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Objective: Nanog is expressed in adult endothelial cells (ECs) at a low-level, however, its functional significance is not known. The goal of our study was to elucidate the role of Nanog in adult ECs using a genetically engineered mouse model system. Approach and Results: Biochemical analyses showed that Nanog is expressed in both adult human and mouse tissues. Primary ECs isolated from adult mice showed detectable levels of Nanog, Tert (telomerase reverse transcriptase), and eNos (endothelial nitric oxide synthase). Wnt3a (Wnt family member 3A) increased the expression of Nanog and hTERT (human telomerase reverse transcriptase) in ECs and increased telomerase activity in these cells. In a chromatin immunoprecipitation experiment, Nanog directly bound to thehTERT andeNOS promoter/enhancer DNA elements, thereby regulating their transcription. Administration of low-dose tamoxifen toROSA mT/mG ::Nanog fl/+ ::Cdh5 CreERT2 mice induced deletion of a singleNanog allele, simultaneously labeling ECs with green fluorescent protein and resulting in decreased Tert and eNos levels. Histological and morphometric analyses of heart tissue sections prepared from these mice revealed cell death, microvascular rarefaction, and increased fibrosis in cardiac vessels. Accordingly, EC-specificNanog -haploinsufficiency resulted in impaired EC homeostasis and angiogenesis. Conversely, re-expression of cDNA encoding thehTERT in Nanog-depleted ECs, in part, restored the effect of loss of Nanog.Conclusions: We showed that low-level Nanog expression is required for normal EC homeostasis and angiogenesis in adulthood.

Low-Level Nanog Expression in the Regulation of Quiescent Endothelium