肿瘤细胞治疗研究
英文原题：Valproic acid increases CAR T cell cytotoxicity against acute myeloid leukemia.
CD123和CLL-1是AML CAR-T细胞治疗的有前景的靶点。
B细胞恶性肿瘤的治疗随着免疫疗法的引入，尤其是CAR-T（CAR-T）细胞疗法，发生了巨大变化。然而，在急性髓系白血病（AML）中观察到的疗效有限。在本研究中，我们检测了复发/难治性AML（R/R AML）患者白血病细胞上CD123和CLL-1的表达。随后，我们构建了具有不同共刺激结构域（CD28或4-1BB）的抗CD123 CAR和CLL-1 CAR，并检测了它们的抗AML效果。为了提高CAR-T细胞疗法的疗效，我们在体内和体外测试了不同策略，包括联合应用检查点抑制剂和组蛋白去乙酰化酶抑制剂（HDACi）。我们发现CD123和CLL-1在AML细胞上高表达。与健康供者相比，AML患者中参与抗肿瘤或抗炎过程的T细胞亚群和NK细胞比例显著降低。CD123 CAR和CLL-1 CAR在体外均显示出特异性抗AML效果。为了提高CAR-T细胞的裂解效果，我们将CAR-T细胞疗法与不同药物联合使用。PD-1/PD-L1抗体仅略微提高了CAR-T细胞疗法的效力（CD123 CAR-T 60.92% 2.9087% vs. 65.43% 2.1893%，60.92% 2.9087% vs. 67.43% 3.4973%；37.37% 3.908% vs. 41.89% 5.1568%，37.37% 3.908% vs. 42.84% 4.2635%）。然而，一种HDACi（丙戊酸[VPA]）显著提高了CAR-T细胞对AML细胞的效力（CLL-1 CAR-T 34.97% 0.3051% vs. 88.167% 1.5327%，p < 0.0001；CD123 CAR-T 26.87% 2.7010% vs. 82.56% 3.086%，p < 0.0001 in MV411；CLL-1 CAR-T 78.77% 1.2061% vs. 93.743% 1.2333%，p < 0.0001；CD123 CAR-T 64.10% 1.5130% vs. 94.427% 0.142%，p = 0.0001 in THP-1）。联合治疗较单用CD123 CAR-T细胞治疗延长了小鼠的总生存期（中位生存期：180天 vs. 未随访到）。可能的机制是活化的CD8+T细胞上调自然杀伤组2成员D（NKG2D），而VPA上调AML细胞中NKG2D配体的表达，从而促进CAR-T细胞通过NKG2D介导的对肿瘤细胞的细胞毒性作用。总之，CD123和CLL-1是AML CAR-T细胞治疗有前景的靶点。VPA预处理与CAR-T联合治疗AML表现出协同效应。
The treatment of B cell malignancies has dramatically changed with the introduction of immunotherapy, especially chimeric antigen receptor T (CAR-T) cell therapy. However, only limited efficacy is observed in acute myeloid leukaemia (AML). In the study, We detected CD123 and CLL-1 expression on leukaemia cells from Relapsed/Refractory AML (R/R AML) patients. Then, we constructed anti-CD123 CAR and CLL-1 CAR with different co-stimulation domains (CD28 or 4-1BB) and detected their anti-AML effects. To increase the efficacy of CAR-T cell therapy, we tested different strategies, including application of combined checkpoint inhibitors and histone deacetylase inhibitors (HDACi) in vivo and in vitro We found CD123 and CLL-1 were highly expressed on AML cells. The proportions of T cell subsets and NK cells involved in anti-tumour or anti-inflammation processes in AML patients significantly decreased when compared with healthy donors. Both CD123 CAR and CLL-1 CAR displayed specific anti-AML effects in vitro To improve the lysis effects of CAR-T cells, we combined CAR-T cell therapy with different agents. PD-1/PD-L1 antibodies only slightly improved the potency of CAR-T cell therapy (CD123 CAR-T 60.92% 2.9087% vs. 65.43% 2.1893%, 60.92% 2.9087% vs. 67.43% 3.4973%; 37.37% 3.908% vs. 41.89% 5.1568%, 37.37% 3.908% vs. 42.84% 4.2635%). However, one HDACi (valproic acid [VPA]) significantly improved CAR-T cell potency against AML cells (CLL-1 CAR-T 34.97% 0.3051% vs. 88.167% 1.5327%, p < 0.0001; CD123 CAR-T 26.87% 2.7010% vs. 82.56% 3.086%, p < 0.0001 in MV411; CLL-1 CAR-T 78.77% 1.2061% vs. 93.743% 1.2333%, p < 0.0001; CD123 CAR-T 64.10% 1.5130% vs. 94.427% 0.142%, p = 0.0001 in THP-1). Combination therapy prolonged the overall survival of mice when compared with single CD123 CAR-T cell therapy (median survival: 180 days vs. unfollowed). A possible mechanism is that activated CD8+T cells upregulate natural-killer group 2 member D (NKG2D), and VPA upregulates NKG2D ligand expression in AML cells, contributing to NKG2D-mediated cytotoxicity of CAR-T cells against tumour cells. In conclusion, CD123 and CLL-1 are promising targets for AML CAR-T cell therapy. A combination of VPA pre-treatment and CAR-T against AML exhibits synergic effects.
MEMBER ACCOUNT
登录成功会直接打开下一页。
