Neuroborreliosis in human neuronal and brain models.

National Institute Of Allergy And Infectious Diseases

Investigators

Abstract

Scope: Lyme disease, caused by several Borrelia species, is the most common tick-borne disease in North America and Europe, affecting over 500,000 people annually. The acute manifestations include skin rash (erythema migrans) at the tick bite site, as well as fever, headache, and fatigue. If untreated, spirochetes can disseminate into deep tissues where infected individuals may develop neurological, cardiac, chronic skin, or articular indications. Approximately 15% of Lyme patients will experience some level of neurological involvement, termed Lyme neuroborreliosis (LNB), such as meningitis, cranial neuritis, radiculoneuritis, parenchymal inflammation of the brain or spinal cord, and/or encephalopathy. Acute Lyme disease has been extensively studied in several animal models, but little progress has been made in our understanding of LNB in the context of the human central nervous system. Our research aims to generate human neuronal and organoid models to address this deficit. Research materials, equipment and methods: Our research predominantly uses cell-based techniques to investigate changes in single cells or in cell networks. We have generated human stem cell models, including differentiation of forebrain neurons and cerebral organoids. To investigate specific functional and disease-related pathways we use a combination of protein, RNA, live cell function and microscopy analyses. The equipment we hold for these analyses include a chip/multiwell/in vivo MEA system, a Seahorse analyser, two plate readers of differing functional capacity, an automated fluorescence microscope, a chemiluminescence/fluorescence imaging system, a 10x chromium controller and Fluent Biosciences PIPseq apparatus. Research accomplishments: We initially reported the generation of a human model of neuroborreliosis using forebrain neuronal cultures and brain organoids from human induced pluripotent stem cells that were exposed to Borrelia isolates. In this financial year, we did not receive any funding for this work so our primary accomplishments were analyzing data submitted for assay with core technologies during the last financial year and preparing this data for publication, and applying for extramural funding to support this research into the future. Of the new data received this year, RNA sequencing analysis revealed several exciting pathways that were changed over time and in a Borrelia isolate specific manner, as well as some changes that were common to all isolates tested. Outstanding core data that we are waiting on include metabolomic and lipidomic analyses.