Riksen, Joost, Patrian, Marta, Garcia Perez, Elena, Wilbers, Ruud, Kammenga, Jan, Bunte, Myrna, Schots, Arjen
[
International Worm Meeting,
2021]
The attachment of phosphorylcholine on carbohydrates (PC-glycans) is a common modification for nematodes including C. elegans. The presence of PC appears to be important for the development of nematodes and for immunomodulation by parasitic nematodes. PC-glycoprotein ES-62 of Acanthoceilonema vitae directs immune cells towards an anti-inflammatory phenotype and this immunomodulatory property appears to be dependent on the presence of PC-glycans. This makes glycoproteins with PC-glycans interesting therapeutic agents to combat immune disorders such as rheumatoid arthritis, systemic lupus erythematosus and asthma. However, the biosynthetic pathway of PC-glycans is not completely understood, where especially the identity of the PC-transferring enzyme, or PC-transferase, is not fully elucidated. There are strong indications suggesting that fukutin-related genes potentially encode for this PC-transferase. In this study, we examined whether four selected fukutin-related genes of C. elegans (W02B3.4, T07A5.1, T07D3.4 and Y22D7AL.11) are involved in the biosynthetic pathway of PC glycans. With CRISPR/Cas9 technology we created C. elegans knock-out lines for each fukutin-related gene, but no significant reduction of PC was observed. Interestingly, one mutant line of W02B3.4 showed an increase of PC due to a potentially introduced signal peptide, indicating that the W02B3.4 gene could encode for a PC-transferase. Currently, we are combining mutant lines into double/triple/quadruple knock-out lines, which may provide more insight whether these fukutin-related genes encode for PC-transferases. These findings will contribute to our understanding of the pathway for PC-glycan biosynthesis, offering potential opportunities for design and synthesis of PC-glycan therapeutics.