Sustained correction of disease in naive and AAV2-pretreated hemophilia B dogs: AAV2/8-mediated, liver-directed gene therapy

Wang, Lili; Calcedo, Roberto; Nichols, Timothy C.; Bellinger, Dwight A.; Dillow, Aaron; Verma, Inder M.; Wilson, James M.

doi:10.1182/blood-2004-10-3867

Cited by 160 publications

(153 citation statements)

References 31 publications

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“…In vitro NAb assays, including those performed in this study, have demonstrated a lack of cross-reactivity between different types (22,23,26). The utilization of other types of AAV vectors to overcome AAV2 NAb activity has been performed in several animal models (27,55,64). Consistent with these studies, AAV1, AAV4, and AAV5 were found to transduce skeletal muscle regardless of preimmunization with AAV2 (Fig.…”

Section: Discussionsupporting

confidence: 71%

Single Amino Acid Modification of Adeno-Associated Virus Capsid Changes Transduction and Humoral Immune Profiles

DiPrimio

Bowles

et al. 2012

J Virol

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Adeno-associated virus (AAV) vectors have the potential to promote long-term gene expression. Unfortunately, humoral immunity restricts patient treatment and in addition provides an obstacle to the potential option of vector readministration. In this study, we describe a comprehensive characterization of the neutralizing antibody (NAb) response to AAV type 1 (AAV1) through AAV5 both in vitro and in vivo. These results demonstrated that NAbs generated from one AAV type are unable to neutralize the transduction of other types. We extended this observation by demonstrating that a rationally engineered, muscle-tropic AAV2 mutant containing 5 amino acid substitutions from AAV1 displayed a NAb profile different from those of parental AAV2 and AAV1. Here we found that a single insertion of Thr from AAV1 into AAV2 capsid at residue 265 preserved high muscle transduction, while also changing the immune profile. To better understand the role of Thr insertion at position 265, we replaced all 20 amino acids and evaluated both muscle transduction and the NAb response. Of these variants, 8 mutants induced higher muscle transduction than AAV2. Additionally, three classes of capsid NAb immune profile were defined based on the ability to inhibit transduction from AAV2 or mutants. While no relationship was found between transduction, amino acid properties, and NAb titer or its cross-reactivity, these studies map a critical capsid motif involved in all steps of AAV infectivity. Our results suggest that AAV types can be utilized not only as templates to generate mutants with enhanced transduction efficiency but also as substrates for repeat administration.A deno-associated virus (AAV) vectors have been safely and successfully used in phase I clinical trials (39,40,54). In particular, therapeutic effects have been achieved in patients with Leber's congenital amaurosis and hemophilia B. Among 30 patients with Leber's congenital amaurosis, 28 demonstrated remarkably improved eyesight after AAV type 2 (AAV2) delivery of the rpe65 gene to the retina. Regarding the recent hemophilia B trial, all six patients had therapeutic factor IX (FIX) expression within a beneficial 1% to 8% of normal levels as long as 18 months after intravenous delivery of an AAV vector encoding an optimized FIX cassette (54).AAV is a nonenveloped, single-stranded DNA virus that requires a helper virus, such as adenovirus (Ad) or herpes simplex virus, for efficient replication. Despite the lack of inflammation or other AAV-associated complications following administration of AAV2 vectors in several organs, neutralizing antibody (NAb) titers against the AAV2 capsid were found to be significantly increased following vector administration, particularly in the lung, muscle, and liver (7, 8, 43-45, 49, 54, 63). NAbs in circulation are able to block AAV transduction after systemic administration. Recently, Manno et al. (44) performed a phase I study of AAV-mediated FIX transgene delivery in patients with hemophilia B, in which one patient with a higher preexisting NAb ti...

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Section: Discussionsupporting

confidence: 71%

Single Amino Acid Modification of Adeno-Associated Virus Capsid Changes Transduction and Humoral Immune Profiles

DiPrimio

Bowles

et al. 2012

J Virol

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“…1,8,23 The most frequently used hepatocyte-specific promoters, with or without fusion of one or two heterologous enhancer elements, are the human AT promoter, 4,12,26,27 the albumin promoter 28,29 and the LSP promoter. 3,7,9 In this direct comparative study using hydrodynamic and adenoviral gene transfer, we show that the DC172 promoter, consisting of a 890 bp human AT promoter and two copies of the 160 bp a 1 -microglobulin enhancer, combined with two or four copies of the HCR-1 3 0 of the transgene, constitutes the most potent expression cassette. The a 1 -microglobulin/bikunin precursor promoter is weak and ubiquitously active, but expression in PC is potently increased by a hepatocytespecific enhancer located 2.7 kb downstream of the promoter.…”

Section: Discussionmentioning

confidence: 99%

“…Transcriptional targeting by use of hepatocyte-specific expression cassettes may lead to immunological ignorance or tolerance for the transgene product. [1][2][3] Expression cassettes for hepatocyte-directed transfer have been improved by using new promoterenhancer combinations, [3][4][5][6][7][8][9] inclusion of introns 5,[10][11][12][13] and inclusion of additional transcriptional sequences like scaffold matrix-attached regions (SMAR) and hepatic control regions (HCR). 12,[14][15][16][17][18] However, because different gene transfer vectors, different doses and different transgenes were used in these studies, direct comparative data for different expression cassettes are often lacking and hamper the interpretation of existing literature.…”

Section: Introductionmentioning

confidence: 99%

Direct comparison of hepatocyte-specific expression cassettes following adenoviral and nonviral hydrodynamic gene transfer

et al. 2008

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Hepatocytes are a key target for treatment of inborn errors of metabolism, dyslipidemia and coagulation disorders. The development of potent expression cassettes is a critical target to improve the therapeutic index of gene transfer vectors.Here we evaluated 22 hepatocyte-specific expression cassettes containing a human apo A-I transgene following hydrodynamic transfer of plasmids or adenoviral transfer with E1E3E4-deleted vectors in C57BL/6 mice. The DC172 promoter consisting of a 890 bp human a 1 -antitrypsin promoter and two copies of the 160 bp a 1 -microglobulin enhancer results in superior expression levels compared to constructs containing the 1.5 kb human a 1 -antitrypsin promoter, the 790 bp synthetic liver-specific promoter or the DC190 promoter containing a 520 bp human albumin promoter and two copies of the 99 bp prothrombin enhancer. The most potent expression cassette consists of the DC172 promoter upstream of the transgene and two copies of the hepatic control region-1. Minicircles containing this expression cassette induce persistent physiological human apo A-I or human factor IX levels after hydrodynamic transfer. In conclusion, in this comparative study of 22 hepatocyte-specific expression cassettes, the DC172 promoter in combination with two copies of the hepatic control region-1 induces the highest expression levels following hydrodynamic and adenoviral transfer.

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“…Among several recently isolated serotypes, AAV8 possesses high transduction rate in liver and low preexisting immunity in human population, therefore, is an attractive vector for liver gene delivery. [29][30][31][32][33] Another major advancement in AAV vector development was the discovery that AAV with self-complementary doublestranded genome possessed 10-to 100-fold higher transduction rate in liver than conventional singlestranded AAV. 34,35 We, therefore, decided to construct double-stranded AAV2/8 pseudotyped vector (dsAAV2/8) to deliver shRNAs and to investigate its inhibition effects in HBV transgenic mice.…”

Section: Introductionmentioning

confidence: 99%

Long-term inhibition of hepatitis B virus in transgenic mice by double-stranded adeno-associated virus 8-delivered short hairpin RNA

Chen

et al. 2006

Gene Ther

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RNA interference (RNAi) was reported to block hepatitis B virus (HBV) gene expression and replication in vitro and in vivo. However, it remains a technical challenge for RNAibased therapy to achieve long-term and complete inhibition effects in chronic HBV infection, which presumably requires more extensive and uniform transduction of the whole infected hepatocytes. To increase the in vivo transfection efficiency in liver, we used a double-stranded adenoassociated virus 8-pseudotyped vector (dsAAV2/8) to deliver shRNA. HBV transgenic mice were used as an animal model to evaluate the inhibition effects of the RNAi-based gene therapy. A single administration of dsAAV2/8 vector, carrying HBV-specific shRNA, effectively suppressed the steady level of HBV protein, mRNA and replicative DNA in liver of HBV transgenic mice, leading to up to 2-3 log 10 decrease in HBV load in the circulation. Significant HBV suppression sustained for at least 120 days after vector administration. The therapeutic effect of shRNA was target sequence dependent and did not involve activation of interferon. These results underscore the potential for developing RNAi-based therapy by dsAAV2/8 vector to treat HBV chronic infection, and possibly other persistent liver infections as well.

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Sustained correction of disease in naive and AAV2-pretreated hemophilia B dogs: AAV2/8-mediated, liver-directed gene therapy

Cited by 160 publications

References 31 publications

Single Amino Acid Modification of Adeno-Associated Virus Capsid Changes Transduction and Humoral Immune Profiles

Single Amino Acid Modification of Adeno-Associated Virus Capsid Changes Transduction and Humoral Immune Profiles

Direct comparison of hepatocyte-specific expression cassettes following adenoviral and nonviral hydrodynamic gene transfer

Long-term inhibition of hepatitis B virus in transgenic mice by double-stranded adeno-associated virus 8-delivered short hairpin RNA

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