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T Cells Directed against the Metastatic Driver Chondromodulin-1 in Ewing Sarcoma: Comparative Engineering with CRISPR/Cas9 vs. Retroviral Gene Transfer for Adoptive Transfer
SIMPLE SUMMARY: The canonical methods of TCR gene delivery in pre-clinical and clinical applications are based on viral transduction of full-coding sequences, including α- and β-chains recognizing tumor-specific antigens and tumor-associated antigens. As the transduced α- and β-chains may mispair wi...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
MDPI
2022
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9688113/ https://www.ncbi.nlm.nih.gov/pubmed/36428578 http://dx.doi.org/10.3390/cancers14225485 |
Sumario: | SIMPLE SUMMARY: The canonical methods of TCR gene delivery in pre-clinical and clinical applications are based on viral transduction of full-coding sequences, including α- and β-chains recognizing tumor-specific antigens and tumor-associated antigens. As the transduced α- and β-chains may mispair with the endogenous α- and β-chains, the resultant new antigen specificities may cause auto-reactivity, potentially leading to graft-versus-host disease. The mispaired TCRs may also lose their function. We assessed the feasibility of endogenous TCR orthotopic replacement with a TCR containing a CHM1 targeting sequence via CRISPR/Cas9, evaluated tumor recognition and cytotoxicity function of the CRISPR/Cas9-engineered T cells; compared the prevention of endogenous TCR expression in CRISPR/Cas9 vs. retrovirally engineered T cells. We show that both engineered T cell products specifically recognize tumor cells and elicit cytotoxicity in vitro, with CRISPR/Cas9 engineered T cells providing a more prolonged cytotoxic activity. ABSTRACT: Ewing sarcoma (EwS) is a highly malignant sarcoma of bone and soft tissue with early metastatic spread and an age peak in early puberty. The prognosis in advanced stages is still dismal, and the long-term effects of established therapies are severe. Efficacious targeted therapies are urgently needed. Our previous work has provided preliminary safety and efficacy data utilizing T cell receptor (TCR) transgenic T cells, generated by retroviral gene transfer, targeting HLA-restricted peptides on the tumor cell derived from metastatic drivers. Here, we compared T cells engineered with either CRISPR/Cas9 or retroviral gene transfer. Firstly, we confirmed the feasibility of the orthotopic replacement of the endogenous TCR by CRISPR/Cas9 with a TCR targeting our canonical metastatic driver chondromodulin-1 (CHM1). CRISPR/Cas9-engineered T cell products specifically recognized and killed HLA-A*02:01+ EwS cell lines. The efficiency of retroviral transduction was higher compared to CRISPR/Cas9 gene editing. Both engineered T cell products specifically recognized tumor cells and elicited cytotoxicity, with CRISPR/Cas9 engineered T cells providing prolonged cytotoxic activity. In conclusion, T cells engineered with CRISPR/Cas9 could be feasible for immunotherapy of EwS and may have the advantage of more prolonged cytotoxic activity, as compared to T cells engineered with retroviral gene transfer. |
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