CYTOKINE AND CHEMOKINE PROFILES IN ALS PATIENTS UNDERGOING STEM CELL EXPERIMENTAL TREATMENT

Judyta Juranek, Joanna Czarzasta, Barczewska Monika, Tomasz Siwek, Marcin Jozwik, Wojciech Maksymowicz and Joanna Wojtkiewicz

Department of Pathophysiology, Faculty of Medical Sciences, University of Warmia, Olsztyn, Poland

Abstract

Introduction: Amyotrophic Lateral Sclerosis (ALS) is an age-related progressive neurodegenerative disease characterized by the selective loss of motor neurons in the cortex, brainstem and spinal cord, neurogenic atrophy of muscle and lateral scarring. In Europe, on average, 2 new cases per 100,000 are noted annually, accounting for approximately 15,000 new occurrences every year. The onset of ALS is between 40-70 years old resulting in most cases in death within 5 years of diagnosis; the markedly abrupt rate of personal decline and unknown etiology make ALS one of the most devastating and perplexing of the neurodegenerative diseases. Although the pathogenesis of the disease is largely unknown, growing evidence suggests that one of key factors contributing to the progress of the disease is an increased activation of microglia and an excessive production of cyto- and chemokines. Here, in this study we aimed to establish cytokine and chemokine expression profiles of serum and cerebrospinal fluid (CSF) in ALS patients undergoing experimental treatment employing Whorton jelly derived stem cells.

Materials and methods: Seven ALS and seven control patients were enrolled in the study. All patients provided a written consent to participate in the study. Stem cells were derived using previously approved methodology established in laboratories of Prof. Maksymowicz and Prof. Wojtkiewicz. The therapy design and treatment regimen were approved by the Ethic Committee of University of Warmia and Mazury (UWM) in Olsztyn, and the trial has been registered as NCT02881489. Serum and cerebrospinal fluid was collected 3, 6 and 9 months post the treatment. Cytokine and chemokine expression levels were studied using Human XL Cytokine Array Kit (R&D Systems, Minneapolis, MN) customized for the detection of following 36 substances: C5a, IL-1a, IL-1, IL-1ra, IL-2, IL-5, IL-6, IL-8, IL-10, IL12, IL-13, IL-16, IL-17, IL-17E, IL-18, IL-23, IL-27, IL-32a, CXCL10/IP-10, CXCL11/I-TAC, CCL2/MCP-1, CCL3/MIP-11a,, CCL4/MIP-1β, CXCL1/GRO a ,CCL1/I-309, CCL5/RANTES, CXCL12/SDF-1, Serpin E1/PAI-1 TREM-1, IFN?, ICAM-1 TNFa, CD40 ligand, G-CSF, GM-CSF, MIF. Statistical evaluation of the data was performed by one-way ANOVA with post-hoc Bonferroni test using GraphPad Instat (GraphPad), p< 0.05. Results Preliminary analysis of serum and CSF samples collected 3 months post treatment showed a statistically significant increase in Il-18 and CXCL1 and decrease in IL-13 and IL-16 serum levels as well as an increase in CXCL10 levels in CSF in ALS group as compared to the controls.

Conclusions This is the first study demonstrating changes in cytokine/chemokine expression levels in ALS patients following the experimental stem cell treatment. The observed results lay background for further studies focused on clinical evaluation of the progression of the disease and treatment, providing prognostic tools and potential biomarkers in ALS diagnosis and treatment.

Keywords: Cytokine, chemokine, ALS, stem cell, experimental treatment.