Journal ArticleDOI
Monitoring Drug Target Engagement in Cells and Tissues Using the Cellular Thermal Shift Assay
Daniel Martinez Molina,Rozbeh Jafari,Marina Ignatushchenko,Takahiro Seki,E. Andreas Larsson,Chen Dan,Lekshmy Sreekumar,Yihai Cao,Yihai Cao,Pär Nordlund,Pär Nordlund +10 more
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TLDR
This cellular thermal shift assay (CETSA) is based on the biophysical principle of ligand-induced thermal stabilization of target proteins and validated drug binding for a set of important clinical targets and monitored processes of drug transport and activation, off-target effects and drug resistance in cancer cell lines, as well as drug distribution in tissues.Abstract:
The efficacy of therapeutics is dependent on a drug binding to its cognate target. Optimization of target engagement by drugs in cells is often challenging, because drug binding cannot be monitored inside cells. We have developed a method for evaluating drug binding to target proteins in cells and tissue samples. This cellular thermal shift assay (CETSA) is based on the biophysical principle of ligand-induced thermal stabilization of target proteins. Using this assay, we validated drug binding for a set of important clinical targets and monitored processes of drug transport and activation, off-target effects and drug resistance in cancer cell lines, as well as drug distribution in tissues. CETSA is likely to become a valuable tool for the validation and optimization of drug target engagement.read more
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Drug repurposing: progress, challenges and recommendations
Sudeep Pushpakom,Francesco Iorio,Patrick A. Eyers,K. Jane Escott,Shirley Hopper,Andrew D. Wells,Andrew J. Doig,Tim Guilliams,Joanna Latimer,Christine J. McNamee,Alan Norris,Philippe Sanseau,David Cavalla,Munir Pirmohamed +13 more
TL;DR: Approaches used for drug repurposing (also known as drug repositioning) are presented, the challenges faced by the repurpose community are discussed, and innovative ways by which these challenges could be addressed are recommended to help realize the full potential of drugRepurposing.
Journal ArticleDOI
The cellular thermal shift assay for evaluating drug target interactions in cells
Rozbeh Jafari,Helena Almqvist,Hanna Axelsson,Marina Ignatushchenko,Thomas Lundbäck,Pär Nordlund,Daniel Martinez Molina +6 more
TL;DR: The cellular thermal shift assay (CETSA) allows studies of target engagement of drug candidates in a cellular context, herein exemplified with experimental data on the human kinases p38α and ERK1/2.
Journal ArticleDOI
Tracking cancer drugs in living cells by thermal profiling of the proteome
Mikhail M. Savitski,Friedrich B M Reinhard,Holger Franken,Thilo Werner,Maria Fälth Savitski,Dirk Eberhard,Daniel Martinez Molina,Rozbeh Jafari,Rebecca Dovega,Susan Klaeger,Susan Klaeger,Bernhard Kuster,Bernhard Kuster,Pär Nordlund,Pär Nordlund,Marcus Bantscheff,Gerard Drewes +16 more
TL;DR: Thermal profiling of cellular proteomes enables the differential assessment of protein ligand binding and other protein modifications, providing an unbiased measure of drug-target occupancy for multiple targets and facilitating the identification of markers for drug efficacy and toxicity.
Journal ArticleDOI
Drugging the p53 pathway: understanding the route to clinical efficacy
TL;DR: The current state of the development of p53 pathway modulators and new pathway targets that have emerged are described.
Journal ArticleDOI
The promise and peril of chemical probes.
Cheryl H. Arrowsmith,James E. Audia,Christopher M. Austin,Jonathan B. Baell,Jonathan Bennett,Julian Blagg,C. Bountra,Paul Brennan,Peter Brown,Mark E. Bunnage,Carolyn Buser-Doepner,Robert M. Campbell,Adrian Carter,Philip Cohen,Robert A. Copeland,Ben Cravatt,Jayme L. Dahlin,Dashyant Dhanak,Aled M. Edwards,Mathias Frederiksen,Stephen V. Frye,Nathanael S. Gray,Charles E. Grimshaw,David Hepworth,Trevor Howe,Kilian Huber,Jian Jin,Stefan Knapp,Joanne Kotz,Ryan G. Kruger,Derek B. Lowe,Mary M. Mader,Brian D. Marsden,Anke Mueller-Fahrnow,Susanne Müller,Ronan C. O'Hagan,John P. Overington,Dafydd R. Owen,Saul H Rosenberg,Bryan L. Roth,Ruth A. Ross,Matthieu Schapira,Stuart L. Schreiber,Brian K. Shoichet,Michael Sundström,Giulio Superti-Furga,Jack Taunton,Leticia Toledo-Sherman,Chris Walpole,Michael A. Walters,Timothy M. Willson,Paul Workman,Robert N. Young,William J. Zuercher +53 more
TL;DR: A community-driven wiki resource to improve quality and convey current best practice on chemical probes, and to help address shortcomings of poor quality or that are used incorrectly generate misleading results.
References
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TL;DR: The origins, challenges and solutions of NIH Image and ImageJ software are discussed, and how their history can serve to advise and inform other software projects.
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Cell cycle, CDKs and cancer: a changing paradigm
TL;DR: Genetic evidence suggests that tumour cells may also require specific interphase CDKs for proliferation, and selective CDK inhibition may provide therapeutic benefit against certain human neoplasias.
Journal ArticleDOI
Clinical efficacy of a RAF inhibitor needs broad target blockade in BRAF -mutant melanoma
Gideon Bollag,Peter Hirth,James Tsai,Jiazhong Zhang,Prabha N. Ibrahim,Hanna Cho,Wayne Spevak,Chao Zhang,Ying Zhang,Gaston Habets,Elizabeth A. Burton,Bernice Wong,Garson Tsang,Brian L. West,Ben Powell,Rafe Shellooe,Adhirai Marimuthu,Hoa Nguyen,Kam Y. J. Zhang,Dean R. Artis,Joseph Schlessinger,Fei Su,Brian Higgins,Raman Mahadevan Iyer,Kurt D'Andrea,Astrid Koehler,Michael Stumm,Paul S. Lin,Richard J. Lee,Joseph F. Grippo,Igor Puzanov,Kevin B. Kim,Antoni Ribas,Grant A. McArthur,Jeffrey A. Sosman,Paul B. Chapman,Keith T. Flaherty,Xiaowei Xu,Katherine L. Nathanson,K. B. Nolop +39 more
TL;DR: The structure-guided discovery of PLX4032 (RG7204), a potent inhibitor of oncogenic B-RAF kinase activity, and a remarkably high 81% response rate in metastatic melanoma patients treated at an oral dose of 960 mg twice daily are described, demonstrating that BRAF-mutant melanomas are highly dependent on B- RAF kinases activity.
Journal ArticleDOI
Principles of Cancer Therapy: Oncogene and Non-oncogene Addiction
TL;DR: Evidence is presented for a large class of non-oncogenes that are essential for cancer cell survival and present attractive drug targets and theoretical considerations for combining orthogonal cancer therapies are provided.