Facile target validation in an animal model with intracellularly expressed monobodies

Nature Chemical Biology
Ankit GuptaShohei Koide

Abstract

Rapidly determining the biological effect of perturbing a site within a potential drug target could guide drug discovery efforts, but it remains challenging. Here, we describe a facile target validation approach that exploits monobodies, small synthetic binding proteins that can be fully functionally expressed in cells. We developed a potent and selective monobody to WDR5, a core component of the mixed lineage leukemia (MLL) methyltransferase complex. The monobody bound to the MLL interaction site of WDR5, the same binding site for small-molecule inhibitors whose efficacy has been demonstrated in cells but not in animals. As a genetically encoded reagent, the monobody inhibited proliferation of an MLL-AF9 cell line in vitro, suppressed its leukemogenesis and conferred a survival benefit in an in vivo mouse leukemia model. The capacity of this approach to readily bridge biochemical, structural, cellular characterization and tests in animal models may accelerate discovery and validation of druggable sites.

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Citations

Dec 28, 2018·Chembiochem : a European Journal of Chemical Biology·Marcus J C LongYimon Aye
Nov 23, 2019·Breast Cancer Research : BCR·Simona PunziLuisa Lanfrancone
Apr 29, 2019·Cellular and Molecular Life Sciences : CMLS·Sascha RöthGopal P Sapkota
Oct 16, 2019·Proceedings of the National Academy of Sciences of the United States of America·Hejia Henry Wang, Andrew Tsourkas
Aug 15, 2020·Nature Communications·César Carrasco-LópezJosé L Avalos
Mar 8, 2020·Current Opinion in Structural Biology·Oliver HantschelShohei Koide
May 7, 2021·ACS Sensors·Lauren M CastleAriel L Furst
May 13, 2021·Nature Communications·Kai Wen TengShohei Koide
Apr 27, 2019·ACS Chemical Biology·Nadine Eliane SchmitOliver Hantschel

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Methods Mentioned

BETA
phage display
flow cytometry
pull-down
transfection
transgenic
size-exclusion chromatography
immunoprecipitation
PCR
ChIP

Software Mentioned

PROCHECK
Molprobity
PHENIX
ImageLab
MOLREP
COOT

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