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Transcription factor combinations that define human astrocyte identity encode significant variation of maturity and function.

Published version
Peer-reviewed

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Authors

Baranes, Koby 
Rahman, Saifur 
Poulin, Noah 
Tavares, Joana M 

Abstract

Increasing evidence indicates that cellular identity can be reduced to the distinct gene regulatory networks controlled by transcription factors (TFs). However, redundancy exists in these states as different combinations of TFs can induce broadly similar cell types. We previously demonstrated that by overcoming gene silencing, it is possible to deterministically reprogram human pluripotent stem cells directly into cell types of various lineages. In the present study we leverage the consistency and precision of our approach to explore four different TF combinations encoding astrocyte identity, based on previously published reports. Analysis of the resulting induced astrocytes (iAs) demonstrated that all four cassettes generate cells with the typical morphology of in vitro astrocytes, which expressed astrocyte-specific markers. The transcriptional profiles of all four iAs clustered tightly together and displayed similarities with mature human astrocytes, although maturity levels differed between cells. Importantly, we found that the TF cassettes induced iAs with distinct differences with regards to their cytokine response and calcium signaling. In vivo transplantation of selected iAs into immunocompromised rat brains demonstrated long term stability and integration. In conclusion, all four TF combinations were able to induce stable astrocyte-like cells that were morphologically similar but showed subtle differences with respect to their transcriptome. These subtle differences translated into distinct differences with regards to cell function, that could be related to maturation state and/or regional identity of the resulting cells. This insight opens an opportunity to precision-engineer cells to meet functional requirements, for example, in the context of therapeutic cell transplantation.

Description

Funder: Addenbrooke's Charitable Trust (ACT)


Funder: bit bio LTD


Funder: Cambridge Brain Injury MIC


Funder: Ferblanc Foundation


Funder: Rosetrees Trust; Id: http://dx.doi.org/10.13039/501100000833

Keywords

astrocytes, human induced pluripotent stem cells, reprogramming, transcription factors, transplantation, Rats, Animals, Humans, Transcription Factors, Astrocytes, Gene Expression Regulation, Neural Stem Cells, Transcriptome, Cell Differentiation

Journal Title

Glia

Conference Name

Journal ISSN

0894-1491
1098-1136

Volume Title

Publisher

Wiley
Sponsorship
Medical Research Council (MR/S005528/1)
Addenbrooke's Charitable Trust (ACT) (Minute 38/19 B3)
Ferblanc Foundation (Unknown)
Rosetrees Trust (Seedcorn2020\100084)
Stoneygate Trust (Seedcorn2020\100084)
Addenbrooke's Charitable Trust (ACT) (900287, Minute 58/20A)
Addenbrooke's Charitable Trust (ACT) (900288, Minute 58/20B)
Department of Health (via National Institute for Health Research (NIHR)) (CS-2015-15-023)