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Epigenetic modulators of B cell fate identified through coupled phenotype-transcriptome analysis

High-throughput methodologies are the cornerstone of screening approaches to identify novel compounds that regulate immune cell function. To identify novel targeted therapeutics to treat immune disorders and haematological malignancies, there is a need to integrate functional cellular information wi...

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Autores principales: Kong, Isabella Y., Trezise, Stephanie, Light, Amanda, Todorovski, Izabela, Arnau, Gisela Mir, Gadipally, Sreeja, Yoannidis, David, Simpson, Kaylene J., Dong, Xueyi, Whitehead, Lachlan, Tempany, Jessica C., Farchione, Anthony J., Sheikh, Amania A., Groom, Joanna R., Rogers, Kelly L., Herold, Marco J., Bryant, Vanessa L., Ritchie, Matthew E., Willis, Simon N., Johnstone, Ricky W., Hodgkin, Philip D., Nutt, Stephen L., Vervoort, Stephin J., Hawkins, Edwin D.
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Nature Publishing Group UK 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9751284/
https://www.ncbi.nlm.nih.gov/pubmed/35831623
http://dx.doi.org/10.1038/s41418-022-01037-5
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author Kong, Isabella Y.
Trezise, Stephanie
Light, Amanda
Todorovski, Izabela
Arnau, Gisela Mir
Gadipally, Sreeja
Yoannidis, David
Simpson, Kaylene J.
Dong, Xueyi
Whitehead, Lachlan
Tempany, Jessica C.
Farchione, Anthony J.
Sheikh, Amania A.
Groom, Joanna R.
Rogers, Kelly L.
Herold, Marco J.
Bryant, Vanessa L.
Ritchie, Matthew E.
Willis, Simon N.
Johnstone, Ricky W.
Hodgkin, Philip D.
Nutt, Stephen L.
Vervoort, Stephin J.
Hawkins, Edwin D.
author_facet Kong, Isabella Y.
Trezise, Stephanie
Light, Amanda
Todorovski, Izabela
Arnau, Gisela Mir
Gadipally, Sreeja
Yoannidis, David
Simpson, Kaylene J.
Dong, Xueyi
Whitehead, Lachlan
Tempany, Jessica C.
Farchione, Anthony J.
Sheikh, Amania A.
Groom, Joanna R.
Rogers, Kelly L.
Herold, Marco J.
Bryant, Vanessa L.
Ritchie, Matthew E.
Willis, Simon N.
Johnstone, Ricky W.
Hodgkin, Philip D.
Nutt, Stephen L.
Vervoort, Stephin J.
Hawkins, Edwin D.
author_sort Kong, Isabella Y.
collection PubMed
description High-throughput methodologies are the cornerstone of screening approaches to identify novel compounds that regulate immune cell function. To identify novel targeted therapeutics to treat immune disorders and haematological malignancies, there is a need to integrate functional cellular information with the molecular mechanisms that regulate changes in immune cell phenotype. We facilitate this goal by combining quantitative methods for dissecting complex simultaneous cell phenotypic effects with genomic analysis. This combination strategy we term Multiplexed Analysis of Cells sequencing (MAC-seq), a modified version of Digital RNA with perturbation of Genes (DRUGseq). We applied MAC-seq to screen compounds that target the epigenetic machinery of B cells and assess altered humoral immunity by measuring changes in proliferation, survival, differentiation and transcription. This approach revealed that polycomb repressive complex 2 (PRC2) inhibitors promote antibody secreting cell (ASC) differentiation in both murine and human B cells in vitro. This is further validated using T cell-dependent immunization in mice. Functional dissection of downstream effectors of PRC2 using arrayed CRISPR screening uncovered novel regulators of B cell differentiation, including Mybl1, Myof, Gas7 and Atoh8. Together, our findings demonstrate that integrated phenotype-transcriptome analyses can be effectively combined with drug screening approaches to uncover the molecular circuitry that drives lymphocyte fate decisions.
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spelling pubmed-97512842022-12-16 Epigenetic modulators of B cell fate identified through coupled phenotype-transcriptome analysis Kong, Isabella Y. Trezise, Stephanie Light, Amanda Todorovski, Izabela Arnau, Gisela Mir Gadipally, Sreeja Yoannidis, David Simpson, Kaylene J. Dong, Xueyi Whitehead, Lachlan Tempany, Jessica C. Farchione, Anthony J. Sheikh, Amania A. Groom, Joanna R. Rogers, Kelly L. Herold, Marco J. Bryant, Vanessa L. Ritchie, Matthew E. Willis, Simon N. Johnstone, Ricky W. Hodgkin, Philip D. Nutt, Stephen L. Vervoort, Stephin J. Hawkins, Edwin D. Cell Death Differ Article High-throughput methodologies are the cornerstone of screening approaches to identify novel compounds that regulate immune cell function. To identify novel targeted therapeutics to treat immune disorders and haematological malignancies, there is a need to integrate functional cellular information with the molecular mechanisms that regulate changes in immune cell phenotype. We facilitate this goal by combining quantitative methods for dissecting complex simultaneous cell phenotypic effects with genomic analysis. This combination strategy we term Multiplexed Analysis of Cells sequencing (MAC-seq), a modified version of Digital RNA with perturbation of Genes (DRUGseq). We applied MAC-seq to screen compounds that target the epigenetic machinery of B cells and assess altered humoral immunity by measuring changes in proliferation, survival, differentiation and transcription. This approach revealed that polycomb repressive complex 2 (PRC2) inhibitors promote antibody secreting cell (ASC) differentiation in both murine and human B cells in vitro. This is further validated using T cell-dependent immunization in mice. Functional dissection of downstream effectors of PRC2 using arrayed CRISPR screening uncovered novel regulators of B cell differentiation, including Mybl1, Myof, Gas7 and Atoh8. Together, our findings demonstrate that integrated phenotype-transcriptome analyses can be effectively combined with drug screening approaches to uncover the molecular circuitry that drives lymphocyte fate decisions. Nature Publishing Group UK 2022-07-13 2022-12 /pmc/articles/PMC9751284/ /pubmed/35831623 http://dx.doi.org/10.1038/s41418-022-01037-5 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open Access This article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in a credit line to the material. If material is not included in the article’s Creative Commons license and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) .
spellingShingle Article
Kong, Isabella Y.
Trezise, Stephanie
Light, Amanda
Todorovski, Izabela
Arnau, Gisela Mir
Gadipally, Sreeja
Yoannidis, David
Simpson, Kaylene J.
Dong, Xueyi
Whitehead, Lachlan
Tempany, Jessica C.
Farchione, Anthony J.
Sheikh, Amania A.
Groom, Joanna R.
Rogers, Kelly L.
Herold, Marco J.
Bryant, Vanessa L.
Ritchie, Matthew E.
Willis, Simon N.
Johnstone, Ricky W.
Hodgkin, Philip D.
Nutt, Stephen L.
Vervoort, Stephin J.
Hawkins, Edwin D.
Epigenetic modulators of B cell fate identified through coupled phenotype-transcriptome analysis
title Epigenetic modulators of B cell fate identified through coupled phenotype-transcriptome analysis
title_full Epigenetic modulators of B cell fate identified through coupled phenotype-transcriptome analysis
title_fullStr Epigenetic modulators of B cell fate identified through coupled phenotype-transcriptome analysis
title_full_unstemmed Epigenetic modulators of B cell fate identified through coupled phenotype-transcriptome analysis
title_short Epigenetic modulators of B cell fate identified through coupled phenotype-transcriptome analysis
title_sort epigenetic modulators of b cell fate identified through coupled phenotype-transcriptome analysis
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9751284/
https://www.ncbi.nlm.nih.gov/pubmed/35831623
http://dx.doi.org/10.1038/s41418-022-01037-5
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