Stem cell models of ARCAs to understand pathogenic, therapeutically tractable mechanisms

- Generate mutations of a wide variety of ARCA genes in H9 stem cells. These cells will provide a resource which can be shared collaboratively with other laboratories for downstream analysis.


The underlying molecular pathogenic mechanisms in many ARCAs remain unclear. Recent technological advances have enabled the differentiation of pluripotent human stem cells into a variety of neuronal sub-types (including cerebellum) that can be studied in vitro, avoiding invasive human testing or using animal models. Patient cells (usually fibroblasts) containing a mutation must be reprogrammed (iPSCs) or wild type iPSCs have mutations introduced. These can be mutation specific, but are relatively expensive and labour intensive options. An alternative is to use a human embryonic stem cell (hESC) line, and introduce mutations using CRISPR/CAS9 gene editing.

We have developed a protocol that can generate knock-out mutations in H9 hES cells using high-fidelity CAS9 in a matter of weeks. This protocol provides a cost-effective, rapid and scalable method for screening the cellular phenotypes of recessive ataxias and for screening of potential therapeutic agents. We illustrate this work with pilot data in which we have generated several knock-out cell lines of ITPR1 using this protocol.

Our aim is to work with other labs to generate other cell lines which can be a resource for further downstream analysis.

Cohorts used All ARCAs
Funding available? To be sought
Trial readiness category 1:   basic prerequisites for trial readiness

Contact persons:
Andrea Németh
University of Oxford, Oxford, United Kingdom

Further project partners:
Ricardo Parolin Schnekenberg
University of Oxford, Oxford, United Kingdom