Long-term reversal of hypercholesterolemia in low density lipoprotein receptor (LDLR)-deficient mice by adenovirus-mediated LDLR gene transfer combined with CD154 blockade.
BACKGROUND
Deficiency of the low density lipoprotein receptor (LDLR) results in abnormal elevation of cholesterol within the intermediate and low density plasma lipoproteins (IDL/LDL), and predisposes to early onset atherosclerosis. Cholesterol reduction after adenovirus-mediated LDLR gene transfer to LDLR-deficient animals is transient, due to the elicitation of an immune response.


METHODS
The LDLR-deficient mouse was used as a model to investigate adenovirus-mediated LDLR gene transfer combined with short-term immunosuppression as a cholesterol lowering therapy. Mice were infused with replication-deficient recombinant adenovirus encoding LDLR under control of the cytomegalovirus promoter/enhancer (AdLDLR), and injected with a blocking antibody directed against CD154 (CD40 ligand) to suppress immune responses against the vector and foreign transgene product.


RESULTS
Mice given AdLDLR and treated with anti-CD154 expressed LDLR on hepatocytes and maintained cholesterol levels below or within normal range for at least 92 days. In contrast, without adjunct immunosuppression LDLR expression was transient, corresponding to temporary decline in cholesterol levels. Analysis of cholesterol concentrations in fractionated plasma showed remarkable reduction in all lipoprotein fractions at early time-points after gene transfer. At later time-points, non-immunosuppressed control mice regained the disease profile with elevated IDL/LDL cholesterol, while profiles of anti-CD154-treated mice were similar to normal. LDLR mRNA transcripts were present in livers of the anti-CD154-treated mice but not controls, 93 days after AdLDLR injection. However, vector DNA was detected in livers of both groups. These results suggest that loss of LDLR expression in the non anti-CD154-treated mice was in part due to immune-mediated promoter silencing, and that anti-CD154 prevented this effect.


CONCLUSIONS
Treatment with anti-CD154 antibody inhibits immune-mediated loss of transgene expression, enabling long-term reduction in cholesterol levels after AdLDLR gene transfer to LDLR-deficient mice.
Stein
C S
CS
Department of Internal Medicine, University of Iowa College of Medicine, Iowa City 52242, USA.
Martins
I
I
Davidson
B L
BL
D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D000911:Antibodies, Monoclonal / Q000494:pharmacology*; D023201:CD40 Ligand; D002784:Cholesterol / Q000378:metabolism; D004274:DNA, Recombinant / Q000235:genetics; D018014:Gene Transfer Techniques; D015316:Genetic Therapy; D005822:Genetic Vectors / Q000235:genetics; D006937:Hypercholesterolemia / Q000628:therapy*; D007150:Immunohistochemistry; D007165:Immunosuppression Therapy; D007274:Injections, Intraperitoneal; D008099:Liver / Q000737:chemistry / Q000378:metabolism; D008297:Male; D008562:Membrane Glycoproteins / Q000276:immunology*; D051379:Mice; D008810:Mice, Inbred C57BL; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis; D011973:Receptors, LDL / Q000172:deficiency / Q000235:genetics* / Q000378:metabolism
D000911:Antibodies, Monoclonal; D004274:DNA, Recombinant; D008562:Membrane Glycoproteins; D012333:RNA, Messenger; D011973:Receptors, LDL; D023201:CD40 Ligand; D002784:Cholesterol