Designed Ankyrin Repeat Protein; RAS; cancer; drug development; oncogene; small GTPase; Molecular Medicine; Oncology; Genetics; Cancer Research
Abstract :
[en] The KRAS oncoprotein is a frequent tumor driver in lung, pancreatic, and colorectal cancers and has proven to be a challenging pharmaceutical target. The first KRAS-targeted therapeutics are now being tested in clinical trials but the consequences of preferentially targeting the GDP or GTP state of KRAS and the relevance of RAS nanoclustering have remained unclear. Here we report a Designed Ankyrin Repeat Protein (DARPin) that recognizes the RAS switch I/II region with low nm affinity, independently of the nucleotide bound (GDP- or GTP state). This DARPin, termed '784_F5', occupies the effector recognition lobe, resulting in interference with SOS-mediated activation, RAS downstream effector interactions, and KRAS nanoclustering. Consequently, this anti-RAS DARPin potently blocks downstream signaling, leading to a strong reduction in proliferation and anchorage-independent growth in RAS-dependent cell lines. We showed that the expression of '784_F5', the pan-RAS, nucleotide-independent DARPin can lead to tumor regression in a colorectal xenograft model which may hold promise for further investigation and development.
Disciplines :
Oncology
Author, co-author :
Kapp, Jonas N; Department of Biochemistry, University of Zurich, Switzerland
Verdurmen, Wouter P R; Department of Biochemistry, University of Zurich, Switzerland
Schaefer, Jonas V; Department of Biochemistry, University of Zurich, Switzerland
Kopra, Kari ; Department of Chemistry, University of Turku, Finland
Nagy-Davidescu, Gabriela; Department of Biochemistry, University of Zurich, Switzerland
Richard, Elodie; Bordeaux Institute of Oncology (BRIC), INSERM U1312, University of Bordeaux, France
Nokin, Marie-Julie ; Université de Liège - ULiège > Département des sciences biomédicales et précliniques ; ACTION Laboratory, IECB, INSERM U1218, University of Bordeaux, France
Ernst, Patrick; Department of Biochemistry, University of Zurich, Switzerland
Tamaskovic, Rastislav; Department of Biochemistry, University of Zurich, Switzerland
Schwill, Martin; Department of Biochemistry, University of Zurich, Switzerland
Degen, Ralph; Department of Biochemistry, University of Zurich, Switzerland
Scholl, Claudia; Division of Applied Functional Genomics, German Cancer Research Center (DKFZ), Heidelberg, Germany ; National Center for Tumor Diseases (NCT), Heidelberg, Germany
Santamaria, David; ACTION Laboratory, IECB, INSERM U1218, University of Bordeaux, France ; Centro de Investigación del Cáncer, CSIC-Universidad de Salamanca, Spain
Plückthun, Andreas ; Department of Biochemistry, University of Zurich, Switzerland
Research Council of Finland KFS - Swiss Cancer Research Foundation
Funding text :
This research was funded by Swiss Cancer Research foundation, grant number KFS 4147\u201002\u20102017 and KFS\u20105290\u201002\u20102021\u2010R to AP and the Research Council of Finland (296225/KK, 323433/KK, 329012/KK, and 353324/KK). We thank Susanne M\u00FCller\u2010Knapp from the Structural Genomics Consortium (SGC) in Oxford for providing the KRAS protein for the selection. We further wish to express our appreciation to Dr William Gillette and his team in the RAS Initiative at the Frederick National Laboratory, Frederick MD USA for providing the SOS and RAF1\u2010RBD proteins. The authors would like to thank Marie Groth for training and scientific discussion. The Figs 2A,B and 5A were created with BioRender.com .
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