Therapeutic Potential of Gene Transfer to Testis; Myth or Reality?

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.

Download Full-text

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.

Download Full-text

Therapeutic potential of bleomycin plus suicide or interferon-β gene transfer combination for spontaneous feline and canine melanoma

Oncoscience ◽

10.18632/oncoscience.387 ◽

2017 ◽

Vol 4 (11-12) ◽

pp. 199-214 ◽

Cited By ~ 2

Author(s):

Lucrecia Agnetti ◽

Chiara Fondello ◽

Marcela S. Villaverde ◽

Gerardo C. Glikin ◽

Liliana M.E. Finocchiaro

Keyword(s):

Gene Transfer ◽

Therapeutic Potential ◽

Interferon Β ◽

Canine Melanoma

Download Full-text

Abstract 113: SUMO1 Gene Transfer Rescues Cardiac Function in a Large Animal Model of Heart Failure

Circulation Research ◽

10.1161/res.111.suppl_1.a113 ◽

2012 ◽

Vol 111 (suppl_1) ◽

Author(s):

Lisa M Tilemann ◽

Kiyotake Ishikawa ◽

Changwon Kho ◽

Ahyoung Lee ◽

Jaime Aguero ◽

...

Keyword(s):

Heart Failure ◽

Gene Transfer ◽

Cardiac Function ◽

Therapeutic Potential ◽

Critical Role ◽

Balloon Occlusion ◽

Large Animal Model ◽

Swine Model ◽

Large Animal ◽

Cardiac Sarcoplasmic Reticulum

Recently, small ubiquitin-related modifier 1 (SUMO1) was found to enhance the activity and stability of the cardiac sarcoplasmic reticulum Ca2+ ATPase, SERCA2a. In both, human and rodent models of heart failure (HF), the total amount of myocardial SUMO1 is decreased and its knock down results in severe HF. Adeno-associated vector (AAV) mediated SUMO1 gene transfer significantly improves cardiac function in murine models of HF. As a critical step towards clinical translation, we evaluated the effects of SUMO1 gene transfer in a swine model of ischemic heart failure. One month after balloon occlusion of the proximal LAD, 21 animals were randomized to receive either AAV1.SUMO1 at two doses, AAV1.SERCA2a, AAV1.SUMO1+AAV1.SERCA2a, or saline via antegrade coronary infusion. In addition, three pigs served as controls and underwent sham procedures. The ejection fraction and the maximum dP/dt significantly increased after gene transfer of SUMO1 at both doses, SERCA2a and the combination of SUMO1 and SERCA2a (p=0.034, p=0.028) compared to saline infusion. The increase in maximum dP/dt was most pronounced in the group that received both SUMO1 and SERCA2a. Furthermore, the increase in end-systolic and end-diastolic volumes was normalized in the treatment groups, while they further deteriorated in the saline group (p=0.001, p=0.022). SUMO1 and SERCA2a gene transfer significantly improved cardiac function and concomitant gene delivery of SUMO1 and SERCA2a had a synergistic effect on improving these parameters in the HF animals. These results strongly support the critical role of SUMO1 for SERCA2a function and underline the therapeutic potential in heart failure patients.

Download Full-text

Reducing Lung ATP Levels and Alleviating Asthmatic Airway Inflammation through Adeno-Associated Viral Vector-Mediated CD39 Expression

Biomedicines ◽

10.3390/biomedicines9060656 ◽

2021 ◽

Vol 9 (6) ◽

pp. 656

Author(s):

Yung-An Huang ◽

Jeng-Chang Chen ◽

Chih-Ching Wu ◽

Chia-Wei Hsu ◽

Albert Min-Shan Ko ◽

...

Keyword(s):

Gene Transfer ◽

Airway Inflammation ◽

Viral Vector ◽

Therapeutic Potential ◽

Adeno Associated Virus ◽

Asthmatic Airway ◽

Potent Inflammatory Mediator ◽

Anti Inflammation ◽

Draining Lymph Nodes

Asthma is a chronic respiratory inflammatory disease. Patients usually suffer long-term symptoms and high medical expenses. Extracellular ATP (eATP) has been identified as a danger signal in innate immunity and serves as a potent inflammatory mediator for asthma. Hydrolyzing eATP in lungs might be a potential approach to alleviate asthmatic inflammation. Recombinant adeno-associated virus (rAAV) vectors that contain tissue-specific cap protein have been demonstrated to efficiently transfer exogenous genes into the lung tissues. To test anti-inflammation efficacy of rAAV-mediated CD39 gene transfer, rAAV-CD39 was generated and applied to OVA-mediated asthmatic mice. BALB/c mice were sensitized intraperitoneally and challenged intratracheally with OVA and treated with rAAV-CD39. At the end of procedure, some inflammatory features were examined. rAAV-CD39 treatment downregulated the levels of pulmonary eATP by the rescued expression of CD39. Several asthmatic features, such as airway hyperresponsiveness, eosinophilia, mucin deposition, and IL-5/IL-13 production in the lungs were decreased in the rAAV-CD39-treated mice. Reduced IL-5/IL-13 production and increased frequency of CD4+FoxP3+ regulatory T cells were detected in draining lymph nodes of rAAV-CD39 treated mice. This evidence suggested that rAAV-mediated CD39 gene transfer attenuated the asthmatic airway inflammation locally. The results suggest that rAAV-CD39 might have therapeutic potential for asthma.

Download Full-text

Proteasome activity restricts lentiviral gene transfer into hematopoietic stem cells and is down-regulated by cytokines that enhance transduction

Blood ◽

10.1182/blood-2005-10-4047 ◽

2006 ◽

Vol 107 (11) ◽

pp. 4257-4265 ◽

Cited By ~ 55

Author(s):

Francesca Romana Santoni de Sio ◽

Paolo Cascio ◽

Anna Zingale ◽

Mauro Gasparini ◽

Luigi Naldini

Keyword(s):

Gene Therapy ◽

Gene Transfer ◽

Cell Cycle Progression ◽

Therapeutic Potential ◽

Ex Vivo ◽

Proteasome Activity ◽

Hematopoietic Stem ◽

Vector Integration ◽

Cytokine Stimulation

AbstractThe therapeutic potential of hematopoietic stem cell (HSC) gene therapy can be fully exploited only by reaching efficient gene transfer into HSCs without compromising their biologic properties. Although HSCs can be transduced by HIV-derived lentiviral vectors (LVs) in short ex vivo culture, they display low permissivity to the vector, requiring cytokine stimulation to reach high-frequency transduction. Using stringent assays of competitive xenograft repopulation, we show that early-acting cytokines synergistically enhanced human HSC gene transfer by LVs without impairing engraftment and repopulation capacity. Using S-phase suicide assays, we show that transduction enhancement by cytokines was not dependent on cell cycle progression and that LVs can transduce quiescent HSCs. Pharmacologic inhibition of the proteasome during transduction dramatically enhanced HSC gene transfer, allowing the reach of very high levels of vector integration in their progeny in vivo. Thus, LVs are effectively restricted at a postentry step by the activity of this proteolytic complex. Unexpectedly, cytokine stimulation rapidly and substantially down-regulated proteasome activity in hematopoietic progenitors, highlighting one mechanism by which cytokines may enhance permissiveness to LV gene transfer. These findings demonstrate that antiviral responses ultimately mediated by proteasomes strongly limit the efficiency of HSC transduction by LVs and establish improved conditions for HSC-based gene therapy.

Download Full-text