Intramuscular Administration of AAV1-Lipoprotein Lipase
            <sup>S447X</sup>
            Lowers Triglycerides in Lipoprotein Lipase–Deficient Patients

Stroes, Erik S.G.; Nierman, Melchior C.; Meulenberg, J.J.M.; Franssen, Remco; Twisk, Jaap; Henny, C. P.; Maas, Mario; Zwinderman, Aeilko H.; Ross, Colin J.D.; Aronica, Eleonora; High, Katherine A.; Levi, Marcel; Hayden, Michael R.; Kastelein, John J.P.; Kuivenhoven, Jan Albert

doi:10.1161/atvbaha.108.175620

Cited by 190 publications

(136 citation statements)

References 3 publications

(2 reference statements)

Supporting

Mentioning

132

Contrasting

Unclassified

Order By: Relevance

“…It is possible that the more abundant impurities in vectors prepared by the standard process coat the surface of vector particles, and thereby inhibit target cell binding. The use of AAV vector with increased transduction efficiency in liver and muscle is relevant because these tissues are important targets of gene transfer studies for inherited and metabolic diseases 9,12,[19][20][21] as well as the need for high levels of transgene expression for some applications to achieve therapeutic efficacy. 20,22 Increasing vector purity and potency is predicted to reduce the risk of deleterious, efficacy-limiting immune responses after administration of recombinant AAV to human subjects, a critical consideration for an inherently immunogenic (that is viral) biologic product that will depend on avoidance of certain immune responses to achieve long-term efficacy.…”

Section: Discussionmentioning

confidence: 99%

“…9,10 Additionally, evidence for dosedependent activation of capsid-specific T cells was also reported by our group in the context of AAV-1 intramuscular gene transfer for lipoprotein lipase deficiency. 11,12 In contrast, in clinical studies in which smaller amounts of AAV vectors were introduced into immunoprivileged body compartments, limited or no immune responses to the AAV capsid were observed. [13][14][15][16] These data support that the route of administration and the total antigen load (that is total vector capsid dose) determine the kinetics and the nature of the immune response against the AAV capsid.…”

Section: Introductionmentioning

confidence: 99%

See 1 more Smart Citation

High AAV vector purity results in serotype- and tissue-independent enhancement of transduction efficiency

et al. 2009

View full text Add to dashboard Cite

The purity of adeno-associated virus (AAV) vector preparations has important implications for both safety and efficacy of clinical gene transfer. Early-stage screening of candidates for AAV-based therapeutics ideally requires a purification method that is flexible and also provides vectors comparable in purity and potency to the prospective investigational product manufactured for clinical studies. The use of cesium chloride (CsCl) gradient-based protocols provides the flexibility for purification of different serotypes; however, a commonly used first-generation CsCl-based protocol was found to result in AAV vectors containing large amounts of protein and DNA impurities and low transduction efficiency in vitro and in vivo. Here, we describe and characterize an optimized, secondgeneration CsCl protocol that incorporates differential precipitation of AAV particles by polyethylene glycol, resulting in higher yield and markedly higher vector purity that correlated with better transduction efficiency observed with several AAV serotypes in multiple tissues and species. Vectors purified by the optimized CsCl protocol were found to be comparable in purity and functional activity to those prepared by more scalable, but less flexible serotype-specific purification processes developed for manufacture of clinical vectors, and are therefore ideally suited for pre-clinical studies supporting translational research.

show abstract

Section: Discussionmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

High AAV vector purity results in serotype- and tissue-independent enhancement of transduction efficiency

et al. 2009

View full text Add to dashboard Cite

show abstract

“…8,22 Among the rAAV serotypes analyzed to date, rAAV1 and rAAV8 are among the most efficient for muscle transduction. 8,[23][24][25][26] After IM administration, it was demonstrated that rAAV DNA resides as episomal circles [27][28][29] in a chromatin structure; 30 and from a biosafety perspective, the inefficient integration of rAAV into the host genome is an attractive feature for the legitimate use of this gene transfer system.…”

Section: Introductionmentioning

confidence: 99%

Longevity of rAAV vector and plasmid DNA in blood after intramuscular injection in nonhuman primates: implications for gene doping

Guiner

Gernoux

et al. 2011

Gene Ther

View full text Add to dashboard Cite

Legitimate uses of gene transfer technology can benefit from sensitive detection methods to determine vector biodistribution in pre-clinical studies and in human clinical trials, and similar methods can detect illegitimate gene transfer to provide sports-governing bodies with the ability to maintain fairness. Real-time PCR assays were developed to detect a performance-enhancing transgene (erythropoietin, EPO) and backbone sequences in the presence of endogenous cellular sequences. In addition to developing real-time PCR assays, the steps involved in DNA extraction, storage and transport were investigated. By real-time PCR, the vector transgene is distinguishable from the genomic DNA sequence because of the absence of introns, and the vector backbone can be identified by heterologous gene expression control elements. After performance of the assays was optimized, cynomolgus macaques received a single dose by intramuscular (IM) injection of plasmid DNA, a recombinant adeno-associated viral vector serotype 1 (rAAV1) or a rAAV8 vector expressing cynomolgus macaque EPO. Macaques received a high plasmid dose intended to achieve a significant, but not life-threatening, increase in hematocrit. rAAV vectors were used at low doses to achieve a small increase in hematocrit and to determine the limit of sensitivity for detecting rAAV sequences by single-step PCR. DNA extracted from white blood cells (WBCs) was tested to determine whether WBCs can be collaterally transfected by plasmid or transduced by rAAV vectors in this context, and can be used as a surrogate marker for gene doping. We demonstrate that IM injection of a conventional plasmid and rAAV vectors results in the presence of DNA that can be detected at high levels in blood before rapid elimination, and that rAAV genomes can persist for several months in WBCs.

show abstract

“…Recently a study by Stroes et al [42] ingested AAV1-LPL with short of two C-terminal amino acids, which is produced due to S447X variant in LPL gene in LPL deficient patients, and observed decreased level of TG.…”

Section: Discussionmentioning

confidence: 99%

Study of Common Genetic Variant S447X in Lipoprotein Lipase and Its Association with Lipids and Lipoproteins in Type 2 Diabetic Patients

Momin¹,

Bankar

Bhoite

2015

Ind J Clin Biochem

View full text Add to dashboard Cite

Elevated plasma triglyceride and non-esterified fatty acid concentrations may cause insulin resistance and type 2 diabetes mellitus. Lipoprotein lipase (LPL) is a ratedetermining enzyme in lipid metabolism. A variant in the LPL gene has been identified which alters the penultimate amino acid Serine at 447 to a stop codon (S447X), and results in a truncated LPL molecule lacking the C-terminal dipeptide Ser-Gly. The present study was designed to evaluate the frequency of S447X variant in the LPL gene and its effect on the lipid and lipoprotein levels in type 2 diabetic subjects. The genotype frequency distributions of type 2 diabetes patients and controls were in HardyWeinberg equilibrium. Comparison of the genotype and allelic frequencies of S447X in subjects with type 2 diabetics compared to controls demonstrated no significant difference. In subjects with type 2 diabetics having hypertriglyceridemia (TG C 150 mg/dl) compared to diabetics with TG level \150 mg/dl, significant difference in genotype frequency was found among these groups, while allelic frequency of X was significantly differed. Logistic regression analysis showed the negative association of LPL S447X variant with TG and VLDL cholesterol, while no association with total cholesterol, HDL cholesterol and LDL cholesterol was found. The lipid levels except for HDL cholesterol were found to be significantly lower in carriers for S447X than wild type in diabetes group. The decreased level of TG and TG rich lipoprotein in subjects with SNP S447X in LPL, predicts anti-atherogenic activity of carriers for S447X variant in general population as well as type 2 diabetic patients.

show abstract

Intramuscular Administration of AAV1-Lipoprotein Lipase ^S447X Lowers Triglycerides in Lipoprotein Lipase–Deficient Patients

Cited by 190 publications

References 3 publications

High AAV vector purity results in serotype- and tissue-independent enhancement of transduction efficiency

High AAV vector purity results in serotype- and tissue-independent enhancement of transduction efficiency

Longevity of rAAV vector and plasmid DNA in blood after intramuscular injection in nonhuman primates: implications for gene doping

Study of Common Genetic Variant S447X in Lipoprotein Lipase and Its Association with Lipids and Lipoproteins in Type 2 Diabetic Patients

Contact Info

Product

Resources

About

Intramuscular Administration of AAV1-Lipoprotein Lipase S447X Lowers Triglycerides in Lipoprotein Lipase–Deficient Patients

Cited by 190 publications

References 3 publications

High AAV vector purity results in serotype- and tissue-independent enhancement of transduction efficiency

High AAV vector purity results in serotype- and tissue-independent enhancement of transduction efficiency

Longevity of rAAV vector and plasmid DNA in blood after intramuscular injection in nonhuman primates: implications for gene doping

Study of Common Genetic Variant S447X in Lipoprotein Lipase and Its Association with Lipids and Lipoproteins in Type 2 Diabetic Patients

Contact Info

Product

Resources

About

Intramuscular Administration of AAV1-Lipoprotein Lipase ^S447X Lowers Triglycerides in Lipoprotein Lipase–Deficient Patients