Determination of Serum Visinin Like Protein-1 and Its Potential for the Diagnosis of Brain Injury Due to the Stroke - A Pilot Study

Abstract

Background. The current diagnosis of stroke relies on clinical examination by a physician supplemented by various neuroimaging techniques. A single set or multiple sets of blood biomarkers that could be used in acute settings to diagnosis stroke, differentiate between stroke types, and ideally predict an initial/recurring stroke would be extremely valuable. The diagnosis of stroke is currently hampered by delay due to lack of a suitable tool for rapid, accurate and analytically sensitive biomarker -based testing. There is a clear need for further assay development and clinical validation in this area (acute stroke setting) in order to improve patient outcomes and quality of life. Visinin like protein 1 (VILIP-1) is a newly discovered CNS-abundant protein which has shown promise in experimental studies, for early stroke diagnosis. However, to date there is no clinical study that has measured VILIP-1 in sera as a marker of stroke.Aim. To develop an assay for the determination of VILIP-1 in human serum, and to investigate its clinical relevance as a marker of ischemic stroke.Design and methods. A new sandwich ELISA was developed, introduced and clinically tested. Mean spiking recovery was 98%. The mean recovery for dilution linearity was 93%. The limit of detection of the assay was 0.01 mcg/l; the intraassay and interassay coefficient of variation (CV) were always less than 10%. The study was approved by the Ethics Commission of the Hospital Šternberk, Czech Republic. A total of 17 healthy individuals (9 men and 8 women, age 64.0 ± 13.0) and 16 individuals with ischemic stroke (10 men and 6 women, age 63.0±11.5) were recruited for our study. The criteria of stroke were proposed by the National Czech Standard. All individuals had blood samples drawn, and VILIP-1 analysis and CT and/or MRI were performed.Results. VILIP-1 serum level significantly differentiated healthy subjects from patients with stroke (P<0.01). All individuals with stroke had VILIP-1 serum values higher than > 0.05 mcg/l, healthy had values below this value. The diagnostic efficacy of serum VILIP-1 was very significant (sensitivity 100%, specificity 100% at 0.093 mcg/l VILIP-1 serum values, AUC 1.0 (CI 0.93-1.0, P<0.01), Chi-squared in the frequency table was 33 (P<0.01).Conclusion. We have introduced a new analytical tool for the study of VILIP-1. Our results support the hypothesis that serum VILIP-1 may be associated with ischemic stroke. The ELISA VILIP-1 assay offers a new research tool for the diagnosis and pathophysiology of stroke and other CNS diseases.

Protein Profile and Morphological Alterations in Penumbra after Focal Photothrombotic Infarction in the Rat Cerebral Cortex

“…After [Ca 2+ ] i increase, VILIP1 moves to the cellular membranes and regulates vesicular trafficking and recycling of receptors and ion channels [46]. Upon stroke, it is released from damaged neuronal cells into the cerebrospinal fluid and blood serum being a stroke marker [47].…”

CSF levels of the neuronal injury biomarker visinin‐like protein‐1 in Alzheimer's disease and dementia with Lewy bodies

“…; Stejskal et al . ). Furthermore, VILIP‐1 plays a critical role in linking calcium‐mediated neurotoxicity and Alzheimer‐type pathological changes (Bernstein et al .…”

“…Perturbation of cellular calcium homeostasis is an important mechanism involved in the progression of neurodegeneration and cognitive impairment (Tymianski and Tator 1996;Mattson 2007). Visinin-like protein-1 (VILIP-1) is a member of the neuronal calcium sensor protein family, and has been implicated in calcium-mediated neuronal injury (Spilker and Braunewell 2003;Laterza et al 2006;Stejskal et al 2011). Furthermore, VILIP-1 plays a critical role in linking calcium-mediated neurotoxicity and Alzheimer-type pathological changes (Bernstein et al 1999;Schnurra et al 2001).…”

Current and Future Bioanalytical Approaches for Stroke Assessment

“…Serum and CSF levels of VILIP-1 were found to be elevated in IS patients approximately 3 h after stroke onset compared with healthy subjects. While the clinical sensitivity and specificity for stroke diagnosis were favorable, the assay and biomarker still requires further validation [128]. Another ELISA-based study using organ samples of mouse models enabled the measurement of VILIP-1 concentrations smaller than 100 ng/l and identified it as a potential candidate as a blood-borne biomarker of IS [129].…”