SRP326550 PRJNA743129 GSE179308 Identification of new TCF21 targets in adrenocortical carcinoma cells Transcription factor 21 (TCF21) directly binds and regulates SF1 in tumor and normal adrenocortical cells, and both are involved in the development and steroidogenesis of the adrenal cortex. TCF21 is a tumor suppressor gene and its expression is reduced in malignant tumors. In adrenocortical tumors, it is less expressed in adrenocortical carcinomas (ACC) than in adrenocortical adenomas (ACA) and normal tissue. However, a comprehensive analysis to identify TCF21 targets have not yet been conducted in any type of cancer. In this study, we performed Chromatin Immunoprecipitation and Sequencing (ChIP-Seq) in adrenocortical carcinoma cell line (NCI-H295R) overexpressing TCF21, with the aim of identifying TCF21 new targets. The five most frequently identified sequences corresponded to the PRDM7, CNTNAP2, CACNA1B, PTPRN2 and KCNE1B genes. Validation experiments showed that, in NCI-H295R cells, TCF21 regulates gene expression positively in PRDM7 and negatively in CACNA1B. Recently, it was observed that the N-type calcium channel v2.2 (Cav2.2) encoded by CACNA1B gene is important in Angiotensin II signal transduction for corticosteroid biosynthesis in NCI-H295R adrenocortical carcinoma cells. Indeed, TCF21 inhibits CACNA1B and Cav2.2 expression in NCI-H295R. In addition, in a cohort of 55 adult patients with adrenocortical tumor, CACNA1B expression was higher in ACC than ACA, and was related to poor disease-free survival in ACC patients. These results suggest a mechanism of steroidogenesis control by TCF21 in adrenocortical tumor cells, in addition to the control observed through SF1 inhibition. Importantly, steroid production could impair tumor immunogenicity, contributing to the immune resistance described in adrenal cancer. Overall design: Fragmented DNA without immunoprecipitation was used as a positive control, and two replicates of NCI-H295R pCMVMycPOD1 cells were sequenced. Reads were mapped using GRCh38 Homo sapiens (human) genome assembly. The Phred Quality Score (Q>30) of the bases was verified, low-quality and duplicate readings were removed, and Peak calling was performed using MACS2 (v2.1.1) (p<0.01) wrapped in AQUAS ChIP-Seq pipeline (https://github.com/NHLBI-BCB/TF_chipseq_pipeline, accessed on 22/Nov/2020) (Zhang et al., 2008). The IDR <0.05 (Irreproducible Discovery Rate) was calculated to identify consistent peaks, which guarantees the reproducibility of the experiment (Li et al., 2011). All readings considered as noise by the blacklist of the Encyclopedia of DNA Elements (ENCODE) were excluded (Amemiya et al., 2019). GSE179308 pubmed 35603952