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Learn MoreRNA sequencing data of SPINK7 gene silencing (SPINK7) as compared to Non silencing control (NSC) treatment in EPC2 cells differentiated in ALI culture for 14 days and RNA sequencing data of NSC cells before differentiation.; We show that a serine protease inhibitor SPINK7, is part of the differentiation program of human esophageal squamous epithelium and that SPINK7 depletion occurs in a human allergic, inflammatory, esophageal condition termed eosinophilic esophagitis (EoE). Herein, we employed experimental manipulation strategies designed to reduce SPINK7 in vitro in order to assess the transcriptional changes that are associates with the loss of SPINK7. A whole-transcriptome sequencing analysis of EPC2 cells after ALI differentiation revealed 270 genes that were differentially expressed in the SPINK7-deficient cells compared to NSC-treated cells (p < 0.05, fold change >2, RPKM >1). The modified genes were enriched for those involved in epidermal differentiation, inflammation and skin physiology including known differentiation markers such as FLG, FLG2, LOR, and keratins. These data suggest that loss of SPINK7 promote an un-differentiated phenotype. We further analyzed the transcription profile of differentiated cells as compared to un-differentiated cells. This analysis revealed 3225 differentially expressed genes (p < 0.05, fold change >2, RPKM >1). The majority of the genes modified by SPINK7 deficiency (77%) were also modulated during epithelial cell differentiation. Importantly, the genes that were most overexpressed during differentiation were downregulated after SPINK7 silencing, indicating that loss of SPINK7 promotes transcriptional changes that result in an undifferentiated epithelial phenotype.; We propose that loss of SPINK7 is a key checkpoint in regulating mucosal differentiation, barrier function, and inflammatory esophageal responses.; mRNA profile of SPINK7 silenced cells and control cells before and after differentiation in the ALI differentiation SOURCE: Nurit,Pereg,Azouz (nurit.azouz@cchmc.org) - Rothenberg Cincinnati Childrens Hospital Medical center
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