Improving cell lines radiotolerance through heterologous expression of a bdelloid rotifer dna ligase [Offre de technologie]

Method for increasing tolerance of a cell or an organism to stress factors such as radiation. The technology provides cell lines with improved radiotolerance applicable in a broad range of fields spanning from space research to cancer therapy.

Some organisms known as the extremophiles are tolerant to extremely harsh environment. Besides bacteria such as Deinococcus radiodurans, some animals also exhibit a high DNA damage tolerance when experiencing desiccation or exposure to high doses of ionizing radiations. Among those, bdelloid rotifers present striking tolerance to ionizing radiation. The Adineta vaga species can for instance survive doses up to 5 kGy of X-ray (Front. Microbiol, 11: 1792). At such high radiation doses, bdelloid rotifers experience considerable DNA damages including double-strand breaks. Those organisms however manage to recover DNA integrity after irradiation through an active DNA repair process.

Adineta vaga 
size: 100-200 µm 

The inventors identified an atypical and horizontally acquired DNA ligase of Adineta vaga (A. vaga) with a structural core similar to prokaryotic DNA ligase E (AvLigE) that strongly contributes to the DNA ligation activity of A. vaga upon DNA damage induction and is thereby key to the radiotolerance of this species. Heterologous expression of AvLigE within human cell lines significantly improved their stress response, in particular their survival upon exposure to ionizing radiation. It could as well be demonstrated that AvLigE expression increased DNA ligation activity of the corresponding CRISPR/Cas9 modified stable cell lines.
The present technology provides methods to genetically engineer cells or transgenic animals to increase their tolerance to stress factors, in particular their radiotolerance.

• Engineered cell lines and/or model animals as biological tools for space research considering the higher radiation exposure in space than on the surface of earth.
• Possibility to engineer T Cells to improve their radio-tolerance for the development of emerging cancer treatments combining CAR T Cells and radio-therapy.

• Recombinant polynucleotide for heterologous expression of a DNA ligase in any type of living cells to improve their tolerance to exogenous stress
• Efficient molecular repair system for maintenance of genome integrity in cells
• Human cell lines with improved radio-resistance readily available

TRL-3 Proof of concept established in vitro on human cell lines.

 After a post-doctoral stay at Harvard University and a position of associate professor at UNamur, Pr Karine Van Doninck founded the research unit of Molecular Biology and Evolution at ULB in 2020 to pursue fundamental research at the interface of molecular and evolutionary biology. Central to her research are micro-organisms called bdelloid rotifers which exhibit unique adaptation mechanisms to survive the most hostile environments (freezing, desiccation, ionizing radiation and outer space). She and her research team study the underlying mechanisms of their evolution, while also disentangling the processes behind their adaptations to extreme environments through interdisciplinary approaches. She was awarded with a European Research Council (ERC) Consolidator grant in 2017 and a European Space Agency project, both related to her research on rotifers. Two experiments have already been launched by SpaceX to the International Space Station, with a third planned in 2024.

1. Iron Ladies – How Desiccated Asexual Rotifer Adineta vaga Deal With X-Rays and Heavy Ions?, Hespeels B., Penninckx S., Cornet V., Bruneau L., Bopp C., Baumlé V., Redivo B., Heuskin A. C., Moeller R., Fujimori A., Lucas S., Van Doninck K., Front. Microbiol, 11: 1792, July 2020. DOI: 10.3389/fmicb.2020.01792
2. Chromosome-level genome assembly reveals homologous chromosomes and recombination in asexual rotifer Adineta vaga, Simion P., Narayan J., Houtain A., Derzelle A., Baudry L., Nicolas E., Arora R., Cariou M., Cruaud C., Gaudray F. R., Gilbert C., Guiglielmoni N., Hespeels B., Kozlowski D. K. L., Labadie K., Limasset A., Llirós M., Marbouty M., Terwagne M., Virgo J., Cordaux R., Danchin E. G. J., Hallet B., Koszul R., Lenormand T., Flot J. F., Van Doninck K., Science Advances, 7 (41): eabg4216, October 2021. DOI: 10.1126/

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