Yap/Taz inhibit goblet cell fate to maintain lung epithelial homeostasis (RNA-seq)

Proper lung function relies on precisely balanced numbers of specialized epithelial cell types that work together and are maintained in homeostasis. In this study we have described essential roles for the transcriptional regulators YAP and TAZ, which are key effectors of Hippo pathway signaling, in maintaining lung epithelial homeostasis. Phenotypes associated with Yap/Taz deletion include alveolar defects and a striking development of goblet cell metaplasia throughout the airways. Knockdown of YAP and TAZ in HBECs similarly drives mucin expression. The TEAD family of transcription factors are well characterized partners of the YAP/TAZ transcriptional effectors, and we have found that knockdown of TEAD1-4 similarly drives elevated mucin expression. In order to further understand the role of the TEAD transcription factors in human lung epithelial cell fate, we conducted TEAD Chromatin Immunopreciptitation (ChIP)-Sequencing. Overall design: HBECs were cultured in PneumaCult-Ex Plus and transfected with a pool of 3 unique controls or dual Yap/Taz targeting siRNAs. All experiments were performed in triplicate and cells were collected 48 hours after transfection. Total RNA was extracted using the RNeasy Mini Kit (Qiagen). Libraries were prepared using the Illumina TruSeq RNA Sample Preparation Kit v2. The libraries from individual samples were pooled in groups of 15 for cluster generation on the Illumina cBot using Illumina TruSeq Single-Read Cluster Kit and then sequenced on an Illumina NextSeq 500 instrument.