ACE2 protein expression within isogenic cell lines is heterogeneous and associated with distinct transcriptomes

Abstract The membrane protein angiotensin-converting enzyme 2 (ACE2) is a physiologic regulator of the renin-angiotensin system and the cellular receptor for the SARS-CoV-2 virus. Prior studies of ACE2 expression have primarily focused on mRNA abundance, with investigation at the protein level limit...

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Autores principales:	Emily J. Sherman, Brian T. Emmer
Formato:	article
Lenguaje:	EN
Publicado:	Nature Portfolio 2021
Materias:	Medicine R Science Q
Acceso en línea:	https://doaj.org/article/7145808995e74ee79c0e061525cf7b93
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spelling	oai:doaj.org-article:7145808995e74ee79c0e061525cf7b932021-12-02T16:35:32ZACE2 protein expression within isogenic cell lines is heterogeneous and associated with distinct transcriptomes10.1038/s41598-021-95308-92045-2322https://doaj.org/article/7145808995e74ee79c0e061525cf7b932021-08-01T00:00:00Zhttps://doi.org/10.1038/s41598-021-95308-9https://doaj.org/toc/2045-2322Abstract The membrane protein angiotensin-converting enzyme 2 (ACE2) is a physiologic regulator of the renin-angiotensin system and the cellular receptor for the SARS-CoV-2 virus. Prior studies of ACE2 expression have primarily focused on mRNA abundance, with investigation at the protein level limited by uncertain specificity of commercial ACE2 antibodies. Here, we report our development of a sensitive and specific flow cytometry-based assay for cellular ACE2 protein abundance. Application of this approach to multiple cell lines revealed an unexpected degree of cellular heterogeneity, with detectable ACE2 protein in only a subset of cells in each isogenic population. This heterogeneity was mediated at the mRNA level by transcripts predominantly initiated from the ACE2 proximal promoter. ACE2 expression was heritable but not fixed over multiple generations of daughter cells, with gradual drift toward the original heterogeneous background. RNA-seq profiling identified distinct transcriptomes of ACE2-expressing relative cells to non-expressing cells, with enrichment in functionally related genes and transcription factor target sets. Our findings provide a validated approach for the specific detection of ACE2 protein at the surface of single cells, support an epigenetic mechanism of ACE2 gene regulation, and identify specific pathways associated with ACE2 expression in HuH7 cells.Emily J. ShermanBrian T. EmmerNature PortfolioarticleMedicineRScienceQENScientific Reports, Vol 11, Iss 1, Pp 1-13 (2021)
institution	DOAJ
collection	DOAJ
language	EN
topic	Medicine R Science Q
spellingShingle	Medicine R Science Q Emily J. Sherman Brian T. Emmer ACE2 protein expression within isogenic cell lines is heterogeneous and associated with distinct transcriptomes
description	Abstract The membrane protein angiotensin-converting enzyme 2 (ACE2) is a physiologic regulator of the renin-angiotensin system and the cellular receptor for the SARS-CoV-2 virus. Prior studies of ACE2 expression have primarily focused on mRNA abundance, with investigation at the protein level limited by uncertain specificity of commercial ACE2 antibodies. Here, we report our development of a sensitive and specific flow cytometry-based assay for cellular ACE2 protein abundance. Application of this approach to multiple cell lines revealed an unexpected degree of cellular heterogeneity, with detectable ACE2 protein in only a subset of cells in each isogenic population. This heterogeneity was mediated at the mRNA level by transcripts predominantly initiated from the ACE2 proximal promoter. ACE2 expression was heritable but not fixed over multiple generations of daughter cells, with gradual drift toward the original heterogeneous background. RNA-seq profiling identified distinct transcriptomes of ACE2-expressing relative cells to non-expressing cells, with enrichment in functionally related genes and transcription factor target sets. Our findings provide a validated approach for the specific detection of ACE2 protein at the surface of single cells, support an epigenetic mechanism of ACE2 gene regulation, and identify specific pathways associated with ACE2 expression in HuH7 cells.
format	article
author	Emily J. Sherman Brian T. Emmer
author_facet	Emily J. Sherman Brian T. Emmer
author_sort	Emily J. Sherman
title	ACE2 protein expression within isogenic cell lines is heterogeneous and associated with distinct transcriptomes
title_short	ACE2 protein expression within isogenic cell lines is heterogeneous and associated with distinct transcriptomes
title_full	ACE2 protein expression within isogenic cell lines is heterogeneous and associated with distinct transcriptomes
title_fullStr	ACE2 protein expression within isogenic cell lines is heterogeneous and associated with distinct transcriptomes
title_full_unstemmed	ACE2 protein expression within isogenic cell lines is heterogeneous and associated with distinct transcriptomes
title_sort	ace2 protein expression within isogenic cell lines is heterogeneous and associated with distinct transcriptomes
publisher	Nature Portfolio
publishDate	2021
url	https://doaj.org/article/7145808995e74ee79c0e061525cf7b93
work_keys_str_mv	AT emilyjsherman ace2proteinexpressionwithinisogeniccelllinesisheterogeneousandassociatedwithdistincttranscriptomes AT briantemmer ace2proteinexpressionwithinisogeniccelllinesisheterogeneousandassociatedwithdistincttranscriptomes
_version_	1718383697269358592

ACE2 protein expression within isogenic cell lines is heterogeneous and associated with distinct transcriptomes

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