Enhanced lipolysis in normal mice expressing liver-derived human lipoprotein lipase after adenoviral gene transfer.
The authors previously demonstrated that the gene for human lipoprotein lipase (hLPL), an enzyme crucial to the breakdown of triglyceride (TG)-rich dietary fats, corrects the hypertriglyceridemia in lipoprotein lipase (LPL)-deficient knockout mice after adenoviral (Ad)-mediated LPL gene transfer. They have now extended their observations to primary cultured mouse hepatocytes and intact animals of normal LPL genotype, and confirm effective overexpression of hLPL from the liver and a sustained TG-lowering effect in plasma over 60 days. A typical first-generation Ad-vector containing the hLPL cDNA (Ad-LPL) resulted in efficient gene transfer into isolated mouse hepatocytes and significant de novo synthesis of active hLPL protein. In this experiment, 5 x 10(9) viral particles (5 x 10(7) pfu) of either Ad-LPL or an Ad-LacZ control vector were injected into CD1 mice of normal LPL genotype. Hepatic expression of hLPL was confirmed at Day 7 postinjection by in situ hybridization and direct measurement of LPL in the liver. This correlated with a total LPL activity (human + mouse) in postheparin plasma (PHP) of 1020.5 standard deviation [SD] 93.6 mU/mL, versus 479.5 SD 129.7 mU/mL (p < 0.001) in Ad-LacZ controls at Day 7. Respective hLPL activity comprised 49% of the total. Significantly raised levels of hLPL protein mass persisted until Day 60. Corresponding plasma TGs decreased to 39% of Ad-LacZ controls at Day 7, and, despite absent hLPL activity from Day 28 on, serum TGs remained significantly lower in Ad-LPL mice up to Day 42. Fast phase liquid chromatography analysis showed a dramatic depletion in TG-rich lipoproteins, mainly very low density lipoproteins (VLDL) and chylomicron fractions. Therefore, Ad-mediated overexpression of hepatic LPL was found to significantly decrease plasma TG levels unrelated to primary LPL deficiency.
Liu
G
G
Department of Medical Genetics, University of British Columbia, Vancouver.
Excoffon
K J
KJ
Wilson
J E
JE
McManus
B M
BM
Miao
L
L
Benoit
P
P
Duverger
N
N
Branellec
D
D
Denefle
P
P
Hayden
M R
MR
Lewis
M E
ME
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D002478:Cells, Cultured; D002914:Chylomicrons / Q000097:blood; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D018014:Gene Transfer Techniques; D006801:Humans; D017403:In Situ Hybridization; D008066:Lipolysis / Q000235:genetics*; D008071:Lipoprotein Lipase / Q000235:genetics* / Q000378:metabolism; D008074:Lipoproteins / Q000097:blood; D008099:Liver / Q000201:enzymology* / Q000821:virology; D008297:Male; D051379:Mice; D008815:Mice, Inbred Strains; D014280:Triglycerides / Q000097:blood
D002914:Chylomicrons; D008074:Lipoproteins; D014280:Triglycerides; D008071:Lipoprotein Lipase