Department of Hematology, Pukou CLL Center, the First Affiliated Hospital of Nanjing Medical University, Jiangsu Province Hospital, Nanjing, China.
Key Laboratory of Hematology of Nanjing Medical University, Nanjing Medical University, Nanjing, China.
Clin Transl Med. 2022 May;12(5):e798. doi: 10.1002/ctm2.798.
Diffuse large B-cell lymphoma (DLBCL) is the most common subtype of non-Hodgkin lymphoma. Histone deacetylase inhibitors (HDACis) have been widely applied in multiple tumours, but the expected efficacy was not observed in DLBCL. Therefore, this study is aimed to explore superior HDACis and optimise a relative combinational therapeutic strategy.
The antitumour effects of the drug were evaluated by Cell Counting Kit-8 (CCK-8) assay and apoptosis analysis. Single-cell RNA sequencing (scRNA-Seq) was used to analyse the intratumoural heterogeneity of DLBCL cells. Whole-exome sequencing and RNA sequencing were performed to analyse the genetic and transcriptional features. Western blotting, qRT-PCR, protein array, immunohistochemistry, and chromatin immunoprecipitation assays were applied to explore the involved pathways. The antitumour effects of the compounds were assessed using subcutaneous xenograft tumour models.
LAQ824 was screened and confirmed to kill DLBCL cells effectively. Using scRNA-Seq, we characterised the heterogeneity of DLBCL cells under different drug pressures, and c-Fos was identified as a critical factor in the survival of residual tumour cells. Moreover, we demonstrated that combinatorial treatment with LAQ824 and a c-Fos inhibitor more potently inhibited tumour cells both in vitro and in vivo.
Altogether, we found an HDACi, LAQ824, with high efficacy in DLBCL and provided a promising HDACi-based combination therapy strategy.
弥漫性大 B 细胞淋巴瘤（DLBCL）是非霍奇金淋巴瘤中最常见的亚型。组蛋白去乙酰化酶抑制剂（HDACi）已广泛应用于多种肿瘤，但在 DLBCL 中并未观察到预期的疗效。因此，本研究旨在探索更有效的 HDACi 并优化相关的联合治疗策略。
通过细胞计数试剂盒（CCK-8）检测和凋亡分析评估药物的抗肿瘤作用。单细胞 RNA 测序（scRNA-Seq）用于分析 DLBCL 细胞的肿瘤内异质性。全外显子测序和 RNA 测序用于分析遗传和转录特征。Western blot、qRT-PCR、蛋白质芯片、免疫组化和染色质免疫沉淀实验用于探索相关通路。采用皮下移植瘤模型评估化合物的抗肿瘤作用。
筛选并证实 LAQ824 可有效杀伤 DLBCL 细胞。通过 scRNA-Seq，我们在不同药物压力下对 DLBCL 细胞的异质性进行了特征描述，并鉴定出 c-Fos 是残留肿瘤细胞存活的关键因素。此外，我们证明 LAQ824 与 c-Fos 抑制剂联合治疗在体外和体内更有效地抑制肿瘤细胞。
总之，我们发现了一种在 DLBCL 中具有高效的 HDACi（LAQ824），并提供了一种有前景的基于 HDACi 的联合治疗策略。
