Interplay between NEuroactiVe stEroids and endoplasmic Reticulum-MitOchondRia intEraction: a novel therapeutic horizon in chemotherapy-induced peripheral neurotoxicity (NEVERMORE)

Chemotherapy-induced peripheral neurotoxicity (CIPN) is a common dose limiting side effect of several chemotherapeutic (CTs) agents. Growing body of evidence suggests that the effect of different chemotherapies’ families converge on a common mitochondrial dysfunction program. Specifically, it has been proposed that a possible target for several CTs is represented by the mitochondria-ER contact sites (MERCS), given by strong association of processes occurring at MERCS and mitochondria. Furthermore, since neuroactive steroids (NAS) hold promise for anti-CIPN therapy, it is conceivable that MERCS-mitochondria axis may be a target for NAS protective effects. The main goal of the NEVERMORE proposal is to test the hypothesis of the involvement of MERCS-mitochondria axis in CIPN and elucidate the role of steroid hormones in mitigating of CT-induced cellular dysfunction both in sensory neurons and glial cells. The project will exploit an innovative approach based on a funnel-like strategy and multidisciplinary approach, and will combine in vitro, ex-vivo and in vivo models of CIPN with different combinations of neuroactive steroids in order to identify novel mechanisms and targets to counteract CIPN. First, multiple CT/NAS combinations will be tested on several cell lines in a high throughput screening for their effects on MERCS-mitochondrial morphology and functions to identify the most efficacious combinations. Next, using a battery of omics (transcriptomics, proteomics and lipidomics) and functional (Ca2+ signaling, bioenergetics, proteostasis) assays we will test shortlisted CT/NAS combinations on selected neuronal and glial cell lines to dissect pathways and identify molecules, related to MERCS-mitochondrial physiology, affected by CT and rescued by NAS. Last, the identified molecules and pathways will be tested in vivo in animals treated with selected CT/NAS combination to correlate CIPN behavioral alterations with MERCS-mitochondrial cellular pathology in DRG sensory neurons, satellite glial cells and Schwann cells. Therefore, the NEVERMORE project will shed light on the role of MERCS-mitochondria in CIPN etiopathogenesis and protective mechanisms of NAS, useful to develop a solid therapeutic approach. In this project three laboratories with complementary expertise and well-established collaboration will work under the leadership of young PI and co-PI, under 40, with an internationally recognized expertise in CIPN and neuroprotection fields.