Strong Expansion of Human Regulatory T Cells for Adoptive Cell Therapy Results in Epigenetic Changes Which May Impact Their Survival and Function

Adoptive transfer of regulatory T cells (Treg) is a promising new therapeutic option to treat detrimental inflammatory conditions after transplantation and during autoimmune disease. To reach sufficient cell yield for treatment, ex vivo isolated autologous or allogenic Tregs need to be expanded exte...

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Autores principales: Kristy Ou, Dania Hamo, Anne Schulze, Andy Roemhild, Daniel Kaiser, Gilles Gasparoni, Abdulrahman Salhab, Ghazaleh Zarrinrad, Leila Amini, Stephan Schlickeiser, Mathias Streitz, Jörn Walter, Hans-Dieter Volk, Michael Schmueck-Henneresse, Petra Reinke, Julia K. Polansky
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Publicado: Frontiers Media S.A. 2021
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spelling oai:doaj.org-article:7b260d54dd1e425b9ebf78e9599970072021-11-18T09:51:20ZStrong Expansion of Human Regulatory T Cells for Adoptive Cell Therapy Results in Epigenetic Changes Which May Impact Their Survival and Function2296-634X10.3389/fcell.2021.751590https://doaj.org/article/7b260d54dd1e425b9ebf78e9599970072021-11-01T00:00:00Zhttps://www.frontiersin.org/articles/10.3389/fcell.2021.751590/fullhttps://doaj.org/toc/2296-634XAdoptive transfer of regulatory T cells (Treg) is a promising new therapeutic option to treat detrimental inflammatory conditions after transplantation and during autoimmune disease. To reach sufficient cell yield for treatment, ex vivo isolated autologous or allogenic Tregs need to be expanded extensively in vitro during manufacturing of the Treg product. However, repetitive cycles of restimulation and prolonged culture have been shown to impact T cell phenotypes, functionality and fitness. It is therefore critical to scrutinize the molecular changes which occur during T cell product generation, and reexamine current manufacturing practices. We performed genome-wide DNA methylation profiling of cells throughout the manufacturing process of a polyclonal Treg product that has proven safety and hints of therapeutic efficacy in kidney transplant patients. We found progressive DNA methylation changes over the duration of culture, which were donor-independent and reproducible between manufacturing runs. Differentially methylated regions (DMRs) in the final products were significantly enriched at promoters and enhancers of genes implicated in T cell activation. Additionally, significant hypomethylation did also occur in promoters of genes implicated in functional exhaustion in conventional T cells, some of which, however, have been reported to strengthen immunosuppressive effector function in Tregs. At the same time, a set of reported Treg-specific demethylated regions increased methylation levels with culture, indicating a possible destabilization of Treg identity during manufacturing, which was independent of the purity of the starting material. Together, our results indicate that the repetitive TCR-mediated stimulation lead to epigenetic changes that might impact functionality of Treg products in multiple ways, by possibly shifting to an effector Treg phenotype with enhanced functional activity or by risking destabilization of Treg identity and impaired TCR activation. Our analyses also illustrate the value of epigenetic profiling for the evaluation of T cell product manufacturing pipelines, which might open new avenues for the improvement of current adoptive Treg therapies with relevance for conventional effector T cell products.Kristy OuDania HamoAnne SchulzeAndy RoemhildDaniel KaiserGilles GasparoniAbdulrahman SalhabGhazaleh ZarrinradLeila AminiLeila AminiStephan SchlickeiserMathias StreitzJörn WalterHans-Dieter VolkHans-Dieter VolkMichael Schmueck-HenneressePetra ReinkeJulia K. PolanskyJulia K. PolanskyFrontiers Media S.A.articleregulatory T cellsadvanced therapy medicinal productsDNA methylationbiomarkeradoptive cell therapygood manufacturing practiceBiology (General)QH301-705.5ENFrontiers in Cell and Developmental Biology, Vol 9 (2021)
institution DOAJ
collection DOAJ
language EN
topic regulatory T cells
advanced therapy medicinal products
DNA methylation
biomarker
adoptive cell therapy
good manufacturing practice
Biology (General)
QH301-705.5
spellingShingle regulatory T cells
advanced therapy medicinal products
DNA methylation
biomarker
adoptive cell therapy
good manufacturing practice
Biology (General)
QH301-705.5
Kristy Ou
Dania Hamo
Anne Schulze
Andy Roemhild
Daniel Kaiser
Gilles Gasparoni
Abdulrahman Salhab
Ghazaleh Zarrinrad
Leila Amini
Leila Amini
Stephan Schlickeiser
Mathias Streitz
Jörn Walter
Hans-Dieter Volk
Hans-Dieter Volk
Michael Schmueck-Henneresse
Petra Reinke
Julia K. Polansky
Julia K. Polansky
Strong Expansion of Human Regulatory T Cells for Adoptive Cell Therapy Results in Epigenetic Changes Which May Impact Their Survival and Function
description Adoptive transfer of regulatory T cells (Treg) is a promising new therapeutic option to treat detrimental inflammatory conditions after transplantation and during autoimmune disease. To reach sufficient cell yield for treatment, ex vivo isolated autologous or allogenic Tregs need to be expanded extensively in vitro during manufacturing of the Treg product. However, repetitive cycles of restimulation and prolonged culture have been shown to impact T cell phenotypes, functionality and fitness. It is therefore critical to scrutinize the molecular changes which occur during T cell product generation, and reexamine current manufacturing practices. We performed genome-wide DNA methylation profiling of cells throughout the manufacturing process of a polyclonal Treg product that has proven safety and hints of therapeutic efficacy in kidney transplant patients. We found progressive DNA methylation changes over the duration of culture, which were donor-independent and reproducible between manufacturing runs. Differentially methylated regions (DMRs) in the final products were significantly enriched at promoters and enhancers of genes implicated in T cell activation. Additionally, significant hypomethylation did also occur in promoters of genes implicated in functional exhaustion in conventional T cells, some of which, however, have been reported to strengthen immunosuppressive effector function in Tregs. At the same time, a set of reported Treg-specific demethylated regions increased methylation levels with culture, indicating a possible destabilization of Treg identity during manufacturing, which was independent of the purity of the starting material. Together, our results indicate that the repetitive TCR-mediated stimulation lead to epigenetic changes that might impact functionality of Treg products in multiple ways, by possibly shifting to an effector Treg phenotype with enhanced functional activity or by risking destabilization of Treg identity and impaired TCR activation. Our analyses also illustrate the value of epigenetic profiling for the evaluation of T cell product manufacturing pipelines, which might open new avenues for the improvement of current adoptive Treg therapies with relevance for conventional effector T cell products.
format article
author Kristy Ou
Dania Hamo
Anne Schulze
Andy Roemhild
Daniel Kaiser
Gilles Gasparoni
Abdulrahman Salhab
Ghazaleh Zarrinrad
Leila Amini
Leila Amini
Stephan Schlickeiser
Mathias Streitz
Jörn Walter
Hans-Dieter Volk
Hans-Dieter Volk
Michael Schmueck-Henneresse
Petra Reinke
Julia K. Polansky
Julia K. Polansky
author_facet Kristy Ou
Dania Hamo
Anne Schulze
Andy Roemhild
Daniel Kaiser
Gilles Gasparoni
Abdulrahman Salhab
Ghazaleh Zarrinrad
Leila Amini
Leila Amini
Stephan Schlickeiser
Mathias Streitz
Jörn Walter
Hans-Dieter Volk
Hans-Dieter Volk
Michael Schmueck-Henneresse
Petra Reinke
Julia K. Polansky
Julia K. Polansky
author_sort Kristy Ou
title Strong Expansion of Human Regulatory T Cells for Adoptive Cell Therapy Results in Epigenetic Changes Which May Impact Their Survival and Function
title_short Strong Expansion of Human Regulatory T Cells for Adoptive Cell Therapy Results in Epigenetic Changes Which May Impact Their Survival and Function
title_full Strong Expansion of Human Regulatory T Cells for Adoptive Cell Therapy Results in Epigenetic Changes Which May Impact Their Survival and Function
title_fullStr Strong Expansion of Human Regulatory T Cells for Adoptive Cell Therapy Results in Epigenetic Changes Which May Impact Their Survival and Function
title_full_unstemmed Strong Expansion of Human Regulatory T Cells for Adoptive Cell Therapy Results in Epigenetic Changes Which May Impact Their Survival and Function
title_sort strong expansion of human regulatory t cells for adoptive cell therapy results in epigenetic changes which may impact their survival and function
publisher Frontiers Media S.A.
publishDate 2021
url https://doaj.org/article/7b260d54dd1e425b9ebf78e959997007
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