Efficient Expression of Retroviral Vector-transduced Human Low Density Lipoprotein (LDL) Receptor in LDL Receptor-deficient Rabbit Fibroblasts in Vitro

Overview

Journal Proc Natl Acad Sci U S A

Specialty Science

Date 1988 Sep 1

PMID 2842777

Citations 7

Authors

A Miyanohara

M F Sharkey

J L Witztum

D Steinberg

T Friedmann

Affiliations

Soon will be listed here.

Abstract

Familial hypercholesterolemia is caused by a genetic deficiency of the low density lipoprotein (LDL) receptor. The Watanabe heritable hyperlipidemic (WHHL) rabbit, which is also defective in LDL receptor activity, provides an excellent animal model of homozygous familial hypercholesterolemia. As a step toward development of effective gene therapy for familial hypercholesterolemia, we have constructed a transmissible retroviral vector containing a full-length human cDNA for the LDL receptor. WHHL fibroblasts infected in vitro expressed the human receptor efficiently, as indicated by RNA and ligand blotting studies. Infected fibroblasts bound and degraded a monoclonal antibody specific for the human LDL receptor (IgGC7) in a manner comparable to that seen with normal human fibroblasts. Human LDL was also degraded by infected WHHL cells and promoted cholesterol esterification to the same degree as seen in normal human fibroblasts. Although technical problems remain to be solved, these studies show that, in principle, gene therapy may be possible for familial hypercholesterolemia patients.

Citing Articles

Scavenger receptors and their potential as therapeutic targets in the treatment of cardiovascular disease.

Stephen S, Freestone K, Dunn S, Twigg M, Homer-Vanniasinkam S, Walker J Int J Hypertens. 2010; 2010:646929.

PMID: 20981357 PMC: 2958427. DOI: 10.4061/2010/646929.

Long-term physiologically regulated expression of the low-density lipoprotein receptor in vivo using genomic DNA mini-gene constructs.

Hibbitt O, McNeil E, Lufino M, Seymour L, Channon K, Wade-Martins R Mol Ther. 2009; 18(2):317-26.

PMID: 19861949 PMC: 2839284. DOI: 10.1038/mt.2009.249.

Retroviral infection and expression of cationic amino acid transporters in rodent hepatocytes.

Closs E, Borel Rinkes I, Bader A, Yarmush M, Cunningham J J Virol. 1993; 67(4):2097-102.

PMID: 8383231 PMC: 240299. DOI: 10.1128/JVI.67.4.2097-2102.1993.

Transfer of 15-lipoxygenase gene into rabbit iliac arteries results in the appearance of oxidation-specific lipid-protein adducts characteristic of oxidized low density lipoprotein.

Yla-Herttuala S, Luoma J, Viita H, Hiltunen T, Sisto T, Nikkari T J Clin Invest. 1995; 95(6):2692-8.

PMID: 7769108 PMC: 295952. DOI: 10.1172/JCI117971.

Retroviral transfer of HSV1-TK gene into human lung cancer cell line.

Hasegawa Y, Emi N, Shimokata K J Mol Med (Berl). 1995; 73(3):107-12.

PMID: 7633946 DOI: 10.1007/BF00198237.

References

Southern P, Berg P . Transformation of mammalian cells to antibiotic resistance with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet. 1982; 1(4):327-41. View

Pittman R, Carew T, Attie A, Witztum J, Watanabe Y, Steinberg D . Receptor-dependent and receptor-independent degradation of low density lipoprotein in normal rabbits and in receptor-deficient mutant rabbits. J Biol Chem. 1982; 257(14):7994-8000. View

Daniel T, Schneider W, Goldstein J, Brown M . Visualization of lipoprotein receptors by ligand blotting. J Biol Chem. 1983; 258(7):4606-11. View

Dietschy J, Kita T, Suckling K, Goldstein J, Brown M . Cholesterol synthesis in vivo and in vitro in the WHHL rabbit, an animal with defective low density lipoprotein receptors. J Lipid Res. 1983; 24(4):469-80. View

Curtiss L, Witztum J . A novel method for generating region-specific monoclonal antibodies to modified proteins. Application to the identification of human glucosylated low density lipoproteins. J Clin Invest. 1983; 72(4):1427-38. PMC: 370427. DOI: 10.1172/JCI111099. View

Yamamoto T, Davis C, Brown M, Schneider W, CASEY M, Goldstein J . The human LDL receptor: a cysteine-rich protein with multiple Alu sequences in its mRNA. Cell. 1984; 39(1):27-38. DOI: 10.1016/0092-8674(84)90188-0. View

Miller A, Law M, Verma I . Generation of helper-free amphotropic retroviruses that transduce a dominant-acting, methotrexate-resistant dihydrofolate reductase gene. Mol Cell Biol. 1985; 5(3):431-7. PMC: 366734. DOI: 10.1128/mcb.5.3.431-437.1985. View

Semenkovich C, Ostlund Jr R, Yang J, Reaban M . Demonstration of functional low-density lipoprotein receptors by protein blotting in fibroblasts from a subject with homozygous receptor-negative familial hypercholesterolemia. J Lab Clin Med. 1985; 106(1):47-52. View

Brown M, Goldstein J . A receptor-mediated pathway for cholesterol homeostasis. Science. 1986; 232(4746):34-47. DOI: 10.1126/science.3513311. View

10.

Yamamoto T, BISHOP R, Brown M, Goldstein J, Russell D . Deletion in cysteine-rich region of LDL receptor impedes transport to cell surface in WHHL rabbit. Science. 1986; 232(4755):1230-7. PMC: 4451858. DOI: 10.1126/science.3010466. View

11.

Demetriou A, WHITING J, Feldman D, LEVENSON S, CHOWDHURY N, Moscioni A . Replacement of liver function in rats by transplantation of microcarrier-attached hepatocytes. Science. 1986; 233(4769):1190-2. DOI: 10.1126/science.2426782. View

12.

Miller A, Buttimore C . Redesign of retrovirus packaging cell lines to avoid recombination leading to helper virus production. Mol Cell Biol. 1986; 6(8):2895-902. PMC: 367857. DOI: 10.1128/mcb.6.8.2895-2902.1986. View

13.

Goldstein J, Brown M, Anderson R, Russell D, Schneider W . Receptor-mediated endocytosis: concepts emerging from the LDL receptor system. Annu Rev Cell Biol. 1985; 1:1-39. DOI: 10.1146/annurev.cb.01.110185.000245. View

14.

Wolff J, Yee J, Skelly H, Moores J, Respess J, Friedmann T . Expression of retrovirally transduced genes in primary cultures of adult rat hepatocytes. Proc Natl Acad Sci U S A. 1987; 84(10):3344-8. PMC: 304866. DOI: 10.1073/pnas.84.10.3344. View

15.

Anson D, Hock R, Austen D, Smith K, Brownlee G, Verma I . Towards gene therapy for hemophilia B. Mol Biol Med. 1987; 4(1):11-20. View

16.

Morgan J, Barrandon Y, Green H, Mulligan R . Expression of an exogenous growth hormone gene by transplantable human epidermal cells. Science. 1987; 237(4821):1476-9. DOI: 10.1126/science.3629250. View

17.

Dzierzak E, Papayannopoulou T, Mulligan R . Lineage-specific expression of a human beta-globin gene in murine bone marrow transplant recipients reconstituted with retrovirus-transduced stem cells. Nature. 1988; 331(6151):35-41. DOI: 10.1038/331035a0. View

18.

Hofmann S, Russell D, Brown M, Goldstein J, Hammer R . Overexpression of low density lipoprotein (LDL) receptor eliminates LDL from plasma in transgenic mice. Science. 1988; 239(4845):1277-81. DOI: 10.1126/science.3344433. View

19.

Goldstein J, Brown M . Binding and degradation of low density lipoproteins by cultured human fibroblasts. Comparison of cells from a normal subject and from a patient with homozygous familial hypercholesterolemia. J Biol Chem. 1974; 249(16):5153-62. View

20.

Goldstein J, Dana S, Brown M . Esterification of low density lipoprotein cholesterol in human fibroblasts and its absence in homozygous familial hypercholesterolemia. Proc Natl Acad Sci U S A. 1974; 71(11):4288-92. PMC: 433867. DOI: 10.1073/pnas.71.11.4288. View