Potentiation of Gene Transfer to the Mouse Lung by Complexes of Adenovirus Vector and Polycations Improves Therapeutic Potential

Pulmonary hypertension (PH) is characterized by increase in pulmonary vascular resistance, and narrowing and loss of pulmonary microvasculare. There is an indispensable need to develop innovative approaches for its control since PH becomes refractory to current therapies in later stages. Recent discovery of angiotensin converting enzyme 2 (ACE2), its involvement in cardiac remodeling, coupled with the limited success of ACE inhibitors in PH has led us to hypothesize that shifting the balance of renin-angiotensin system (RAS) to vasoprotective ACE2-Ang1–7- mas receptor axis would result the beneficial outcome in PH. We tested this hypothesis with the use of ACE2 overexpression in lungs by lentiviral vector-mediated gene transfer. Lentiviral vector particle(3x10^8 TU) containing murine ACE2 (letni-ACE2) were injected into 6 weeks old C57BL/6 mice prior to induction of PH by administration of weekly 600 mg/kg of monocrotaline (MCT) for 8 weeks for prevention studies. In addition, lenti-ACE2 was delivered following 6 weeks MCT treatment in reversal studies. Right ventricle systolic pressure (RVSP), Real-time RT-PCR, immunohisitochemistory of ACE2 and Ang (1–7) and histology of lungs in control and lent-ACE2 treated mice were carried out to evaluated the outcome on PH. Delivery of lenti-ACE2 resulted in a long-term increase in ACE2 expression in the lungs. A 60% and 100 % increases in protein and mRNA levels for ACE2 were observed. ACE2 and Ang (1–7) immunoreactivity were observed in epithelial and alveolar cells and alveolar macrophages. MCT treatment increased in RVSP (MCT 44.5+/−5.7 mmHg, control 24+/−1.0mmHg), RV hypertrophy (RV/LV+Sp ratio; 0.31+/−0.01), and wall thickness of pulmonary vessels. ACE2 gene transfer prevented increases in RVSP (26.1+/− 1.1mmHg), and RV hypertrophy (0.26+/−0.1), and reduced vessel wall thickness. In addition, ACE2 overexpression resulted in a significant reversal of RVSP (23.5+/−0.6mmHg). Futhermore, ACE2 overexpression in mice exhibited better general appearance and gained weight compared to MCT-treated mice. ACE2 gene transfer to lungs prevents and reverses vascular remodeling and PH in MCT model of PH. These observations suggest that targeting of pulmonary ACE2 holds novel therapeutic potential for PH.

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290. Improved Gene Transfer into Renal Carcinoma Cells Using an Adenovirus Vector

Molecular Therapy ◽

10.1016/s1525-0016(16)38648-8 ◽

2009 ◽

Vol 17 ◽

pp. S113-S114

Keyword(s):

Gene Transfer ◽

Renal Carcinoma ◽

Adenovirus Vector ◽

Carcinoma Cells ◽

Renal Carcinoma Cells

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Laser Photocoagulation and, to a Lesser Extent, Photodynamic Therapy Target and Enhance Adenovirus Vector–Mediated Gene Transfer in the Rat Retina

Investigative Opthalmology & Visual Science ◽

10.1167/iovs.05-0593 ◽

2005 ◽

Vol 46 (10) ◽

pp. 3883 ◽

Cited By ~ 3

Author(s):

Kaname Anzai ◽

Shin Yoneya ◽

Peter L. Gehlbach ◽

Daisuke Imai ◽

Lisa L. Wei ◽

...

Keyword(s):

Photodynamic Therapy ◽

Gene Transfer ◽

Laser Photocoagulation ◽

Adenovirus Vector ◽

Rat Retina ◽

Therapy Target

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Adrenomedullin: angiogenesis and gene therapy

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.00662.2004 ◽

2005 ◽

Vol 288 (6) ◽

pp. R1432-R1437 ◽

Cited By ~ 62

Author(s):

Noritoshi Nagaya ◽

Hidezo Mori ◽

Shinsuke Murakami ◽

Kenji Kangawa ◽

Soichiro Kitamura

Keyword(s):

Gene Transfer ◽

Cell Biology ◽

Therapeutic Potential ◽

Lattice Structure ◽

Rabbit Model ◽

Mitogen Activated Protein Kinase ◽

Tissue Ischemia ◽

Hypoxic Conditions

Adrenomedullin (AM) is a potent, long-lasting vasodilator peptide that was originally isolated from human pheochromocytoma. AM signaling is of particular significance in endothelial cell biology since the peptide protects cells from apoptosis, promotes angiogenesis, and affects vascular tone and permeability. The angiogenic effect of AM is mediated by activation of Akt, mitogen-activated protein kinase/extracellular signal-regulated kinase 1/2, and focal adhesion kinase in endothelial cells. Both AM and its receptor, calcitonin receptor-like receptor, are upregulated through a hypoxia-inducible factor-1-dependent pathway under hypoxic conditions. Thus AM signaling plays an important role in the regulation of angiogenesis in hypoxic conditions. Recently, we have developed a nonviral vector, gelatin. Positively charged gelatin holds negatively charged plasmid DNA in its lattice structure. DNA-gelatin complexes can delay gene degradation, leading to efficient gene transfer. Administration of AM DNA-gelatin complexes induces potent angiogenic effects in a rabbit model of hindlimb ischemia. Thus gelatin-mediated AM gene transfer may be a new therapeutic strategy for the treatment of tissue ischemia. Endothelial progenitor cells (EPCs) play an important role in endothelial regeneration. Interestingly, EPCs phagocytose ionically linked DNA-gelatin complexes in coculture, which allows nonviral gene transfer into EPCs. AM gene transfer into EPCs inhibits cell apoptosis and induces proliferation and migration, suggesting that AM gene transfer strengthens the therapeutic potential of EPCs. Intravenous administration of AM gene-modified EPCs regenerate pulmonary endothelium, resulting in improvement of pulmonary hypertension. These results suggest that in vivo and in vitro transfer of AM gene using gelatin may be applicable for intractable cardiovascular disease.

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Human menstrual blood-derived stem cells mitigate bleomycin-induced pulmonary fibrosis through anti-apoptosis and anti-inflammatory effects

Stem Cell Research & Therapy ◽

10.1186/s13287-020-01926-x ◽

2020 ◽

Vol 11 (1) ◽

Cited By ~ 1

Author(s):

Xin Chen ◽

Yi Wu ◽

Yanling Wang ◽

Lijun Chen ◽

Wendi Zheng ◽

...

Keyword(s):

Stem Cells ◽

Pulmonary Fibrosis ◽

Therapeutic Potential ◽

Lung Fibroblasts ◽

Menstrual Blood ◽

Inflammatory Factors ◽

Mouse Lung ◽

Control Group ◽

Antibody Array

Abstract Background Idiopathic pulmonary fibrosis is a kind of diffuse interstitial lung disease, the pathogenesis of which is unclear, and there is currently a lack of good treatment to improve the survival rate. Human menstrual blood-derived mesenchymal stem cells (MenSCs) have shown great potential in regenerative medicine. This study aimed to explore the therapeutic potential of MenSCs for bleomycin-induced pulmonary fibrosis. Methods We investigated the transplantation of MenSCs in a pulmonary fibrosis mouse model induced by BLM. Mouse was divided into three groups: control group, BLM group, MenSC group. Twenty-one days after MenSC transplantation, we examined collagen content, pathological, fibrosis area in the lung tissue, and the level of inflammatory factors of serum. RNA sequence was used to examine the differential expressed gene between three groups. Transwell coculture experiments were further used to examine the function of MenSCs to MLE-12 cells and mouse lung fibroblasts (MLFs) in vitro. Results We observed that transplantation of MenSCs significantly improves pulmonary fibrosis mouse through evaluations of pathological lesions, collagen deposition, and inflammation. Transwell coculturing experiments showed that MenSCs suppress the proliferation and the differentiation of MLFs and inhibit the apoptosis of MLE-12 cells. Furthermore, antibody array results demonstrated that MenSCs inhibit the apoptosis of MLE-12 cells by suppressing the expression of inflammatory-related cytokines, including RANTES, Eotaxin, GM-CSF, MIP-1γ, MCP-5, CCL1, and GITR. Conclusions Collectively, our results suggested MenSCs have a great potential in the treatment of pulmonary fibrosis, and cytokines revealed in antibody array are expected to become the target of future therapy of MenSCs in clinical treatment of pulmonary fibrosis.

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Gene Transfer for Experimental Saccular Aneurysms

Interventional Neuroradiology ◽

10.1177/15910199010070s119 ◽

2001 ◽

Vol 7 (1_suppl) ◽

pp. 131-135

Author(s):

T. Okamoto ◽

S. Miyachi ◽

M. Negoro ◽

O. Suzuki ◽

G. Otsuka ◽

...

Keyword(s):

Gene Therapy ◽

Experimental Study ◽

Gene Transfer ◽

Cerebral Aneurysms ◽

Adenovirus Vector ◽

Aneurysm Rupture ◽

Systemic Complications ◽

Aneurysmal Wall ◽

Carotid Wall ◽

Saccular Aneurysms

Reinforcing an aneurysmal wall is one possible way to prevent from aneurysm rupture. We preliminarily tried focal gene transfer against the wall of experimental aneurysms to aim the transgene remodeling of aneurysmal wall. Two experimental saccular aneurysms were created on canine common carotid artery with an artificial dissecting method, which resemble clinical aneurysms. Adenovirus vector (AxCALacZ, 108 pfu) was slowly injected into the aneurysm cavity for over 30 minutes under the condition of in-traaneurysmal flow arrest using balloon-assisted neck-plasty technique. The arteries and aneurysms were evaluated 48 hours after the transduction with X-gal staining, and β-galac-tosidase expression was detected mainly in the intima in both cases. No adverse effects on the normal carotid wall and no systemic complications were observed after the procedure. This experimental study suggests the possibility of gene therapy for cerebral aneurysms.

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