Poly(cationic lipid)-mediated in vivo gene delivery to mouse liver

Abstract

We have previously demonstrated that liposomes generated from poly(cationic lipid) (PCL) and cholesterol (Chol) have low cytotoxicity, are serum resistant, and display a transfection efficiency in vitro similar to commercially available cationic liposomes. Our in vivo experiments demonstrated that PCL-Chol liposomes bound much less avidly to serum proteins than did liposomes composed of 1,2-bis(dioleoyloxy)-3-(trimethylamonio)propane (DOTAP)-Chol or DO-TAP-L-a dioleoyl phosphatidylethanolamine (DOPE). Injection of the lipoplexes (PCL-Chol+DNA) through the portal vein after partial hepatectomy (PH) led to much higher reporter gene expression (luciferase) in the liver than did naked DNA injection. Marked green fluorescent protein expression was visualized in almost all hepatocytes in the liver of mice receiving lipoplex injection, even in the absence of PH. Subcutaneous injection of thyroid hormone triiodothyromine (T 3 ) significantly promoted hepatocyte regeneration and markedly enhanced PCL-Chol-mediated gene transfer in mouse liver when the lipoplex was administrated through either portal or tail vein. With T 3 pretreatment, PCL-Chol exerted a better gene transfer efficacy in mouse liver than DOTAP-Chol or DOTAP-DOPE. Two injections of lipoplexes through an indwelling catheter in the portal vein extended the transgene expression at a high level when T 3 injection was repeated. In conclusion, our findings demonstrate that the polymerized cationic liposomes are very stable in the blood and are effective agents for in vivo gene delivery, and that thyroid hormone administration offers a noninvasive approach to enhance liposome-mediated liver gene delivery.

Cyclic Arg-Gly-Asp Peptide-Labeled Liposomes for Targeting Drug Therapy of Hepatic Fibrosis in Rats

“…Thus, developing drug carriers that possess much more delivering capacity and that are clinically applicable is a critical step toward selective drug delivery for the treatment of liver fibrosis. Based on our experience in targeting drug and gene delivery in the treatment of liver injury (Wu et al, 1998a(Wu et al, , 2004Liu et al, 2003) and in the establishment of liposomal formulations (Pan et al, 2006), we established a new formulation of sterically stable liposomes, which are approximately 100 nm in diameter and which contain PEG as a spacer for the cyclic peptide linkage. The entrapping rate for IFN-␣1b was very high, and the liposome size is in the range of the fenestrae of SECs for easily crossing through the fenestrae to reach the Disse (30,000ϫ).…”

Hedgehog signaling pathway affects the sensitivity of hepatoma cells to drug therapy through the ABCC1 transporter

“…Firefly and renilla luciferase activity in transfected cells was determined by a dual-luciferase reporter system (Promega). 19 Assays of Cell Viability CD133 + /EpCAM + , CD133 − /EpCAM − , Huh-7-DN, and Huh-7-trans subpopulations were seeded at 10 4 /well in 96-well plates and allowed to adhere overnight. The cells were treated with Sorafenib (Selleck, TX, USA), Itraconazole (Biovision, Milpitas, CA, USA) and LDE225 (Cellagen Technology, San Diego, CA, USA) at various concentrations for 24 h. The cell viability was assayed with thiazolyl blue tetrazolium bromide (MTT) assay.…”

Aberrant hedgehog signaling is responsible for the highly invasive behavior of a subpopulation of hepatoma cells

“…17 Luciferase activity in transfected cells was determined 48 h after transfection as we previously reported. 36 RNA isolation and quantitative RT-PCR Total RNA was extracted from various cell types or subpopulations using RNeasy Mini Kit (QIAGEN, Mainz, Germany). Quantitative RT-PCR was performed using the ABI RT-PCR system.…”