Validation of serum IGF-I as a biomarker to monitor the bioactivity of exogenous growth hormone agonists and antagonists in rabbits.
Validation of serum IGF-I as a biomarker to monitor the bioactivity of exogenous growth hormone agonists and antagonists in rabbits.
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DOI：
10.1242/dmm.016519
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发表时间：
2014-11
影响因子：
4.3
通讯作者：
Bidlingmaier M
中科院分区：
文献类型：
--
作者：
Bielohuby M;Zarkesh-Esfahani SH;Manolopoulou J;Wirthgen E;Walpurgis K;Toghiany Khorasgani M;Aghili ZS;Wilkinson IR;Hoeflich A;Thevis M;Ross RJ;Bidlingmaier M
新的生长激素（GH）激动剂和生长激素拮抗剂（GHAs）的开发需要用于临床前测试的动物模型。理想情况下，使用与后来在临床实践中使用的药效学标志物相同的药效学标志物监测治疗效果。然而，完整的啮齿类动物用于此目的的价值有限，因为血清IGF-I，GH在人体中作用的最敏感的药效学标志物，显示对重组人GH治疗无反应，并且几乎没有证据表明GHA的作用，除非以非常高的剂量给药或过度表达。作为一种替代的，更合适的模型，我们探讨了完整的家兔GH作用的药效学标志物。我们对用于分析兔血清的IGF-I测定法进行了首次验证，并使用自动化人IGF-I测定法（IDS-1）检测了精密度、灵敏度、线性和回收率。此外，在不同品系、年龄组和性别的兔中测量IGF-I，并监测IGF-I对重组人GH或GHA Pegvisomant治疗的反应。对于一部分样本，我们使用LC-MS/MS测量IGF-I，并使用定量蛋白质配体印迹分析IGF结合蛋白（IGFBPs）。虽然重组兔IGF-I在人IGF-I测定中的回收率仅为50%，但我们的结果表明，在兔样品中，灵敏度、精密度（1.7-3.3%变异系数）和线性（90.4-105.6%）都很好。正如预期的那样，性别，年龄和遗传背景是兔IGF-I浓度的主要决定因素。单次和多次注射重组人GH后，IGF-I和IGFBP-2水平升高（IGF-I：286±22 vs 434±26 ng/ml; P<0.01），且高度相关（P<0.0001）。从第四次注射开始，GHA治疗降低了IGF-I水平（P<0.01）。总之，我们证明了IDS-IGF-I免疫测定法可用于家兔。与啮齿类动物相似，兔子显示IGF-I的变化取决于性别，年龄和遗传背景。与啮齿类动物不同，IGF-I对重组人GH或GHA治疗的反应与人体中观察到的药效学非常相似，表明兔是检测人GH激动剂和拮抗剂的合适新模型。
The development of new growth hormone (GH) agonists and growth hormone antagonists (GHAs) requires animal models for pre-clinical testing. Ideally, the effects of treatment are monitored using the same pharmacodynamic marker that is later used in clinical practice. However, intact rodents are of limited value for this purpose because serum IGF-I, the most sensitive pharmacodynamic marker for the action of GH in humans, shows no response to treatment with recombinant human GH and there is little evidence for the effects of GHAs, except when administered at very high doses or when overexpressed. As an alternative, more suitable model, we explored pharmacodynamic markers of GH action in intact rabbits. We performed the first validation of an IGF-I assay for the analysis of rabbit serum and tested precision, sensitivity, linearity and recovery using an automated human IGF-I assay (IDS-iSYS). Furthermore, IGF-I was measured in rabbits of different strains, age groups and sexes, and we monitored IGF-I response to treatment with recombinant human GH or the GHA Pegvisomant. For a subset of samples, we used LC-MS/MS to measure IGF-I, and quantitative western ligand blot to analyze IGF-binding proteins (IGFBPs). Although recovery of recombinant rabbit IGF-I was only 50% in the human IGF-I assay, our results show that the sensitivity, precision (1.7–3.3% coefficient of variation) and linearity (90.4–105.6%) were excellent in rabbit samples. As expected, sex, age and genetic background were major determinants of IGF-I concentration in rabbits. IGF-I and IGFBP-2 levels increased after single and multiple injections of recombinant human GH (IGF-I: 286±22 versus 434±26 ng/ml; P<0.01) and were highly correlated (P<0.0001). Treatment with the GHA lowered IGF-I levels from the fourth injection onwards (P<0.01). In summary, we demonstrated that the IDS-iSYS IGF-I immunoassay can be used in rabbits. Similar to rodents, rabbits display variations in IGF-I depending on sex, age and genetic background. Unlike in rodents, the IGF-I response to treatment with recombinant human GH or a GHA closely mimics the pharmacodynamics seen in humans, suggesting that rabbits are a suitable new model to test human GH agonists and antagonists.
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影响因子：
40.5
作者：
Kargi, Atil Y.;Merriam, George R.
通讯作者：
Merriam, George R.
影响因子：
2.9
作者：
HOSSENLOPP, P;SEURIN, D;BINOUX, M
通讯作者：
BINOUX, M
影响因子：
4.8
作者：
Costa, C;Solanes, G;Bosch, F
通讯作者：
Bosch, F
影响因子：
3.2
作者：
Kassem, M;Brixen, K;Flyvbjerg, A
通讯作者：
Flyvbjerg, A
影响因子：
4.2
作者：
Katznelson, Laurence;Atkinson, John L. D.;Miller, Karen K.
通讯作者：
Miller, Karen K.