Department of Neurosurgery, Center for Brain Injury and Repair, University of Pennsylvania School of Medicine, Philadelphia, Pennsylvania 19104, USA.
J Neurotrauma. 2009 Nov;26(11):1867-77. doi: 10.1089/neu.2009.0882.
Surrogate markers have enormous potential for contributing to the diagnosis, prognosis, and therapeutic evaluation of acute brain damage, but extensive prior study of individual candidates has not yielded a biomarker in widespread clinical practice. We hypothesize that a panel of neuron-enriched proteins measurable in cerebrospinal fluid (CSF) and blood should vastly improve clinical evaluation and therapeutic management of acute brain injuries. Previously, we developed such a panel based initially on the study of protein release from degenerating cultured neurons, and subsequently on rodent models of traumatic brain injury (TBI) and ischemia, consisting of 14-3-3beta, 14-3-3zeta, three distinct phosphoforms of neurofilament H, ubiquitin hydrolase L1, neuron-specific enolase, alpha-spectrin, and three calpain- and caspase-derived fragments of alpha-spectrin. In the present study, this panel of 11 proteins was evaluated as CSF and serum biomarkers for severe TBI in humans. By quantitative Western blotting and sandwich immunoassays, the CSF protein levels were near or below the limit of detection in pre-surgical and most normal pressure hydrocephalus (NPH) controls, but following TBI nine of the 11 were routinely elevated in CSF. Whereas different markers peaked coordinately, the time to peak varied across TBI cases from 24-96 h post-injury. In serum, TBI increased all four members of the marker panel for which sandwich immunoassays are currently available: a calpain-derived NH(2)-terminal alpha-spectrin fragment and the three neurofilament H phosphoforms. Our results identify neuron-enriched proteins that may serve as a panel of CSF and blood surrogate markers for the minimally invasive detection, management, mechanistic, and therapeutic evaluation of human TBI.
替代标志物在急性脑损伤的诊断、预后和治疗评估方面具有巨大的潜力,但对个别候选标志物的广泛前期研究并未在广泛的临床实践中产生生物标志物。我们假设,一组可在脑脊液 (CSF) 和血液中测量的富含神经元的蛋白质应该极大地改善急性脑损伤的临床评估和治疗管理。此前,我们基于从退化培养神经元中研究蛋白质释放的初始研究,并随后基于创伤性脑损伤 (TBI) 和缺血的啮齿动物模型,开发了这样的一个面板,其中包括 14-3-3beta、14-3-3zeta、三种不同的神经丝 H 磷酸化形式、泛素水解酶 L1、神经元特异性烯醇化酶、α- spectrin 和三种钙蛋白酶和半胱氨酸蛋白酶衍生的α- spectrin 片段。在本研究中,该面板中的 11 种蛋白质被评估为人类严重 TBI 的 CSF 和血清生物标志物。通过定量 Western 印迹和夹心免疫测定,在术前和大多数正常压力脑积水 (NPH) 对照中,CSF 蛋白水平接近或低于检测限,但在 TBI 后,11 种中有 9 种在 CSF 中常规升高。虽然不同的标志物协调地达到峰值,但 TBI 病例之间的峰值时间从损伤后 24-96 小时不等。在血清中,TBI 增加了当前可用夹心免疫测定的标志物面板中的所有四个成员:钙蛋白酶衍生的 NH(2)-末端α- spectrin 片段和三种神经丝 H 磷酸化形式。我们的研究结果确定了富含神经元的蛋白质,它们可能作为 CSF 和血液替代标志物的一个面板,用于微创检测、管理、机制和治疗评估人类 TBI。