@article{0c4d3a2884b34e63aae7dd6297589fc5,
title = "Chemical Biology Toolkit for DCLK1 Reveals Connection to RNA Processing",
abstract = "Doublecortin-like kinase 1 (DCLK1) is critical for neurogenesis, but overexpression is also observed in multiple cancers and is associated with poor prognosis. Nevertheless, the function of DCLK1 in cancer, especially the context-dependent functions, are poorly understood. We present a “toolkit” that includes the DCLK1 inhibitor DCLK1-IN-1, a complementary DCLK1-IN-1-resistant mutation G532A, and kinase dead mutants D511N and D533N, which can be used to investigate signaling pathways regulated by DCLK1. Using a cancer cell line engineered to be DCLK1 dependent for growth and cell migration, we show that this toolkit can be used to discover associations between DCLK1 kinase activity and biological processes. In particular, we show an association between DCLK1 and RNA processing, including the identification of CDK11 as a potential substrate of DCLK1 using phosphoproteomics.",
keywords = "cancer, doublecortin-like kinase 1, kinase inhibitor, mass spectrometry, phosphoproteomics, signal transduction, structural biology",
author = "Yan Liu and Ferguson, {Fleur M.} and Lianbo Li and Miljan Kuljanin and Mills, {Caitlin E.} and Kartik Subramanian and Wayne Harshbarger and Sudershan Gondi and Jinhua Wang and Sorger, {Peter K.} and Mancias, {Joseph D.} and Gray, {Nathanael S.} and Westover, {Kenneth D.}",
note = "Funding Information: We gratefully acknowledge Dr. Steven Gygi for use of CORE for mass spectrometry data analysis software. This research has been supported by a grant from the Hale Family Center for Pancreatic Cancer Research to J.D.M. and N.S.G. American Cancer Society Award 132205-RSG-18-039-01-DMC (to K.D.W.); Cancer Prevention and Research Institute of Texas RP170373 (to K.D.W); Welch Foundation Grant I1829 (to K.D.W.); 2017 AACR -Bayer Innovation and Discovery grant no. 17-80-44-GRAY (to N.S.G.); DF/HCC GI SPORE Developmental Research Project Award P50CA127003 (to N.S.G.); NIH grant 5U01CA207160 (to S.J.C., E.N.M., and A.T.); NIH U54-HL127365 and U24-DK116204 (to P.K.S.). We thank Samantha Westover for drawings in the graphical abstract. We thank Isabelle Lucet and colleagues for discussions regarding the G532V mutation. Funding Information: F.M.F. and N.S.G. are inventors on a patent application related to the DCLK1 inhibitors described in this manuscript (WO/2018/075608). K.D.W. has received consulting fees from Sanofi Oncology and is a member of the SAB for Vibliome Therapeutics. K.D.W. declares that none of these relationships are directly or indirectly related to the content of this manuscript. N.S.G. is a Scientific Founder, member of the SAB, and equity holder in C4 Therapeutics, Syros, Soltego (board member), B2S, Aduro Gatekeeper, and Petra Pharmaceuticals. The Gray lab receives or has received research funding from Novartis , Takeda , Astellas , Taiho , Janssen , Kinogen , Voroni , Her2llc , Deerfield , and Sanofi . P.K.S. is a member of the SAB or Board of Directors of Applied Biomath and RareCyte, Inc., and has equity in these companies. P.K.S. has received research funding from Novartis and Merck in the last 5 years. P.K.S. declares that none of these relationships are directly or indirectly related to the content of this manuscript. P.K.S. is a current employee of Bristol-Myers Squibb. Funding Information: We gratefully acknowledge Dr. Steven Gygi for use of CORE for mass spectrometry data analysis software. This research has been supported by a grant from the Hale Family Center for Pancreatic Cancer Research to J.D.M. and N.S.G. American Cancer Society Award 132205-RSG-18-039-01-DMC (to K.D.W.); Cancer Prevention and Research Institute of Texas RP170373 (to K.D.W); Welch Foundation Grant I1829 (to K.D.W.); 2017 AACR-Bayer Innovation and Discovery grant no. 17-80-44-GRAY (to N.S.G.); DF/HCC GI SPORE Developmental Research Project Award P50CA127003 (to N.S.G.); NIH grant 5U01CA207160 (to S.J.C. E.N.M. and A.T.); NIH U54-HL127365 and U24-DK116204 (to P.K.S.). We thank Samantha Westover for drawings in the graphical abstract. We thank Isabelle Lucet and colleagues for discussions regarding the G532V mutation. K.D.W. and Y.L. conceived and led the study. Y.L. F.M.F. J.D.M. L.L. M.K. W.H. S.G. J.W. C.E.M. K.S. P.K.S. N.S.G. and W.H. designed and conducted the experiments, analyzed and interpreted the data. J.D.M. K.D.W. and N.S.G. supervised aspects of the project. K.D.W. and Y.L. wrote the manuscript, with edits from F.M.F. J.D.M, and N.S.G. All authors read and approved the manuscript. F.M.F. and N.S.G. are inventors on a patent application related to the DCLK1 inhibitors described in this manuscript (WO/2018/075608). K.D.W. has received consulting fees from Sanofi Oncology and is a member of the SAB for Vibliome Therapeutics. K.D.W. declares that none of these relationships are directly or indirectly related to the content of this manuscript. N.S.G. is a Scientific Founder, member of the SAB, and equity holder in C4 Therapeutics, Syros, Soltego (board member), B2S, Aduro Gatekeeper, and Petra Pharmaceuticals. The Gray lab receives or has received research funding from Novartis, Takeda, Astellas, Taiho, Janssen, Kinogen, Voroni, Her2llc, Deerfield, and Sanofi. P.K.S. is a member of the SAB or Board of Directors of Applied Biomath and RareCyte, Inc. and has equity in these companies. P.K.S. has received research funding from Novartis and Merck in the last 5 years. P.K.S. declares that none of these relationships are directly or indirectly related to the content of this manuscript. P.K.S. is a current employee of Bristol-Myers Squibb. Publisher Copyright: {\textcopyright} 2020 Elsevier Ltd",
year = "2020",
month = oct,
day = "15",
doi = "10.1016/j.chembiol.2020.07.011",
language = "English (US)",
volume = "27",
pages = "1229--1240.e4",
journal = "Cell Chemical Biology",
issn = "2451-9448",
publisher = "Elsevier Inc.",
number = "10",
}