Low Expression of the Apolipoprotein B mRNA–Editing Transgene in Mice Reduces LDL Levels but Does Not Cause Liver Dysplasia or Tumors

From the Gladstone Institute of Cardiovascular Disease (X.Q., M.E.B., J.B., I.L., T.L.I.), the Cardiovascular Research Institute (X.Q., J.B., T.L.I.), and the Department of Pathology (T.L.I.), University of California, San Francisco; and the Department of Pharmacology (S.Y.), Osaka City University Medical School, Osaka, Japan.

Correspondence to Xiaobing Qian, MD, PhD, Gladstone Institute of Cardiovascular Disease, 365 Vermont St, San Francisco, CA 94103. E-mail xiaobing_qian{at}quickmail.ucsf.edu

Abstract—Hepatic expression of apolipoprotein (apo) B mRNA–editing enzyme catalytic polypeptide 1 (APOBEC-1) has been proposed as a gene therapy approach for lowering plasma low density lipoprotein (LDL) levels. However, high-level expression of APOBEC-1 in transgenic mouse and rabbit livers causes liver dysplasia and hepatocellular carcinoma. To determine the physiological and pathological effects of low-level hepatic expression of APOBEC-1, we used a 52-kb rat APOBEC-1 genomic clone (RE4) to generate transgenic mice expressing low levels of APOBEC-1 (2 to 5 times those in nontransgenic mice). Liver function, liver histology, editing of apoB mRNA at the normal editing site (C⁶⁶⁶⁶), and abnormal editing at multiple sites (hyperediting) in these mice were compared with those in transgenic mice expressing intermediate (I-20) or high (I-28) levels of APOBEC-1 in the liver. Hyperediting of mRNA coding for the novel APOBEC-1 target 1 (NAT1) was also examined. In the high-expressing I-28 line, 50% of the mice had palpable tumors at 15 weeks of age, whereas in the intermediate-expressing I-20 line, 50% of the mice had evidence of liver tumors after 1 year. In contrast, low-expressing RE4 mice had normal liver function and histology and did not develop liver tumors when examined at 3 to 17 months of age. Moreover, hyperediting of apoB and NAT1 mRNA in the liver was robust in the I-20 mice but barely detectable in the RE4 mice. The low-level expression resulted in sufficient APOBEC-1 to edit essentially all apoB mRNA at the normal editing site, virtually eliminating apoB-100 and LDL in the plasma of RE4 mice. When RE4 mice were crossed with human apoB transgenic mice, which possess high plasma LDL concentrations, plasma LDL levels in the offspring were reduced to very low levels. These results indicates that long-term hepatic expression of APOBEC-1 at low levels sufficient to eliminate LDL does not cause apparent liver damage or liver tumors in transgenic mice. RE4 APOBEC-1 transgenic mice should prove valuable for studying the roles of apoB-containing lipoproteins in lipid metabolism and atherosclerosis.

Key Words: APOBEC-1 • cholesterol • NAT1 • RNA editing • tumorigenesis

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