<i>shp-2</i> gene knockout upregulates CAR-driven cytotoxicity of YT NK cells
In Russia, cancer is the second leading cause of death following cardiovascular diseases. Adoptive transfer of NK cells is a promising approach to fight cancer; however, for their successful use in cancer treatment, it is necessary to ensure their robust accumulation at tumor foci, provide resistanc...
Saved in:
| Main Authors: | , , , , , , , |
|---|---|
| Format: | Article |
| Language: | English |
| Published: |
Siberian Branch of the Russian Academy of Sciences, Federal Research Center Institute of Cytology and Genetics, The Vavilov Society of Geneticists and Breeders
2020-03-01
|
| Series: | Вавиловский журнал генетики и селекции |
| Subjects: | |
| Online Access: | https://vavilov.elpub.ru/jour/article/view/2483 |
| Tags: |
Add Tag
No Tags, Be the first to tag this record!
|
| _version_ | 1832575131293057024 |
|---|---|
| author | V. G. Subrakova S. V. Kulemzin T. N. Belovezhets A. N. Chikaev N. A. Chikaev O. A. Koval A. A. Gorchakov A. V. Taranin |
| author_facet | V. G. Subrakova S. V. Kulemzin T. N. Belovezhets A. N. Chikaev N. A. Chikaev O. A. Koval A. A. Gorchakov A. V. Taranin |
| author_sort | V. G. Subrakova |
| collection | DOAJ |
| description | In Russia, cancer is the second leading cause of death following cardiovascular diseases. Adoptive transfer of NK cells is a promising approach to fight cancer; however, for their successful use in cancer treatment, it is necessary to ensure their robust accumulation at tumor foci, provide resistance to the immunosuppressive tumor microenvironment, and to engineer them with higher cytotoxic activity. NK lymphocytes are known to kill cancer cells expressing a number of stress ligands; and the balance of signals from inhibitory and activating receptors on the surface of the NK cell determines whether a cytotoxic reaction is triggered. We hypothesized that stronger cytotoxicity of NK cells could be achieved via gene editing aimed at enhancing the activating signaling cascades and/or weakening the inhibitory ones, thereby shifting the balance of signals towards NK cell activation and target cell lysis. Here, we took advantage of the CRISPR/Cas9 system to introduce mutations in the coding sequence of the shp-2 (PTPN11) gene encoding the signaling molecule of inhibitory pathways in NK cells. These shp-2 knock-out NK cells were additionally transduced to express a chimeric antigen receptor (CAR) that selectively recognized the antigen of interest on the target cell surface and generated an activating signal. We demonstrate that the combination of shp-2 gene knockout and CAR expression increases the cytotoxicity of effector NK-like YT cells against human prostate cancer cell line Du-145 with ectopic expression of PSMA protein, which is specifically targeted by the CAR. |
| format | Article |
| id | doaj-art-00ec533aee1649809b97f8a9cd4bceba |
| institution | Kabale University |
| issn | 2500-3259 |
| language | English |
| publishDate | 2020-03-01 |
| publisher | Siberian Branch of the Russian Academy of Sciences, Federal Research Center Institute of Cytology and Genetics, The Vavilov Society of Geneticists and Breeders |
| record_format | Article |
| series | Вавиловский журнал генетики и селекции |
| spelling | doaj-art-00ec533aee1649809b97f8a9cd4bceba2025-02-01T09:58:08ZengSiberian Branch of the Russian Academy of Sciences, Federal Research Center Institute of Cytology and Genetics, The Vavilov Society of Geneticists and BreedersВавиловский журнал генетики и селекции2500-32592020-03-01241808610.18699/VJ20.5981025<i>shp-2</i> gene knockout upregulates CAR-driven cytotoxicity of YT NK cellsV. G. Subrakova0S. V. Kulemzin1T. N. Belovezhets2A. N. Chikaev3N. A. Chikaev4O. A. Koval5A. A. Gorchakov6A. V. Taranin7Institute of Molecular and Cellular Biology, Siberian Branch of the Russian Academy of Sciences; Novosibirsk State UniversityInstitute of Molecular and Cellular Biology, Siberian Branch of the Russian Academy of SciencesInstitute of Molecular and Cellular Biology, Siberian Branch of the Russian Academy of Sciences; Novosibirsk State UniversityInstitute of Molecular and Cellular Biology, Siberian Branch of the Russian Academy of SciencesInstitute of Molecular and Cellular Biology, Siberian Branch of the Russian Academy of SciencesInstitute of Molecular and Cellular Biology, Siberian Branch of the Russian Academy of Sciences; Institute of Chemical Biology and Fundamental Medicine, Siberian Branch of the Russian Academy of SciencesInstitute of Molecular and Cellular Biology, Siberian Branch of the Russian Academy of Sciences; Novosibirsk State UniversityInstitute of Molecular and Cellular Biology, Siberian Branch of the Russian Academy of Sciences; Novosibirsk State UniversityIn Russia, cancer is the second leading cause of death following cardiovascular diseases. Adoptive transfer of NK cells is a promising approach to fight cancer; however, for their successful use in cancer treatment, it is necessary to ensure their robust accumulation at tumor foci, provide resistance to the immunosuppressive tumor microenvironment, and to engineer them with higher cytotoxic activity. NK lymphocytes are known to kill cancer cells expressing a number of stress ligands; and the balance of signals from inhibitory and activating receptors on the surface of the NK cell determines whether a cytotoxic reaction is triggered. We hypothesized that stronger cytotoxicity of NK cells could be achieved via gene editing aimed at enhancing the activating signaling cascades and/or weakening the inhibitory ones, thereby shifting the balance of signals towards NK cell activation and target cell lysis. Here, we took advantage of the CRISPR/Cas9 system to introduce mutations in the coding sequence of the shp-2 (PTPN11) gene encoding the signaling molecule of inhibitory pathways in NK cells. These shp-2 knock-out NK cells were additionally transduced to express a chimeric antigen receptor (CAR) that selectively recognized the antigen of interest on the target cell surface and generated an activating signal. We demonstrate that the combination of shp-2 gene knockout and CAR expression increases the cytotoxicity of effector NK-like YT cells against human prostate cancer cell line Du-145 with ectopic expression of PSMA protein, which is specifically targeted by the CAR.https://vavilov.elpub.ru/jour/article/view/2483nk cellscrispr/cas9car-nkshp-2 |
| spellingShingle | V. G. Subrakova S. V. Kulemzin T. N. Belovezhets A. N. Chikaev N. A. Chikaev O. A. Koval A. A. Gorchakov A. V. Taranin <i>shp-2</i> gene knockout upregulates CAR-driven cytotoxicity of YT NK cells Вавиловский журнал генетики и селекции nk cells crispr/cas9 car-nk shp-2 |
| title | <i>shp-2</i> gene knockout upregulates CAR-driven cytotoxicity of YT NK cells |
| title_full | <i>shp-2</i> gene knockout upregulates CAR-driven cytotoxicity of YT NK cells |
| title_fullStr | <i>shp-2</i> gene knockout upregulates CAR-driven cytotoxicity of YT NK cells |
| title_full_unstemmed | <i>shp-2</i> gene knockout upregulates CAR-driven cytotoxicity of YT NK cells |
| title_short | <i>shp-2</i> gene knockout upregulates CAR-driven cytotoxicity of YT NK cells |
| title_sort | i shp 2 i gene knockout upregulates car driven cytotoxicity of yt nk cells |
| topic | nk cells crispr/cas9 car-nk shp-2 |
| url | https://vavilov.elpub.ru/jour/article/view/2483 |
| work_keys_str_mv | AT vgsubrakova ishp2igeneknockoutupregulatescardrivencytotoxicityofytnkcells AT svkulemzin ishp2igeneknockoutupregulatescardrivencytotoxicityofytnkcells AT tnbelovezhets ishp2igeneknockoutupregulatescardrivencytotoxicityofytnkcells AT anchikaev ishp2igeneknockoutupregulatescardrivencytotoxicityofytnkcells AT nachikaev ishp2igeneknockoutupregulatescardrivencytotoxicityofytnkcells AT oakoval ishp2igeneknockoutupregulatescardrivencytotoxicityofytnkcells AT aagorchakov ishp2igeneknockoutupregulatescardrivencytotoxicityofytnkcells AT avtaranin ishp2igeneknockoutupregulatescardrivencytotoxicityofytnkcells |