Scale bar: 100 microns. Given that genetic deletion of and at initiation completely suppressed mucinous gastric differentiation, we evaluated expression of both transcription factors in AdSCCs. between tumor cells sorted from mice (K, n?=?3 mice) and mice (KN, n?=?3 mice) elife-38579-supp4.xlsx (8.9M) DOI:?10.7554/eLife.38579.019 Supplementary file 5: List of normal murine tissues used and their source. elife-38579-supp5.xlsx (12K) DOI:?10.7554/eLife.38579.020 Supplementary file 6: Cosine similarity table quantitating similarity between single cell clusters and each normal tissue evaluated. elife-38579-supp6.xlsx (9.2K) Patchouli alcohol DOI:?10.7554/eLife.38579.021 Transparent reporting form. elife-38579-transrepform.docx (246K) DOI:?10.7554/eLife.38579.022 Data Availability StatementAll data generated or analysed during this study are included in the manuscript and supporting files. Sequencing data will be deposited in GEO under accession codes “type”:”entrez-geo”,”attrs”:”text”:”GSE115901″,”term_id”:”115901″GSE115901. The following dataset was generated: Snyder E. 2018. FoxA1 and FoxA2 are required for gastric differentiation in NKX2-1-unfavorable lung adenocarcinoma. NCBI Gene Expression Omnibus. GSE115901 Abstract Changes in malignancy cell identity can alter malignant potential and therapeutic response. Loss of the pulmonary lineage specifier NKX2-1 augments the growth of KRAS-driven lung adenocarcinoma and causes pulmonary to gastric transdifferentiation. Here, we show that this transcription factors FoxA1 and FoxA2 are required for Rabbit Polyclonal to APOBEC4 initiation of mucinous NKX2-1-unfavorable lung adenocarcinomas in the mouse and for activation of their gastric differentiation program. deletion severely impairs tumor initiation and causes a proximal shift in cellular identity, yielding tumors expressing markers of the squamocolumnar junction of the gastrointestinal tract. In contrast, we observe downregulation of FoxA1/2 expression in the squamous component of both murine and human lung adenosquamous carcinoma. Using sequential in vivo recombination, we find that FoxA1/2 loss in established KRAS-driven neoplasia originating from SPC-positive alveolar cells induces keratinizing squamous cell carcinomas. Thus, NKX2-1, FoxA1 and FoxA2 coordinately regulate the growth and identity of lung malignancy in a context-specific manner. deletion in established tumors causes malignancy cells to shed their pulmonary identity and adopt a gastric-like differentiation state characterized by considerable mucin production and expression of multiple gastrointestinal markers, including HNF4 and Gastrokine 1. These tumors morphologically resemble a subtype of human lung cancer called invasive mucinous adenocarcinoma (IMA), which also expresses gastrointestinal markers and is predominantly driven by mutations (Guo et al., 2017). Approximately 10C15% of human lung adenocarcinomas express HNF4 with no detectable NKX2-1 (9), including both IMAs and more moderately differentiated tumors. In many of these tumors, the gene appears to be silenced by genetic and/or epigenetic mechanisms (Hwang et al., 2016; Matsubara et al., 2017). Aside from NKX2-1 itself, the Polycomb Repressive Complex 2 (PRC2) appears to play a role in suppressing mucinous differentiation in KRAS-driven, p53-deficient lung adenocarcinoma (Serresi et al., 2016). However, the precise mechanisms by which a gastric gene expression program is activated in NKX2-1-deficient tumors remain to be fully elucidated. Many of the gastrointestinal transcripts expressed in IMA are known targets of the forkhead box transcription factors FoxA1 and FoxA2 (FoxA1/2). These transcription factors govern the development of a variety of tissues and are expressed in both the adult lung and GI tract (examined in Golson and Kaestner, 2016). FoxA1/2 are also expressed in both murine and human IMA (Physique 1A and Physique 1figure product 1ACB). We previously found that deletion in autochthonous lung Patchouli alcohol tumors caused FoxA1/2 to re-localize from your regulatory elements of pulmonary-specific genes (such as (Gao et al., 2008) and (Sund et al., 2000) to abrogate their function in an autochthonous mouse model of NKX2-1-unfavorable lung adenocarcinoma. We found that FoxA1/2 are crucial and redundant regulators of both the gastric differentiation program and growth of NKX2-1-unfavorable tumors. Moreover, we found that the cellular identity adopted by tumors was highly dependent Patchouli alcohol on the context in which FoxA1/2 activity is usually lost, suggesting that a cells baseline epigenetic state can influence the identity it adopts in response to changes in lineage specifier expression. Open.