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Mitigation of chromosome loss in clinical CRISPR-Cas9-engineered T cells [CROPSeq_Data]

ABSTRACT: Mitigation of chromosome loss in clinical CRISPR-Cas9-engineered T cells [CROPSeq_Data]

PROVIDER: PRJNA1010684 | ENA |

REPOSITORIES: ENA

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			Action	DRS
	SRR25788800_1.fastq.gz	Fastqsanger.gz
	SRR25788800_2.fastq.gz	Fastqsanger.gz
	SRR25788801_1.fastq.gz	Fastqsanger.gz
	SRR25788801_2.fastq.gz	Fastqsanger.gz
	SRR25788802_1.fastq.gz	Fastqsanger.gz

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Project description:CRISPR-Cas9 genome editing has enabled advanced T cell therapies, but occasional loss of the targeted chromosome remains a safety concern. To investigate whether Cas9-induced chromosome loss is a universal phenomenon and evaluate its clinical significance, we conducted a systematic analysis in primary human T cells. Arrayed and pooled CRISPR screens revealed that chromosome loss was generalizable across the genome and resulted in partial and entire loss of the chromosome, including in pre-clinical chimeric antigen receptor T cells. T cells with chromosome loss persisted for weeks in culture, implying the potential to interfere with clinical use. A modified cell manufacturing process, employed in our first-in-human clinical trial of Cas9-engineered T cells, 1 dramatically reduced chromosome loss while largely preserving genome editing efficacy. Expression of p53 correlated with protection from chromosome loss observed in this protocol, suggesting both a mechanism and strategy for T cell engineering that mitigates this genotoxicity in the clinic.

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