The difficulty to model Huntington\u2019s disease in vitro using striatal medium spiny neurons differentiated from human induced pluripotent stem cells

Kim Le Cann,Petra Hautvast,Corinna Rösseler,Sebastian Giesselmann,Ingo Kurth,Geraldine Zimmer-Bensch,Angelika Lampert,Stephan Von Hörsten,Virginia B Mattis,Andelain Erickson,Tim Clarner,Alec Foerster,Daniel Pensold,Markus Rothermel,Jannis Meents,Bernd Denecke

doi:10.1038/s41598-021-85656-x

Abstract

Huntington’s disease (HD) is an autosomal dominant neurodegenerative disorder caused by an expanded polyglutamine repeat in the huntingtin gene. The neuropathology of HD is characterized by the decline of a specific neuronal population within the brain, the striatal medium spiny neurons (MSNs). The origins of this extreme vulnerability remain unknown. Human induced pluripotent stem cell (hiPS cell)-derived MSNs represent a powerful tool to study this genetic disease. However, the differentiation protocols published so far show a high heterogeneity of neuronal populations in vitro. Here, we compared two previously published protocols to obtain hiPS cell-derived striatal neurons from both healthy donors and HD patients. Patch-clamp experiments, immunostaining and RT-qPCR were performed to characterize the neurons in culture. While the neurons were mature enough to fire action potentials, a majority failed to express markers typical for MSNs. Voltage-clamp experiments on voltage-gated sodium (Nav) channels revealed a large variability between the two differentiation protocols. Action potential analysis did not reveal changes induced by the HD mutation. This study attempts to demonstrate the current challenges in reproducing data of previously published differentiation protocols and in generating hiPS cell-derived striatal MSNs to model a genetic neurodegenerative disorder in vitro.

Highlights

Huntington’s disease (HD) is an autosomal dominant neurodegenerative disorder caused by an expanded polyglutamine repeat in the huntingtin gene
Using the Stanslowsky protocol, we found in two control cell lines (Ctrl[1] and Ctrl3) and in one HD cell line (HD72) a higher percentage of electrically active neurons (i.e. = neurons firing at least one action potential) at DIV55 than at DIV40 (50%, 53.8% and 58.8% for Ctrl[1], Ctrl[3] and HD72 respectively at DIV40 vs 65.2%, 71.8% and 70.4% respectively at DIV55) (Fig. 3A)
As medium spiny neurons (MSNs) seemed to be the minority of the cells derived by the differentiation protocols in our hands, we investigated which other neuronal populations were present in our preparations

Summary

Introduction

Huntington’s disease (HD) is an autosomal dominant neurodegenerative disorder caused by an expanded polyglutamine repeat in the huntingtin gene. Human induced pluripotent stem cell (hiPS cell)-derived MSNs represent a powerful tool to study this genetic disease. This study attempts to demonstrate the current challenges in reproducing data of previously published differentiation protocols and in generating hiPS cell-derived striatal MSNs to model a genetic neurodegenerative disorder in vitro. Significant species differences between rodent and human cells limit the use of HD rodent models to accurately represent the disease and to predict the electrical activity in human striatal MSNs. As an alternative, patient-derived human induced pluripotent stem (hiPS) cells have emerged as a powerful tool to decipher mechanisms underlying MSN degeneration and to investigate their firing p roperties[12]. These essential properties gave rise to the establishment of many hiPS cell-derived MSN differentiation protocols

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