Adenovirus vectors (Ad) are the most common viral vector type used in clinical studies worldwide. Upon intravascular delivery, Ad elicits multifaceted host innate immune and inflammatory responses that drastically compromise both the safety and the efficacy of the Ad-based therapy due to a dose-limiting systemic toxicity. The molecular and cellular mechanisms governing this systemic anti-Ad inflammatory response remain poorly understood. By analyzing the initiation of inflammation in vivo after the injection of capsid-modified Ad vectors into mice knockout for critical inflammatory mediators, we have made an original observation that Ad interaction with 23 integrins and macrophage cell-surface receptors triggers a unique inflammatory pathway mediated by IL-11. Activation of this pathway initiates a downstream cytokine and chemokine cascade that depends on functional IL-1RI signaling. However, our studies also indicate that, in addition to this cytokine production, intravenous Ad administration induces a rapid pro-inflammatory MF death and the influx of pro-inflammatory leukocytes into affected sites.
The Specific Aims of this proposal are designed to define the contribution of each of the known components of Ad-induced inflammation into the clinically relevant systemic toxicity observed after the intravenous Ad administration.
In Specific Aim 1, we will investigate the molecular mechanisms involved in mediating Ad-induced pro- inflammatory macrophage cell death in vivo, and its role in activating systemic anti-Ad inflammatory response.
In Specific Aim 2, we will analyze the phenotypic markers and functional activation states of inflammatory leukocytes, accumulating in the spleen and liver after intravenous Ad injection, and their role in mediating the acute systemic anti-Ad inflammatory response.
In Specific Aim 3, we will construct novel Ad vectors that would avoid interaction with 23 integrins and analyze their gene delivery and systemic toxicity profiles in vivo. These studies will improve our understanding of the fundamental mechanisms of host defense against viral pathogens and may ultimately lead to the development of safe and effective Ad vectors for the therapy of a wide range of inborn and acquired human diseases.

Public Health Relevance

Adenovirus vectors (Ad) are the most common viral vector type used in clinical studies worldwide. Despite extensive preclinical data on the use of Ad in gene transfer applications, Ad's use for gene therapy in humans is severely limited due to life-threatening innate immune and inflammatory responses that arise after intravascular delivery of the high (and potentially therapeutic) virus doses. This proposal is to fill the major void in our understanding of Ad-induced inflammation in vivo triggered after intravascular virus delivery. These studies will provide new insights into fundamental mechanisms of the host defense against viral pathogens, and may ultimately lead to the development of safe and effective Ad vectors for the therapy of a wide range of inborn and acquired human diseases.

Agency
National Institute of Health (NIH)
Institute
National Institute of Allergy and Infectious Diseases (NIAID)
Type
Research Project (R01)
Project #
5R01AI065429-08
Application #
8242001
Study Section
Gene and Drug Delivery Systems Study Section (GDD)
Program Officer
Palker, Thomas J
Project Start
2005-07-01
Project End
2015-03-31
Budget Start
2012-04-01
Budget End
2013-03-31
Support Year
8
Fiscal Year
2012
Total Cost
$386,100
Indirect Cost
$138,600
Name
University of Washington
Department
Internal Medicine/Medicine
Type
Schools of Medicine
DUNS #
605799469
City
Seattle
State
WA
Country
United States
Zip Code
98195
Di Paolo, Nelson C; Baldwin, Lisa K; Irons, Eric E et al. (2014) IL-1? and complement cooperate in triggering local neutrophilic inflammation in response to adenovirus and eliminating virus-containing cells. PLoS Pathog 10:e1004035
Di Paolo, Nelson C; Shayakhmetov, Dmitry M (2014) The analysis of innate immune response to adenovirus using antibody arrays. Methods Mol Biol 1089:133-41
Irons, Eric E; Flatt, Justin W; Doronin, Konstantin et al. (2013) Coagulation factor binding orientation and dimerization may influence infectivity of adenovirus-coagulation factor complexes. J Virol 87:9610-9
Di Paolo, Nelson C; Doronin, Konstantin; Baldwin, Lisa K et al. (2013) The transcription factor IRF3 triggers "defensive suicide" necrosis in response to viral and bacterial pathogens. Cell Rep 3:1840-6
Di Paolo, Nelson C; Shayakhmetov, Dmitry M (2013) Interleukin-1 receptor 2 keeps the lid on interleukin-1?. Immunity 38:203-5
Shayakhmetov, Dmitry M; Di Paolo, Nelson C; Mossman, Karen L (2010) Recognition of virus infection and innate host responses to viral gene therapy vectors. Mol Ther 18:1422-9
Di Paolo, Nelson C; van Rooijen, Nico; Shayakhmetov, Dmitry M (2009) Redundant and synergistic mechanisms control the sequestration of blood-born adenovirus in the liver. Mol Ther 17:675-84
Di Paolo, Nelson C; Miao, Edward A; Iwakura, Yoichiro et al. (2009) Virus binding to a plasma membrane receptor triggers interleukin-1 alpha-mediated proinflammatory macrophage response in vivo. Immunity 31:110-21
Di Paolo, Nelson C; Shayakhmetov, Dmitry M (2009) Adenovirus de-targeting from the liver. Curr Opin Mol Ther 11:523-31
Kalyuzhniy, O; Di Paolo, N C; Silvestry, M et al. (2008) Adenovirus serotype 5 hexon is critical for virus infection of hepatocytes in vivo. Proc Natl Acad Sci U S A 105:5483-8

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