Bacterial rhamnolipids (RLs) in saliva of Alzheimer's disease and Mild Cognitive Impairment patients and correlation with neuroinflammation and cognitive state

Eleni G. Andreadou; Georgios Katsipis; Magda Tsolaki; Anastasia A. Pantazaki

doi:10.30574/gscarr.2021.6.3.0062

Authors

Eleni G. Andreadou Laboratory of Biochemistry, Department of Chemistry, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece.
Georgios Katsipis Laboratory of Biochemistry, Department of Chemistry, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece.
Magda Tsolaki First Neurology Department, “AHEPA” University General Hospital of Thessaloniki, Aristotle University of Thessaloniki, 54124 Thessaloniki, Greece.
Anastasia A. Pantazaki Center for Interdisciplinary Research and Innovation, Laboratory of Neurodegenerative Diseases (LND), 57001 Thermi, Thessaloniki, Greece.

DOI:

https://doi.org/10.30574/gscarr.2021.6.3.0062

Keywords:

Biomarkers, Alzheimer’s disease, Mild Cognitive Impairment, Saliva, Rhamnolipids, Inflammation

Abstract

Alzheimer’s disease (AD) is increasingly affecting the aging population and the estimated prevalence reaches 50 million people worldwide. The need for the discovery of new biomarkers for AD diagnosis is urgent and especially in biological fluids other than cerebrospinal fluid (CSF), as its collection is invasive. Arguments are numerous that chronic bacterial infections might be considered as one of the possible causes of AD. Rhamnolipids (RLs) are bacterial virulence factors, suspicious for dysfunctions and disorders including AD. The aim of this pilot trial was to investigate RLs levels in saliva of Mild Cognitive Impairment (MCI) and AD patients with indirect ELISA. Specifically, salivary RLs were determined in 30 AD patients, 24 MCI patients and 15 cognitively healthy individuals and were found elevated in AD and MCI patients compared to those of the control group. The established biomarkers of AD, tau and Aβ₄₂ amyloid, and the inflammatory markers cyclooxygenases (COX-1 and COX-2) were also determined, to evaluate their possible interdependence from RLs levels. Levels of RLs positively correlate with COX-2 levels and negatively with the mental state according to Mini–Mental State Examination (MMSE) score of donors. Multilinear regression verified the tight interrelation of RLs with COX-2 in saliva of MCI and AD patients. The results of this study stand by the hypothesis of inflammatory involvement in AD and indicate that RLs could be suggested as eventual biomarkers for AD diagnosis using saliva as biological fluid.

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References

Lopez OL, McDade E, Riverol M, et al. Evolution of the diagnostic criteria for degenerative and cognitive disorders. Curr Opin Neurol 2011; 24:532–541.

Liang D, Lu H. Salivary biological biomarkers for Alzheimer’s disease. Arch Oral Biol 2019; 105:5–12.

Menéndez-González M. Routine lumbar puncture for the early diagnosis of Alzheimer’s disease. Is it safe? Front Aging Neurosci 2014; 6:65.

Schepici G, Silvestro S, Trubiani O, et al. Salivary Biomarkers: Future Approaches for Early Diagnosis of Neurodegenerative Diseases. Brain Sci 2020; 10:245.

Tvarijonaviciute A, Zamora C, Ceron JJ, et al. Salivary biomarkers in Alzheimer ’ s disease. 2020; 3437–3444.

Gleerup HS, Hasselbalch SG, Simonsen AH. Biomarkers for Alzheimer’s disease in saliva: A systematic review. Dis Markers; 2019. Epub ahead of print 2019. DOI: 10.1155/2019/4761054.

Ashton NJ, Ide M, Zetterberg H, et al. Salivary Biomarkers for Alzheimer’s Disease and Related Disorders. Neurol Ther 2019; 8:83–94.

Yoshizawa JM, Schafer CA, Schafer JJ, et al. Salivary Biomarkers: Toward Future Clinical and Diagnostic Utilities. Clin Microbiol Rev 2013; 26:781–791.

Kamer AR, Craig RG, Niederman R, et al. Periodontal disease as a possible cause for Alzheimer’s disease. Periodontol 2000 2020; 83:242–271.

Dioguardi M, Crincoli V, Laino L, et al. The Role of Periodontitis and Periodontal Bacteria in the Onset and Progression of Alzheimer’s Disease: A Systematic Review. J Clin Med 2020; 9:495.

Sureda A, Daglia M, Argüelles Castilla S, et al. Oral microbiota and Alzheimer’s disease: Do all roads lead to Rome? Pharmacol Res 2020; 151:104582.

Sochocka M, Donskow-Łysoniewska K, Diniz BS, et al. The Gut Microbiome Alterations and Inflammation-Driven Pathogenesis of Alzheimer’s Disease-a Critical Review. Mol Neurobiol 2019; 56:1841–1851.

Holmes C. Review: Systemic inflammation and Alzheimer’s disease. Neuropathol Appl Neurobiol 2013; 39:51–68.

Akiyama H, Barger S, Barnum S, et al. Inflammation and Alzheimer’s disease. Neurobiol Aging 2000; 21:383–421.

François M, Bull CF, Fenech MF, et al. Current State of Saliva Biomarkers for Aging and Alzheimer’s Disease. Curr Alzheimer Res 2019; 16:56–66.

Paraskevaidi M, Allsop D, Karim S, et al. Diagnostic Biomarkers for Alzheimer’s Disease Using Non-Invasive Specimens. J Clin Med; 9. Epub ahead of print June 2020. DOI: 10.3390/jcm9061673.

Bouftas M A Systematic Review on the Feasibility of Salivary Biomarkers for Alzheimer’s Disease. J Prev Alzheimer’s Dis 2021; 8:84–91.

Batista CRA, Gomes GF, Candelario-Jalil E, et al. Lipopolysaccharide-Induced Neuroinflammation as a Bridge to Understand Neurodegeneration. Int J Mol Sci 2019; 20:2293.

Marizzoni M, Cattaneo A, Mirabelli P, et al. Short-Chain Fatty Acids and Lipopolysaccharide as Mediators Between Gut Dysbiosis and Amyloid Pathology in Alzheimer’s Disease. J Alzheimers Dis 2020; 78:683–697.

Zakaria R, Wan Yaacob WM, Othman Z, et al. Lipopolysaccharide-induced memory impairment in rats: a model of Alzheimer’s disease. Physiol Res 2017; 66:553–565.

Andreadou E, Pantazaki AA, Daniilidou M, et al. Rhamnolipids, Microbial Virulence Factors, in Alzheimer’s Disease. J Alzheimers Dis 2017; 59:209–222.

Chong H, Li Q. Microbial production of rhamnolipids: opportunities, challenges and strategies. Microb Cell Fact 2017; 16:137.

Soberón-Chávez G, Lépine F, Déziel E. Production of rhamnolipids by Pseudomonas aeruginosa. Appl Microbiol Biotechnol 2005; 68:718–25.

Abdel-Mawgoud AM, Lépine F, Déziel E. Rhamnolipids: diversity of structures, microbial origins and roles. Appl Microbiol Biotechnol 2010; 86:1323–1336.

Zulianello L, Canard C, Köhler T, et al. Rhamnolipids are virulence factors that promote early infiltration of primary human airway epithelia by Pseudomonas aeruginosa. Infect Immun 2006; 74:3134–47.

Pantazaki AA, Choli-Papadopoulou T. On the Thermus thermophilus HB8 potential pathogenicity triggered from rhamnolipids secretion: Morphological alterations and cytotoxicity induced on fibroblastic cell line. Amino Acids 2012; 42:1913–1926.

Andreadou E, Moschopoulou A, Simou O, et al. T. thermophilus Rhamnolipids Induce Cytogenetic Damage on Human Lymphocytes and Bind DNA in vitro. Br Biotechnol J 2016; 10:1–12.

Rahimi K, Lotfabad TB, Jabeen F, et al. Cytotoxic effects of mono- and di-rhamnolipids from Pseudomonas aeruginosa MR01 on MCF-7 human breast cancer cells. Colloids Surfaces B Biointerfaces 2019; 181:943–952.

Williams CS, Mann M, DuBois RN. The role of cyclooxygenases in inflammation, cancer, and development. Oncogene 1999; 18:7908–16.

Aïd S, Bosetti F. Targeting cyclooxygenases-1 and -2 in neuroinflammation: Therapeutic implications. Biochimie 2011; 93:46–51.

Fu JY, Masferrer JL, Seibert K, et al. The induction and suppression of prostaglandin H2 synthase (cyclooxygenase) in human monocytes. J Biol Chem 1990; 265:16737–40.

Lee SH, Soyoola E, Chanmugam P, et al. Selective expression of mitogen-inducible cyclooxygenase in macrophages stimulated with lipopolysaccharide. J Biol Chem 1992; 267:25934–8.

O’Sullivan MG, Huggins EM, McCall CE. Lipopolysaccharide-induced expression of prostaglandin H synthase-2 in alveolar macrophages is inhibited by dexamethasone but not by aspirin. Biochem Biophys Res Commun 1993; 191:1294–300.

Giagkas D, Choli-Papadopoulou T, Pantazaki A. Development of an Antibody for Detection of Rhamnolipids Characterized as a Major Bacterial Virulence Factor. Antibodies 2013; 2:501–516.

Tabaraud F, Leman JP, Milor AM, et al. Alzheimer CSF biomarkers in routine clinical setting. Acta Neurol Scand 2012; 125:16–423.

McKhann G, Drachman D, Folstein M, et al. Clinical diagnosis of alzheimer’s disease: Report of the NINCDS-ADRDA work group⋆ under the auspices of department of health and human services task force on alzheimer’s disease. Neurology 1984; 34:939–944.

Hoo ZH, Candlish J, Teare D. What is an ROC curve? Emerg Med J 2017; 34:357–359.

Kownatzki R, Tümmler B, Döring G. Rhamnolipid of Pseudomonas Aeruginosa in Sputum of Cystic Fibrosis Patients. Lancet 1987; 329:1026–1027.

Shoemark DK, Allen SJ. The Microbiome and Disease: Reviewing the Links between the Oral Microbiome, Aging, and Alzheimer’s Disease. J Alzheimer’s Dis 2014; 43:725–738.

Li B, He Y, Ma J, et al. Mild cognitive impairment has similar alterations as Alzheimer’s disease in gut microbiota. Alzheimer’s Dement 2019; 15:1357–1366.

Martande SS, Pradeep AR, Singh SP, et al. Periodontal Health Condition in Patients With Alzheimer’s Disease. Am J Alzheimer’s Dis Other Dementiasr 2014; 29:498–502.

Balin BJ, Gérard HC, Arking EJ, et al. Identification and localization of Chlamydia pneumoniae in the Alzheimer’s brain. Med Microbiol Immunol 1998; 187:23–42.

Miklossy J. Alzheimer’s disease - a neurospirochetosis. Analysis of the evidence following Koch’s and Hill’s criteria. J Neuroinflammation 2011; 8:90.

Dominy SS, Lynch C, Ermini F, et al. Porphyromonas gingivalis in Alzheimer’s disease brains: Evidence for disease causation and treatment with small-molecule inhibitors. Sci Adv 2019; 5:eaau3333.

e Silva SS, Carvalho JWP, Aires CP, et al. Disruption of Staphylococcus aureus biofilms using rhamnolipid biosurfactants. J Dairy Sci 2017; 100:7864–7873.

Nickzad A, Déziel E. The involvement of rhamnolipids in microbial cell adhesion and biofilm development - an approach for control? Lett Appl Microbiol 2014; 58:447–453.

Jensen PØ, Bjarnsholt T, Phipps R, et al. Rapid necrotic killing of polymorphonuclear leukocytes is caused by quorum-sensing-controlled production of rhamnolipid by Pseudomonas aeruginosa. Microbiology 2007; 153: 1329–1338.

Andersen KK, Vad BS, Kjær L, et al. Pseudomonas aeruginosa rhamnolipid induces fibrillation of human α‐synuclein and modulates its effect on biofilm formation. FEBS Lett 2018; 592:1484–1496.

McElroy SJ, Hobbs S, Kallen M, et al. Transactivation of EGFR by LPS induces COX-2 expression in enterocytes. PLoS One 2012; 7:e38373.

Ho L, Purohit D, Haroutunian V, et al. Neuronal Cyclooxygenase 2 Expression in the Hippocampal Formation as a Function of the Clinical Progression of Alzheimer Disease. Arch Neurol; 58. Epub ahead of print 1 March 2001. DOI: 10.1001/archneur.58.3.487.

Garcia-Bueno B, Serrats J, Sawchenko PE. Cerebrovascular Cyclooxygenase-1 Expression, Regulation, and Role in Hypothalamic-Pituitary-Adrenal Axis Activation by Inflammatory Stimuli. J Neurosci 2009; 29:12970–12981.

Cao L-L, Guan P-P, Liang Y-Y, et al. Cyclooxygenase-2 is Essential for Mediating the Effects of Calcium Ions on Stimulating Phosphorylation of Tau at the Sites of Ser 396 and Ser 404. J Alzheimers Dis 2019; 68:1095–1111.

Wang Y, Guan P-P, Yu X, et al. COX-2 metabolic products, the prostaglandin I2 and F2α, mediate the effects of TNF-α and Zn2+ in stimulating the phosphorylation of Tau. Oncotarget 2017; 8:99296–99311.

Najarzadeh Z, Pedersen JN, Christiansen G, et al. Bacterial amphiphiles as amyloid inducers: Effect of Rhamnolipid and Lipopolysaccharide on FapC fibrillation. Biochim Biophys Acta - Proteins Proteomics 2019; 1867:140263.

Reiss AB, Arain HA, Stecker MM, et al. Amyloid toxicity in Alzheimer’s disease. Rev Neurosci 2018; 29:613–627.

Heneka MT, Carson MJ, Khoury J El, et al. Neuroinflammation in Alzheimer’s disease. Lancet Neurol 2015; 14: 388–405.

Dinan TG, Stilling RM, Stanton C, et al. Collective unconscious: how gut microbes shape human behavior. J Psychiatr Res 2015; 63:1–9.