Periodic Reporting for period 1 - AlgaeNet4AV (Training network for exploring microalgae diversity for the development of novel antiviral compounds)
Reporting period: 2023-01-01 to 2024-12-31
Microalgae are very diverse, and adapted to a broad variety of environmental conditions, the chances to find novel and unexplored bioactive metabolites with properties of interest for biotechnological and biomedical applications are high. The potential of exploiting the microalgae-based ingredients with antiviral properties has already been documented (Falaise et al., 2016; Carbone et al 2021; Romero et al., 2021), however, due to the lack of systematic R&D efforts, these compounds are unexploited. Lectins and polyphenols from microalgae are compounds that display strong antiviral activities, that haven’t been exploited so far. Lectins are ubiquitous proteins/glycoproteins of non-immune origin that bind reversibly to carbohydrates in a non-covalent and highly specific manner. These lectin-glycan interactions could be exploited for the establishment of novel therapeutics, targeting the adherence stage of viruses and thus helpful in eliminating widespread viral infections (Singh et al., 2018; Lee et al., 2019; Car-bone et al 2021; Romero et al., 2021). They have a unique and
rapid mode-of-action holding several advantages compared to conventional drugs. For ex-ample, their ability to bind viral surface sugars in seconds, makes them much faster than the existing drugs. Lectins resistance has not been observed among virus from diverse sources and resistant strains can generally not be selected during in vitro experiments.
As we are still unable to predict with confidence the progress of the coronavirus pandemic and considering that the next pandemic is most likely to be caused by influenza, the discovery of antiviral compounds continues to be the priority public health threat in the world (Mani et al., 2020; Martinez et al., 2020). Most importantly, we need to have a range of available antiviral tools that can respond rapidly and effectively to emergencies. For practical application, non-drug antiviral compounds must be available in massive amounts and produced cost-effectively.
These requirements might eliminate recombinant proteins from consideration as the time and costs of production in cell-based fermentation systems are prohibitively high. The main objective of the AlgaeNet4AV project is the characterization, and exploitation of microalgae biodiversity as a source for bio-based cosmetics with antiviral potential towards influenza and coronavirus pandemic strains. These products aims to be used as non-drug interventions for eliminating the propagation of harmful viruses and directly contribute to reduce the risk and speed of transmission, acting as first-line weapons towards pan-demic outbreaks.
rapid mode-of-action holding several advantages compared to conventional drugs. For ex-ample, their ability to bind viral surface sugars in seconds, makes them much faster than the existing drugs. Lectins resistance has not been observed among virus from diverse sources and resistant strains can generally not be selected during in vitro experiments.
As we are still unable to predict with confidence the progress of the coronavirus pandemic and considering that the next pandemic is most likely to be caused by influenza, the discovery of antiviral compounds continues to be the priority public health threat in the world (Mani et al., 2020; Martinez et al., 2020). Most importantly, we need to have a range of available antiviral tools that can respond rapidly and effectively to emergencies. For practical application, non-drug antiviral compounds must be available in massive amounts and produced cost-effectively.
These requirements might eliminate recombinant proteins from consideration as the time and costs of production in cell-based fermentation systems are prohibitively high. The main objective of the AlgaeNet4AV project is the characterization, and exploitation of microalgae biodiversity as a source for bio-based cosmetics with antiviral potential towards influenza and coronavirus pandemic strains. These products aims to be used as non-drug interventions for eliminating the propagation of harmful viruses and directly contribute to reduce the risk and speed of transmission, acting as first-line weapons towards pan-demic outbreaks.
So far we performed a) develop transcriptomics and metabolomics resources from microalgae diversity as a valuable source for the discovery novel lectins and polyphenols;
b) characterize their spectrum of bioactivity and toxicity and provide structure-function relationships;
c) develop and optimize application-based microalgae culture systems and downstream processing strategies for higher production rate of the desired compounds;
d) develop, formulate and evaluate lectins and polyphenols-based final cosmetics products.
Over the past two years, a total of 38 secondments have been completed, 13 deliverables have been submitted, and both milestones have been successfully reached. The majority of the planned actions and scientific objectives for the first reference period have been accomplished. Various microalgae species were grow and characterized under different environmental conditions, including UV exposure, temperature, pH, and nutrient variations. Genomic DNA and RNA sequencing of Jaagichlorella luteoviridis from UV and temperature experiments has been conducted, along with genomic DNA sequencing of two provisionally classified Arthrospira/Limnospira spp. Metabolic analysis is underway to determine optimal growth conditions for enhanced lectin and polyphenol production in each strain. The identified conditions will be shared with industrial partners for large-scale production by NECTON and Allmicroalgae. Novel lectin sequences have been identified, cloned, purified and characterized. In addition, polyphenol analysis allowed the selection of strains the display antiviral and antioxidant activity. Selected microalgae extracts have been provided to the industrial partners BIONOS for toxicological analysis and bioactivity assessing using tissue culture experiments. Also, microalgae extracts have been provided to Fresh Line for cosmetic formulation and stability test analysis.
Additionally, the project’s website (http://algaenet4av.eu/(opens in new window)) has been released for project dissemination, knowledge transfer, and training among academic and commercial partners. The consortium has actively participated in outreach activities, including Researchers' Night and the MSCA-NET Final Conference.
Due to minor delays and challenges in the scientific work, we have submitted an amendment to include two new partners, ensuring the effective achievement of all project objectives.
b) characterize their spectrum of bioactivity and toxicity and provide structure-function relationships;
c) develop and optimize application-based microalgae culture systems and downstream processing strategies for higher production rate of the desired compounds;
d) develop, formulate and evaluate lectins and polyphenols-based final cosmetics products.
Over the past two years, a total of 38 secondments have been completed, 13 deliverables have been submitted, and both milestones have been successfully reached. The majority of the planned actions and scientific objectives for the first reference period have been accomplished. Various microalgae species were grow and characterized under different environmental conditions, including UV exposure, temperature, pH, and nutrient variations. Genomic DNA and RNA sequencing of Jaagichlorella luteoviridis from UV and temperature experiments has been conducted, along with genomic DNA sequencing of two provisionally classified Arthrospira/Limnospira spp. Metabolic analysis is underway to determine optimal growth conditions for enhanced lectin and polyphenol production in each strain. The identified conditions will be shared with industrial partners for large-scale production by NECTON and Allmicroalgae. Novel lectin sequences have been identified, cloned, purified and characterized. In addition, polyphenol analysis allowed the selection of strains the display antiviral and antioxidant activity. Selected microalgae extracts have been provided to the industrial partners BIONOS for toxicological analysis and bioactivity assessing using tissue culture experiments. Also, microalgae extracts have been provided to Fresh Line for cosmetic formulation and stability test analysis.
Additionally, the project’s website (http://algaenet4av.eu/(opens in new window)) has been released for project dissemination, knowledge transfer, and training among academic and commercial partners. The consortium has actively participated in outreach activities, including Researchers' Night and the MSCA-NET Final Conference.
Due to minor delays and challenges in the scientific work, we have submitted an amendment to include two new partners, ensuring the effective achievement of all project objectives.
Thus far the project in the first two years of its life has progressed significantly beyond the state of the art
1) Cultivation of various microalgae species, including Nostoc muscorum, Tetradesmus obliquus, Kirchneriella lunaris, Euglena cantabrica, Dunaliella salina, Jaagichlorella luteoviridis, Chlorella vulgaris, Nannochloropsis oceanica and Spirulina spp., was achieved under different growth conditions such as UV exposure, temperature variations, pH levels, and nutrient availability. Their polyphenolic content, along with their antimicrobial and antiviral properties, were investigated.
2) Extremophilic and robust strains (e.g. Arthrospira platensis, Dunaliella salina, C. calcitrans, C. fusiformis and N. shiloi) were cultivated and their polyphenols, antimicrobial and antiviral properties are being investigated.
3) The genomic DNA and RNA from Jaagichlorella luteoviridis have been sequenced following UV and temperature experiments, and we are currently analyzing the results alongside genomic DNA samples extracted from two provisionally classified Arthrospira/Limnospira spp.
4) Metabolic analyses are being conducted on microalgae grown under varying conditions to determine the optimal parameters for enhanced lectin and polyphenol production in each strain. The established protocols will be provided to industrial partners for large-scale production of the best performing strains.
5) Transcriptomic and bioinformatics analysis allowed the identification of nine putative novel lectin sequences that have been successfully cloned, expressed in E. coli and purified. The sugar-binding specificity and the antiviral activity of recombinant lectins are currently under investigation.
6) Selected microalgae extracts, rich in polyphenols with high antioxidant capacity and antiaging properties have been characterized and supplied to Fresh Line for cosmetic formulation. Stability analysis of cosmetic formulas is currently being assessed. Also, the same extracts are being evaluated by BIONOS for toxicological analysis and bioactivity assessment using tissue culture experiments.
1) Cultivation of various microalgae species, including Nostoc muscorum, Tetradesmus obliquus, Kirchneriella lunaris, Euglena cantabrica, Dunaliella salina, Jaagichlorella luteoviridis, Chlorella vulgaris, Nannochloropsis oceanica and Spirulina spp., was achieved under different growth conditions such as UV exposure, temperature variations, pH levels, and nutrient availability. Their polyphenolic content, along with their antimicrobial and antiviral properties, were investigated.
2) Extremophilic and robust strains (e.g. Arthrospira platensis, Dunaliella salina, C. calcitrans, C. fusiformis and N. shiloi) were cultivated and their polyphenols, antimicrobial and antiviral properties are being investigated.
3) The genomic DNA and RNA from Jaagichlorella luteoviridis have been sequenced following UV and temperature experiments, and we are currently analyzing the results alongside genomic DNA samples extracted from two provisionally classified Arthrospira/Limnospira spp.
4) Metabolic analyses are being conducted on microalgae grown under varying conditions to determine the optimal parameters for enhanced lectin and polyphenol production in each strain. The established protocols will be provided to industrial partners for large-scale production of the best performing strains.
5) Transcriptomic and bioinformatics analysis allowed the identification of nine putative novel lectin sequences that have been successfully cloned, expressed in E. coli and purified. The sugar-binding specificity and the antiviral activity of recombinant lectins are currently under investigation.
6) Selected microalgae extracts, rich in polyphenols with high antioxidant capacity and antiaging properties have been characterized and supplied to Fresh Line for cosmetic formulation. Stability analysis of cosmetic formulas is currently being assessed. Also, the same extracts are being evaluated by BIONOS for toxicological analysis and bioactivity assessment using tissue culture experiments.