Safety and efficacy of optimized AAV3 vectors in murine and NHP livers

Summary

Principal Investigator: Arun Srivastava
Abstract: DESCRIPTION (provided by applicant): The main aims of this proposal are to develop recombinant adeno-associated virus 3 (AAV3) serotype vectors for high-efficiency transduction of hepatocytes, and to evaluate the safety and efficacy of the optimized AAV3 serotype vectors in non-human primates, with the long-term goal of their potential use in liver-directed gene therapy in humans. We have observed that of the 10 most commonly used AAV serotypes, AAV3 vectors transduce human liver cancer cell lines extremely efficiently. AAV3 vectors also transduce primary human hepatocytes efficiently, as they utilize human hepatocyte growth factor receptor (HGFR) as a cellular co-receptor. Based on our recent studies on the development of the next generation of AAV2 vectors containing mutations in the surface-exposed tyrosine residues, which transduce murine hepatocytes exceedingly well at low doses, we have also generated tyrosine-mutant AAV3 serotype vectors, and identified an optimized vector that efficiently transduces human liver tumors in a murine xenograft model in vivo. We now wish to parlay some of this information to explore whether optimized AAV3 vectors are the ideal serotype for high-efficiency transduction of human hepatocytes. We propose to test the following hypotheses: a. High-efficiency transduction of primary human hepatocytes can be achieved by optimized AAV3 serotype vectors in a mouse xenograft model in vivo. b. The safety and efficacy of the systemically delivered optimized AAV3 serotype vectors, prior to their potential use in human gene therapy, can be evaluated in a non-human primate model for liver-directed gene transfer in vivo. The following two Specific Aims will be pursued: Specific Aim 1: Development of optimized surface-exposed tyrosine-, serine and threonine-mutant AAV serotype 3 vectors for high-efficiency transduction of human hepatocytes. Specific Aim 2: Biodistribution and safety of optimized AAV3 serotype vectors in non-human primates following systemic delivery. A better understanding of the AAV3-hepatocyte interactions at the molecular level is likely to lead to development of safe and effective vectors for their potential use in gen therapy of human liver diseases in general, and hemophilia B in particular.
Funding Period: 2012-07-01 - 2014-06-30
more information: NIH RePORT

Top Publications

  1. ncbi Lignin nanotubes as vehicles for gene delivery into human cells
    Elena Ten
    Department of Microbiology and Cell Science, Genetics Institute, Department of Pediatrics, and Powell Gene Therapy Center, University of Florida, Gainesville, Florida 32610, United States
    Biomacromolecules 15:327-38. 2014

Research Grants

  1. Hypoxic Adenosine Responses
    Michael R Blackburn; Fiscal Year: 2013
  2. GENE AND PHARMACOLOGICAL THERAPIES FOR CYSTIC FIBROSIS
    William B Guggino; Fiscal Year: 2013
  3. Gene Transfer for Galactosialidosis - Resub
    Arthur W Nienhuis; Fiscal Year: 2013
  4. AAV-Mediated Gene Therapy for Hemophilia
    Andrew M Davidoff; Fiscal Year: 2013
  5. Preclinical &Phase I/II Trials of AAV-AAT Vectors
    Terence R Flotte; Fiscal Year: 2013
  6. Development of AAV vectors for the phenotypic correction of sickle cell disease
    Angela Rivers; Fiscal Year: 2013
  7. Colorado Nutrition Obesity Research Center
    James O Hill; Fiscal Year: 2013
  8. The Role of Bioactive Lipids in Inflammation and Cancer
    RAYMOND NELSON DUBOIS; Fiscal Year: 2013
  9. Lung Endothelial Cell Phenotypes
    Troy Stevens; Fiscal Year: 2013
  10. Multi functional studies of candidate dyslexia susceptibility genes in the rat
    ALBERT MARK GALABURDA; Fiscal Year: 2013

Detail Information

Publications2

  1. ncbi Lignin nanotubes as vehicles for gene delivery into human cells
    Elena Ten
    Department of Microbiology and Cell Science, Genetics Institute, Department of Pediatrics, and Powell Gene Therapy Center, University of Florida, Gainesville, Florida 32610, United States
    Biomacromolecules 15:327-38. 2014
    ..These combined features of LNTs make LNTs attractive as smart delivery vehicles of DNA without the cytotoxicity associated with CNTs or the immunogenicity of viral vectors. ..

Research Grants30

  1. Hypoxic Adenosine Responses
    Michael R Blackburn; Fiscal Year: 2013
    ..Three Component Projects, Two Scientific Cores and an Administrative Core are planned to facilitate the research goals and interactions of this PPG. ..
  2. GENE AND PHARMACOLOGICAL THERAPIES FOR CYSTIC FIBROSIS
    William B Guggino; Fiscal Year: 2013
    ..Project IV will focus on the Biology of AAV. Finally, there are three cores, an Expression, a Vector Core, and an Administration Core. ..
  3. Gene Transfer for Galactosialidosis - Resub
    Arthur W Nienhuis; Fiscal Year: 2013
    ..The tril will be monitored for evidence of gene transfer, for biochemical evidence of PPCA production and for correction of the clinical manifestations of the disorder. ..
  4. AAV-Mediated Gene Therapy for Hemophilia
    Andrew M Davidoff; Fiscal Year: 2013
    ..In addition, results from these studies will be of great utility not only for hemophilia A gene therapy but also other disorders that are potentially amenable to AAV-mediated, liver-targeted gene transfer. ..
  5. Preclinical &Phase I/II Trials of AAV-AAT Vectors
    Terence R Flotte; Fiscal Year: 2013
    ..In addition, the issues to be investigated in aims 3 and 4, i.e., the immune response questions, will also be essential in developing the appropriate strategy for later clinical trials. ..
  6. Development of AAV vectors for the phenotypic correction of sickle cell disease
    Angela Rivers; Fiscal Year: 2013
    ..The environment at the UF will maximize the applicant's potential to establish a career as an independent translational investigator. ..
  7. Colorado Nutrition Obesity Research Center
    James O Hill; Fiscal Year: 2013
    ..We look forward to continuing to enhance nutrition research within Colorado over the next 5 years. ..
  8. The Role of Bioactive Lipids in Inflammation and Cancer
    RAYMOND NELSON DUBOIS; Fiscal Year: 2013
    ..Information gained from these studies will be important for achieving the long-term goal of reducing the cancer burden and revealing more effective ways to prevent cancer. ..
  9. Lung Endothelial Cell Phenotypes
    Troy Stevens; Fiscal Year: 2013
    ..abstract_text> ..
  10. Multi functional studies of candidate dyslexia susceptibility genes in the rat
    ALBERT MARK GALABURDA; Fiscal Year: 2013
    ..abstract_text> ..
  11. Hepatic Gene Transfer for Treatment of Hemophilias A &B
    Mark A Kay; Fiscal Year: 2013
    ..Our studies will advance gene therapeutic approaches towards achieving a cure for the hemophilias. ..
  12. OXIDATIVE STRESS IN THE KIDNEY IN HYPERTENSION
    Christopher S Wilcox; Fiscal Year: 2013
    ..These are supported by the Administrative, Animal and Bioanalytical Cores. ..
  13. Neutralizing Antibody &AAV FIX Gene Therapy
    Richard J Samulski; Fiscal Year: 2013
    ..The long-term objective of this PPG is to advance basic understanding of vector-cell-animal model interactions for safe gene delivery. ..
  14. Cardiac Myosin Binding Protein-C: Structure, Function, and Regulation
    David M Warshaw; Fiscal Year: 2013
    ..abstract_text> ..
  15. Immunology of Factor IX Gene Transfer to Liver
    Roland W Herzog; Fiscal Year: 2013
    ..IX in hemophilia B mice;and v) use of a codon-optimized F.VIII cDNA and transient B cell depletion facilitate tolerance induction to F.VIII by liver gene transfer in hemophilia A mice. ..
  16. Penn Center for Study of Epigenetics in Reproduction
    Marisa S Bartolomei; Fiscal Year: 2013
    ..abstract_text> ..
  17. AAV Gene Therapy for AAT deficiency
    Richard J Samulski; Fiscal Year: 2013
    ..The long-term goal of the proposal is to design safer and more effective AAV vectors for gene therapy in patients with AAT deficiency. ..
  18. Next Generation of Recombinant AAV Serotype Vectors for Gene Therapy
    Sergei Zolotukhin; Fiscal Year: 2013
    ....
  19. VASODEPRESSOR MECHANISMS IN HYPERTENSION
    Carlos M Ferrario; Fiscal Year: 2013
    ..Understanding the regulation of this system is vital to the prevention of cardiovascular disease as well as in the design of effective therapies to reduce the consequences of the disease. ..
  20. rAAV-CNGB3 Gene Therapy for Achromatopsia: Translational Research Studies
    William W Hauswirth; Fiscal Year: 2013
    ..Based on these data, in Aim 6 we will prepare and submit an IND to the FDA. The IND will include a protocol for a Phase 1/2 clinical trial whose design will be guided by results from the previous five Specific Aims. ..
  21. Molecular Biology of Recombinant AAV Genomes
    Weidong Xiao; Fiscal Year: 2013
    ..To study the molecular status of recombinant AAV genomes in the cytoplasm. Completion of specific aims in this application will markedly enhance the way of rAAV vectors to be used in the human gene therapy field. ..
  22. Clinical trial of self complementary AAV8-mediated gene transfer for hemophilia B
    Andrew M Davidoff; Fiscal Year: 2013
    ..abstract_text> ..
  23. Immune Responses To AAV-Mediated FIX Gene Transfer
    Hildegund C J Ertl; Fiscal Year: 2013
    ..HC ErtI): T Cells to AAV and AAV-Encoded Transgene Products Project 3 (RW Herzog, C Terliorst): Pathways Towards Immune Tolerance to Coagulation Factors Core A (HC ErtI): Administrative Core Core B (S Zliou): Vector Core ..
  24. CTL response to AAV Vector
    Richard J Samulski; Fiscal Year: 2013
    ....