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Differences in the phenotypes and transcriptomic signatures of chimeric antigen receptor T lymphocytes manufactured via electroporation or lentiviral transfection

ABSTRACT: Differences in the phenotypes and transcriptomic signatures of chimeric antigen receptor T lymphocytes manufactured via electroporation or lentiviral transfection

PROVIDER: PRJNA873765 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR24086037_1.fastq.gz	Fastqsanger.gz
	SRR24086037_2.fastq.gz	Fastqsanger.gz
	SRR24086038_1.fastq.gz	Fastqsanger.gz
	SRR24086038_2.fastq.gz	Fastqsanger.gz
	SRR24086039_1.fastq.gz	Fastqsanger.gz

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Differences in the phenotypes and transcriptomic signatures of chimeric antigen receptor T lymphocytes manufactured via electroporation or lentiviral transfection

Project description:Chimeric antigen receptor (CAR)-T cell therapy is an innovative treatment for CD19-expressing lymphomas. CAR-T cells are primarily manufactured via lentivirus transfection or transposon electroporation. While anti-tumor efficacy comparisons between the two methods have been conducted, there is a current dearth of studies investigating the phenotype and transcriptome alterations induced in T cells by the two distinct manufacturing methods. Here, we established CAR-T signatures using fluorescent imaging, flow cytometry, and RNA-sequencing. A small fraction of CAR-T cells that were produced using the PiggyBac transposon (PB CAR-T cells) exhibited much higher expression of CAR than those produced using a lentivirus (Lenti CAR-T cells). PB and Lenti CAR-T cells contained a more cytotoxic T cell subsets than control T cells, and Lenti CAR-T cells presented a more pronounced memory phenotype. RNA-sequencing further revealed vast disparities between the two CAR-T cell groups, with PB CAR-T cells exhibiting greater upregulation of cytokines, chemokines, and their receptors. Intriguingly, PB CAR-T cells singularly expressed IL-9 and fewer cytokine release syndrome-associated cytokines when activated by target cells. In addition, PB CAR-T cells exerted faster in vitro cytotoxicity against CD19-expressing K562 cells but similar in vivo anti-tumor efficacy with Lenti CAR-T. Taken together, these data provide insights into the phenotypic alterations induced by lentiviral transfection or transposon electroporation and will attract more attention to the clinical influence of different manufacturing procedures.

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Differences in the phenotypes and transcriptomic signatures of chimeric antigen receptor T lymphocytes manufactured <i>via</i> electroporation or lentiviral transfection.

Project description:Chimeric antigen receptor (CAR)-T cell therapy is an innovative treatment for CD19-expressing lymphomas. CAR-T cells are primarily manufactured via lentivirus transfection or transposon electroporation. While anti-tumor efficacy comparisons between the two methods have been conducted, there is a current dearth of studies investigating the phenotypes and transcriptome alterations induced in T cells by the two distinct manufacturing methods. Here, we established CAR-T signatures using fluorescent imaging, flow cytometry, and RNA-sequencing. A small fraction of CAR-T cells that were produced using the PiggyBac transposon (PB CAR-T cells) exhibited much higher expression of CAR than those produced using a lentivirus (Lenti CAR-T cells). PB and Lenti CAR-T cells contained more cytotoxic T cell subsets than control T cells, and Lenti CAR-T cells presented a more pronounced memory phenotype. RNA-sequencing further revealed vast disparities between the two CAR-T cell groups, with PB CAR-T cells exhibiting greater upregulation of cytokines, chemokines, and their receptors. Intriguingly, PB CAR-T cells singularly expressed IL-9 and fewer cytokine release syndrome-associated cytokines when activated by target cells. In addition, PB CAR-T cells exerted faster in vitro cytotoxicity against CD19-expressing K562 cells but similar in vivo anti-tumor efficacy with Lenti CAR-T. Taken together, these data provide insights into the phenotypic alterations induced by lentiviral transfection or transposon electroporation and will attract more attention to the clinical influence of different manufacturing procedures.

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2023-03-10 | GSE205507 | GEO

Gene expression changes in human cord blood-derived hematopoietic stem and progenitor cells upon Centrifugation enhanced Nanostraw Transfection or Conventional Electroporation

Project description:Gene expression changes in human cord blood-derived hematopoietic stem and progenitor cells upon Centrifugation enhanced Nanostraw Transfection or Conventional Electroporation

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Liposome transfection of miR-145 induces immune response via RIG-I in mesenchymal stem cells

Project description:Liposomal transfection reagents are frequenly used in gene deliverytransfer experiments. The validity of these experiments depends on the absence of changes in genes other than the target gene. Here we report that transfection of synthetic microRNA-145 using liposomes, but not electroporation, induced the upregulation of a range of immune-related genes in human mesenchymal stem cells. The immune response was dependent on the binding of microRNA-145 to retinoic acid inducible gene-I, and was independent on endosome functionality and toll-like receptors. Immune response was not observed after transfection of any of nine other small RNAs, suggesting a sequence restricted response. One of the upregulated mRNAs encode interferon β, which presumably in turn induced the upregulation of another group of immune associated genes. Interestingly, exposure of liposomes alone led to the upregulation of some immune genes, one of them retinoic acid inducible gene-I mRNA.

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Generation of diffuse intrinsic pontine glioma mouse models by brainstem targeted in utero electroporation

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