpose of the Study:
- To determine which NANOG family members are transcribed in t(4;11) leukemia cells.
- To characterize novel NANOG variants and develop a method to distinguish NANOG transcripts.
- To investigate the functional equivalence and role of NANOG1 and NANOG2 in leukemia.
Main Methods:
- 5'-RACE (Rapid Amplification of cDNA Ends) experiments to identify novel 5' exons.
- Development of a novel PCR-based method for transcript discrimination.
- Recombinant expression of NANOG1 and NANOG2 variants in 293 cells for functional testing.
Main Results:
- Novel 5' exons for NANOG1 and NANOG2 were identified, leading to multiple protein variants.
- A PCR method successfully distinguished transcripts from NANOG1, NANOG2, and pseudogenes.
- NANOG2 transcripts were detected in human hematopoietic stem cells and various leukemia cells.
- Both NANOG1 and NANOG2 protein variants were functionally equivalent, activating stem cell genes.
Conclusions:
- Transcriptional activation of NANOG2 in leukemia may represent a 'gain-of-stem cell function'.
- NANOG2 plays a significant role in the stem cell-like properties observed in acute leukemia.
- The findings provide insights into the molecular mechanisms driving leukemogenesis.
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