دورية أكاديمية
Reversing T Cell Exhaustion by Converting Membrane PD-1 to Its Soluble form in Jurkat Cells; Applying The CRISPR/Cas9 Exon Skipping Strategy
العنوان: | Reversing T Cell Exhaustion by Converting Membrane PD-1 to Its Soluble form in Jurkat Cells; Applying The CRISPR/Cas9 Exon Skipping Strategy |
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المؤلفون: | Zeinab Yousefi-Najafabadi, Zohreh Mehmandoostli, Yazdan Asgari, Saeed Kaboli, Reza Falak, Gholam Ali Kardar |
المصدر: | Cell Journal, Vol 25, Iss 9, Pp 633-644 (2023) |
بيانات النشر: | Royan Institute (ACECR), Tehran, 2023. |
سنة النشر: | 2023 |
المجموعة: | LCC:Medicine LCC:Science |
مصطلحات موضوعية: | crispr-cas systems, exhaustion, exons, pd-1-pd-l1 blockade, programmed cell death 1 receptor, Medicine, Science |
الوصف: | Objective: T-cells express two functional forms of the programmed cell death protein 1 (PD-1): membrane (mPD-1) andsoluble (sPD-1). The binding of mPD-1 and its ligand (PD-L1) on tumor cells could lead activated lymphocytes towardexhaustion. Selective deletion of the transmembrane domain via alternative splicing of exon-3 in PD-1 mRNA couldgenerate sPD-1. Overexpression of sPD-1 could disrupt the mPD-1/PD-L1 interaction in tumor-specific T cells. Weinvestigated the effect of secreted sPD-1 from pooled engineered and non-engineered T cell supernatant on survivaland proliferation of lymphocytes in the tumor microenvironment (TME).Materials and Methods: In this experimental study, we designed two sgRNA sequences upstream and downstream ofexon-3 in the PDCD1 gene. The lentiCRISPRv2 puro vector was used to clone the dual sgRNAs and produce lentiviralparticles to transduce Jurkat T cells. Analysis assays were used to clarify the change in PD-1 expression pattern in thepooled (engineered and non-engineered) Jurkat cells. Co-culture conditions were established with PD-L1+ cancer cellsand lymphocytes.Results: CRISPR/Cas9 could delete exon-3 of the PDCD1 gene in the engineered cells based on the tracking of indelsby decomposition (TIDE) and interference of CRISPR edit (ICE) sequencing analysis reports. Our results showed a12% reduction in mPD-1 positive cell population after CRISPR manipulation and increment in sPD-1 concentration inthe supernatant. The increased sPD-1 confirmed its positive effect on proliferation of lymphocytes co-cultured with PDL1+cancer cells. The survival percent of lymphocytes co-cultured with the pooled cells supernatant was 12.5% morethan the control.Conclusion: The CRISPR/Cas9 exon skipping approach could be used in adoptive cell immunotherapies to changePD-1 expression patterns and overcome exhaustion. |
نوع الوثيقة: | article |
وصف الملف: | electronic resource |
اللغة: | English |
تدمد: | 2228-5806 2228-5814 |
Relation: | https://www.celljournal.org/article_705477_e7bc442402e593b16d83783f0b5931bf.pdf; https://doaj.org/toc/2228-5806; https://doaj.org/toc/2228-5814 |
DOI: | 10.22074/cellj.2023.1999548.1269 |
URL الوصول: | https://doaj.org/article/2dbb8b5c5e5743ccb6d644fb7bd0df0f |
رقم الأكسشن: | edsdoj.2dbb8b5c5e5743ccb6d644fb7bd0df0f |
قاعدة البيانات: | Directory of Open Access Journals |
تدمد: | 22285806 22285814 |
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DOI: | 10.22074/cellj.2023.1999548.1269 |