Proteome and Network Analysis Provides Novel Insights Into Developing and Established Chemotherapy-Induced Peripheral Neuropathy

Larissa De Clauser,Christin Kappert,Julia R Sondermann,David Gomez-Varela,Manuela Schmidt,Sarah J L Flatters

doi:10.3389/fphar.2022.818690

Abstract

Chemotherapy-induced peripheral neuropathy (CIPN) is a debilitating side-effect of cancer therapies. So far, the development of CIPN cannot be prevented, neither can established CIPN be reverted, often leading to the cessation of necessary chemotherapy. Thus, there is an urgent need to explore the mechanistic basis of CIPN to facilitate its treatment. Here we used an integrated approach of quantitative proteome profiling and network analysis in a clinically relevant rat model of paclitaxel-induced peripheral neuropathy. We analysed lumbar rat DRG at two critical time points: (1) day 7, right after cessation of paclitaxel treatment, but prior to neuropathy development (pre-CIPN); (2) 4 weeks after paclitaxel initiation, when neuropathy has developed (peak-CIPN). In this way we identified a differential protein signature, which shows how changes in the proteome correlate with the development and maintenance of CIPN, respectively. Extensive biological pathway and network analysis reveals that, at pre-CIPN, regulated proteins are prominently implicated in mitochondrial (dys)function, immune signalling, neuronal damage/regeneration, and neuronal transcription. Orthogonal validation in an independent rat cohort confirmed the increase of β-catenin (CTNNB1) at pre-CIPN. More importantly, detailed analysis of protein networks associated with β-catenin highlights translationally relevant and potentially druggable targets. Overall, this study demonstrates the enormous value of combining animal behaviour with proteome and network analysis to provide unprecedented insights into the molecular basis of CIPN. In line with emerging approaches of network medicine our results highlight new avenues for developing improved therapeutic options aimed at preventing and treating CIPN.

Highlights

Chemotherapy-induced peripheral neuropathy (CIPN) is the major dose-limiting side-effect of several widely-used chemotherapeutics, affecting up to 70% of patients following standard chemotherapy regimens (Flatters et al, 2017; Seretny et al, 2014)
We used an integrated workflow from rat behaviour via quantitative proteomics to network analysis to comprehensively study proteome dynamics upon paclitaxel treatment in rat dorsal root ganglia (DRG) (Figure 1)
In accordance with Ingenuity Pathway Analysis (IPA)-based analysis, KEGG (Figure 3D) and Reactome (Figure 3E) pathway analysis suggested that, at pre-CIPN, regulated proteins were associated with major cellular pathways such as those related to the extracellular matrix (ECM), metabolism, mitochondrial processes, and endocytosis

Summary

INTRODUCTION

Chemotherapy-induced peripheral neuropathy (CIPN) is the major dose-limiting side-effect of several widely-used chemotherapeutics, affecting up to 70% of patients following standard chemotherapy regimens (Flatters et al, 2017; Seretny et al, 2014). Peak-CIPN, i.e., around day 28, when neuropathy and paclitaxel-induced mechanical hypersensitivity are fully developed despite the fact that paclitaxel treatment finished at day 6 - a phenomenon, which is referred to as “coasting.” To enhance the translational significance of our work (i) we chose a CIPN model induced by a relatively low dose of paclitaxel (Flatters and Bennett, 2006; Duggett et al, 2016; Duggett et al, 2017; Griffiths et al, 2018); (ii) we mimicked clinically used cycles of chemotherapy by repetitive paclitaxel application; (iii) we prepared a clinically relevant formulation of paclitaxel (please see Methods for details); (iv) we present relevant symptoms of mechanical hypersensitivity as well as coasting; and (v) we performed all animal studies in a blinded and fully randomised manner.

EXPERIMENTAL PROCEDURES

RESULTS

DISCUSSION

ETHICS STATEMENT