Concerted cell and in vivo screen for pancreatic ductal adenocarcinoma (PDA) chemotherapeutics

• Published in: Scientific reports 2020-11, Vol.10 (1), p.20662, Article 20662
• Main Authors: Layeghi-Ghalehsoukhteh, Somayeh, Pal Choudhuri, Shreoshi, Ocal, Ozhan, Zolghadri, Yalda, Pashkov, Victor, Niederstrasser, Hanspeter, Posner, Bruce A., Kantheti, Havish S., Azevedo-Pouly, Ana C., Huang, Huocong, Girard, Luc, MacDonald, Raymond J., Brekken, Rolf A., Wilkie, Thomas M.
• Format: Article
• Language: English
• Subjects: 631/67; 631/67/1059; 631/67/1059/99; 631/67/1504/1713; 631/67/70; Acinar Cells - drug effects; Acinar Cells - metabolism; Acinar Cells - pathology; Adenocarcinoma; Adenocarcinoma - drug therapy; Adenocarcinoma - metabolism; Adenocarcinoma - pathology; Animal models; Animals; Antineoplastic Agents - pharmacology; Calcium (intracellular); Calcium - metabolism; Calcium signalling; Carcinogenesis - drug effects; Carcinogenesis - metabolism; Carcinogenesis - pathology; Carcinoma, Pancreatic Ductal - drug therapy; Carcinoma, Pancreatic Ductal - metabolism; Carcinoma, Pancreatic Ductal - pathology; Cell culture; Cell Dedifferentiation - drug effects; Cell Transformation, Neoplastic - drug effects; Cell Transformation, Neoplastic - metabolism; Cells, Cultured; Ceruletide - metabolism; Cytotoxicity; Deoxycytidine - analogs & derivatives; Deoxycytidine - pharmacology; Disease Progression; Epigenetics; Gemcitabine; Genetic engineering; GTP-Binding Proteins - metabolism; Histone deacetylase; Histone Deacetylase Inhibitors - pharmacology; Humanities and Social Sciences; Intracellular signalling; Lesions; Mice; multidisciplinary; Mutation; Pancreas; Pancreatic cancer; Pancreatic Ducts - drug effects; Pancreatic Ducts - metabolism; Pancreatic Neoplasms; Pancreatic Neoplasms - drug therapy; Pancreatic Neoplasms - metabolism; Pancreatic Neoplasms - pathology; Pancreatitis; Pancreatitis - drug therapy; Pancreatitis - metabolism; Point mutation; Proto-Oncogene Proteins p21(ras) - metabolism; RGS Proteins - metabolism; Science; Science (multidisciplinary); Signal Transduction - drug effects; Tumor suppressor genes
• ISSN: 2045-2322; 2045-2322
• DOI: 10.1038/s41598-020-77373-8

PDA is a major cause of US cancer-related deaths. Oncogenic Kras presents in 90% of human PDAs. Kras mutations occur early in pre-neoplastic lesions but are insufficient to cause PDA. Other contributing factors early in disease progression include chronic pancreatitis, alterations in epigenetic regulators, and tumor suppressor gene mutation. GPCRs activate heterotrimeric G-proteins that stimulate intracellular calcium and oncogenic Kras signaling, thereby promoting pancreatitis and progression to PDA. By contrast, Rgs proteins inhibit Gi/q-coupled GPCRs to negatively regulate PDA progression. Rgs16::GFP is expressed in response to caerulein-induced acinar cell dedifferentiation, early neoplasia, and throughout PDA progression. In genetically engineered mouse models of PDA, Rgs16::GFP is useful for pre-clinical rapid in vivo validation of novel chemotherapeutics targeting early lesions in patients following successful resection or at high risk for progressing to PDA. Cultured primary PDA cells express Rgs16::GFP in response to cytotoxic drugs. A histone deacetylase inhibitor, TSA, stimulated Rgs16::GFP expression in PDA primary cells, potentiated gemcitabine and JQ1 cytotoxicity in cell culture, and Gem + TSA + JQ1 inhibited tumor initiation and progression in vivo. Here we establish the use of Rgs16::GFP expression for testing drug combinations in cell culture and validation of best candidates in our rapid in vivo screen.