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Description
Hepatitis B and C viruses infect the human liver, triggering persistent inflammation and eventually generating a form of cancer which is the second leading cause of cancer-related mortality worldwide. The main objective of the project is production in premiere of novel antigens with improved properties. In this context, our project proposal aims to: a) build upon our experience and knowledge accumulated in our previous research collaboration on Hepatitis B and C viruses (GreenVac project 2014-2017 funded by the EEA Norway-Romania Program) and produce high yields of novel Hepatitis B and C antigens with superior properties based on innovative molecular design; b) take this experience a step forward and establish in premiere an advanced biotechnological platform for production of Hepatitis B and C antigens in algae; c) take advantage of the multiple recombinant protein expression systems and perform a systematic, comparative characterization of the biochemical and functional properties of the new antigens; d) develop institutional networking around innovative plant and microalgae biotechnologies and train human resources in protein science in applied research in Romania and Norway.
Summary of project results
Hepatitis B (HBV) and C viruses (HCV) cause 80% of all liver cancer, killing about 1.4 million people every year. More than 500 million individuals are chronically infected with HBV or HCV worldwide, and at high risk of developing end-stage liver disease. WHO indicates that by 2030, “an estimated 4.5 million premature deaths due to HBV/HCV infections could be prevented in low- and middle-income countries through vaccination, diagnostic tests, medicines and education campaigns”. The ultimate goal of prophylactic treatment in viral hepatitis is virus eradication, achievable through universal vaccination programs. These programs are financially very challenging and countries confronted with the highest rates of infection, cannot afford them. Production of cost-effective vaccines would alleviate the economic burden on public health systems and increase availability to vulnerable societies. In the case of HBV, more immunogenic antigens are needed to increase efficiency of the immune response in poor-responders to current vaccines. For HCV, no vaccine is yet available.
Our study aimed to develop alternative technologies for production of innovative vaccines at low-cost in plants and algae, following rational design and computer modelling. Thus, by addressing these major challenges in HBV/HCV vaccine development, our project has joined the international fight against viral hepatitis.
Our project proposed (i) an innovative HBV/HCV antigen design, based on combination of relevant epitopes (HBV) and stabilization of immunogenic protein structures (HCV) and, (ii) evaluation of plants and algae as cost-efficient production platforms for the novel antigens. Plants and mammalian cells (the physiological host of viral proteins) have slightly different protein processing mechanisms. Plants may change proteins by adding structures that can be allergenic to humans, making production of recombinant proteins of human medical use, such as antigens and antibodies, very challenging. To tackle this issue, we successfully used genome-edited plants with “humanized” N-glycosylation pathway for production of glyco-engineered viral antigens. Moreover we demonstrated in premiere that very high amounts of antigens can be obtained in green and red algae, with similar properties to those produced in mammalian cells. Of the list of antigens generated in our study, two vaccine candidates with best properties (one HBV and one HCV) were selected for further development, produced in mammalian cells, plants and algae and tested in animal trials. The antigens produced in both “humanized” plants and red microalgae induced a significant virus-neutralization immune response, validating the improved design and the novel production hosts for complex viral proteins.
To combat viral diseases relevant for human health, continuous development of improved vaccines is the ONLY effective solution. Our SmartVac project has delivered the scientific community novel research data regarding the development of improved HBV/HCV vaccines, thus responding to the challenges of the WHO call “to eliminate viral hepatitis by 2030”. Our project has advanced the field of molecular farming, by proposing plants with “humanized” N-glycosylation and microalgae, as novel hosts for cost-effective production of complex viral proteins. Upscaling of marine algae is very feasible when compared with fermenter-based traditional pharmaceutical production systems, thus, our results also pave the way for the development of edible marine algae as promising candidates for oral vaccine production in the future.
In our study, we used HBV/HCV envelope proteins as model antigens to investigate these new expression systems, however, the knowledge we have generated, together with the experimental protocols, can be easily adapted to any protein of medical interest, for rapid production in case of emergency. Provided these vaccine candidates will successfully make their way towards vaccine development, they will be available to all individuals at risk of developing HBV/HCV infections, regardless of their race, gender, sexuality, or abilities.
Summary of bilateral results
By tackling a challenge of global importance, the SmartVac project has also created and ideal frame for international cooperation, knowledge exchange and capacity building in biotechnologies for human health, in both Romania and Norway. This fruitful collaboration will continue beyond the project life-span through (i) collaborative project applications including the Romanian and Norwegian partners that have already been initiated; (ii) further development of the HBV/HCV vaccine candidates generated in the SmartVac project, in the frame of a national project supported by The European Regional Development Fund; (iii) extending the current consortium by recruiting other European teams and apply in highly competitive EU programs, to advance current state-of-the art in plant and microalgae biotechnologies and develop novel medical applications.