The patent has been filed, now the drug is being developed. Italian research has made available to the scientific community a technique for precision treatment capable of hindering the infection of the coronavirus and its rapid spread between cells. The study, published in the journal Pharmacological Research, which will pave the way for a new therapeutic approach to prevent severe Covid-19 infection, is a collaboration between the Italian Institute of Technology, the Scuola Universitaria Superiore Sant'Anna Pisa, and the University of Milan.
The three research groups, led by Paolo Ciana from the University of Milan, Angelo Reggiani from Istituto Italiano di Tecnologia (IIT) and Vincenzo Lionetti from Scuola Superiore Sant'Anna Pisa, have shifted the focus from the characteristics of the virus to the gateway it uses to enter human cells.
Why did you make this choice?
What were the development phases of the study?
To find out more, we interviewed the scientists behind the patent that fights the coronavirus and its variants. The word to: Angelo Reggiani fromItalian Institute of Technology, Paolo Ciana fromUniversità di Milano e Vincenzo Lionetti from Sant'Anna School of Advanced Studies Pisa.
How did the idea for the project and the collaboration between the 3 institutions come about?
The idea was born at the beginning of the first lockdown when the world was dealing with Covid-19 disease for the first time. These were days when thousands of infected people became ill with Covid-19 and died far from their families, due to severe acute respiratory failure often associated with multi-organ dysfunction. This emergency rushed researchers around the world to suspend their research activities and collaborate to find a therapeutic solution now.
We knew that the Sars-CoV-2 beta-coronavirus spike protein binds to the human ACE2 protein, which is an important enzyme that catalyses the hydrolysis of angiotensin II, a protein hormone that acts in the cardiovascular and renal system and is involved in inflammation. This enzyme is already present on the membrane of most of our cells, and its levels increase as we age and take certain drugs. The real challenge was of finding a the way to block the coronavirus from using ACE2 without damaging the cell and minimising the risk of side effects.
Hence the idea of using a DNA aptamerwhich is known to be very effective at selectively binding proteins but also poorly immunogenic. To test the idea quickly, a multidisciplinary approach was needed, bringing together different skills, using the latest technology and drawing on the resources available at the time. For the three of us - an anaesthetist, a biotechnologist and a pharmacologist - it was not difficult to work together as we already knew each other and had clear ideas on how to proceed, albeit in a short space of time.
Training
What does the study consist of and what were the stages of development?
The study we recently published in the journal Pharmacological Research follows the deposit of a patent application and is a very important first step towards drug development. Initially, we searched for the right aptamer among countless potential candidates and eventually found two DNA aptamers capable of effectively binding the K353 residue of human ACE2 that is so attracted to the viral spike. We then tested them to assess their binding affinity to human ACE2 expressed by live cells.
We then tested whether aptamer binding to residue K353 of ACE2 was sufficient to prevent infection by the virus. As a final step, we tested whether the same dose of aptamer prevented other coronavirus variants from entering the same cell.
Did each of you deal with a specific phase?
The study and the patent were the result of extraordinary teamwork. The three of us met at least once a week and shared the work plan and strategy for each phase of the study. During our meetings, we planned each experiment together, analysed and discussed the results and drew up alternative strategies when we encountered obstacles.
The severe restrictions limiting access to our laboratories slowed us down, but did not prevent us from pooling our efforts to achieve the result.
You decided to shift the focus from the characteristics of the virus to the gateway it uses to enter human cells. Why and what led you to take this direction?
Sars-CoV-2 is an RNA virus and as such tends to mutate very often. This is evidenced by the numerous variants of the virus that have been isolated since the beginning of this pandemic. However, we now know that humans are equally infected by any variant because the coronavirus uses the same gateway to enter cells.
The severity of the disease mediated by an enormous inflammatory response depends on the availability of ACE2 to bind the viral spike, thus making possible primary infection but also its spread within the body by passing the virus from one cell to another. Like any virus, the coronavirus also needs a cell to proliferate and increase its infectious load.
How can be stopped the viral spread? Many researchers immediately decided to focus on developing strategies that annihilate the virus. We, on the other hand, decided to focus on ACE2 by developing an approach that is as effective as it is safe to make it unavailable to Sars-CoV-2 and its variants.
It is a very important element in this epidemiological phase given the growing number of mutations of the virus, is that so?
It is so. The growing number of virus mutations may require further efforts to update the effectiveness of an anti-viral drug. This risk does not exist if we decide to act on a stable cellular target such as ACE2.
What were the most critical issues you encountered?
Having to work during the lockdown was the biggest challenge, but we overcame it as best we could.
Patent
The precision technique developed by the three research groups has been patented. Work is now under way on the drug.
This study will make it possible to develop a new precision therapeutic approach to prevent severe Covid-19 infection. What are the advantages over monoclonal antibodies?
Aptamers are synthetic molecules structured as single-stranded DNA or the RNA oligonucleotides that can are able to mim because they mimic the functional properties of monoclonal antibodies. Unlike the latter, aptamers are not immunogenic molecules and are safer.
To date, neither activation of the immune system nor complement activation by aptamers has been reported. There is no evidence that they cause off-target side effects, as is described in the case of monoclonal antibodies.
The value of Italian scientific research
What does this important result mean for you and for Italian and international scientific research?
It is not today that we discover the great value of Italian scientific research. The international scientific community, to which Italian researchers belong, has taught us that collaboration between research groups is the only way to achieve great results quickly, especially in times of emergency such as the current one.
This result is important for everyone because it gives doctors the hope of working with a therapy that will help prevent serious forms of the disease and people the hope of receiving a cure. We were only an instrument to achieve a first result that will require further development.
Are there further developments for the future?
Our collaboration is aimed at accompanying the aptamer through to production.
We are trying, but the funds needed to transform the extraordinary results of our study into a useful drug for humans are very large. Without the necessary support, including financial support, we will never succeed on our own.
What is your message for young Italian researchers in Italy and abroad?
Young Italian researchers participated in our study. We invite them, even when working abroad, to continue to pursue their dreams through their work. Not to be afraid of sharing an idea.
Seeking collaborations is always a great opportunity to find quick solutions to a problem, as well as being moments of personal and professional growth.
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