Glyco-engineering Human O-Glycosylation in plant cells

Human mucins are large heavily O-glycosylated glycoproteins (>200 kDa) in mucus layers, which hydrate, lubricate and protect cells from proteases as well as from pathogens. O-linked mucin glycans are truncated in many cancers, e.g. yielding the truncated cancer specific epitope Tn (a single GalNAc sugar attached to Ser or Thr on the protein backbone).

 

In collaboration with Henrik Clausen and Eric P. Bennett (Copenhagen Center for Glycomics, http://icmm.ku.dk/forskergrupper/ccg/) we have engineered plant cells to produce the Tn mucin-type protein O-glycosylation, which basically involves: 





1. Engineering O-glycosylation capacity: expression of Golgi-localized human GalNAc transferases (GalNAc-T2 and -T4) and a cytoplasmic UDP-GlcNAc C4-epimerase to produce UDP-GalNAc, which is not present in plants. 
 


 

2. Expression of human target proteins (e.g. Mucin 1, interferon alpha 2 B, podoplanin, … ) in the O-glycosylation capacity background.

 

 

Applications of the general glycosylation platform include production of cancer glyco-specific vaccines, increased compatibility with immune system in man and half-life in blood stream of glycoprotein pharmaceuticals.
 
Selected publications:


Yang Z, Bennett EP, Arigi EA, Drew DP, Levery SB, Ulvskov P, Clausen H, Petersen BL (2011) “Methods for Glyco-engineering Controlled Human O-Glycosylation in plants" - US App 13/070,248 (23/3-2011)

Tarp MA, Sørensen AL, Mandel U, Paulsen H, Burchell J, Taylor-Papadimitriou J, Clausen H
(2007) Glycobiology 17(2):197-209

 

Wandall HH, Blixt O, Tarp MA, Pedersen JW, Bennett EP, Mandel U, Ragupathi G,  Livingston PO, Hollingsworth MA, Taylor-Papadimitriou J, Burchell J, Clausen H (2010) Cancer Biomarkers Defined by Autoantibody Signatures to Aberrant O-Glycopeptide Epitopes. Cancer Res 70(4):