Adeno-associated virus vector-mediated interleukin-10 gene transfer inhibits atherosclerosis in apolipoprotein E-deficient mice

Abstract

Inflammation is a major contributor to atherosclerosis by its effects on arterial wall biology and lipoprotein metabolism. Interleukin-10 (IL-10) is an anti-inflammatory cytokine that may modulate the atherosclerotic disease process. We investigated the effects of adeno-associated virus (AAV) vector-mediated gene transfer of IL-10 on atherogenesis in apolipoprotein E (ApoE)-deficient mice. A murine myoblast cell line, C2C12, transduced with AAV encoding murine IL-10 (AAV2-mIL10) secreted substantial amounts of IL-10 into conditioned medium. The production of monocyte chemoattractant protein-1 (MCP-1) by the murine macrophage cell line, J774, was significantly inhibited by conditioned medium from AAV2-mIL10-transduced C2C12 cells. ApoE-deficient mice were injected with AAV5-mIL10 into their anterior tibial muscle at 8 weeks of age. The expression of MCP-1 in the vascular wall of the ascending aorta and serum MCP-1 concentration were decreased in AAV5-mIL10-transduced mice compared with AAV5-LacZ-transduced mice. Oil red-O staining of the ascending aorta revealed that IL-10 gene transfer resulted in a 31% reduction in plaque surface area. Serum cholesterol concentrations were also significantly reduced in AAV5-mIL10-transduced mice. To understand the cholesterol-lowering mechanism of IL-10, we measured the cellular cholesterol level in HepG2 cells, resulting in its significant decrease by the addition of IL-10 in a dosedependent manner. Furthermore, IL-10 suppressed HMGCoA reductase expression in the HepG2 cells. These observations suggest that intramuscular injection of AAV5-mIL10 into ApoE-deficient mice inhibits atherogenesis through anti-inflammatory and cholesterol-lowering effects.

Adenoassociated Virus–Mediated Prostacyclin Synthase Expression Prevents Pulmonary Arterial Hypertension in Rats

“…[3][4][5] In contrast, the AAV vector used in this study achieved PGIS expression with a single intramuscular injection, and this expression was sustained for 1 year. 7 Furthermore, gene transfer was believed to be safer when performed via an intramuscular approach as opposed to the intratracheal or intrahepatic approaches. 6 Currently, researchers are using adenoviral gene transfer in most clinical trials because of its high efficiency for gene expression.…”

“…We reported previously that a stable serum concentration of a secretory protein was achieved over a 1-year period by using a single intramuscular injection of several AAV vector (AAV2 and AAV5) serotypes in mice. 7 Currently, AAV1 is one of the most efficient serotypes for muscle transduction. 8,9 Single subcutaneous injection of a pyrrolizidine alkaloid, namely, monocrotaline (MCT), produces severe PAH and PA remodeling in rats.…”

Systemic human Netrin-1 gene delivery by adeno-associated virus type 8 alters leukocyte accumulation and atherogenesis in vivo

“…For example, general immune suppressors interleukin-10 and tumor growth factor-b1 have been shown to inhibit atherosclerosis in mouse models. [8][9][10][11] The manipulation of the immune system may be achieved through cytokines or other genes that selectively inhibit T-helper cell type 1 response or other arms of the immune system, or to increase CD4(+), CD25(+) regulatory T cells (Tregs). 12,13 Atherogenesis has also been inhibited by altering lipid transport by the delivery of apolipoprotein E or apolipoprotein A1 Milano.…”

Systemic delivery of IL-10 by an AAV vector prevents vascular remodeling and end-organ damage in stroke-prone spontaneously hypertensive rat

“…7 The decreased levels of glucose and triglyceride that appeared in the control group may be associated with malnutrition after stroke episodes. Decreased serum albumin levels in this group might result from proteinuria caused by hypertensive renal dysfunction.…”

“…[4][5][6] We also reported that in ApoE-deficient mice, a single intramuscular adenoassociated virus (AAV) vector injection resulted in sustained IL-10 expression for over 6 months and improved atherosclerosis. 7 Several evidences suggest that IL-10 regulates endothelial NOS, endothelin-I and heme oxygenase-1 that substantially associate with blood pressure and vascular remodeling of resistance artery in a genetic model of hypertension. Although the overall contribution of inflammation to vascular damage in patients with hypertension remains to be clarified, several forms of experimental hypertension have been used to demonstrate monocyte/macrophage infiltration into the vessels of target organs, such as the brain and kidney.…”