Abstract/Details

Investigation of adenoviral gene transfer and associated complement-dependent inflammatory responses


2006 2006

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Abstract (summary)

Glycogen Storage Disease type II (GSDII) is caused by the lack of human acid-α-glucosidase (hGAA) activity to effectively reduce the lysosomal glycogen accumulation in striated muscle cells. Adenoviral (Ad) gene transfer vectors show potential for GSDII therapy. Previous studies demonstrated that a single intravenous injection of high-dose AdhGAA vector was able to correct glycogen accumulation, purely by hepatic transduction and secretion of hGAA to affected muscle groups. However, viral promoter shutdown and strong adaptive immune responses to hGAA curtailed the extent of efficacy. We sought to investigate whether the use of a fully deleted (FD) Ad vector expressing hGAA from liver specific promoter/enhancer elements could afford greater efficacy. Intravenous injection of this vector into GAA tolerant or nontolerant GAA-KO mice resulted in persistent, high-level hepatic GAA secretion, and complete glycogen correction in multiple muscle tissues for minimally 6 and 10 months respectively.

Despite the long-term efficacy and reduced adaptive immune response profile, toxicities resulting from Ad capsid interactions with the innate immune system limit Ad gene transfer in vivo. Ad vectors are able to activate complement pathways, which interact with Kupffer cells, platelets, and cytokines/chemokines. Since these same systems have been correlated with Ad associated toxicities, we hypothesized if these toxicities may be significantly diminished by preemptive complement system blockade.

High dose Ad injection into Ad wild-type (WT) and complement deficient (C3-KO) mice resulted in similar Ad transduction levels of liver hepatocytes, but altered the innate toxicity of Ads. The profound thrombocytopenia and elevated plasma cytokine/chemokine concentrations (KC, IL-5, IL-6, G-CSF, and GM-CSF) typically induced after Ad injections into WT mice were minimized after identical injections into C3-KO mice. Microarray analysis revealed that Ad injected WT mice livers as compared to Ad injected C3-KO mice livers, were marked by profound upregulation of genes in coagulation, metabolic, immune response, apoptosis related pathways. Our results confirm that complement inhibition strategies initiated prior to intravenous Ad vector administration should be investigated as a potential means to preemptively abrogate numerous Ad capsid induced toxicities and facilitate the known benefits of Ad mediated gene therapy and/or immunization applications.

Indexing (details)


Subject
Genetics;
Virology
Classification
0369: Genetics
0720: Virology
Identifier / keyword
Biological sciences; Acid-alpha-glucosidase; Adenovirus; Gene transfer; Glycogen storage disease; Inflammatory responses
Title
Investigation of adenoviral gene transfer and associated complement-dependent inflammatory responses
Author
Kiang, Anne
Number of pages
129
Publication year
2006
Degree date
2006
School code
0066
Source
DAI-B 68/08, Dissertation Abstracts International
Place of publication
Ann Arbor
Country of publication
United States
ISBN
9780549163084
Advisor
Amalfitano, Andrea
University/institution
Duke University
University location
United States -- North Carolina
Degree
Ph.D.
Source type
Dissertations & Theses
Language
English
Document type
Dissertation/Thesis
Dissertation/thesis number
3276001
ProQuest document ID
305329881
Copyright
Database copyright ProQuest LLC; ProQuest does not claim copyright in the individual underlying works.
Document URL
http://search.proquest.com/docview/305329881
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