Human ALS/FTD brain organoid slice cultures display distinct early astrocyte and targetable neuronal pathology

Kornélia Szebényi,Léa M D Wenger,Gabriel Balmus,Elena Conci,George M Gibbons,Susanna B Mierau,Alexander W E Dunn,Ole Paulsen,Yu Sun,András Lakatos,Colleen A Limegrover

doi:10.1038/s41593-021-00923-4

Abstract

Amyotrophic lateral sclerosis overlapping with frontotemporal dementia (ALS/FTD) is a fatal and currently untreatable disease characterized by rapid cognitive decline and paralysis. Elucidating initial cellular pathologies is central to therapeutic target development, but obtaining samples from presymptomatic patients is not feasible. Here, we report the development of a cerebral organoid slice model derived from human induced pluripotent stem cells (iPSCs) that recapitulates mature cortical architecture and displays early molecular pathology of C9ORF72 ALS/FTD. Using a combination of single-cell RNA sequencing and biological assays, we reveal distinct transcriptional, proteostasis and DNA repair disturbances in astroglia and neurons. We show that astroglia display increased levels of the autophagy signaling protein P62 and that deep layer neurons accumulate dipeptide repeat protein poly(GA), DNA damage and undergo nuclear pyknosis that could be pharmacologically rescued by GSK2606414. Thus, patient-specific iPSC-derived cortical organoid slice cultures are a reproducible translational platform to investigate preclinical ALS/FTD mechanisms as well as novel therapeutic approaches.

Highlights

Amyotrophic lateral sclerosis overlapping with frontotemporal dementia (ALS/FTD) is a fatal and currently untreatable disease characterized by rapid cognitive decline and paralysis
Diverse cell populations segregate in human iPSC (hiPSC)-derived ALI-cortical organoid (CO)
We examined whether C9 ALI-COs develop similar cell-type diversity and cytoarchitecture to their healthy control patient-derived counterparts to assess their relevance to disease-affected phenotypes

Summary

Introduction

Amyotrophic lateral sclerosis overlapping with frontotemporal dementia (ALS/FTD) is a fatal and currently untreatable disease characterized by rapid cognitive decline and paralysis. We report the development of a cerebral organoid slice model derived from human induced pluripotent stem cells (iPSCs) that recapitulates mature cortical architecture and displays early molecular pathology of C9ORF72 ALS/FTD. Brain organoids have been grown from cells derived from patients with Parkinson’s disease and from patients with Alzheimer’s disease for 30 or 84 days in vitro (DIV), respectively[8,9] This has been a considerable step forward in disease model development; the relatively short longevity and variable cortical-cell-type composition[10,11] may limit the fidelity for observing a broad spectrum of pathology. Similar advances are a prerequisite for patient-specific iPSC-derived organoids for precise pathomechanistic discoveries This is warranted for ALS/FTD, an untreatable neurodegenerative disease with rapid cognitive decline and paralysis. Our results demonstrate that hiPSC-derived ALI-COs provide a reproducible platform with the necessary longevity and maturity for investigating ALS/FTD, thereby revealing

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Conclusion