Lentiviral-Mediated Transgene Expression Can Potentiate Intestinal Mesenchymal-Epithelial Signaling

Biological Procedures Online

Adria D Dismuke²  Aimee D Kohn⁴  Randall T Moon³  Melissa H Wong¹ 

[1] Departments of Dermatology; Cell and Developmental Biology; Knight Cancer Institute, Oregon Health & Science University, Portland, OR, USA

[2] Department of Molecular and Medical Genetics, Oregon Health & Science University, Portland, OR, USA

[3] Howard Hughes Medical Institute, Division of Hematology, Department of Pharmacology, and the Center for Developmental Biology, Institute for Stem Cell and Regenerative Medicine, University of Washington School of Medicine, Seattle, WA, USA

[4] Cascade Cancer Center, Kirkland, WA, USA

关键词: mouse;    Wnt signaling;    transgene expression;    intestine;    mesenchymal-epithelial signaling;    lentivirus;   

Others  :  797161
DOI  :  10.1007/s12575-009-9014-z

 received in 2009-04-15, accepted in 2009-06-26,  发布年份 2009

【 摘 要 】

Mesenchymal-epithelial signaling is essential for the development of many organs and is often disrupted in disease. In this study, we demonstrate the use of lentiviral-mediated transgene delivery as an effective approach for ectopic transgene expression and an alternative to generation of transgenic animals. One benefit to this approach is that it can be used independently or in conjunction with established transgenic or knockout animals for studying modulation of mesenchymal-epithelial interactions. To display the power of this approach, we explored ectopic expression of a Wnt ligand in the mouse intestinal mesenchyme and demonstrate its functional influence on the adjacent epithelium. Our findings highlight the efficient use of lentiviral-mediated transgene expression for modulating mesenchymal-epithelial interactions in vivo.

【 授权许可】

   
2009 Dismuke et al.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

【 参考文献 】

• [1]Fevr T, Robine S, Louvard D, Huelsken J: Wnt/beta-catenin is essential for intestinal homeostasis and maintenance of intestinal stem cells. Mol Cell Biol 2007, 27(21):7551-7559.
• [2]Gregorieff A, Pinto D, Begthel H, Destree O, Kielman M, Clevers H: Expression pattern of Wnt signaling components in the adult intestine. Gastroenterology 2005, 129(2):626-638.
• [3]McMahon AP, Bradley A: The Wnt-1 (int-1) proto-oncogene is required for development of a large region of the mouse brain. Cell 1990, 62(6):1073-1085.
• [4]Takada S, Stark KL, Shea MJ, Vassileva G, McMahon JA, McMahon AP: Wnt-3a regulates somite and tailbud formation in the mouse embryo. Genes Dev 1994, 8(2):174-189.
• [5]Yamaguchi TP, Bradley A, McMahon AP, Jones S: A Wnt5a pathway underlies outgrowth of multiple structures in the vertebrate embryo. Development 1999, 126(6):1211-1223.
• [6]Dull T, Zufferey R, Kelly M, Mandel RJ, Nguyen M, Trono D, et al.: A third-generation lentivirus vector with a conditional packaging system. J Virol 1998, 72(11):8463-8471.
• [7]Naldini L: Lentiviruses as gene transfer agents for delivery to non-dividing cells. Curr Opin Biotechnol 1998, 9(5):457-463.
• [8]Zufferey R, Dull T, Mandel RJ, Bukovsky A, Quiroz D, Naldini L, et al.: Self-inactivating lentivirus vector for safe and efficient in vivo gene delivery. J Virol 1998, 72(12):9873-9880.
• [9]Fassler R: Lentiviral transgene vectors. EMBO Rep 2004, 5(1):28-29.
• [10]Skarsgard ED, Huang L, Reebye SC, Yeung AY, Jia WW: Lentiviral vector-mediated, in vivo gene transfer to the tracheobronchial tree in fetal rabbits. J Pediatr Surg 2005, 40(12):1817-1821.
• [11]Seppen J, van der Rijt R, Looije N, van Til NP, Lamers WH, Oude Elferink RP: Long-term correction of bilirubin UDPglucuronyltransferase deficiency in rats by in utero lentiviral gene transfer. Mol Ther 2003, 8(4):593-599.
• [12]Batlle E, Henderson JT, Beghtel H, van den Born MM, Sancho E, Huls G, et al.: Beta-catenin and TCF mediate cell positioning in the intestinal epithelium by controlling the expression of EphB/ephrinB. Cell 2002, 111(2):251-263.
• [13]Clevers H, Batlle E: EphB/EphrinB receptors and Wnt signaling in colorectal cancer. Cancer Res 2006, 66(1):2-5.
• [14]Rizvi AZ, Wong MH: Epithelial stem cells and their niche: there's no place like home. Stem cells (Dayt, Ohio) 2005, 23(2):150-165.
• [15]DasGupta R, Fuchs E: Multiple roles for activated LEF/TCF transcription complexes during hair follicle development and differentiation. Development 1999, 126(20):4557-4568.
• [16]Willenbring H, Grompe M: Delineating the hepatocyte's hematopoietic fusion partner. Cell Cycle 2004, 3(12):1489-1491.
• [17]Davies PS, Dismuke AD, Powell AE, Carroll KH, Wong MH: Wnt-reporter expression pattern in the mouse intestine during homeostasis. BMC Gastroenterol 2008, 8:57. BioMed Central Full Text
• [18]Weiser MM: Intestinal epithelial cell surface membrane glycoprotein synthesis. II. Glycosyltransferases and endogenous acceptors of the undifferentiated cell surface membrane. J Biol Chem 1973, 248(7):2542-2548.
• [19]Wong MH, Saam JR, Stappenbeck TS, Rexer CH, Gordon JI: Genetic mosaic analysis based on Cre recombinase and navigated laser capture microdissection. Proc Natl Acad Sci USA 2000, 97(23):12601-12606.
• [20]Sambrook J, Russell DW: Molecular cloning: a laboratory manual. 3rd edition. Cold Spring Harbor Laboratory Press, Cold Spring Harbor, NY; 2001.
• [21]Wong MH, Rubinfeld B, Gordon JI: Effects of forced expression of an NH2-terminal truncated beta-Catenin on mouse intestinal epithelial homeostasis. J Cell Biol 1998, 141(3):765-777.
• [22]McKee F, Stewart W: Passage of radioactive erythrocytes from the peritoneal cavity into the blood stream during experimental ascites. J Exp Med 1950, 91(6):599-605.
• [23]Olmedo DG, Tasat D, Guglielmotti MB, Cabrini RL: Titanium transport through the blood stream. An experimental study on rats. J Mater Sci 2003, 14(12):1099-1103.
• [24]Harada N, Tamai Y, Ishikawa T, Sauer B, Takaku K, Oshima M, et al.: Intestinal polyposis in mice with a dominant stable mutation of the beta-catenin gene. EMBO J 1999, 18(21):5931-5942.
• [25]Sansom OJ, Reed KR, Hayes AJ, Ireland H, Brinkmann H, Newton IP, et al.: Loss of Apc in vivo immediately perturbs Wnt signaling, differentiation, and migration. Genes Dev 2004, 18(12):1385-1390.
• [26]Huang S, Chen Y, Podsypanina K, Li Y: Comparison of expression profiles of metastatic versus primary mammary tumors in MMTV-Wnt-1 and MMTV-Neu transgenic mice. Neoplasia 2008, 10(2):118-124.
• [27]Herold MJ, van den Brandt J, Seibler J, Reichardt HM: Inducible and reversible gene silencing by stable integration of an shRNA-encoding lentivirus in transgenic rats. Proc Natl Acad Sci USA 2008, 105(47):18507-18512.