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Full-Text Articles in Genetics and Genomics
Gene Transfer In The Lung Using Recombinant Adeno-Associated Virus, Alisha Gruntman, Christian Mueller, Terence Flotte, Guangping Gao
Gene Transfer In The Lung Using Recombinant Adeno-Associated Virus, Alisha Gruntman, Christian Mueller, Terence Flotte, Guangping Gao
Christian Mueller
Adeno-associated virus (AAV) is a small replication-deficient DNA virus belonging to the Parvovirinae family. It has a single-stranded ∼4.7-kb genome. Recombinant AAV (rAAV) is created by replacing the viral rep and cap genes with the transgene of interest along with promoter and polyadenylation sequences. The short viral inverted terminal repeats must remain intact for replication and packaging in production, as well as vector genome processing and persistence in the transduction process. The AAV capsid (serotype) determines the tissue tropism of the rAAV vector. In this unit we will discuss serotype selection for lung targeting along with the factors effecting efficient …
The Pros And Cons Of Immunomodulatory Il-10 Gene Therapy With Recombinant Aav In A Cftr-/- -Dependent Allergy Mouse Model, Christian Mueller, Sofia Braag, A. Martino, Qiushi Tang, M. Campbell-Thompson, Terence Flotte
The Pros And Cons Of Immunomodulatory Il-10 Gene Therapy With Recombinant Aav In A Cftr-/- -Dependent Allergy Mouse Model, Christian Mueller, Sofia Braag, A. Martino, Qiushi Tang, M. Campbell-Thompson, Terence Flotte
Christian Mueller
Cystic fibrosis (CF) patients have decreased levels of lung epithelial interleukin (IL)-10 and increased levels of proinflammatory cytokines (tumor necrosis factor-alpha, IL-4, IL-8 and IL-6). This has also been documented in Cftr (cystic fibrosis transmembrane conductance regulator)-deficient mice (Cftr 489X(-/-), FABP-hCFTR(+/+)). Our laboratory has recently characterized a peculiar hyper-IgE phenotype in these mice, in response to Aspergillus fumigatus crude protein extract (Af-cpe). Thus, we hypothesized that sustained systemic circulating IL-10 levels achieved through skeletal muscle transduction with recombinant adeno-associated vectors expressing IL-10 (rAAV1-IL-10) would serve to downregulate Th1 and Th2 cytokine production. This in turn would dampen the allergic response …
Partial Correction Of The Cftr-Dependent Abpa Mouse Model With Recombinant Adeno-Associated Virus Gene Transfer Of Truncated Cftr Gene, Christian Mueller, Daniel Torrez, Sofia Braag, Ashley Martino, Tracy Clarke, Martha Campbell-Thompson, Terence Flotte
Partial Correction Of The Cftr-Dependent Abpa Mouse Model With Recombinant Adeno-Associated Virus Gene Transfer Of Truncated Cftr Gene, Christian Mueller, Daniel Torrez, Sofia Braag, Ashley Martino, Tracy Clarke, Martha Campbell-Thompson, Terence Flotte
Christian Mueller
Recently, we have developed a model of airway inflammation in a CFTR knockout mouse utilizing Aspergillus fumigatus crude protein extract (Af-cpe) to mimic allergic bronchopulmonary aspergillosis (ABPA) 1, an unusual IgE-mediated hypersensitivity syndrome seen in up to 15% of cystic fibrosis (CF) patients and rarely elsewhere. We hypothesized that replacement of CFTR via targeted gene delivery to airway epithelium would correct aberrant epithelial cytokine signaling and ameliorate the ABPA phenotype in CFTR-deficient (CFTR 489X - /-, FABP-hCFTR + / +) mice. CFTR knockout mice underwent intra-tracheal (IT) delivery of recombinant adeno-associated virus serotype 5 (rAAV5Delta-264CFTR) or rAAV5-GFP at 2.58 x …
In Vivo Post-Transcriptional Gene Silencing Of Alpha-1 Antitrypsin By Adeno-Associated Virus Vectors Expressing Sirna, Pedro Cruz, Christian Mueller, Travis Cossette, Alexandra Golant, Qiushi Tang, Stuart Beattie, Mark Brantly, Martha Campbell-Thompson, Keith Blomenkamp, Jeffrey Teckman, Terence Flotte
In Vivo Post-Transcriptional Gene Silencing Of Alpha-1 Antitrypsin By Adeno-Associated Virus Vectors Expressing Sirna, Pedro Cruz, Christian Mueller, Travis Cossette, Alexandra Golant, Qiushi Tang, Stuart Beattie, Mark Brantly, Martha Campbell-Thompson, Keith Blomenkamp, Jeffrey Teckman, Terence Flotte
Christian Mueller
alpha-1 Antitrypsin (AAT) deficiency is one of the most common genetic diseases in North America, with a carrier frequency of approximately 4% in the US population. Homozygosity for the most common mutation (Glu342Lys, PI(*)Z) leads to the synthesis of a mutant protein, which accumulates and polymerizes within hepatocytes rather than being efficiently secreted. This lack of secretion causes severe serum deficiency predisposing to chronic lung disease. Twelve to fifteen percent of patients with PI(*)ZZ also develop liver disease, which can be severe, even in infancy. This is thought to be due to toxic effects of the accumulated mutant Z-AAT within …
In Vitro And In Vivo Functional Characterization Of Gutless Recombinant Sv40-Derived Cftr Vectors, Christian Mueller, M. Strayer, Jeffrey Sirninger, Sofia Braag, Francisco Branco, Jean-Pierre Louboutin, Terence Flotte, David Strayer
In Vitro And In Vivo Functional Characterization Of Gutless Recombinant Sv40-Derived Cftr Vectors, Christian Mueller, M. Strayer, Jeffrey Sirninger, Sofia Braag, Francisco Branco, Jean-Pierre Louboutin, Terence Flotte, David Strayer
Christian Mueller
In cystic fibrosis (CF), respiratory failure caused by progressive airway obstruction and tissue damage is primarily a result of the aberrant inflammatory responses to lung infections with Pseudomonas aeruginosa. Despite considerable improvement in patient survival, conventional therapies are mainly supportive. Recent progress toward gene therapy for CF has been encouraging; however, several factors such as immune response and transduced cell turnover remain as potential limitations to CF gene therapy. As alternative gene therapy vectors for CF, we examined the feasibility of using recombinant SV40-derived vectors (rSV40s), which may circumvent some of these obstacles. To accommodate the large cystic fibrosis transmembrane …
Dual Reporter Comparative Indexing Of Raav Pseudotyped Vectors In Chimpanzee Airway, Terence Flotte, Anne Fischer, Jason Goetzmann, Christian Mueller, Liudmila Cebotaru, Ziying Yan, Lilli Wang, James Wilson, William Guggino, John Engelhardt
Dual Reporter Comparative Indexing Of Raav Pseudotyped Vectors In Chimpanzee Airway, Terence Flotte, Anne Fischer, Jason Goetzmann, Christian Mueller, Liudmila Cebotaru, Ziying Yan, Lilli Wang, James Wilson, William Guggino, John Engelhardt
Christian Mueller
Selecting the most efficient recombinant adeno-associated virus (rAAV) serotype for airway gene therapy has been difficult due to cross-specific differences in tropism and immune response between humans and animal models. Chimpanzees--the closest surviving genetic relative of humans--provide a valuable opportunity to select the most effective serotypes for clinical trials in humans. However, designing informative experiments using this protected species is challenging due to limited availability and experimental regulations. We have developed a method using Renilla luciferase (RL) and firefly luciferase (FL) reporters to directly index the relative transduction and immune response of two promising rAAV serotypes following lung coinfection. Analysis …
Gene Therapy For Cystic Fibrosis, Christian Mueller, Terence Flotte
Gene Therapy For Cystic Fibrosis, Christian Mueller, Terence Flotte
Christian Mueller
Cystic Fibrosis (CF) is an autosomal recessive disorder due to mutations in the CF transmembrane conductance regulator (CFTR) gene that lead to defective ion transport in the conducting pulmonary airways and exocrine glands. Through a process that is not fully understood, CFTR defects predispose affected patients to chronic endobronchial infections with organisms such as Pseudomonas aeruginosa and Staphylococcus aureus. Following the discovery of the CFTR gene in 1989, CF became one of the primary targets for gene therapy research. Early enthusiasm surrounded the new field of gene therapy during most of the 1990s and it led academics and clinicians on …
Recombinant Aav Serotype And Capsid Mutant Comparison For Pulmonary Gene Transfer Of Alpha-1-Antitrypsin Using Invasive And Noninvasive Delivery, Rejean Liqun Wang, Thomas Mclaughlin, Travis Cossette, Qiushi Tang, Kevin Foust, Martha Campbell-Thompson, Ashley Martino, Pedro Cruz, Scott Loiler, Christian Mueller, Terence Flotte
Recombinant Aav Serotype And Capsid Mutant Comparison For Pulmonary Gene Transfer Of Alpha-1-Antitrypsin Using Invasive And Noninvasive Delivery, Rejean Liqun Wang, Thomas Mclaughlin, Travis Cossette, Qiushi Tang, Kevin Foust, Martha Campbell-Thompson, Ashley Martino, Pedro Cruz, Scott Loiler, Christian Mueller, Terence Flotte
Christian Mueller
Recombinant adeno-associated viral (rAAV) vectors have been widely used in pulmonary gene therapy research. In this study, we evaluated the transduction and expression efficiencies of several AAV serotypes and AAV2 capsid mutants with specific pulmonary targeting ligands in the mouse lung. The noninvasive intranasal delivery was compared with the traditional intratracheal lung delivery. The rAAV8 was the most efficient serotype at expressing alpha-1-antitrypsin (AAT) in the lung among all the tested serotypes and mutants. A dose of 1 x 10(10) vg of rAAV8-CB-AAT transduced a high percentage of cells in the lung when delivered intratrachealy. The serum and the broncho-alveolar …