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Updated: Jan 18, 2026
Generation of Induced Regulatory T Cells from Primary Human Naïve and Memory T Cells
Published on: April 16, 2012
Latent Regulatory Programs Generate Synthetic T Cell States with Enhanced Therapeutic Potential
Brandon M Pratt1,2, Genevieve N Mullins1,3, Nolan Brown1
1Lineberger Comprehensive Cancer Center, University of North Carolina at Chapel Hill, Chapel Hill, NC, 27599, USA.
Abstract:
Transcription factors (TFs) govern cell fate through coordinated gene-regulatory networks, yet the full potential of these networks to generate non-native, therapeutically advantageous cell states in vivo remains largely unexplored. We hypothesized that systematic gain-of-function (GOF) overexpression of TFs in CD8+ T cells, central mediators of immune protection, could reveal latent, or "hidden," regulatory programs capable of generating synthetic T cell states with therapeutic utility. To test this, we developed single-cell GOF sequencing (scGOF-seq), a multiplexed platform for unbiased, in vivo mapping of GOF effects on T cell fate in immunocompetent mouse models of infection and cancer. scGOF-seq uncovered unexpected regulators of T cell differentiation and accumulation, including SOX2, OCT4, and GATA2, which are normally silenced during T cell differentiation. Notably, outside its native regulatory context, supraphysiologic cMyc GOF reprogrammed CD8+ T cells into a synthetic stem-effector hybrid state, enabling >5,000-fold antigen-dependent expansion and antitumor activity, contrasting sharply with its native function in driving terminal differentiation. scGOF-seq further identified TF modules that cooperate with cMyc GOF to promote robust CD8+ T cell responses in solid tumors. Together, these findings establish GOF perturbation as a powerful strategy for revealing latent immune regulatory programs and engineering synthetic immune states with therapeutic potential.
Insights
Systematic gain-of-function (GOF) screening of transcription factors in CD8+ T cells revealed hidden programs. This approach engineered synthetic T cell states with potent therapeutic potential, including significant expansion and antitumor activity.
Area of Science:
- Immunology
- Molecular Biology
- Systems Biology
Background:
- Transcription factors (TFs) orchestrate cell fate via gene regulatory networks.
- The potential of these networks to create novel, therapeutically beneficial cell states in vivo is underexplored.
- CD8+ T cells are key immune mediators, making them ideal candidates for therapeutic engineering.
Purpose of the Study:
- To investigate if systematic gain-of-function (GOF) overexpression of TFs in CD8+ T cells can uncover latent regulatory programs.
- To generate synthetic T cell states with therapeutic utility.
- To develop and apply a novel screening platform for in vivo TF analysis.
Main Methods:
- Development of single-cell GOF sequencing (scGOF-seq), a multiplexed platform for unbiased in vivo mapping of TF GOF effects.
- Application of scGOF-seq in immunocompetent mouse models of infection and cancer.
- Analysis of TF effects on CD8+ T cell fate, differentiation, and accumulation.
Main Results:
- scGOF-seq identified SOX2, OCT4, and GATA2 as unexpected regulators of T cell differentiation and accumulation.
- Supraphysiologic cMyc GOF reprogrammed CD8+ T cells into a synthetic stem-effector hybrid state.
- This engineered state demonstrated >5,000-fold antigen-dependent expansion and significant antitumor activity.
- TF modules cooperating with cMyc GOF were identified to enhance CD8+ T cell responses in solid tumors.
Conclusions:
- Gain-of-function (GOF) perturbation is a powerful strategy for uncovering latent immune regulatory programs.
- scGOF-seq enables the engineering of synthetic immune cell states with therapeutic potential.
- This study opens new avenues for developing advanced cell-based immunotherapies.
