Romidepsin Enhances the Killing Ability of NKG2D-CAR-T Cells through Enhanced Expression of NKG2DL against Ovarian Cancer Cells

Background: Upregulating tumor cell targeting antigens and improving the cytotoxicity of chimeric antigen receptor T cell (CAR-T) are expected to facilitate better treatment efficacy for solid cancers represented by ovarian cancer. Methods: Killer cell lectin-like receptor subfamily K member 1 ligan...

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Main Authors: Liang Wang, Xite Lin, Lirui Yu, Pengming Sun
Format: Article
Language:English
Published: IMR Press 2022-09-01
Series:Clinical and Experimental Obstetrics & Gynecology
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Online Access:https://www.imrpress.com/journal/CEOG/49/10/10.31083/j.ceog4910227
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author Liang Wang
Xite Lin
Lirui Yu
Pengming Sun
author_facet Liang Wang
Xite Lin
Lirui Yu
Pengming Sun
author_sort Liang Wang
collection DOAJ
description Background: Upregulating tumor cell targeting antigens and improving the cytotoxicity of chimeric antigen receptor T cell (CAR-T) are expected to facilitate better treatment efficacy for solid cancers represented by ovarian cancer. Methods: Killer cell lectin-like receptor subfamily K member 1 ligands (NKG2DL) are the target antigens for ovarian cancer. NKG2D-CAR-T cells were constructed for NKG2D ligand on the ovarian cancer cell surface. We used flow cytometry to evaluate the expression of NKG2DL on SKOV3 (a human ovarian cancer adenocarcinoma cell line). Innovatively, when combined with romidepsin to treat ovarian cancer cell SKOV3, to evaluate the killing ability of the combined strategy, we verified the cytotoxicity of CAR-T cells by lactate dehydrogenase (LDH) release test and determined the secretion of cytokines by enzyme-linked immuno sorbent assay (ELISA). Results: The results of flow cytometry showed effective activation of the NKG2D-CAR-T cells, and romidepsin treatment resulted in increased expression of NKG2DL on the surface of SKOV3. Cytotoxicity test showed that romidepsin could enhance the killing ability of NKG2D-CAR-T cells against ovarian cancer cells by regulating their NKG2DL expression (p < 0.05). The killing effects and secretion of interferon-γ (IFN-γ) increased synchronously (p < 0.05). Levels of interleukin-2 (IL-2) and Pore-forming protein (PFP) were statistically significant at a low target ratio but programmed cell death protein 1 (PD-1) remained unaffected (p ≥ 0.05). Conclusions: Enhancing the expression of tumor target antigen is a solution to improve the limited application of CAR-T cells in solid cancers.
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spelling doaj-art-d8d6d41c9c1243b2b2abaa6e0a9710822025-08-20T03:52:02ZengIMR PressClinical and Experimental Obstetrics & Gynecology0390-66632022-09-01491022710.31083/j.ceog4910227S0390-6663(22)01919-4Romidepsin Enhances the Killing Ability of NKG2D-CAR-T Cells through Enhanced Expression of NKG2DL against Ovarian Cancer CellsLiang Wang0Xite Lin1Lirui Yu2Pengming Sun3College of Clinical Medicine for Obstetrics & Gynecology and Pediatrics, Department of Obstetrics and Gynecology, Fujian Maternity and Child Health Hospital, Fujian Medical University, 350000 Fuzhou, Fujian, ChinaCollege of Clinical Medicine for Obstetrics & Gynecology and Pediatrics, Department of Obstetrics and Gynecology, Fujian Maternity and Child Health Hospital, Fujian Medical University, 350000 Fuzhou, Fujian, ChinaCollege of Clinical Medicine for Obstetrics & Gynecology and Pediatrics, Department of Obstetrics and Gynecology, Fujian Maternity and Child Health Hospital, Fujian Medical University, 350000 Fuzhou, Fujian, ChinaCollege of Clinical Medicine for Obstetrics & Gynecology and Pediatrics, Department of Obstetrics and Gynecology, Fujian Maternity and Child Health Hospital, Fujian Medical University, 350000 Fuzhou, Fujian, ChinaBackground: Upregulating tumor cell targeting antigens and improving the cytotoxicity of chimeric antigen receptor T cell (CAR-T) are expected to facilitate better treatment efficacy for solid cancers represented by ovarian cancer. Methods: Killer cell lectin-like receptor subfamily K member 1 ligands (NKG2DL) are the target antigens for ovarian cancer. NKG2D-CAR-T cells were constructed for NKG2D ligand on the ovarian cancer cell surface. We used flow cytometry to evaluate the expression of NKG2DL on SKOV3 (a human ovarian cancer adenocarcinoma cell line). Innovatively, when combined with romidepsin to treat ovarian cancer cell SKOV3, to evaluate the killing ability of the combined strategy, we verified the cytotoxicity of CAR-T cells by lactate dehydrogenase (LDH) release test and determined the secretion of cytokines by enzyme-linked immuno sorbent assay (ELISA). Results: The results of flow cytometry showed effective activation of the NKG2D-CAR-T cells, and romidepsin treatment resulted in increased expression of NKG2DL on the surface of SKOV3. Cytotoxicity test showed that romidepsin could enhance the killing ability of NKG2D-CAR-T cells against ovarian cancer cells by regulating their NKG2DL expression (p < 0.05). The killing effects and secretion of interferon-γ (IFN-γ) increased synchronously (p < 0.05). Levels of interleukin-2 (IL-2) and Pore-forming protein (PFP) were statistically significant at a low target ratio but programmed cell death protein 1 (PD-1) remained unaffected (p ≥ 0.05). Conclusions: Enhancing the expression of tumor target antigen is a solution to improve the limited application of CAR-T cells in solid cancers.https://www.imrpress.com/journal/CEOG/49/10/10.31083/j.ceog4910227car-t cellsovarian cancernkg2dlromidepsincytotoxicity
spellingShingle Liang Wang
Xite Lin
Lirui Yu
Pengming Sun
Romidepsin Enhances the Killing Ability of NKG2D-CAR-T Cells through Enhanced Expression of NKG2DL against Ovarian Cancer Cells
Clinical and Experimental Obstetrics & Gynecology
car-t cells
ovarian cancer
nkg2dl
romidepsin
cytotoxicity
title Romidepsin Enhances the Killing Ability of NKG2D-CAR-T Cells through Enhanced Expression of NKG2DL against Ovarian Cancer Cells
title_full Romidepsin Enhances the Killing Ability of NKG2D-CAR-T Cells through Enhanced Expression of NKG2DL against Ovarian Cancer Cells
title_fullStr Romidepsin Enhances the Killing Ability of NKG2D-CAR-T Cells through Enhanced Expression of NKG2DL against Ovarian Cancer Cells
title_full_unstemmed Romidepsin Enhances the Killing Ability of NKG2D-CAR-T Cells through Enhanced Expression of NKG2DL against Ovarian Cancer Cells
title_short Romidepsin Enhances the Killing Ability of NKG2D-CAR-T Cells through Enhanced Expression of NKG2DL against Ovarian Cancer Cells
title_sort romidepsin enhances the killing ability of nkg2d car t cells through enhanced expression of nkg2dl against ovarian cancer cells
topic car-t cells
ovarian cancer
nkg2dl
romidepsin
cytotoxicity
url https://www.imrpress.com/journal/CEOG/49/10/10.31083/j.ceog4910227
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AT liruiyu romidepsinenhancesthekillingabilityofnkg2dcartcellsthroughenhancedexpressionofnkg2dlagainstovariancancercells
AT pengmingsun romidepsinenhancesthekillingabilityofnkg2dcartcellsthroughenhancedexpressionofnkg2dlagainstovariancancercells