Covid-19 research projects and special fund

Establishing a prospective register of patients with Covid-19 infection to describe clinically relevant case studies (hospitalized patients).

Objectives:

• characterize patients hospitalized with Covid-19 in Italy, with a focus on early markers of clinical deterioration in order to identify the predictors of the need for invasive mechanical ventilation
• define the clinical outcome of short-term patients with a focus on those with chronic obstructive pulmonary disease (COPD)
• define the role of diagnostic methods in SARS-CoV-2 pneumonia, including bronchoscopy and new imaging techniques such as lung ultrasound
• quickly generate hypotheses on the effectiveness of different types of respiratory support or drug therapies available, to be confirmed by interventional studies
• analyse medium- and long-term consequences and the residual disability of surviving patients 
• highlight the key issues of clinical difficulties in the management of the Covid-19 pandemic as a model to enhance NHS resilience to any future pandemics.

Principal Investigator: Giuseppe Francesco Sferrazza Papa

The project objective is to identify antigenic determinants for the serological diagnosis of SARS-CoV-2 infection. The project will develop an ease-of-use and cheap prototype assay for the specific dosage of antibodies to SARS-CoV-2 suitable for common clinical laboratories. The assay will allow to confirm past and ongoing infection with the pathogen. Moreover, the project will help elucidating the antibody response possibly correlated with the protective immunity to the pathogen.

Responsabile scientifico: Sergio Abrignani

The aim of the project is to identify dysregulated glycosphingolipids in the plasma of COVID-19 patients and their correlation with clinical evolution.  

The study will include the determination of the quali/quantitative levels of ceramide, dihydroceramide, sphingomyelin, dihydrosphingomyelin and hexosylceramide by LC MS / MS. While conducting quantitative analyses, we will obtain a qualitative profile.

The specific quantitative determination of S1P by targeted LC-MS/MS will provide data on the evolution of the disease.  In asymptomatic subjects we expect comparable S1P levels, while a decrease in S1P levels and a concomitant increase in ceramide levels would be related to the onset of respiratory problems.

The lowest levels are expected in ICU patients. Obese individuals will start by default with lower levels, predisposing them to a worse evolution.

Principal Investigator:  Cecilia Gelfi 

Based on data univocally identifying ACE2 as a specific cellular target for Covid-19 entering pneumocytes, it is possible to conceive molecules that selectively prevent interaction with the virus, thus blocking its infection. 

The aim of the study is to develop an aptamer to be nebulized in the airways that is capable of binding to alveolar and bronchial ACE2, and preventing the SPIKE protein of Covid-19 from using ACE2 to enter cells.

Compared to monoclonal antibodies, peptides or small molecules aimed at destroying the same interaction, the aptamer-based therapeutic strategy would be equally specific and effective without stimulating the immune system (as therapeutic proteins) or inducing potential systemic side effects (as small synthetic molecules). 

Principal Investigator: Paolo Ciana

Given the serious situation produced by the spread of COVID-19 contagion, and the long time required for vaccine or ad-hoc drug development, we must implement strategies for potential therapeutic applications, though sub-optimal, in a short or very short time.

In this context, in silico approaches can help identify opportunities for the repositioning of known molecules (drugs, natural organic substances, endogenous ligands), in order to evaluate their ability to interfere with COVID-19 viral proteins.

The strategy described above will be implemented using the SPILLO-PBSS* software in an innovative way, for the purpose of identifying, in the context of easily available molecules, specific opportunities for drug repositioning not identifiable by traditional in silico approaches.

To this end, the protein structures of COVID-19 (starting from COVID-19 main protease), already available in public databases, will be used.

Principal Investigator:  Paolo Magni

This multicentre research aims to assess the risk of fungal co-infection in COVID-19 by involving clinical microbiology laboratories throughout the country, in order to provide epidemiological data on fungal infections in COVID patients, and to study the incidence and dynamics of fungal infections during the pandemic, especially in patients hospitalized in intensive or sub-intensive care units, and infectious disease wards.  

Objectives:  

• Determine the incidence of fungal infections in hospitalized COVID patients  
• Assess mortality in COVID patients with fungal infection compared with COVID patients not affected by fungal infection  
• Collect and analyse epidemiological data on risk factors related to the onset of fungal infections in COVID patients  
• Assess the incidence of fungal infections in COVID patients hospitalized in 2020 compared to the incidence of the same infections in patients hospitalized with severe flu in the 2018/2019 season  
• Collect and analyse data on in vitro antifungal sensitivities of strains of Candida and other yeasts isolated from COVID patient haemocultures in order to monitor changes in susceptibility profiles during the pandemic  
• Collect the strains of Candida that proved resistant for studies on the mechanisms underlying resistance 
• Collect strains of Aspergillus isolated from different biological samples of COVID patients and test their in vitro sensitivity to azole antifungals to detect any resistances, investigate the molecular mechanism related to resistance and estimate the prevalence of azole-resistance 

Principal Investigator: Luisa Romano

SARS‐CoV‐2 uses ACE2 as entry receptor. ACE2 gene displays several sequence variations that may have an impact on the variability of disease severity, as like on the sex-biased death rate, being ACE2 an X-linked locus.

We aim to develop a rapid genetic test that will include the genetic variants of ACE2 and other genes associated with the severity of the COVID-19 infection, useful for calculating a predictive score for the severity basis of the infection. 

Principal Investigator: Monica Rosa Miozzo

The study is aimed at isolating SARS-CoV-2 (from bronchial aspiration, nasopharyngeal swab and saliva) and defining its genotypic characteristics. Covalent inhibitors of the 3CL hydrolase protein of SARS-CoV-2 will be designed and synthesized.

The CoV inhibitors will be subsequently tested in vitro with distinct mechanisms of action, and their toxicity (therapeutic index) and phenotypic susceptibility (IC50 and IC90) will be studied.

We will then study the additive/synergistic activity of the same compounds, and finally quantize the in vitro production of pro-inflammatory cytokines in the culture supernatant.

The experimental strategy will include several steps with different milestones.

Principal Investigator: Gianguglielmo Zehender
 

The ACE2 gene encodes the angiotensin converting enzyme 2, which has been shown to be the receptor for coronavirus (SARS-CoV) and novel coronavirus 2019-nCoV/SARS-CoV-2. 

Previous studies have shown a correlation between ACE2 expression and SARS-CoV infection in vitro. The different expression of ACE2 and conformation of the receptor or its signaling could play an important role in explaining the susceptibility, symptoms and outcome of Covid-19 infection.

The regulation of ACE2 expression is controlled by miRNA. Upregulation of miR-200c-3p expression in H5N1 influenza results in severe pneumonia in patients and is associated with reduced ACE2 levels, increased angiotensin II levels and consequently with lung damage. A similar picture can be observed in patients suffering from Covid-19 and it is possible to hypothesize a similar pathogenetic mechanism. 

The ACE2 receptor has a large number of genetic polymorphisms, and the association between these polymorphisms and pathological pictures has already been identified. In particular, 3 polymorphisms (rs2074192, rs233575, and rs2158083) have been associated in the Caucasian population with hypertension, which is a condition that predisposes to a more severe course of COVID-19 infection.

Objective: Evaluate the role of polymorphisms of the ACE2 gene and miRNA 200c-3p on the susceptibility, course and outcome of COVID-19 infection.
100 patients with a severe course of COVID infection will be compared with 100 patients with an asymptomatic or pauci-symptomatic course.

Material: peripheral blood with DNA extraction for the analysis of polymorphisms by PCR method and dosage of miRNA 200c-3p on plasma

Principal Investigator: Massimo Galli – Agostino Riva

This project aims to disclose the immunological basis of COVID-19 disease, by solving one of the molecular mechanisms that is mainly responsible for the onset of inflammatory response in SARS-CoV-2 infected patients: inflammasome activation.

Functional assay performed on PBMCs isolated from SARS-CoV-2 infected patients will allow to establish a correlation between harmful inflammation and symptom severity allowing to hypotize new therapeutic strategies to be validated in animal models and possibly translated to COVID-19 clinical practise. Altogether these results will allow to identify new players in the pathogenesis of SARS-CoV-2 infection that will be useful to: 

1. identify new molecular marker of SARS-CoV-2 infection and improve the diagnosis 
2. identify new target to improve the therapeutic approach of affected patients.  

Responsabile scientifico: Daria Trabattoni

The general objective of the QuoViRuS project is to combine research in the virological field with the environmental sector by investigating the presence of SARS-CoV-2 genetic material within rural surface waters receiving overflow water from some urban centres of the metropolitan city of Milan.  
Specifically, the project aims to:

• develop a robust and scalable methodological approach to water sampling, concentration and conservation  
• conduct appropriate molecular investigations aimed at viral RNA determination, using polymerase chain reaction (PCR) methods; experimental contamination of certain water samples with inactivated virus portions will allow validation of both the water concentration method and molecular assays  
• compare RNA levels in rural surface waters before and after overflow events 
• evaluate viral RNA concentrations at certain points in the rural grid at different times of the year, with different hydrological regimes  
• analyse the data obtained, and liaise with the local health agency (ATS) for a comparison with the available epidemiological data relating to COVID-19 cases in the population of the geographical area in question  
• evaluate the survival and thus the infectivity of any virus identified in the different collection areas, at different times of the year.  

Principal Investigator:   Prof. Elisabetta Tanzi
 

The study inquires how public opinion changed during the Covid-19 health emergency, in order to assess the social impact of the pandemic in Lombardy and the rest of Italy. The effects of the pandemic on social relations, mental and physical well-being, and the economy may lead to permanent transformations in the social fabric at the local and national level. Understanding these effects is essential to define appropriate, effective and efficient policy responses to the crisis in the medium and long term.  

Objective:  The study starts from a daily survey of a national population sample, which was conducted by the Department from 6 April to 8 July 2020. The opinions and behaviours of Italians were surveyed using the Rolling Cross Section technique (daily sampling) from the most severe phase of the pandemic through to the introduction of so-called phase 3. With the resumption of contagion, it will be useful to resume monitoring in a longer time frame, in order to understand adaptation and resilience strategies, as well as suffering and stress in individuals and society as a whole.  

We will monitor four key dimensions (perception of risk; compliance with government measures against Covid-19; perceived physical and mental well-being; changes in individual behaviour in the private and working spheres) and study their relations with socio-demographic characteristics; previous attitudes that may influence the individual response to the crisis; place of residence, with a focus on local infection trends.  

The second group of variables includes: horizontal (in other people) and institutional trust; conformism; perception of the individual-community relationship; solidarity; authoritarianism.  
The questions assess the potential bases of crisis-recovery public-health, social, and economic policies, and any habit-forming and / or intolerance phenomena due to the ongoing health, economic and social crisis.  

Given the wide sample, these questions may be used for studying evolution over time, as well as for a disaggregated analysis by geographical areas, up to the regional level for Lombardy (about 2,797 cases in the previous survey) and for the 8 regions with over 4 million inhabitants (over 1,000 each in the previous survey).  
 
Principal Investigator: Antonio M. Chiesi 
 

The UNICORN voluntary study asks non-symptomatic staff of the University of Milan to undergo a swab test for SARS-CoV-2 and a blood test for immunoglobulins (IgG and IgM) for the same virus, in order to:

1. estimate the current prevalence of the virus in an asymptomatic population, through swab analysis
2. identify whether a share of the population has already developed immunity to the virus, through immunoglobulin analysis.

Principal Investigators: Valentina Bollati and Gregorio Milani