ChIP‐on‐chip analysis reveals angiopoietin 2 (Ang2, ANGPT2) as a novel target of steroidogenic factor‐1 (SF‐1, NR5A1) in the human adrenal gland

Abstract

The nuclear receptor steroidogenic factor‐1 (SF‐1, NR5A1 ) is a key regulator of adrenal and gonadal biology. Disruption of SF‐1 can lead to disorders of adrenal development, while increased SF‐1 dosage has been associated with adrenocortical tumor‐igenesis. We aimed to identify a novel subset of SF‐1 target genes in the adrenal by using chromatin immunoprecipitation (ChIP) microarrays (ChIP‐on‐chip) combined with systems analysis. SF‐1 ChIP‐on‐chip was performed in NCI‐H295R human adrenocortical cells using promoter tiling arrays, leading to the identification of 445 gene loci where SF‐1‐binding regions were located from 10 kb upstream to 3 kb downstream of a transcriptional start. Network analysis of genes identified as putative SF‐1 targets revealed enrichment for angiogenic process networks. A 1.1‐kb SF‐1‐binding region was identified in the angiopoietin 2 (Ang2, ANGPT2 ) promoter in a highly repetitive region, and SF‐1‐dependent activation was confirmed in luciferase assays. Angiogenesis is paramount in adrenal development and tumorigenesis, but until now a direct link between SF‐1 and vascular remodeling has not been established. We have identified Ang2 as a potentially important novel target of SF‐1 in the adrenal gland, indicating that regulation of angiogenesis might be an important additional mechanism by which SF‐1 exerts its actions in the adrenal gland.—Ferraz‐de‐Souza, B., Lin, L., Shah, S., Jina, N., Hubank, M., Dattani, M. T., Achermann, J. C. ChIP‐on‐chip analysis reveals angiopoietin 2 (Ang2, ANGPT2) as a novel target of steroidogenic fac‐tor‐1 (SF‐1, NR5A1) in the human adrenal gland. FASEBJ. 25, 1166–1175 (2011). www.fasebj.org