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Genome-wide CRISPR screens reveal a Wnt-FZD5 signaling circuit as a druggable vulnerability of RNF43-mutant pancreatic tumors.
- Source :
-
Nature medicine [Nat Med] 2017 Jan; Vol. 23 (1), pp. 60-68. Date of Electronic Publication: 2016 Nov 21. - Publication Year :
- 2017
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Abstract
- Forward genetic screens with CRISPR-Cas9 genome editing enable high-resolution detection of genetic vulnerabilities in cancer cells. We conducted genome-wide CRISPR-Cas9 screens in RNF43-mutant pancreatic ductal adenocarcinoma (PDAC) cells, which rely on Wnt signaling for proliferation. Through these screens, we discovered a unique requirement for a Wnt signaling circuit: engaging FZD5, one of the ten Frizzled receptors encoded in the human genome. Our results uncover an underappreciated level of context-dependent specificity at the Wnt receptor level. We further derived a panel of recombinant antibodies that reports the expression of nine FZD proteins and confirms that FZD5 functional specificity cannot be explained by protein expression patterns. Additionally, antibodies that specifically bind FZD5 and FZD8 robustly inhibited the growth of RNF43-mutant PDAC cells grown in vitro and as xenografts in vivo, providing orthogonal support for the functional specificity observed genetically. Proliferation of a patient-derived PDAC cell line harboring an RNF43 variant was also selectively inhibited by the FZD5 antibodies, further demonstrating their use as a potential targeted therapy. Tumor organoid cultures from colorectal carcinoma patients that carried RNF43 mutations were also sensitive to the FZD5 antibodies, highlighting the potential generalizability of these findings beyond PDAC. Our results show that CRIPSR-based genetic screens can be leveraged to identify and validate cell surface targets for antibody development and therapy.
- Subjects :
- Adenocarcinoma genetics
Adenocarcinoma metabolism
Animals
Carcinoma, Pancreatic Ductal metabolism
Cell Line, Tumor
Clustered Regularly Interspaced Short Palindromic Repeats
Colorectal Neoplasms metabolism
Flow Cytometry
Fluorescent Antibody Technique
Frizzled Receptors metabolism
Humans
Mice
Mice, SCID
Molecular Targeted Therapy
Neoplasm Transplantation
Organoids drug effects
Organoids metabolism
Pancreatic Neoplasms metabolism
RNA, Messenger metabolism
Real-Time Polymerase Chain Reaction
Reverse Transcriptase Polymerase Chain Reaction
Ubiquitin-Protein Ligases
Wnt Signaling Pathway genetics
Antibodies pharmacology
Carcinoma, Pancreatic Ductal genetics
Cell Cycle Checkpoints drug effects
Colorectal Neoplasms genetics
DNA-Binding Proteins genetics
Frizzled Receptors antagonists & inhibitors
Oncogene Proteins genetics
Pancreatic Neoplasms genetics
Wnt Signaling Pathway drug effects
Subjects
Details
- Language :
- English
- ISSN :
- 1546-170X
- Volume :
- 23
- Issue :
- 1
- Database :
- MEDLINE
- Journal :
- Nature medicine
- Publication Type :
- Academic Journal
- Accession number :
- 27869803
- Full Text :
- https://doi.org/10.1038/nm.4219