The IAP antagonist birinapant potentiates bortezomib anti-myeloma activity in vitro and in vivo
The IAP antagonist birinapant potentiates bortezomib anti-myeloma activity in vitro and in vivo
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IAP 拮抗剂 birinapant 增强硼替佐米的体外和体内抗骨髓瘤活性
DOI：
10.1186/s13045-019-0713-x
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发表时间：
2019-03
期刊：
影响因子：
--
通讯作者：
Grant S
中科院分区：
文献类型：
--
作者：
Zhou L;Zhang Y;Leng Y;Dai Y;Kmieciak M;Kramer L;Sharma K;Wang Y;Craun W;Grant S
背景Smac模拟物（SM）与蛋白酶体抑制剂（例如，硼替佐米）在很大程度上是未知的，特别是在多发性骨髓瘤（MM），其中硼替佐米代表治疗的支柱的疾病中。MethodsInteractions临床相关的IAP（细胞凋亡蛋白抑制剂）拮抗剂birinapant（TL 32711）和蛋白酶体抑制剂硼替佐米之间的相互作用在多发性骨髓瘤（MM）细胞系和原代细胞以及体内模型中进行了研究。使用MM细胞系监测细胞凋亡的诱导以及基因和蛋白质表达的变化，并在原代MM细胞群中证实。遗传修饰的细胞（例如，显示shRNA敲低或异位表达）来评价birinapant/硼替佐米诱导的蛋白质水平变化的功能意义。MM异种移植模型被用来评估在体内活动的birinapant/bortezelatin regiment.ResultsBirinapant和bortezelatin协同诱导不同的细胞系，包括硼替佐米耐药细胞（PS-R）的凋亡。该方案强烈下调cIAP 1/2，但不下调经典NF-κB通路，反映了p65磷酸化和核积聚。相反，硼替佐米/birinapant方案上调TRAF 3，下调TRAF 2，并减少p52加工和BCL-XL表达，与非经典NF-κB通路的破坏一致。TRAF 3敲除、异位TRAF 2或BCL-XL表达显著降低了birinapant/硼替佐米毒性。该方案急剧增加了外源性凋亡途径的激活，表达显性阴性FADD或caspase-8的细胞显示出显著降低的birinapant/硼替佐米敏感性。原发性CD 138+（n= 43）和原始MM群体（CD 138 −/19+/20+/27+;n= 31）但非正常CD 34+细胞在联合治疗中表现出显著增强的毒性（P< 0.0001）。该方案在HS-5基质细胞或生长因子（例如，IL-6和VEGF）。最后，在MM异种移植模型中，与单药治疗相比，该方案耐受性良好，存活率显著增加（P< 0.05和P < 0.001）。联合治疗也下调cIAP 1/2和p52，同时增加PARP裂解在MM cells in vivo.ConclusionsOur data suggests birinapant和bortezalide互动协同MM cells，包括那些耐bortezalide，通过失活的非经典NF-κB和激活的外源性凋亡途径在体外和体内。他们还认为，cIAP拮抗剂和蛋白酶体抑制剂相结合的策略值得MM的关注。
BackgroundMechanisms by which Smac mimetics (SMs) interact with proteasome inhibitors (e.g., bortezomib) are largely unknown, particularly in multiple myeloma (MM), a disease in which bortezomib represents a mainstay of therapy.MethodsInteractions between the clinically relevant IAP (inhibitor of apoptosis protein) antagonist birinapant (TL32711) and the proteasome inhibitor bortezomib were investigated in multiple myeloma (MM) cell lines and primary cells, as well as in vivo models. Induction of apoptosis and changes in gene and protein expression were monitored using MM cell lines and confirmed in primary MM cell populations. Genetically modified cells (e.g., exhibiting shRNA knockdown or ectopic expression) were employed to evaluate the functional significance of birinapant/bortezomib-induced changes in protein levels. A MM xenograft model was used to evaluate the in vivo activity of the birinapant/bortezomib regimen.ResultsBirinapant and bortezomib synergistically induced apoptosis in diverse cell lines, including bortezomib-resistant cells (PS-R). The regimen robustly downregulated cIAP1/2 but not the canonical NF-κB pathway, reflected by p65 phosphorylation and nuclear accumulation. In contrast, the bortezomib/birinapant regimen upregulated TRAF3, downregulated TRAF2, and diminished p52 processing and BCL-XLexpression, consistent with disruption of the non-canonical NF-κB pathway. TRAF3 knockdown, ectopic TRAF2, or BCL-XLexpression significantly diminished birinapant/bortezomib toxicity. The regimen sharply increased extrinsic apoptotic pathway activation, and cells expressing dominant-negative FADD or caspase-8 displayed markedly reduced birinapant/bortezomib sensitivity. Primary CD138+(n= 43) and primitive MM populations (CD138−/19+/20+/27+;n= 31) but not normal CD34+cells exhibited significantly enhanced toxicity with combined treatment (P< 0.0001). The regimen was also fully active in the presence of HS-5 stromal cells or growth factors (e.g., IL-6 and VEGF). Finally, the regimen was well tolerated and significantly increased survival (P< 0.05 andP< 0.001) compared to single agents in a MM xenograft model. Combined treatment also downregulated cIAP1/2 and p52 while increasing PARP cleavage in MM cells in vivo.ConclusionsOur data suggest that birinapant and bortezomib interact synergistically in MM cells, including those resistant to bortezomib, through inactivation of the non-canonical NF-κB and activation of the extrinsic apoptotic pathway both in vitro and in vivo. They also argue that a strategy combining cIAP antagonists and proteasome inhibitors warrants attention in MM.
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影响因子：
4
作者：
Dubrez L;Berthelet J;Glorian V
通讯作者：
Glorian V
影响因子：
--
作者：
Fristedt Duvefelt C;Lub S;Agarwal P;Arngården L;Hammarberg A;Maes K;Van Valckenborgh E;Vanderkerken K;Jernberg Wiklund H
通讯作者：
Jernberg Wiklund H
影响因子：
82.9
作者：
Chesi M;Mirza NN;Garbitt VM;Sharik ME;Dueck AC;Asmann YW;Akhmetzyanova I;Kosiorek HE;Calcinotto A;Riggs DL;Keane N;Ahmann GJ;Morrison KM;Fonseca R;Lacy MQ;Dingli D;Kumar SK;Ailawadhi S;Dispenzieri A;Buadi F;Gertz MA;Reeder CB;Lin Y;Chanan-Khan AA;Stewart AK;Fooksman D;Bergsagel PL
通讯作者：
Bergsagel PL
影响因子：
30.5
作者：
通讯作者：
--
DOI：
10.1083/jcb.200904158
发表时间：
2009-12-28
期刊：
The Journal of cell biology
影响因子：
--
作者：
Geserick P;Hupe M;Moulin M;Wong WW;Feoktistova M;Kellert B;Gollnick H;Silke J;Leverkus M
通讯作者：
Leverkus M