Cystatin B is essential for proliferation and interneuron migration in individuals with EPM1 epilepsy.

Francesco Di Matteo,Emilio Ciusani,Angela Chambery,Silvia Cappello,Isabella Tovecci,Rossella Di Giaimo,Fabrizia Pipicelli,Marianna Crispino,Anke Hoffmann,Magdalena Götz,Christina Kyrousi,Eduardo Penna,Ane Cristina Ayo‐Martin,Rosita Russo,Laura Canafoglia,Martina Giordano

doi:10.15252/emmm.201911419

Abstract

Progressive myoclonus epilepsy (PME) of Unverricht–Lundborg type (EPM1) is an autosomal recessive neurodegenerative disorder with the highest incidence of PME worldwide. Mutations in the gene encoding cystatin B (CSTB) are the primary genetic cause of EPM1. Here, we investigate the role of CSTB during neurogenesis in vivo in the developing mouse brain and in vitro in human cerebral organoids (hCOs) derived from EPM1 patients. We find that CSTB (but not one of its pathological variants) is secreted into the mouse cerebral spinal fluid and the conditioned media from hCOs. In embryonic mouse brain, we find that functional CSTB influences progenitors’ proliferation and modulates neuronal distribution by attracting interneurons to the site of secretion via cell‐non‐autonomous mechanisms. Similarly, in patient‐derived hCOs, low levels of functional CSTB result in an alteration of progenitor's proliferation, premature differentiation, and changes in interneurons migration. Secretion and extracellular matrix organization are the biological processes particularly affected as suggested by a proteomic analysis in patients’ hCOs. Overall, our study sheds new light on the cellular mechanisms underlying the development of EPM1.

Highlights

Progressive myoclonus epilepsy (PME) of Unverricht–Lundborg type (EPM1) is an autosomal recessive neurodegenerative disorder with the highest incidence of PME worldwide
cystatin B (CSTB) is the gene that is most often mutated in reported cases of EPM1 disease, and the severity of EPM1 is inversely correlated with the amount of residual functional CSTB protein
Functional MRI studies performed on brains of individuals with EPM1 show atrophic changes in the cerebral cortex and, marginally, in the cerebellum

Summary

Introduction

Progressive myoclonus epilepsy (PME) of Unverricht–Lundborg type (EPM1) is an autosomal recessive neurodegenerative disorder with the highest incidence of PME worldwide. We investigate the role of CSTB during neurogenesis in vivo in the developing mouse brain and in vitro in human cerebral organoids (hCOs) derived from EPM1 patients. We find that CSTB (but not one of its pathological variants) is secreted into the mouse cerebral spinal fluid and the conditioned media from hCOs. In embryonic mouse brain, we find that functional CSTB influences progenitors’ proliferation and modulates neuronal distribution by attracting interneurons to the site of secretion via cell-non-autonomous mechanisms. In patient-derived hCOs, low levels of functional CSTB result in an alteration of progenitor’s proliferation, premature differentiation, and changes in interneurons migration. Secretion and extracellular matrix organization are the biological processes affected as suggested by a proteomic analysis in patients’ hCOs. Overall, our study sheds new light on the cellular mechanisms underlying the development of EPM1

Methods

Results

Conclusion