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-...
| Main Authors: | , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
eLife Sciences Publications Ltd
2020-11-01
|
| Series: | eLife |
| Subjects: | |
| Online Access: | https://elifesciences.org/articles/60188 |
| _version_ | 1811252896551927808 |
|---|---|
| author | Kristen L Wells Corey N Miller Andreas R Gschwind Wu Wei Jonah D Phipps Mark S Anderson Lars M Steinmetz |
| author_facet | Kristen L Wells Corey N Miller Andreas R Gschwind Wu Wei Jonah D Phipps Mark S Anderson Lars M Steinmetz |
| author_sort | Kristen L Wells |
| collection | DOAJ |
| 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. |
| first_indexed | 2024-04-12T16:42:24Z |
| format | Article |
| id | doaj.art-7e6e9a65e4af4c6ea418368160781fcd |
| institution | Directory Open Access Journal |
| issn | 2050-084X |
| language | English |
| last_indexed | 2024-04-12T16:42:24Z |
| publishDate | 2020-11-01 |
| publisher | eLife Sciences Publications Ltd |
| record_format | Article |
| series | eLife |
| spelling | doaj.art-7e6e9a65e4af4c6ea418368160781fcd2022-12-22T03:24:44ZengeLife Sciences Publications LtdeLife2050-084X2020-11-01910.7554/eLife.60188Combined transient ablation and single-cell RNA-sequencing reveals the development of medullary thymic epithelial cellsKristen L Wells0https://orcid.org/0000-0002-7466-8164Corey N Miller1https://orcid.org/0000-0002-1291-0074Andreas R Gschwind2https://orcid.org/0000-0002-0769-6907Wu Wei3Jonah D Phipps4Mark S Anderson5https://orcid.org/0000-0002-3093-4758Lars M Steinmetz6https://orcid.org/0000-0002-3962-2865Department of Genetics, Stanford University School of Medicine, Stanford, United StatesDiabetes Center, University of California, San Francisco, San Francisco, United States; Department of Medicine, University of California San Francisco, San Francisco, United StatesDepartment of Genetics, Stanford University School of Medicine, Stanford, United StatesStanford Genome Technology Center, Stanford University, Palo Alto, United StatesDiabetes Center, University of California, San Francisco, San Francisco, United States; Department of Medicine, University of California San Francisco, San Francisco, United StatesDiabetes Center, University of California, San Francisco, San Francisco, United States; Department of Medicine, University of California San Francisco, San Francisco, United StatesDepartment of Genetics, Stanford University School of Medicine, Stanford, United States; Stanford Genome Technology Center, Stanford University, Palo Alto, United States; Genome Biology Unit, European Molecular Biology Laboratory (EMBL), Heidelberg, GermanyMedullary 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.https://elifesciences.org/articles/60188single-cell transcriptomicsmedullary thymic epithelial cellimmune system |
| spellingShingle | Kristen L Wells Corey N Miller Andreas R Gschwind Wu Wei Jonah D Phipps Mark S Anderson Lars M Steinmetz Combined transient ablation and single-cell RNA-sequencing reveals the development of medullary thymic epithelial cells eLife single-cell transcriptomics medullary thymic epithelial cell immune system |
| 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 | single-cell transcriptomics medullary thymic epithelial cell immune system |
| url | https://elifesciences.org/articles/60188 |
| work_keys_str_mv | AT kristenlwells combinedtransientablationandsinglecellrnasequencingrevealsthedevelopmentofmedullarythymicepithelialcells AT coreynmiller combinedtransientablationandsinglecellrnasequencingrevealsthedevelopmentofmedullarythymicepithelialcells AT andreasrgschwind combinedtransientablationandsinglecellrnasequencingrevealsthedevelopmentofmedullarythymicepithelialcells AT wuwei combinedtransientablationandsinglecellrnasequencingrevealsthedevelopmentofmedullarythymicepithelialcells AT jonahdphipps combinedtransientablationandsinglecellrnasequencingrevealsthedevelopmentofmedullarythymicepithelialcells AT marksanderson combinedtransientablationandsinglecellrnasequencingrevealsthedevelopmentofmedullarythymicepithelialcells AT larsmsteinmetz combinedtransientablationandsinglecellrnasequencingrevealsthedevelopmentofmedullarythymicepithelialcells |