HLA-G gene editing: a novel therapeutic alternative in cancer immunotherapy

Palma, M. B.; Tronik-Le Roux, D.; Amin, G.; Castaneda, S.; Mobbs, A. M.; Scarafia, M. A.; La Greca, A.; Daouya, M.; Poras, I.; Inda, A. M.; Moro, L. N.; Carosella, E. D.; Garcia, M. N.; Miriuka, S. G.

doi:10.1101/2021.01.21.427294

bioRxiv · 10.1101/2021.01.21.427294

HLA-G gene editing: a novel therapeutic alternative in cancer immunotherapy

Palma, M. B.·Tronik-Le Roux, D.·Amin, G.·Castaneda, S.·Mobbs, A. M.·Scarafia, M. A.·La Greca, A.·Daouya, M.·Poras, I.·Inda, A. M.·Moro, L. N.·Carosella, E. D.·Garcia, M. N.·Miriuka, S. G.

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Abstract

Cancer immunotherapies based mainly on the blockade of immune-checkpoint (IC) molecules by anti-IC antibodies offer new alternatives for treatment in oncological diseases. However, a considerable proportion of patients remain unresponsive to them. Hence, the development of novel clinical immunotherapeutic approaches and/or targets are crucial. In this context, targeting the immune-checkpoint HLA-G/ILT2/ILT4 has caused great interest since it is abnormally expressed in several malignancies generating a tolerogenic microenvironment. Here, we used CRISPR/Cas9 gene editing to block the HLA-G expression in two tumor cell lines expressing HLA-G, including a renal cell carcinoma (RCC7) and a choriocarcinoma (JEG-3). Different sgRNA/Cas9 plasmids targeting HLA-G exon 1 and 2 were transfected in both cell lines. Downregulation of HLA-G was reached to different degrees, including complete silencing. Most importantly, HLA-G - cells triggered a higher in vitro response of immune cells with respect to HLA-G + wild type cells. Altogether, we demonstrated for the first time the HLA-G downregulation through gene editing. We propose this approach as a first step to develop novel clinical immunotherapeutic approaches in cancer.

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Palma, M. B., Tronik-Le Roux, D., Amin, G., Castaneda, S., Mobbs, A. M., Scarafia, M. A., La Greca, A., Daouya, M., Poras, I., Inda, A. M., Moro, L. N., Carosella, E. D., Garcia, M. N., Miriuka, S. G.. 2021-01-22. HLA-G gene editing: a novel therapeutic alternative in cancer immunotherapy. https://doi.org/10.1101/2021.01.21.427294

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