Proceedings

Scalable CD45 and PD-1 Gene Knockout Protocols in T cell using CRISPR/Cas9 System for Improved Efficacy and Safety of CAR T cell Therapy

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Chimeric Antigen Receptor (CAR) T cell therapy has revolutionised immunotherapy by boosting T celldriven tumour cell destruction through specific antigen recognition. Despite its success in haematological cancers, significant hurdles in using CAR T cells for solid tumours are the suppression of CAR T cell proliferation and effector functions. This research addresses the pressing challenge in cancer treatment by proposing a strategy to enhance CAR T cell therapy's efficacy in solid tumours. Focusing on CD45 and PD-1, known suppressors in the tumour microenvironment, CRISPR/Cas9 technology was employed to precisely knockout these genes in T cells. The study optimised sgRNA design, nucleotide delivery, and production protocols, successfully generating CD45 and PD-1 negative "enhanced" T cells. Notably, the study also explored the impact of PD-1 knockout on exhaustion markers, revealing stable expressions of LAG3 and TIGIT but dynamic regulation of TIM3. These findings contribute to refining a scalable protocol for integrating immune checkpoint modulation into CAR T cell therapy, potentially improving therapeutic outcomes in solid tumour treatment.

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Detail Information

Series Title

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Call Number

EP BM-008

Publisher

i3L, Jakarta : i3L, Jakarta., 2024

Collation

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Language

English

ISBN/ISSN

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Classification

EP BM-008

Content Type

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Media Type

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Carrier Type

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Edition

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Subject(s)

CD45
PD-1
CAR T cell
CRISPR/Cas9
gene knockout
optimisation

Specific Detail Info

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Statement of Responsibility

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