Low-density lipoprotein (LDL) receptor/transferrin fusion protein: in vivo production and functional evaluation as a potential therapeutic tool for lowering plasma LDL cholesterol.
A soluble form of human low-density lipoprotein receptor (LDL-R) fused in frame with rabbit transferrin (LDL-Rs(hu)/Tf(rab)) is assessed in vivo as a therapeutic tool for lowering plasma LDL cholesterol. The cDNA encoding LDL-Rs(hu)/Tf(rab) is expressed in mice, using a hydrodynamics-based gene transfer procedure. The transgene is transcribed in the liver of transduced animals and the corresponding protein is secreted into the bloodstream. Circulating LDL-Rs(hu)/Tf(rab) binds LDL specifically, thus indicating that it is correctly processed through the cellular compartments in vivo. More importantly, the expression of LDL-Rs(hu)/Tf(rab) allows the removal of injected human (125)I-labeled LDL ((123)I-LDL) from the bloodstream of transduced CD1 mice, which show faster LDL plasma clearance, anticipating by approximately 90 min the same clearance value observed in control animals. A similar effect is observed in transduced LDL-R(-/-) mice, in which the clearance of injected human LDL depends solely on the presence of circulating LDL-Rs(hu) /Tf(rab). In these animals the extent of plasma LDL clearance is directly related to the concentration of LDL-Rs(hu)/Tf(rab) in the blood. Finally, LDL-Rs(hu)/Tf(rab) does not alter the pattern of LDL organ distribution: in transduced animals, as well as in control animals, liver and bladder are the predominantly labeled organs after (123)I-LDL injection. However, LDL-Rs(hu)/Tf(rab) has a quantitative effect on LDL tissue deposition: in treated animals LDL-Rs(hu)/Tf(rab) determines an increase in radioactivity in the liver at early times after (123)I-LDL injection and a progressive labeling of the bladder, starting 20 min after injection.
Razzini
Giorgia
G
Dipartimento di Scienze Biomediche, Università di Modena e Reggio Emilia, Via G. Campi 287, 41100 Modena, Italy.
Parise
Flavia
F
Calebiro
Davide
D
Battini
Renata
R
Bagni
Bruno
B
Corazzari
Tolmino
T
Tarugi
Patrizia
P
Angelelli
Cecilia
C
Molinari
Susanna
S
Falqui
Luca
L
Ferrari
Stefano
S
D000818:Animals; D015153:Blotting, Western; D008078:Cholesterol, LDL / Q000097:blood*; D005260:Female; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D006720:Homozygote; D006801:Humans; D008099:Liver / Q000187:drug effects / Q000378:metabolism; D008657:Metabolic Clearance Rate; D051379:Mice; D018345:Mice, Knockout; D012333:RNA, Messenger / Q000378:metabolism; D011817:Rabbits; D011973:Receptors, LDL / Q000235:genetics*; D011993:Recombinant Fusion Proteins / Q000097:blood / Q000502:physiology*; D014018:Tissue Distribution; D014168:Transferrin / Q000235:genetics*; D019076:Transgenes / Q000502:physiology; D001743:Urinary Bladder / Q000187:drug effects / Q000378:metabolism
D008078:Cholesterol, LDL; D012333:RNA, Messenger; D011973:Receptors, LDL; D011993:Recombinant Fusion Proteins; D014168:Transferrin