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

• Published in: Gene therapy Vol. 18; no. 5; pp. 437 - 444
• Main Authors: KHAN, J. A, CAO, M, KANG, B. Y, LIU, Y, MEHTA, J. L, HERMONAT, P. L
• Format: Journal Article
• Language: English
• Published: London Nature Publishing Group 01.05.2011
• Subjects: Adeno-associated virus; Anesthesia. Intensive care medicine. Transfusions. Cell therapy and gene therapy; Animals; Aorta; Aorta - pathology; Applied cell therapy and gene therapy; Arteries; Arteriosclerosis; Atherogenesis; Atherosclerosis; Atherosclerotic plaque; Biological and medical sciences; Biotechnology; Blood Flow Velocity; CD25 antigen; CD8-Positive T-Lymphocytes - immunology; Chemotactic factors; Cholesterol; Cholesterol - blood; Dependovirus - genetics; Dependoviruses; Foxp3 protein; Fundamental and applied biological sciences. Psychology; Gene therapy; Gene transfer; Gene Transfer Techniques; Genes; Genetic aspects; Genetic Therapy - methods; Health aspects; Health. Pharmaceutical industry; High cholesterol diet; Immunoregulation; Industrial applications and implications. Economical aspects; Inflammation - prevention & control; Inflammatory diseases; Leukocytes; Leukocytes - immunology; Low density lipoprotein receptors; Lymphocytes T; Macrophages; Medical sciences; Mice; Mice, Knockout; Monocytes; Mucin; Neomycin; Nerve Growth Factors - genetics; Netrin-1; Physiological aspects; Plaque, Atherosclerotic - immunology; Plaque, Atherosclerotic - pathology; Plaque, Atherosclerotic - prevention & control; Receptor density; Receptors, LDL - genetics; Risk factors; Statistical analysis; Transfusions. Complications. Transfusion reactions. Cell and gene therapy; Tumor Suppressor Proteins - genetics; United States; Human; Leukocyte; Cardiovascular disease; Systemic; adeno-associated virus; Dependovirus; Vascular disease; Virus; In vivo; Parvoviridae; Atherosclerosis; T-Lymphocyte; chemo-repellent; Gene therapy; suppressor T cell; Parvovirinae; Macrophage
• ISSN: 0969-7128; 1476-5462
• DOI: 10.1038/gt.2010.155

Atherosclerosis is an inflammatory disorder of arteries. Atherosclerotic plaque, in its early to intermediate stages, is composed largely of lipid-engorged foam cells. These foam cells are derived from the trafficking of monocytes (Mo) into the arterial intima, attracted to the site by chemoattractants. Given that foam cells are derived from the trafficking of Mo, the use of Netrin-1, an Mo chemorepellent, may be useful in limiting Mo accumulation and subsequent plaque formation. To investigate the potential of Netrin-1 for limiting atherosclerosis, we systemically delivered its human (h) cDNA by adeno-associated virus type 8 (AAV8, single-stranded structure) delivery into low-density lipoprotein receptor knockout (LDLR-/-) mice and placed the animals on a high cholesterol diet (HCD). Compared with control neomycin resistance (Neo) gene delivery/HCD, hNetrin-1 delivery resulted in a significant reduction in plaque formation, as determined by larger aortic lumen size, thinner intima-media thickness and lower blood velocity than the Neo/HCD control (all statistically significant). Indices of monocyte/macrophage (Mo/MΦ) accumulation, CD68, integrin, alpha M (ITGAM) and egf-like module containing, mucin-like, hormone receptor-like 1 (EMR-1), were reduced in hNetrin-1/HCD-treated animal's aortas and spleens compared with Neo/HCD-treated animals. Unexpectedly, CD25 and foxp3 (regulatory T cells (Tregs)) in the aorta were strongly upregulated. This is the first time the Mo/MΦ chemorepellent approach, and specific Netrin-1 gene delivery, has been performed for the reduction of Mo/MΦ burden and atherosclerosis. In addition, Netrin-1 has never before been linked to altered Treg levels. These data strongly suggest that hNetrin-1 gene delivery can reduce Mo/MΦ accumulation, inflammation and subsequent plaque formation.