Gene Transfer and Gene Therapy for Cardiovascular Disease

Although drug therapy is available for the treatment of cardiovascular disease, cardiovascular morbidity and mortality are poorly controlled throughout the world. Many of the pharmacological agents used to treat cardiovascular disease are expensive and therefore unavailable to poor segments of all societies. The available drugs are transient or short lasting in effect (usually < 24 h), have adverse side effects, and are not highly specific. Pharmacological therapy only partly mitigates...

Considerations for Gene Therapy

An effective and efficient vector system is essential for delivering therapeutic genes. Gene therapy uses a variety of gene transfer techniques. Some gene therapy approaches to cardiovascular disease are reviewed in Chapter 2. Advantages and disadvantages of some of the virus- and nonvirus-mediated gene delivery methods are discussed in this chapter. A vector is chosen according to its delivering efficiency, size, safety, and stability. Of note, only a few of the currently available viral...

Tetracycline Transactivator System in the Heart

Use of tetracycline-inducible systems in the heart has been exclusively with the Tet-Off system. Most studies reported have used the activator line developed by Fishman et al. (165-167) containing the myocardium-specific rat a-MHC promoter (Table 2). The production of the cellular toxin nitric oxide (NO) by immune cells is thought to play a role in controlling invading microorganisms, viruses, and abnormal cells. NO is produced by nitric oxide synthases (NOS). Although NOS has been reported to...

Acute Recording of RSND

Three days after the Ad.nNOS or Ad.p-gal transfection of the PVN as described previously in this chapter, anesthetize the rat with 0.75 g kg urethane ip and 70 mg kg a-chloraiose ip (see Note 22). 2. Catheterize the left femoral artery with polyethylene tubing (PE-50, filled with saline) connected to a Statham pressure transducer. Input the pressure signal from the transducer to a Grass bridge amplifier (S72), and then record the amplified signal with a PowerLab data acquisition system. 3....

Transductional and Transcriptional Targeting

Gene expression following systemic adenoviral vector delivery is affected by the natural tropism of the virus for the coxsackie adenoviral receptor (CAR). CAR is expressed to high levels in the liver, which consequently has a high capacity for nonspecific vector uptake. Transductional targeting alters the natural infection pathway and can be performed by pseudotyping, whereby modulation of the viral envelope allows manipulation of viral vector cell targeting specificity. This has been performed...

P65 AD GAL4 DBDpGene Luc

Sv40 Polya

Diagram of the amplification system. (A) The single plasmid system is the traditional way to express transgene in which a promoter drives a transgene (luciferase) directly in one plasmid. The promoter could be MLC-2v, the HRE SV40 promoter, or other promoters. (B) The double plasmid system can amplify the power of promoters based on the strong transcription activity of the GAL4 p65 fusion protein. The promoter in the transactivator plasmid (pGS) could be replaced with the SV40, HRE...

Lentiviral Vectors Used in Strategies for Conditional Expression

Development of retroviral vectors for reversible Tet regulation have been generated Table 3 with success and great promise. These vectors allow trans-duction of a large number of cell types, require relatively minimal effort to prepare, are stable, and can yield populations of regulatable cells within weeks as opposed to months. Kringstein et al. used both the rtTA Tet-On and tTA Tet-Off systems, carried within a SIN-RetroTet vector 209 that was based on the Moloney murine leukemia virus. Tet...

Transgenic Techniques in Cardiovascular Research

Rapid advances in molecular genetics have heralded a new era of genetic cardiology to study cardiovascular function and disease at the molecular level. Since the first cardiac-specific transgenic mouse was created in 1988 27 , hundreds of transgenic or gene-targeted murine models have been generated for the overexpression, genetic ablation, or site-specific mutation of key proteins governing cardiac structure and function. Transgenic techniques are powerful and valuable tools for cardiovascular...