BRD4 PROTAC degrader ARV-825 inhibits T-cell acute lymphoblastic leukemia by targeting 'Undruggable' Myc-pathway genes.
BRD4 PROTAC degrader ARV-825 inhibits T-cell acute lymphoblastic leukemia by targeting 'Undruggable' Myc-pathway genes.
复制标题
BRD4 PROTAC 降解剂 ARV-825 通过靶向“不可成药”Myc 通路基因抑制 T 细胞急性淋巴细胞白血病
DOI：
10.1186/s12935-021-01908-w
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发表时间：
2021-04-22
影响因子：
5.8
通讯作者：
Hu S
中科院分区：
文献类型：
--
作者：
Wu S;Jiang Y;Hong Y;Chu X;Zhang Z;Tao Y;Fan Z;Bai Z;Li X;Chen Y;Li Z;Ding X;Lv H;Du X;Lim SL;Zhang Y;Huang S;Lu J;Pan J;Hu S
背景
T细胞急性淋巴细胞白血病（T-ALL）是一种侵袭性疾病，具有诱导失败和不良结局的高风险，由于耐药性而复发。最近的研究表明，溴结构域和额外的终端（BET）蛋白抑制剂是有前途的抗癌药物。ARV-825包含与cereblon配体缀合的BET抑制剂，最近开发用于在体外和体内减弱多种肿瘤的生长。然而，ARV-825在T-ALL中的功能和分子机制仍不清楚。本研究旨在探讨ARV-825治疗T-ALL的疗效及可能机制。
方法
在儿科T-ALL样本中测定BRD 4的表达，并通过RNA-seq和定量逆转录聚合酶链反应探索ARV-825治疗后的差异基因表达。ARV-825给药后，通过CCK 8测定法测量T-ALL细胞活力。碘化丙啶（PI）染色检测细胞周期，Annexin V/PI染色检测细胞凋亡。用Western blot检测ARV-825处理的细胞中BRD 4、BRD 3和BRD 2蛋白。在体内分析ARV-825对T-ALL细胞的作用。通过蛋白质印迹和染色质免疫沉淀（ChIP）验证了ARV-825治疗T-ALL的功能和分子途径。
结果
与来自健康供体的T细胞相比，儿科T-ALL样本中的BRD 4表达更高。BRD 4高表达表明预后不良。ARV-825通过阻止细胞周期和诱导细胞凋亡抑制体外细胞增殖，并升高聚ADP核糖聚合酶和切割的caspase 3。BRD 4、BRD 3和BRD 2的降解与T-ALL细胞中cereblon表达的降低一致。ARV-825在T-ALL细胞中的IC 50低于JQ 1、dBET 1和OTX 015。根据RNA-seq和ChIP，ARV-825扰乱了H3 K27 Ac-Myc通路，并降低了T-ALL细胞中的c-Myc蛋白水平。在T-ALL异种移植模型中，ARV-825显著降低了肿瘤生长，并导致Ki 67和裂解的caspase 3失调。此外，ARV-825通过在体外和体内消耗BET和c-Myc蛋白来抑制细胞增殖。
结论
BRD 4提示T-ALL预后不良。BRD 4降解剂ARV-825通过去除BET蛋白和抑制c-Myc，可有效抑制T-ALL细胞增殖并促进其凋亡，为T-ALL的治疗提供了新的策略。
Background
T-cell acute lymphoblastic leukemia (T-ALL) is an aggressive disease with a high risk of induction failure and poor outcomes, with relapse due to drug resistance. Recent studies show that bromodomains and extra-terminal (BET) protein inhibitors are promising anti-cancer agents. ARV-825, comprising a BET inhibitor conjugated with cereblon ligand, was recently developed to attenuate the growth of multiple tumors in vitro and in vivo. However, the functional and molecular mechanisms of ARV-825 in T-ALL remain unclear. This study aimed to investigate the therapeutic efficacy and potential mechanism of ARV-825 in T-ALL.
Methods
Expression of the BRD4 were determined in pediatric T-ALL samples and differential gene expression after ARV-825 treatment was explored by RNA-seq and quantitative reverse transcription-polymerase chain reaction. T-ALL cell viability was measured by CCK8 assay after ARV-825 administration. Cell cycle was analyzed by propidium iodide (PI) staining and apoptosis was assessed by Annexin V/PI staining. BRD4, BRD3 and BRD2 proteins were detected by western blot in cells treated with ARV-825. The effect of ARV-825 on T-ALL cells was analyzed in vivo. The functional and molecular pathways involved in ARV-825 treatment of T-ALL were verified by western blot and chromatin immunoprecipitation (ChIP).
Results
BRD4 expression was higher in pediatric T-ALL samples compared with T-cells from healthy donors. High BRD4 expression indicated a poor outcome. ARV-825 suppressed cell proliferation in vitro by arresting the cell cycle and inducing apoptosis, with elevated poly-ADP ribose polymerase and cleaved caspase 3. BRD4, BRD3, and BRD2 were degraded in line with reduced cereblon expression in T-ALL cells. ARV-825 had a lower IC50 in T-ALL cells compared with JQ1, dBET1 and OTX015. ARV-825 perturbed the H3K27Ac-Myc pathway and reduced c-Myc protein levels in T-ALL cells according to RNA-seq and ChIP. In the T-ALL xenograft model, ARV-825 significantly reduced tumor growth and led to the dysregulation of Ki67 and cleaved caspase 3. Moreover, ARV-825 inhibited cell proliferation by depleting BET and c-Myc proteins in vitro and in vivo.
Conclusions
BRD4 indicates a poor prognosis in T-ALL. The BRD4 degrader ARV-825 can effectively suppress the proliferation and promote apoptosis of T-ALL cells via BET protein depletion and c-Myc inhibition, thus providing a new strategy for the treatment of T-ALL.
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影响因子：
3.6
作者：
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通讯作者：
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