Methods for Assessing Serpins as Neuroprotective Therapeutics

Abstract

As the systematic work on the pathogenesis of the white matter injury in the spinal cord models progresses, it becomes obvious that a severe and extraordinarily protracted, destructive inflammation follows the initial injury. Appropriate anti-inflammatory therapies of sufficient duration should not only inhibit but also lead to the elimination of this destructive inflammation, thus resulting in neuroprotection of the spinal cord tissue and a greater preservation of the neurologic function. While dexamethasone, a powerful, anti-inflammatory steroid analog administered continuously by subdural infusion for 7 days inhibited severe macrophage infiltration in the cavity of injury, the dose used was remarkably toxic. A 2-week-long infusion of lower doses of dexamethasone resulted in dose-dependent inhibition of macrophage infiltration and was better tolerated by the rats, but it became evident that a much longer duration of subdural administration of a powerful anti-inflammatory drug is required to eliminate myelin-rich, necrotic debris from the cavity and synthetic steroids such as dexamethasone, and methylprednisolone may be too toxic for this application. Therefore, nontoxic but powerful anti-inflammatory compounds are required for neuroprotective treatment of the spinal cord injury (SCI) and also brain trauma and stroke where the massive injury to the white matter occurs. Serpins have been associated with neurological damage. The mammalian serpin neuroserpin (SERPINI1) is reported to act in a protective manner after cerebrospinal infarction. The serine protease, tissue-type plasminogen activator (tPA), and the serpin plasminogen activator inhibitor (PAI-1, SERPINE1) are both upregulated at sites of central nervous system damage. In preliminary studies, subdural infusion of the myxomaviral serpin, Serp-1, resulted in the powerful inhibition of the macrophage infiltration of the cavity of injury, comparable to the inhibition by high dose of dexamethasone that has proven to be unduly toxic. Nontoxic, yet powerful neuroprotective, anti-inflammatory effects of Serp-1 may indicate this serpin protein as a potential attractive compound to treat SCI and similar syndromes involving massive injury to the white matter such as brain trauma and stroke. Novel methods of drug delivery, chronic subdural infusion, and novel analytic methods to measure the effectiveness of the neuroprotective serpin treatments are discussed in this chapter.