Speaker
Description
Long COVID refers to persistent symptoms lasting over a year after a SARS-CoV-2 infection, with no other underlying cause. These include neurological symptoms (NeuroCOVID) such as memory disorders and concentration impairment, which disrupt daily life and can persist even after other viral symptoms resolve. The NeuroCOV Consortium was established to address the emerging clinical need to investigate the long-term neurological impacts of COVID-19. As part of this consortium, our aim is to decipher the cellular and molecular mechanisms underlying the diverse NeuroCOVID trajectories using patient-derived immunocompetent brain organoids. As a first step to achieve this goal, we are coupling clinical metadata with transcriptomic profiles at single cell resolution from peripheral blood mononuclear cells (PBMCs) of patients belonging to two European cohorts (from Italy and Germany) to perform a deep phenotyping of the patients to identify key molecular signatures associated with their neurological profiles. Using that information, we are generating a NeuroCOVID specific human induced pluripotent stem cells (hiPSCs) cell bank that will allow us to ensure representation across the spectrum of neurological manifestations when differentiated to brain organoids (BOs). Additionally, we are using a combination of small molecules to fine-tune a previously described protocol in our lab (Caporale et al. 2024) that will allow us to obtain organoids with a more matured phenotype to produce a scalable model applicable to cohorts’ studies. Immunostainings to assess neuronal population markers (SATB2, TBR1, TBR2, among others) were performed on cortical BOs at different timepoints (D25, 38 and 50) and calcium dynamics were assessed by Fluo-4 probe based imaging. Finally, to dissect neuronal and immune factors contributing to NeuroCOVID in a post SARS-CoV-2 infection setting, we are comparing two hiPSC derived microglia protocols to establish their reproducibility and robustness in different cell lines to generate both patient-derived microglia and cortical BOs in order to subsequently perform an integrated omics approach that will allow us to investigate the cellular and molecular mechanisms underlying neuroinflammation and neurodegeneration in Long COVID, with the ultimate goal of identifying potential drivers of NeuroCOVID vulnerability and resilience.
| Author(s) | Luciana Isaja1, Giuseppe Cammarata1, Alessia Valenti1,2,3, Vittorio Aiello1,2,3, Benedetta Muda1,4, Narges Yahyazadeh1,3, Samuele Galimi1, Nicolo Caporale1, Emanuele Villa1, Giuseppe Testa1,3,5 |
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| Affiliation(s) | 1 Human Technopole Foundation, Milan, Italy; 2 SEMM European School of Molecular Medicine, Milan, Italy; 3 Department of Experimental Oncology, University of Milan, Milan, Italy; 4 translational medicine, university of Milan, Milan, Italy 5 Department of Experimental Oncology, European Institute of Oncology, Milan, Italy. |
