SRPKIN-1: A Covalent SRPK1/2 Inhibitor that Potently Converts VEGF from Pro-angiogenic to Anti-angiogenic Isoform

John M Hatcher; Guowei Wu; Chuyue Zeng; Jie Zhu; Fan Meng; Sherrina Patel; Wenqiu Wang; Scott B Ficarro; Alan L Leggett; Chelsea E Powell; Jarrod A Marto; Kang Zhang; Jacky Chi Ki Ngo; Xiang-Dong Fu; Tinghu Zhang; Nathanael S Gray

doi:10.1016/j.chembiol.2018.01.013

SRPKIN-1: A Covalent SRPK1/2 Inhibitor that Potently Converts VEGF from Pro-angiogenic to Anti-angiogenic Isoform

Cell Chem Biol. 2018 Apr 19;25(4):460-470.e6. doi: 10.1016/j.chembiol.2018.01.013. Epub 2018 Feb 22.

Authors

John M Hatcher¹, Guowei Wu², Chuyue Zeng³, Jie Zhu⁴, Fan Meng², Sherrina Patel⁵, Wenqiu Wang⁵, Scott B Ficarro⁶, Alan L Leggett⁷, Chelsea E Powell¹, Jarrod A Marto⁶, Kang Zhang⁵, Jacky Chi Ki Ngo³, Xiang-Dong Fu⁸, Tinghu Zhang⁹, Nathanael S Gray¹⁰

Affiliations

¹ Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA 02115, USA; Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA.
² Department of Cellular and Molecular Medicine, Institute of Genomic Medicine, University of California, San Diego, La Jolla, CA 92093, USA.
³ School of Life Sciences, The Chinese University of Hong Kong, Shatin, Hong Kong Special Administrative Region NA, China.
⁴ Shiley Eye Institute, Institute for Engineering in Medicine, Institute for Genomic Medicine, University of California, San Diego, La Jolla, CA 92093, USA; Guangzhou Women and Children's Medical Center, Guangzhou Medical University, Guangzhou 510623, China.
⁵ Shiley Eye Institute, Institute for Engineering in Medicine, Institute for Genomic Medicine, University of California, San Diego, La Jolla, CA 92093, USA.
⁶ Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA 02115, USA; Department of Oncologic Pathology and Blais Proteomics Center, Dana-Farber Cancer Institute, Boston, MA 02115, USA; Department of Pathology, Brigham and Women's Hospital, Harvard Medical School, Boston, MA 02115, USA.
⁷ Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA 02115, USA.
⁸ Department of Cellular and Molecular Medicine, Institute of Genomic Medicine, University of California, San Diego, La Jolla, CA 92093, USA. Electronic address: xdfu@ucsd.edu.
⁹ Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA 02115, USA; Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA. Electronic address: tinghu_zhang@dfci.harvard.edu.
¹⁰ Department of Cancer Biology, Dana-Farber Cancer Institute, Boston, MA 02115, USA; Department of Biological Chemistry and Molecular Pharmacology, Harvard Medical School, Boston, MA 02115, USA. Electronic address: nathanael_gray@dfci.harvard.edu.

Abstract

The SRPK family of kinases regulates pre-mRNA splicing by phosphorylating serine/arginine (SR)-rich splicing factors, signals splicing control in response to extracellular stimuli, and contributes to tumorigenesis, suggesting that these splicing kinases are potential therapeutic targets. Here, we report the development of the first irreversible SRPK inhibitor, SRPKIN-1, which is also the first kinase inhibitor that forms a covalent bond with a tyrosine phenol group in the ATP-binding pocket. Kinome-wide profiling demonstrates its selectivity for SRPK1/2, and SRPKIN-1 attenuates SR protein phosphorylation at submicromolar concentrations. Vascular endothelial growth factor (VEGF) is a known target for SRPK-regulated splicing and, relative to the first-generation SRPK inhibitor SRPIN340 or small interfering RNA-mediated SRPK knockdown, SRPKIN-1 is more potent in converting the pro-angiogenic VEGF-A165a to the anti-angiogenic VEGF-A165b isoform and in blocking laser-induced neovascularization in a murine retinal model. These findings encourage further development of SRPK inhibitors for treatment of age-related macular degeneration.

Keywords: SRPK1/2; SRPKIN-1; VEGF; alternative splicing; irreversible inhibitor.

Publication types

Research Support, N.I.H., Extramural
Research Support, Non-U.S. Gov't

MeSH terms

Alternative Splicing / drug effects
Angiogenesis Inhibitors / chemistry
Angiogenesis Inhibitors / pharmacology*
Angiogenesis Inhibitors / therapeutic use
Animals
Cell Line
HeLa Cells
Humans
Mice, Inbred C57BL
Molecular Docking Simulation
Neovascularization, Pathologic / drug therapy
Neovascularization, Pathologic / genetics
Neovascularization, Pathologic / metabolism
Protein Isoforms / antagonists & inhibitors
Protein Isoforms / chemistry
Protein Isoforms / metabolism
Protein Kinase Inhibitors / chemistry
Protein Kinase Inhibitors / pharmacology*
Protein Kinase Inhibitors / therapeutic use
Protein Serine-Threonine Kinases / antagonists & inhibitors*
Protein Serine-Threonine Kinases / chemistry
Protein Serine-Threonine Kinases / metabolism
Vascular Endothelial Growth Factor A / genetics
Vascular Endothelial Growth Factor A / metabolism*

Substances

Angiogenesis Inhibitors
Protein Isoforms
Protein Kinase Inhibitors
Vascular Endothelial Growth Factor A
SRPK1 protein, human
Protein Serine-Threonine Kinases
SRPK2 protein, human

Abstract

Publication types

MeSH terms

Substances

Grants and funding