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Endothelial cell‐specific deletion of a single Nanog allele demonstrates its crucial role in the microenvironment of adult tissues
Author(s) -
Baruah Jugajyoti,
Wary Kishore
Publication year - 2017
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.31.1_supplement.1065.11
Subject(s) - homeobox protein nanog , rex1 , biology , chromatin immunoprecipitation , microbiology and biotechnology , gene knockdown , nanog homeobox protein , telomerase reverse transcriptase , embryonic stem cell , cancer research , chemistry , promoter , induced pluripotent stem cell , genetics , telomerase , gene expression , cell culture , gene
Rationale Although Nanog is a known regulator of pluripotency, the functional significance of low level expression of Nanog in endothelial cells (ECs) remains incompletely understood. Methods and results We have created ROSA mT/mG ::Nanog fl/+ ::Cdh5 CreERT2 genetically engineered mouse model (GEMM) system. Administration of tamoxifen (TAM) to these adult mice induced EC‐specific loss of single Nanog allele. GFP lineage tracing combined with high‐resolution microscopy revealed GFP‐positive ECs in the microenvironment of adult tissues. Knockout (KO) or knockdown of single Nanog allele in ECs decreased expression of VEGFR2 and hTERT upon Wnt3a stimulation, suggesting a close relationship between Nanog and hTERT. In a chromatin immunoprecipitation (ChIP) assay, we identified Nanog binding sites on the promoter/enhancer region of the human hTERT in ECs. Elevated expression of Nanog increased hTERT ‐promoter luciferase activity in ECs. Conversely, in Wnt3a stimulated ECs, Nanog‐ depletion decreased hTERT protein which modulates EC proliferation. Importantly, re‐expression of hTERT in Nanog ‐depleted ECs partially restored proliferation and angiogenesis. Summary These novel findings establish a key role for low level expression of Nanog in the regulation of EC homeostasis that is independent from its recognized function as a regulator of pluripotency. Support or Funding Information Studies were supported by the National Institute of Health (HL079356), American Heart Association (GRNT25710129) and by the University of Illinois at Chicago Center for Clinical and Translation Science Award (UL1RR029879) to K.K.W. J.B was supported by AHA pre‐doctoral fellowship mid‐west affiliate (15PRE22760004) and UIC‐CCTS‐PECTS fellowship (#1UL1TR002003).