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Combined transient ablation and single-cell RNA-sequencing reveals the development of medullary thymic epithelial cells

Medullary thymic epithelial cells (mTECs) play a critical role in central immune tolerance by mediating negative selection of autoreactive T cells through the collective expression of the peripheral self-antigen compartment, including tissue-specific antigens (TSAs). Recent work has shown that gene-...

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Autores principales: Wells, Kristen L, Miller, Corey N, Gschwind, Andreas R, Wei, Wu, Phipps, Jonah D, Anderson, Mark S, Steinmetz, Lars M
Formato: Online Artículo Texto
Lenguaje:English
Publicado: eLife Sciences Publications, Ltd 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7771965/
https://www.ncbi.nlm.nih.gov/pubmed/33226342
http://dx.doi.org/10.7554/eLife.60188
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author Wells, Kristen L
Miller, Corey N
Gschwind, Andreas R
Wei, Wu
Phipps, Jonah D
Anderson, Mark S
Steinmetz, Lars M
author_facet Wells, Kristen L
Miller, Corey N
Gschwind, Andreas R
Wei, Wu
Phipps, Jonah D
Anderson, Mark S
Steinmetz, Lars M
author_sort Wells, Kristen L
collection PubMed
description Medullary thymic epithelial cells (mTECs) play a critical role in central immune tolerance by mediating negative selection of autoreactive T cells through the collective expression of the peripheral self-antigen compartment, including tissue-specific antigens (TSAs). Recent work has shown that gene-expression patterns within the mTEC compartment are heterogenous and include multiple differentiated cell states. To further define mTEC development and medullary epithelial lineage relationships, we combined lineage tracing and recovery from transient in vivo mTEC ablation with single-cell RNA-sequencing in Mus musculus. The combination of bioinformatic and experimental approaches revealed a non-stem transit-amplifying population of cycling mTECs that preceded Aire expression. We propose a branching model of mTEC development wherein a heterogeneous pool of transit-amplifying cells gives rise to Aire- and Ccl21a-expressing mTEC subsets. We further use experimental techniques to show that within the Aire-expressing developmental branch, TSA expression peaked as Aire expression decreased, implying Aire expression must be established before TSA expression can occur. Collectively, these data provide a roadmap of mTEC development and demonstrate the power of combinatorial approaches leveraging both in vivo models and high-dimensional datasets.
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spelling pubmed-77719652020-12-30 Combined transient ablation and single-cell RNA-sequencing reveals the development of medullary thymic epithelial cells Wells, Kristen L Miller, Corey N Gschwind, Andreas R Wei, Wu Phipps, Jonah D Anderson, Mark S Steinmetz, Lars M eLife Genetics and Genomics Medullary thymic epithelial cells (mTECs) play a critical role in central immune tolerance by mediating negative selection of autoreactive T cells through the collective expression of the peripheral self-antigen compartment, including tissue-specific antigens (TSAs). Recent work has shown that gene-expression patterns within the mTEC compartment are heterogenous and include multiple differentiated cell states. To further define mTEC development and medullary epithelial lineage relationships, we combined lineage tracing and recovery from transient in vivo mTEC ablation with single-cell RNA-sequencing in Mus musculus. The combination of bioinformatic and experimental approaches revealed a non-stem transit-amplifying population of cycling mTECs that preceded Aire expression. We propose a branching model of mTEC development wherein a heterogeneous pool of transit-amplifying cells gives rise to Aire- and Ccl21a-expressing mTEC subsets. We further use experimental techniques to show that within the Aire-expressing developmental branch, TSA expression peaked as Aire expression decreased, implying Aire expression must be established before TSA expression can occur. Collectively, these data provide a roadmap of mTEC development and demonstrate the power of combinatorial approaches leveraging both in vivo models and high-dimensional datasets. eLife Sciences Publications, Ltd 2020-11-23 /pmc/articles/PMC7771965/ /pubmed/33226342 http://dx.doi.org/10.7554/eLife.60188 Text en © 2020, Wells et al http://creativecommons.org/licenses/by/4.0/ http://creativecommons.org/licenses/by/4.0/This article is distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/4.0/) , which permits unrestricted use and redistribution provided that the original author and source are credited.
spellingShingle Genetics and Genomics
Wells, Kristen L
Miller, Corey N
Gschwind, Andreas R
Wei, Wu
Phipps, Jonah D
Anderson, Mark S
Steinmetz, Lars M
Combined transient ablation and single-cell RNA-sequencing reveals the development of medullary thymic epithelial cells
title Combined transient ablation and single-cell RNA-sequencing reveals the development of medullary thymic epithelial cells
title_full Combined transient ablation and single-cell RNA-sequencing reveals the development of medullary thymic epithelial cells
title_fullStr Combined transient ablation and single-cell RNA-sequencing reveals the development of medullary thymic epithelial cells
title_full_unstemmed Combined transient ablation and single-cell RNA-sequencing reveals the development of medullary thymic epithelial cells
title_short Combined transient ablation and single-cell RNA-sequencing reveals the development of medullary thymic epithelial cells
title_sort combined transient ablation and single-cell rna-sequencing reveals the development of medullary thymic epithelial cells
topic Genetics and Genomics
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7771965/
https://www.ncbi.nlm.nih.gov/pubmed/33226342
http://dx.doi.org/10.7554/eLife.60188
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