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Updated: Jun 3, 2025

A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations

A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations

Published on: October 25, 2018

Simultaneous Epigenetic and Gene Expression Profiling at Single Cell Resolution Uncovers Stem-Like Treg Subsets

Hyo Jeong Nam^1,2, Jeong-Ryeol Gong³, Yong-Hee Kim^1,4

¹Department of Biomedical Sciences, Seoul National University College of Medicine, Seoul 03080, Korea.

|January 13, 2025

Summary

Oligonucleotide expansion generates a unique subset of regulatory T cells (Tregs) with enhanced stem-like and functional properties. This discovery aids in developing Treg cell therapies by improving in vitro Treg expansion methods.

Keywords:

Epigenetics Regulatory T cell Transcriptomics

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A Combinatorial Single-cell Approach to Characterize the Molecular and Immunophenotypic Heterogeneity of Human Stem and Progenitor Populations

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Published on: October 25, 2018

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Generation of Induced Regulatory T Cells from Primary Human Naïve and Memory T Cells

Generation of Induced Regulatory T Cells from Primary Human Naïve and Memory T Cells

Published on: April 16, 2012

Area of Science:

Immunology
Cell Biology
Translational Medicine

Background:

Regulatory T cells (Tregs) are crucial for immune tolerance and hold promise for cell therapy.
Challenges exist in producing functional Tregs in vitro and understanding their behavior during ex vivo expansion.
Previous work indicated oligonucleotide expansion yields FoxP3highHelioshigh Treg subsets.

Purpose of the Study:

To investigate the impact of oligonucleotides on gene expression and epigenetic states in Tregs during in vitro expansion.
To identify and characterize novel Treg subsets generated through oligonucleotide-induced expansion.
To elucidate the molecular mechanisms, including transcription factors, driving the generation of these expanded Tregs.

Main Methods:

Isolation and in vitro expansion of Tregs from healthy individuals.
Single-cell resolution profiling of expanded and non-expanded Tregs.
Analysis of histone modification data and gene regulatory networks.
Identification of specific Treg subsets (seTregs) enriched after oligonucleotide expansion.

Main Results:

Oligonucleotide expansion identified a specific subset of Tregs (seTregs) enriched in expanded populations.
seTregs exhibited enhanced stem-like characteristics and improved functional attributes.
IKZF2 (Helios) was identified as a key transcription factor regulating the generation of these seTregs.

Conclusions:

Oligonucleotide-mediated expansion can generate functionally superior Treg subsets (seTregs).
Understanding the role of IKZF2 (Helios) provides insights into Treg manufacturing for cell therapy.
These findings contribute to optimizing in vitro Treg expansion for clinical applications in cell therapy.