Measuring the pharmacodynamic effects of a novel Hsp90 inhibitor on HER2/neu expression in mice using Zr-DFO-trastuzumab.
Measuring the pharmacodynamic effects of a novel Hsp90 inhibitor on HER2/neu expression in mice using Zr-DFO-trastuzumab.
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DOI：
10.1371/journal.pone.0008859
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发表时间：
2010-01-25
期刊：
影响因子：
3.7
通讯作者：
Lewis JS
中科院分区：
文献类型：
--
作者：
Holland JP;Caldas-Lopes E;Divilov V;Longo VA;Taldone T;Zatorska D;Chiosis G;Lewis JS
使用正电子发射放射性核素89 Zr（t1/2 = 3.17天）制备89 Zr放射性标记的曲妥珠单抗，用作表征HER 2/neu阳性乳腺肿瘤的放射性示踪剂。 此外，还对治疗剂量的PU-H71（热休克蛋白90 [Hsp 90]的特异性抑制剂）的HER 2/neu表达水平进行了药效学研究。曲妥珠单抗用去铁胺B（DFO）官能化，并使用改良的文献方法在室温下用[89 Zr] Zr-草酸盐进行放射性标记。在携带皮下BT-474（HER 2/neu阳性）和/或MDA-MB-468（HER 2/neu阴性）肿瘤异种移植物的雌性无胸腺nu/nu小鼠中进行免疫PET和生物分布实验。通过生物分布研究、蛋白质印迹分析和免疫PET评价了89 Zr-DFO-曲妥珠单抗组织摄取对高比活度和低比活度制剂以及PU-H71联合给药的响应变化。89 Zr-DFO-曲妥珠单抗放射性标记以高放射化学产率和比活度104.3±2.1 MBq/mg（2.82±0.05 mCi/mg mAb）进行。体外测定表明放射化学纯度>99%，免疫反应性分数为0.87±0.07。体内生物分布实验显示，BT-474在24、48和72小时后具有高特异性吸收（分别为64.68±13.06%ID/g; 71.71±10.35%ID/g和85.18±11.10%ID/g），活性保留超过120小时。用PU-H71预处理后进行89 Zr-DFO-曲妥珠单抗的生物分布研究和免疫PET。HER 2/neu的表达水平在给药后的前24和48小时内被调节（分别为29.75±4.43%ID/g和41.42±3.64%ID/g）。通过72小时放射性示踪剂摄入（73.64±12.17%ID/g）和蛋白质印迹分析表明HER 2/neu表达恢复至基线水平。结果表明，89 Zr-DFO-曲妥珠单抗提供了HER 2/neu阳性肿瘤的定量和高度特异性描绘，并且具有用于测量用Hsp 90抑制剂（如PU-H71）长期治疗的功效的潜力，其显示出扩展的药效学特征。
The positron-emitting radionuclide 89Zr (t 1/2 = 3.17 days) was used to prepare 89Zr-radiolabeled trastuzumab for use as a radiotracer for characterizing HER2/neu-positive breast tumors. In addition, pharmacodynamic studies on HER2/neu expression levels in response to therapeutic doses of PU-H71 (a specific inhibitor of heat-shock protein 90 [Hsp90]) were conducted. Trastuzumab was functionalized with desferrioxamine B (DFO) and radiolabeled with [89Zr]Zr-oxalate at room temperature using modified literature methods. ImmunoPET and biodistribution experiments in female, athymic nu/nu mice bearing sub-cutaneous BT-474 (HER2/neu positive) and/or MDA-MB-468 (HER2/neu negative) tumor xenografts were conducted. The change in 89Zr-DFO-trastuzumab tissue uptake in response to high- and low-specific-activity formulations and co-administration of PU-H71 was evaluated by biodistribution studies, Western blot analysis and immunoPET. 89Zr-DFO-trastuzumab radiolabeling proceeded in high radiochemical yield and specific-activity 104.3±2.1 MBq/mg (2.82±0.05 mCi/mg of mAb). In vitro assays demonstrated >99% radiochemical purity with an immunoreactive fraction of 0.87±0.07. In vivo biodistribution experiments revealed high specific BT-474 uptake after 24, 48 and 72 h (64.68±13.06%ID/g; 71.71±10.35%ID/g and 85.18±11.10%ID/g, respectively) with retention of activity for over 120 h. Pre-treatment with PU-H71 was followed by biodistribution studies and immunoPET of 89Zr-DFO-trastuzumab. Expression levels of HER2/neu were modulated during the first 24 and 48 h post-administration (29.75±4.43%ID/g and 41.42±3.64%ID/g, respectively). By 72 h radiotracer uptake (73.64±12.17%ID/g) and Western blot analysis demonstrated that HER2/neu expression recovered to baseline levels. The results indicate that 89Zr-DFO-trastuzumab provides quantitative and highly-specific delineation of HER2/neu positive tumors, and has potential to be used to measure the efficacy of long-term treatment with Hsp90 inhibitors, like PU-H71, which display extended pharmacodynamic profiles.
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影响因子：
9.3
作者：
Dijkers, Eli C. F.;Kosterink, Jos G. W.;Lub-de Hooge, Marjolijn N.
通讯作者：
Lub-de Hooge, Marjolijn N.
DOI：
10.1073/pnas.0903392106
发表时间：
2009-05-19
影响因子：
11.1
作者：
Caldas-Lopes, Eloisi;Cerchietti, Leandro;Chiosis, Gabriela
通讯作者：
Chiosis, Gabriela
评估作为双功能螯合剂的去铁醛在用 Zr-89 标记抗体方面的作用
影响因子：
1.6
作者：
MEIJS, WE;HERSCHEID, JDM;PINEDO, HM
通讯作者：
PINEDO, HM
影响因子：
9.3
作者：
Kim, Jin Su;Lee, Jae Sung;Moon, Dae Hyuk
通讯作者：
Moon, Dae Hyuk
影响因子：
3.1
作者：
Meijs, WE;Haisma, HJ;Herscheid, JDM
通讯作者：
Herscheid, JDM