Galectin-1, The New Player in Staging and Prognosis of COVID-19 Soja Sekulic University of Kragujevac, Faculty of Medical Sciences, Department of Infectious disease Nevena Gajovic University of Kragujevac, Faculty of Medical Sciences, Center for Molecular Medicine and Stem Cell Research, Milena Jurisevic University of Kragujevac, Faculty of Medical Sciences, Department of Clinical pharmacy Marina Jovanovic ( [email protected] ) University of Kragujevac, Faculty of Medical Sciences, Department of Internal medicine Biljana Popovska Jovicic University of Kragujevac, Faculty of Medical Sciences, Department of Infectious disease Nebojsa Arsenijevic University of Kragujevac, Faculty of Medical Sciences, Center for Molecular Medicine and Stem Cell Research, Zeljko Mijailovic University of Kragujevac, Faculty of Medical Sciences, Department of Infectious disease Marina Jovanovic University of Kragujevac, Faculty of Medical Sciences, Department of Otorinolaringology Zana Dolicanin State University of Novi Pazar, Department of Biomedical sciences Ivan Jovanovic University of Kragujevac, Faculty of Medical Sciences, Center for Molecular Medicine and Stem Cell Research, Research Article Keywords: galectin-1, COVID-19, prognostic marker Posted Date: September 3rd, 2021 DOI: https://doi.org/10.21203/rs.3.rs-830345/v1 License: This work is licensed under a Creative Commons Attribution 4.0 International License. Read Full License Galectin-1, the new player in staging and prognosis of COVID-19 Running title: Galectin-1 in COVID-19 progression Sofija Sekulic1, Nevena Gajovic2, Milena Jurisevic3, Marina Jovanovic*4, Biljana Popovska Jovicic1, Nebojsa Arsenijevic2, 5, Zeljko Mijailovic1, Marina Jovanovic6, Zana Dolicanin7, Ivan Jovanovic2 1 University of Kragujevac, Faculty of Medical Sciences, Department of Infectious disease, Kragujevac, 34000, Serbia 2 University of Kragujevac, Faculty of Medical Sciences, Center for Molecular Medicine and Stem Cell Research, Kragujevac, 34000, Serbia 3 University of Kragujevac, Faculty of Medical Sciences, Department of Clinical pharmacy, Kragujevac, 34000, Serbia 4 University of Kragujevac, Faculty of Medical Sciences, Department of Internal medicine, Kragujevac, 34000, Serbia 5 Public health institute Kragujevac, Kragujevac, 34000, Serbia 6 University of Kragujevac, Faculty of Medical Sciences, Department of Otorinolaringology, Kragujevac, 34000, Serbia 7 State University of Novi Pazar, Department of Biomedical sciences, Novi Pazar, 36300, Serbia Correspondence: Marina Jovanovic*, MD PhD University of Kragujevac, Faculty of Medical Sciences, Department of Internal medicine, Kragujevac, 34000, Serbia Svetozara Markovica 69, 34000 Kragujevac, Serbia Tel +38134306800, Fax. +38134306800112 E-mail: marinna034@gmail. com Key words: galectin-1, COVID-19, prognostic marker Abstract Background A new virus from the group of coronaviruses was identified as the cause of atypical pneumonia and called severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) and disease called Corona Virus Disease (COVID-19). During the cytokine storm, the main cause of the death, proinflammatory cytokines are released which stimulate further tissue destruction. Galectin-1 is pleiotropic cytokine involved in many immune and inflammatory processes and its role in COVID-19 is still unknown. Objective The aim of this study was to determine systemic values of galectin-1 and correlations between gal-1 and proinflammatory cytokines and clinical parameters during COVID-19 progression. Design Observational and cross-sectional study. Patients 210 COVID-19 patients were included and divided into mild, severe or critical group according to COVID-19 severity. Main Measures Serum levels of IL-1β, IL-6, IL-10, IL-23, IL- 33 and galectin-1 were measured using sensitive enzyme-linked immunosorbent assay (ELISA) kits. Key Results Systemic levels of IL-1β, IL-6, IL-10, IL-23, IL-33 and galectin-1 were significantly higher in stage III of COVID-19 patients compared to stage I and II. There were no significant differences in ratio between Galectin-1 and IL-10 with proinflammatory cytokines. Positive correlation was detected between Gal-1 and IL-1β, IL6, IL-10, IL-23 and IL-33. Gal-1 positively correlated with chest radiographic finding, dry cough and headache and negatively correlated with normal breathing sound. Conclusions Linear regression model and ROC curve analysis point on Gal-1 as significant predictors for COVID-19 severity. Presented results implicate on Gal-1 and IL-10 dependent immunomodulation. The precise mechanism of Gal-1 effect in COVID-19 and its potential as a stage marker of disease severity is still to be clarified. INTRODUCTION COVID-19 is an infectious respiratory disease caused by the novel coronavirus, named as severe acute respiratory syndrome–corona virus 2 (SARS-CoV2). (1). Briefly after COVID-19 outbreak in China, the disease spread around the world and the World Health Organization declared a pandemic1. The clinical presentation of COVID-19 can vary from asymptomatic to acute respiratory distress syndrome (ARDS) and respiratory insufficiency2, 3. Most common underlying mechanism of respiratory failure in patients with COVID-19 is characterized by rapid activation of proinflammatory cytokines and inflammatory cell recruitment in the lungs (e. g. cytokine storm)4, 5. An increased concentration of IL-6, TNF-α, IL-2, IL-8, IL-1β, G-CSF, IL-17, GM- CSF, IL10, MCP1, MIP1α, chemokines, galectins in serum, was detected in COVID-19 patients 6-8. Interestingly, role of immunosuppressive cytokines, especially IL-10 may also contribute to COVID-19 induced damage of the lungs in latter stages of the disease9. Galectins are oligosaccharides chain complexes located on the cell surface and play an important role in regulating immune reactions and inflammatory responses10, 11, 12. Galectin-1 (Gal-1) was the first identified protein in this family. Even though it is known that Gal-1 has many regulatory functions in viral infections, to date, the role of Gal-1 in immunopathogenesis of the COVID-19 disease has not been clarified. Until now, only one study by Kazancioglu et al investigated potential role of galectins in SARS-CoV-2 induced inflammation. The results imply that levels of galectins (Gal-1 and Gal-3) are elevated in COVID-19 patients in comparison to healthy patients13. The aim of this study was to determine the association of Gal-1 and innate immunity cytokines such as IL-1β, IL-6, IL-10, IL-23, IL-33 with disease severity, alongside with clinical and biochemical features as well as CXR lung findings on disease severity in 210 patients with COVID-19. Results Clinical features of COVID-19 patients All of 210 patients included in the study met the criteria for COVID-1914 and were classified into the three groups according to disease severity. The median age of mild group (54. 9±1. 7) was significantly different from that of severe patients (59. 5±1. 3 ) or critical (61. 3±1. 4) (Table 1). Among all of them, 135 (57, 14%) were male. The analysis revealed a higher percentage of male patients and a lower percentage of female patients in group II and III with statistical significance (p = 0. 041; Table 1). The most frequent auscultatory finding in group I was attenuated breathing sound in 44. 3% of patients, with CXR findings described like interstitial thickening in 51. 4%, focal consolidation in 28. 5% and normal findings in 20%. In group II, 58. 5% of patients had auscultatory attenuated breathing sound, 48. 5% audible crackles diffusely with CXR findings described like multifocal consolidation in 44. 2% of hospitalized patients, focal consolidation in 30% and interstitial thickening in 25. 7%. In group III, auscultatory 90% of patients had audible crackles diffusely, attenuated breathing sound in 85. 7% with diffuse alveolar changes / ARDS on CXR findings. Only 10 (14. 2%) among this group had multifocal consolidation. Compared with mild group, the severe and critical group were more likely to have dyspnea, fatigue, auscultatory audible crackles diffusely. The percentage of patients with these findings was significantly higher than in the mild group (p < 0. 05; Table 1), which is in line with CXR findings of the patients. None of the patients in the severe and critical group had normal CRX finding, interstitial thickening or focal consolidation zones (CRX I, II, III), and none of the patients in the mild group had multifocal consolidation zones or diffuse alveolar changes / ARDS (CRX IV, V) (p=0. 000) (Table 1). In patients with severe and critical features, an increase in neutrophil count (p=0. 001), urea (p=0. 018), AST (p=0. 001), ALT (p=0. 001), BILD (p = 0. 018), CK (p=0. 012), LDH (p=0. 001), D-dimer (p=0. 001), CRP (p=0. 001), PCT (p=0. 001), Ferritin (p=0. 001) was noted, with reduced lymphocyte count (p=0. 001), monocyte count (p=0. 006), decreased levels of albumin (p=0. 001) and lower values of SaO2 (p=0. 001) and pO2 (p=0. 001) (Table 2). There were no statistically significant differences in white blood cell count, erythrocytes and platelets, + + hemoglobin; levels of plasma glucose, creatinine, BILT, K , Na , Fe, pH of blood, pCO2 (Table 2). Patients with the more severe form of COVID-19 had increased systemic values of Gal-1 Serum pro- and anti-inflammatory cytokines of interest were measured in COVID-19 patients. Systemic values of IL-1β (p=0. 001), IL-6 (p=0. 001), IL-10 (p=0. 026), IL-23 (p=0. 001), IL-33 (p=0. 001), and Gal-1 (p=0. 001) were significantly higher in stage III of COVID-19 patients compared to patients in stage I and II (Figure 1). Unchanged flow of ratios of Gal-1 and IL-10 with proinflammatory cytokines during COVID-19 development Analyses of the ratios of Gal-1 and IL-10 with IL-1β, IL-6, IL-23 and IL-33 were done. There were no statistically significant differences in the ratios of Gal-1/IL-1β, Gal-1/IL-6, Gal-1/IL-23, Gal-1/IL-33 and Gal-1/IL-10 as well as IL-10/IL-1β, IL-10/IL-6, IL-10/IL-23, IL-10/IL-33 between defined groups of COVID-19 patients (Figure 2).