Towards a biotechnological platform for the production of human pro-angiogenic growth factors in the green alga Chlamydomonas reinhardtii
| Autor: | Myra N. Chávez, Hans-Günther Machens, Alexandra-Viola Bohne, José T. Egaña, Jörg Nickelsen, Montserrat Jarquín-Cordero, Ursula Hopfner, Carolina Centeno-Cerdas |
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| Rok vydání: | 2019 |
| Předmět: |
Vascular Endothelial Growth Factor A
Proteome Transcription Genetic Transgene Genetic Vectors Chlamydomonas reinhardtii Gene Expression Biology Applied Microbiology and Biotechnology law.invention 03 medical and health sciences law Vector (molecular biology) Gene 030304 developmental biology Tube formation 0303 health sciences Expression vector 030306 microbiology Gene Expression Profiling Endothelial Cells Biological activity General Medicine Proto-Oncogene Proteins c-sis biology.organism_classification Chemokine CXCL12 Recombinant Proteins Cell biology Recombinant DNA Angiogenesis Inducing Agents Biotechnology |
| Zdroj: | Applied microbiology and biotechnology. 104(2) |
| ISSN: | 1432-0614 |
| Popis: | The recent use of photosynthetic organisms such as Chlamydomonas reinhardtii in biomedical applications has demonstrated their potential for the treatment of acute and chronic tissue hypoxia. Moreover, transgenic microalgae have been suggested as an alternative in situ drug delivery system. In this study, we set out to identify the best available combination of strains and expression vectors to establish a robust platform for the expression of human pro-angiogenic growth factors, i.e., hVEGF-165, hPDGF-B, and hSDF-1, in biomedical settings. As a case study, combinations of two expression vectors (pOpt and pBC1) and two C. reinhardtii strains (UVM4 and UVM11) were compared with respect to hVEGF-165 transgene expression by determination of steady-state levels of transgenic transcripts and immunological detection of recombinant proteins produced and secreted by the generated strains. The results revealed the combination of the UVM11 strain with the pBC1 vector to be the most efficient one for high-level hVEGF-165 production. To assess the robustness of this finding, the selected combination was used to create hPDGF-B and hSDF-1 transgenic strains for optimized recombinant protein expression. Furthermore, biological activity and functionality of algal-produced recombinant pro-angiogenic growth factors were assessed by receptor phosphorylation and in vitro angiogenesis assays. The results obtained revealed a potentiating effect in the combinatorial application of transgenic strains expressing either of the three growth factors on endothelial cell tube formation ability, and thus support the idea of using transgenic algae expressing pro-angiogenic growth factors in wound healing approaches. |
| Databáze: | OpenAIRE |
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