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

Abstract

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.

Pyroptosis Tuning in Intestinal Cryptosporidiosis via the Natural Histone Deacetylase Inhibitor Romidepsin

“…In contrast to the cryptosporidial impact, romidepsin was reported to enhance IFN-γ signalling and natural killer T-cell activation in many studies. For example, in Wang et al, the killing effects of killer cells and IFN-γ secretion increased synchronously after exposure to romidepsin [69,70]. This runs parallel to the significant increase in IFN-γ production, systemically and locally after romidepsin administration in our study.…”

Recent advances of benzimidazole as anticancer agents

“…1. Romidepsin is an FDA-approved benzimidazole drug which is a histone deacetylase inhibitor for the treatment of cutaneous T-cell lymphoma and is currently being tested in clinical trials for the treatment of other cancers (Wang et al, 2022). 2.…”

“…These compounds have been discovered to stop the growth of cancer cells through several different processes, including prevention of cell division, inducement of cell death, and suppression of angiogenesis. Some examples of benzimidazole drugs that are present in clinical trials (Purushottamachar et al, 2019; Yadav & Narasimhan, 2016) are as follows: Romidepsin is an FDA‐approved benzimidazole drug which is a histone deacetylase inhibitor for the treatment of cutaneous T‐cell lymphoma and is currently being tested in clinical trials for the treatment of other cancers (Wang et al, 2022). Belinostat is a benzimidazole drug, which is a histone deacetylase inhibitor currently tested in clinical trials for the treatment of several cancer types, especially solid tumors and hematological malignancies (Kenny et al, 2020).…”

CAR-T cell immunotherapy for ovarian cancer: hushing the silent killer

“…A dual CAR-T platform, such as the one developed by Jiang and colleagues, might offer therapeutic benefits for the treatment of metastatic peritoneal tumor lesions proficient in the expression of NKG2DL and PD-L1 ( 129 ). According to another study, Wang and colleagues investigated whether treatment of the ovarian cancer cell line SKOV3 with romidepsin (an anticancer agent utilized in the treatment of T-cell lymphomas) could increase their susceptibility to NKG2DL-redirected CAR-T treatment through increasing NKG2DL expression ( 130 ). The results of the in vitro experiments implicated increased surface expression of NKG2DL in the SKOV3 cells which consequently resulted in the increased tumoricidal effects of the NKG2DL-redirected CAR-Ts against these cells (accompanied by elevated levels of secreted INF-γ) ( 130 ).…”

“…According to another study, Wang and colleagues investigated whether treatment of the ovarian cancer cell line SKOV3 with romidepsin (an anticancer agent utilized in the treatment of T-cell lymphomas) could increase their susceptibility to NKG2DL-redirected CAR-T treatment through increasing NKG2DL expression ( 130 ). The results of the in vitro experiments implicated increased surface expression of NKG2DL in the SKOV3 cells which consequently resulted in the increased tumoricidal effects of the NKG2DL-redirected CAR-Ts against these cells (accompanied by elevated levels of secreted INF-γ) ( 130 ). Ultimately, Wang and colleagues concluded that increasing the expression rate of the CAR-T-targeted antigens could enhance the antitumor efficacy of this platform of immunotherapy; however in-depth clinical evaluations of such strategies are warranted for further elucidation ( 130 ).…”