Glikozilacija IgG-a u žena: Pubertet, trudnoća i menopauza

undergraduate thesis

• Title (english): IgG glycosylation in women: Puberty, pregnancy, and menopause
• Author: Maja Perinović
• Mentor: Vlatka Zoldoš (mentor)
• Granter: University of Zagreb; Faculty of Science; (Department of Biology); Zagreb
• Defense date and country: 2023-09-14, Croatia
• Scientific / art field, discipline and subdiscipline: NATURAL SCIENCES; Biology

Abstract

Imunoglobulin G (IgG) najzastupljenija je izoforma imunoglobulina u krvotoku čovjeka. Nglikani vezani na IgG utječu na efektorske funkcije ovog antitijela i na taj način reguliraju
imunološki odgovor. Kompozicija IgG glikoma značajno se mijenja u pubertetu, trudnoći i
menopauzi kao direktna posljedica promjena u koncentraciji estrogena. Međutim, točni
mehanizmi kojima estrogen dovodi do promjene glikozilacije IgG-a nisu poznati. Cjelogenomske
studije povezanosti su otkrile kako je gen ABCF2 reguliran estrogenom uključen u glikozilaciju
IgG-a. Kako bismo potvrdili ulogu gena ABCF2 u tom procesu, povećali smo njegovu ekspresiju
u staničnoj liniji FreeStyle-293-F koja stabilno eksprimira fuzijski protein dCas9-VPR i luči
molekule IgG u stanični medij. Povećana ekspresija gena ABCF2 nakon manipulacije stanica
alatom CRISPR/dCas9-VPR rezultirala je povećanom razinom galaktoziliranih glikoformi IgG-a
(G1 i G2) i istovremenim smanjenjem agalaktoziliranih glikoformi IgG-a (G0) što sugerira da
ABCF2, gen reguliran estrogenom, najvjerojanije sudjeluje u regulaciji glikozilacije IgG-a.

Abstract (english)

Immunoglobulin G (IgG) is the most abundant immunoglobulin isoform in the human
bloodstream. Different N-glycans attached to IgG directly influence this antibody's effector
functions and regulate the immune response. IgG glycome composition is changed significantly
during puberty, pregnancy, and menopause, as a direct consequence of the change in estrogen
concentration. However, the exact mechanisms by which estrogen modulates IgG glycosylation
are unknown. Genome-wide association studies have shown that the estrogen-regulated gene
ABCF2 is associated with IgG glycosylation. To functionally validate the role of the ABCF2 gene
in this process, I overexpressed it in the FreeStyle-293-F cell line that stably expresses the dCas9-
VPR fusion protein and secretes IgG. Overexpression of the ABCF2 gene resulted in an increased
level of galactosylated IgG glycoforms (G1 and G2) and a simultaneous decrease in
agalactosylated glycoforms of IgG (G0), suggesting that estrogen-regulated gene ABCF2 has a
role in the regulation of IgG glycosylation.

• Keywords: estrogen; glikozilacija proteina; IgG; CRISPR/dCas9; galaktozilacija; gen ABCF2
• Keywords (english): estrogen; protein glycosylation; IgG; CRISPR/dCas9; galactosylation; ABCF2
• Language: croatian
• URN:NBN: urn:nbn:hr:217:185283
• Study programme: Title: Molecular Biology; Study programme type: university; Study level: undergraduate; Academic / professional title: sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) molekularne biologije (sveučilišni/a prvostupnik/prvostupnica (baccalaureus/baccalaurea) molekularne biologije)
• Type of resource: Text
• File origin: Born digital
• Access conditions: Embargoed access; Embargo expiration date: 2025-09-14
• Created on: 2023-09-15 08:54:35