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CTLA-4 tail fusion enhances CAR-T anti-tumor immunity

ABSTRACT: CTLA-4 tail fusion enhances CAR-T anti-tumor immunity

PROVIDER: PRJNA980728 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR24843437_1.fastq.gz	Fastqsanger.gz
	SRR24843437_2.fastq.gz	Fastqsanger.gz
	SRR24843438_1.fastq.gz	Fastqsanger.gz
	SRR24843438_2.fastq.gz	Fastqsanger.gz
	SRR24843439_1.fastq.gz	Fastqsanger.gz

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CTLA-4 tail fusion enhances CAR-T anti-tumor immunity [bulk RNA-seq]

Project description:CTLA-4 tail fusion enhances CAR-T anti-tumor immunity [bulk RNA-seq]

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CTLA-4 tail fusion enhances CAR-T anti-tumor immunity [single-cell RNA-seq]

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CTLA-4 tail fusion enhances CAR-T anti-tumor immunity [bulk RNA-seq]

Project description:To gain further insight into how CCT fusion engineering impacts their function, we characterized CAR-T cells using transcriptome profiling. We performed mRNA-seq from all four groups of CAR-T cells, either without stimulation (baseline) or with 2 rounds of NALM6GL stimulation. For CAR-T cells that were cocultured with 2 rounds of NALM6GL cells, we sorted the pure CAR-T populations (CAR+; GFP-) before subjecting them to RNA sequencing

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CTLA-4 tail fusion enhances CAR-T anti-tumor immunity [single-cell RNA-seq]

Project description:To systematically investigate the effect of CCT engineering on CAR-T cell phenotypes in vivo, we used single-cell RNA sequencing (scRNA-seq) to characterize the full spectrum of engineered CAR-T cells and their transcriptomic profiles

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CTLA-4 tail fusion enhances CAR-T antitumor immunity.

Project description:Chimeric antigen receptor (CAR)-T cells are powerful therapeutics; however, their efficacy is often hindered by critical hurdles. Here utilizing the endocytic feature of the cytotoxic T-lymphocyte-associated antigen-4 (CTLA-4) cytoplasmic tail, we reprogram CAR function and substantially enhance CAR-T efficacy in vivo. CAR-T cells with monomeric, duplex or triplex CTLA-4 cytoplasmic tails (CCTs) fused to the C terminus of CAR exhibit a progressive increase in cytotoxicity under repeated stimulation, accompanied by reduced activation and production of proinflammatory cytokines. Further characterization reveals that CARs with increasing CCT fusion show a progressively lower surface expression, regulated by their constant endocytosis, recycling and degradation under steady state. The molecular dynamics of reengineered CAR with CCT fusion results in reduced CAR-mediated trogocytosis, loss of tumor antigen and improved CAR-T survival. CARs with either monomeric (CAR-1CCT) or duplex CCTs (CAR-2CCT) have superior antitumor efficacy in a relapsed leukemia model. Single-cell RNA sequencing and flow cytometry analysis reveal that CAR-2CCT cells retain a stronger central memory phenotype and exhibit increased persistence. These findings illuminate a unique strategy for engineering therapeutic T cells and improving CAR-T function through synthetic CCT fusion, which is orthogonal to other cell engineering techniques.

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CTLA-4 tail fusion enhances CAR-T anti-tumor immunity

Project description:This SuperSeries is composed of the SubSeries listed below.

2023-06-09 | GSE234307 | GEO

Transfer of antigen target into CAR T cells via trogocytosis

Project description:To characterize transfer of molecules from target cells into CAR T cells via trogocytosis we cultured NALM-6 leukemia cell line expressing a CD19-mCherry fusion protein with CAR T cells. NALM6-CD19-mCherry were loaded with heavy amino acid and cocultured with CAR T cells for 1 hour. CAR T cells were next sorted into two fractions, mCherry-positive (TrogPos), and -negative (TrogNeg). Proteomics analysis revealed the presence of targeted antigen (CD19) in the TrogPos only.

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