(D) Brightfield images of KP4 cell morphology when grown in 2D monolayer following KO of KO cell lines. elife-45313-fig6-data1.xlsx (19K) DOI:?10.7554/eLife.45313.020 Physique 7source data 1: OPN loss impairs growth of basal-like PDA. elife-45313-fig7-data1.xlsx (16K) DOI:?10.7554/eLife.45313.022 Supplementary file 1: Human and mouse primer sequences Nandrolone propionate used in the study. elife-45313-supp1.docx (184K) DOI:?10.7554/eLife.45313.023 Supplementary file 2: Basal-like and classical gene signatures. List of genes associated with the classical and basal-like gene signatures and their expression in the corresponding Moffitt, Collisson and Bailey signatures. elife-45313-supp2.docx (113K) DOI:?10.7554/eLife.45313.024 Transparent reporting form. elife-45313-transrepform.docx (247K) DOI:?10.7554/eLife.45313.025 Data Availability StatementSequencing data from Determine 3 have been deposited in GEO under accession code “type”:”entrez-geo”,”attrs”:”text”:”GSE131222″,”term_id”:”131222″GSE131222. The following dataset was generated: Adams CR, Htwe HH, Marsh T, Wang AL, Montoya ML, Tward AD, Bardeesy N, Perera R. 2019. Gene expression changes associated with induction of GLI2 in human PDA cells. NCBI Gene Expression Omnibus. GSE131222 Abstract Pancreatic ductal adenocarcinoma (PDA) is usually a heterogeneous disease comprised of a basal-like subtype with mesenchymal gene signatures, undifferentiated histopathology and worse prognosis compared to the classical subtype. Despite their prognostic and therapeutic value, the key drivers that establish and control subtype identity remain unknown. Here, we demonstrate that PDA subtypes are not permanently encoded, and identify the GLI2 transcription factor as a grasp regulator of subtype inter-conversion. GLI2 is usually elevated in basal-like PDA lines and patient specimens, and forced GLI2 activation is sufficient to convert classical PDA cells to basal-like. Mechanistically, GLI2 upregulates expression of the pro-tumorigenic secreted protein, Osteopontin (OPN), which is especially critical for metastatic growth in vivo and adaptation to oncogenic KRAS ablation. Accordingly, elevated GLI2 and OPN levels Nandrolone propionate predict shortened overall survival of PDA patients. Thus, the GLI2-OPN circuit is usually a driver of PDA cell plasticity that establishes and maintains an aggressive variant of this disease. in?~95% of PDA and inactivating mutations Nandrolone propionate or deletions of in 50C70% (Jones et al., 2008; Biankin et al., 2012; Ryan et al., 2014; Waddell et al., 2015; Witkiewicz et al., 2015). Recently, transcriptional profiling from resected PDA specimens has identified two main subtypes with distinct molecular features, termed classical and basal-like (Collisson et al., 2011; Moffitt et al., 2015; Bailey et al., 2016). Classical Lamp3 PDA is usually enriched for expression of epithelial differentiation genes, whereas basal-like PDA is usually characterized by laminin and basal keratin gene expression, stem cell and epithelial-to-mesenchymal transition (EMT) markers, analogous to the basal subtypes previously defined in bladder and breast cancers (Perou et al., 2000; Parker et al., 2009; Curtis Nandrolone propionate et al., 2012; Cancer Genome Atlas Research Network, 2014; Damrauer et al., 2014). Importantly, basal-like subtype tumors display poorly differentiated histological features and correlate with markedly worse prognosis (Moffitt et al., 2015; Cancer Genome Atlas Research Network, 2017; Aung et al., 2018). These subtypes are preserved in different experimental models of PDA including organoids (Boj et al., 2015; Huang et al., 2015; Seino et al., 2018), cell line cultures (Collisson et al., 2011; Moffitt et al., 2015; Martinelli et al., 2017), and a genetically engineered mouse (GEM) model of PDA in which ablation of oncogenic Kras resulted in subtype conversion (Kapoor et al., 2014). However, the identity of key factors responsible for establishing and maintaining subtype specificity and how these programs integrate with pathways known to be deregulated in PDA remain largely unknown. The Hedgehog (Hh) pathway is usually activated in PDA and?has been found to play important and complex roles in PDA pathogenesis (Morris et al., 2010). Whereas the developing and normal adult pancreas lack expression of Hh pathway ligands, the Sonic Hedgehog (SHH) and Indian Hedgehog (IHH) ligands are prominently induced in the pancreatic epithelium upon injury and throughout PDA development, from early precursor pancreatic intraepithelial neoplasia (PanIN) to invasive disease (Berman et al., 2003; Thayer et al., 2003; Prasad et al., 2005; Nolan-Stevaux et al., 2009). The neoplastic cells and stromal fibroblasts also express the Hh receptor Smoothened (SMO) and the Glioma-associated oncogene.
