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CRISPRi-mediated knock-down of PRDM1/BLIMP1 programs central memory differentiation in ex vivo-expanded human T cells

Azadbakht, M ; Sharif University of Technology | 2022

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  1. Type of Document: Article
  2. DOI: 10.34172/bi.2021.23522
  3. Publisher: Tabriz University of Medical Sciences , 2022
  4. Abstract:
  5. Introduction: B lymphocyte-induced maturation protein 1 (BLIMP1) encoded by the positive regulatory domain 1 gene (PRDM1), is a key regulator in T cell differentiation in mouse models. BLIMP1-deficiency results in a lower effector phenotype and a higher memory phenotype. Methods: In this study, we aimed to determine the role of transcription factor BLIMP1 in human T cell differentiation. Specifically, we investigated the role of BLIMP1 in memory differentiation and exhaustion of human T cells. We used CRISPR interference (CRISPRi) to knock-down BLIMP1 and investigated the differential expressions of T cell memory and exhaustion markers in BLIMP1-deficient T cells in comparison with BLIMP1-sufficient ex vivo expanded human T cells. Results: BLIMP1-deficiency caused an increase in central memory (CM) T cells and a decrease in effector memory (EM) T cells. There was a decrease in the amount of TIM3 exhaustion marker expression in BLIMP1-deficient T cells; however, there was an increase in PD1 exhaustion marker expression in BLIMP1-deficient T cells compared with BLIMP1-sufficient T cells. Conclusion: Our study provides the first functional evidence of the impact of BLIMP1 on the regulation of human T cell memory and exhaustion phenotype. These findings suggest that BLIMP1 may be a promising target to improve the immune response in adoptive T cell therapy settings. © 2022 The Author(s)
  6. Keywords:
  7. BLIMP1 ; CRISPR interference ; Memory T cell ; PRDM1 ; T cell ; B lymphocyte induced maturation protein 1 ; CD4 antigen ; CD45RA antigen ; CD8 antigen ; Complementary DNA ; CRISPR associated endonuclease Cas9 KRAB fusion protein ; Fusion protein ; L selectin ; Lymphocyte function associated antigen 3 ; Messenger RNA ; Plasmid vector ; Puromycin ; RANTES ; Unclassified drug ; Adoptive immunotherapy ; BHLHE14 gene ; Cell differentiation ; Clustered regularly interspaced short palindromic repeat interference ; Controlled study ; CST7 gene ; Differential expression analysis ; DNA structure ; DNA synthesis ; Effector cell ; ERN1 gene ; Ex vivo study ; Flow cytometry ; Gene ; Gene editing ; Gene expression profiling ; Gene knockdown ; Gene vector ; Genetic transduction ; Human ; Human cell ; Immune response ; MAF gene ; Memory T lymphocyte ; Microarray analysis ; mRNA expression level ; MYO1F gene ; Peripheral blood mononuclear cell ; Phenotype ; Real time polymerase chain reaction ; RNA isolation ; Stem cell ; T cell culture ; T lymphocyte activation ; T lymphocyte subpopulation ; Upregulation
  8. Source: BioImpacts ; Volume 12, Issue 4 , 2022 , Pages 337-347 ; 22285652 (ISSN)
  9. URL: https://pubmed.ncbi.nlm.nih.gov/35975204