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Gastric Cancer
Published in: Gastric Cancer 5/2020

01-09-2020 | Gastric Cancer | Original Article

Phosphoproteomic analysis identifies CLK1 as a novel therapeutic target in gastric cancer

Authors: Niraj Babu, Sneha M. Pinto, Manjusha Biswas, Tejaswini Subbannayya, Manoj Rajappa, Sonali V. Mohan, Jayshree Advani, Pavithra Rajagopalan, Gajanan Sathe, Nazia Syed, Vinod D. Radhakrishna, Oliyarasi Muthusamy, Sanjay Navani, Rekha V. Kumar, Gopal Gopisetty, Thangarajan Rajkumar, Padhma Radhakrishnan, Saravanan Thiyagarajan, Akhilesh Pandey, Harsha Gowda, Pradip Majumder, Aditi Chatterjee

Published in: Gastric Cancer | Issue 5/2020

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Abstract

Background

Phosphorylation is an important regulatory mechanism of protein activity in cells. Studies in various cancers have reported perturbations in kinases resulting in aberrant phosphorylation of oncoproteins and tumor suppressor proteins.

Methods

In this study, we carried out quantitative phosphoproteomic analysis of gastric cancer tissues and corresponding xenograft samples. Using these data, we employed bioinformatics analysis to identify aberrant signaling pathways. We further performed molecular inhibition and silencing of the upstream regulatory kinase in gastric cancer cell lines and validated its effect on cellular phenotype. Through an ex vivo technology utilizing patient tumor and blood sample, we sought to understand the therapeutic potential of the kinase by recreating the tumor microenvironment.

Results

Using mass spectrometry-based high-throughput analysis, we identified 1,344 phosphosites and 848 phosphoproteins, including differential phosphorylation of 177 proteins (fold change cut-off ≥ 1.5). Our data showed that a subset of differentially phosphorylated proteins belonged to splicing machinery. Pathway analysis highlighted Cdc2-like kinase (CLK1) as upstream kinase. Inhibition of CLK1 using TG003 and CLK1 siRNA resulted in a decreased cell viability, proliferation, invasion and migration as well as modulation in the phosphorylation of SRSF2. Ex vivo experiments which utilizes patient’s own tumor and blood to recreate the tumor microenvironment validated the use of CLK1 as a potential target for gastric cancer treatment.

Conclusions

Our data indicates that CLK1 plays a crucial role in the regulation of splicing process in gastric cancer and that CLK1 can act as a novel therapeutic target in gastric cancer.
Appendix
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Metadata
Title
Phosphoproteomic analysis identifies CLK1 as a novel therapeutic target in gastric cancer
Authors
Niraj Babu
Sneha M. Pinto
Manjusha Biswas
Tejaswini Subbannayya
Manoj Rajappa
Sonali V. Mohan
Jayshree Advani
Pavithra Rajagopalan
Gajanan Sathe
Nazia Syed
Vinod D. Radhakrishna
Oliyarasi Muthusamy
Sanjay Navani
Rekha V. Kumar
Gopal Gopisetty
Thangarajan Rajkumar
Padhma Radhakrishnan
Saravanan Thiyagarajan
Akhilesh Pandey
Harsha Gowda
Pradip Majumder
Aditi Chatterjee
Publication date
01-09-2020
Publisher
Springer Singapore
Published in
Gastric Cancer / Issue 5/2020
Print ISSN: 1436-3291
Electronic ISSN: 1436-3305
DOI
https://doi.org/10.1007/s10120-020-01062-8

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