Phosphoproteomics of Retinoblastoma: A Pilot Study Identifies Aberrant Kinases.

Lakshmi Selvan,Subramanian Krishnakumar,Thottethodi Prasad,Harsha Gowda,Sailaja Elchuri,Anil Madugundu,Akhilesh Pandey,Vikas Khetan,Uma Krishnan,Gajanan Sathe,Vinuth Puttamallesh,Pukhraj Rishi,Ravikanth Danda

doi:10.3390/molecules23061454

Abstract

Retinoblastoma is a malignant tumour of the retina which most often occurs in children. Earlier studies on retinoblastoma have concentrated on the identification of key players in the disease and have not provided information on activated/inhibited signalling pathways. The dysregulation of protein phosphorylation in cancer provides clues about the affected signalling cascades in cancer. Phosphoproteomics is an ideal tool for the study of phosphorylation changes in proteins. Hence, global phosphoproteomics of retinoblastoma (RB) was carried out to identify signalling events associated with this cancer. Over 350 proteins showed differential phosphorylation in RB compared to control retina. Our study identified stress response proteins to be hyperphosphorylated in RB which included H2A histone family member X (H2AFX) and sirtuin 1. In particular, Ser140 of H2AFX also known as gamma-H2AX was found to be hyperphosphorylated in retinoblastoma, which indicated the activation of DNA damage response pathways. We also observed the activation of anti-apoptosis in retinoblastoma compared to control. These observations showed the activation of survival pathways in retinoblastoma. The identification of hyperphosphorylated protein kinases including Bromodomain containing 4 (BRD4), Lysine deficient protein kinase 1 (WNK1), and Cyclin-dependent kinase 1 (CDK1) in RB opens new avenues for the treatment of RB. These kinases can be considered as probable therapeutic targets for RB, as small-molecule inhibitors for some of these kinases are already in clinical trials for the treatment other cancers.

Highlights

IntroductionPhosphorylation is one of the major post-translational modifications of proteins that regulates many cellular processes, including cellular communication, proliferation, differentiation, and survival
Phosphorylation is one of the major post-translational modifications of proteins that regulates many cellular processes, including cellular communication, proliferation, differentiation, and survival.This is emphasized by the fact that about one third of all eukaryotic proteins are phosphorylated at some point [1]
Phosphoproteomic analysis of retinoblastoma and retina identified 1393 proteins that were phosphorylated in both tissues

Summary

Introduction

Phosphorylation is one of the major post-translational modifications of proteins that regulates many cellular processes, including cellular communication, proliferation, differentiation, and survival. This is emphasized by the fact that about one third of all eukaryotic proteins are phosphorylated at some point [1]. Protein phosphorylation is governed by the coordinated function of kinases and phosphatases. Dysregulated kinase signalling is observed in many diseases, including cancer [2]. Kinases are the key phosphoproteins, as they are central in various cellular signalling processes. There are 518 kinases identified in the human genome [3], and studying the kinome map under diseased conditions has become a research focus in the last decade

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