The broad objectives of this research are to develop novel adenovirus (Ad) vectors for human gene therapy. Recombinant Ad vectors are promising agents for a number of applications including immunotherapy, cancer treatment, and gene delivery for treatment of acquired or inherited diseases. Of the more than 1,000 gene therapy clinical trials ongoing worldwide, >25% involve Ad vectors. Ad vectors may be broadly grouped into two categories: replication-competent vectors and replication-defectivevectors. Replication-competent vectors include oncolytic Ad vectors. Replication-defective Ad vectors are being used for gene-based immunotherapy, tumor suppressor gene therapy, prophylactic and therapeutic vaccine delivery, and gene therapy of acquired and inherited disease. High capacity Ad (HC-Ad) vectors are replication-defective vectors that lack all viral coding regions. HC-Ad vectors elicit a significantly reduced host immune response and have shown encouraging results in animal models for long-term transgene delivery to a variety of tissues. Large scale production of HC-Ad vectors is difficult and invariably results in contamination with helper virus. HC-Ad vectors are not in clinical trials due, in large part, to the limitations of HC-Ad vector production. Significant improvements in the production of HC-Ad vectors will be required to readily obtain clinical grade preparations. We have developed novel approaches toward optimizing the production of HC- Ad vectors. These include the use of adeno-associatedvirus (AAV) excision elements to generate an HC- Ad vector in a single step and the development of a new helper virus to reduce helper virus contamination of HC-Ad preparations. We also have utilized AAV integration elements to develop Ad vectors capable of site- specific integration. The goals of this renewal application are to significantly improve HC-Ad production using new viral vectors and helper viruses and to examine the capacity of Ad vectors to integrate site- specifically at the AAVS1 locus using AAV integrationelements.

National Institute of Health (NIH)
National Institute of Allergy and Infectious Diseases (NIAID)
Research Project (R01)
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Gene and Drug Delivery Systems Study Section (GDD)
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Park, Eun-Chung
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State University New York Stony Brook
Schools of Medicine
Stony Brook
United States
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Guimet, Diana; Hearing, Patrick (2013) The adenovirus L4-22K protein has distinct functions in the posttranscriptional regulation of gene expression and encapsidation of the viral genome. J Virol 87:7688-99
Wu, Kai; Guimet, Diana; Hearing, Patrick (2013) The adenovirus L4-33K protein regulates both late gene expression patterns and viral DNA packaging. J Virol 87:6739-47
Wu, Kai; Orozco, Diana; Hearing, Patrick (2012) The adenovirus L4-22K protein is multifunctional and is an integral component of crucial aspects of infection. J Virol 86:10474-83
Alba, Raul; Cots, Dan; Ostapchuk, Philomena et al. (2011) Altering the Ad5 packaging domain affects the maturation of the Ad particles. PLoS One 6:e19564
Banerjee, Partha Sarathi; Ostapchuk, Philomena; Hearing, Patrick et al. (2011) Unnatural amino acid incorporation onto adenoviral (Ad) coat proteins facilitates chemoselective modification and retargeting of Ad type 5 vectors. J Virol 85:7546-54
Sitaraman, Varsha; Hearing, Patrick; Ward, Charles B et al. (2011) Computationally designed adeno-associated virus (AAV) Rep 78 is efficiently maintained within an adenovirus vector. Proc Natl Acad Sci U S A 108:14294-9
Ma, Hsin-Chieh; Hearing, Patrick (2011) Adenovirus structural protein IIIa is involved in the serotype specificity of viral DNA packaging. J Virol 85:7849-55
Ostapchuk, Philomena; Almond, Matthew; Hearing, Patrick (2011) Characterization of Empty adenovirus particles assembled in the absence of a functional adenovirus IVa2 protein. J Virol 85:5524-31
Banerjee, Partha Sarathi; Ostapchuk, Philomena; Hearing, Patrick et al. (2010) Chemoselective attachment of small molecule effector functionality to human adenoviruses facilitates gene delivery to cancer cells. J Am Chem Soc 132:13615-7
Ostapchuk, Philomena; Hearing, Patrick (2008) Adenovirus IVa2 protein binds ATP. J Virol 82:10290-4

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