BANK COV. Pathology - tissue repository

In the scenario of an epidemic caused by a new and relatively unknown agent, the autopsy represents a fundamental tool for the investigation of phenomena related to the pathophysiology of the disease and its impact on target and systemic organs. In the case of COVID-19, rare autopsies have been performed so far, due to the risk of contagion by the professionals involved in the process. Minimally invasive autopsy (AMI) has been proposed as an alternative to conventional autopsy (CA) and presents itself as a useful and effective tool in this scenario. The method has been shown to be effective, particularly for the diagnosis of infectious diseases. In addition, the procedure proved to be adequate for collecting samples for molecular studies and for identifying microorganisms (Duarte-Neto et al, 2019). Subproject 1: Post-mortem evaluation of systemic impairment in COVID-19 In this project, we propose to perform the AMI-US of patients with death from COVID-19 with tissue collection through puncture of the following organs and tissue: lungs, heart, liver, kidneys , spleen, CNS, skeletal muscle and skin. Autopsies have the general objective of evaluating the pulmonary and systemic involvement of COVID-19 in critically ill patients, identifying secondary infections associated with a poor prognosis, in addition to creating a biorepository for future studies. The main objectives are: a) Description of the pulmonary and systemic histopathological findings in fatal cases of COVID-19. b) Identification of secondary bacterial and/or fungal infections by histochemical and molecular methods. c) Description of electron microscopy findings in lung parenchyma samples, focusing on the 2019-nCOV virion encounter, septal capillary endothelial cell changes, pneumocyte changes, and pulmonary interstitium. d) Characterization of immunostaining of 2019-nCOV antigens in different tissues by means of immunohistochemistry e) Detection and quantification of 2019-nCOV virus RNA in tissue samples collected post mortem, using the PCR technique. f) Transcriptome analysis of lung tissue samples collected in the post-mortem period of patients with fatal COVID-19, correlating it with biochemical parameters, with the morphological aspect of the lesions under optical microscopy and with the result of in situ immunostaining of antigens , through immunohistochemistry, in order to clarify pathogenic processes associated with the unfavorable evolution of COVID-19. Subproject. Immunopathology of fatal COVID-19 in elderly patients The greater severity and mortality of the disease in the elderly caused by COVID 19 is associated with immune dysregulation in this age group, which promotes a more sustained inflammatory reaction than in young people. We hypothesized that in the lung tissues of elderly patients who did not die from pulmonary causes there will be greater expression of inflammatory markers linked to aging, which will be accentuated in patients with SARS-Cov2. We further hypothesize that pulmonary ACE2 expression will be lower in elderly patients who died of SARS-2. The main objectives are: To phenotype lung tissue injury in tissue obtained by MIA, comparing with lungs of similar age groups, Compare the amount of T and B lymphocyte cell markers, neutrophils, dendritic cells, macrophages and mast cells in adults versus older adults (>65 years) with normal lungs at autopsy and those infected by COVID2; Compare tissue expression of ACE2 and TMPRSS receptors in adults versus older adults (>65 years) with normal lungs at autopsy and those infected by COVID2, Compare expression of TLR-3, IL-6, TNF-alpha, IFN- gamma , MMPs, TGF beta, IL-1B and IL-17 in lung tissue of adults versus older adults (>65 years old), Correlate pathological findings with clinical severity markers and serum inflammatory markers in SARS-Covid2 patients; Compare and validate these data by transcriptome/PCR data from lung tissue from patients with SARS-Cov2.