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Diagnostic and therapeutic applications of bacteriophage and adeno-associated virus technologies in pulmonary emphysema
294
Abstracts / Nanomedicine: Nanotechnology, Biology, and Medicine 2 (2006) 269–312
We recently showed that RNA can be used as a building block for bottom-up assembly in nanotechnology (Nano Letters, 2004, 4:1717). Here we report the assembly of 30–40 nm trimeric RNA particles that can simultaneously codeliver three therapeutic RNAs to specific cancer cells. The RNA building block harboring the therapeutic molecule was subsequently fabricated into a trimer by utilizing engineered right and left interlocking RNA loops. Incubation of such nanoscale devices containing receptor-binding aptamer or other ligands resulted in binding and co-entry of the trivalent therapeutic particles into cells, subsequently modulating the apoptosis of cancer or viral infected cells. The specificity and efficiency were further confirmed in animal trials. Stable RNA nanoparticles were constructed via the use of chemical modification. Toxicity of the deliverable complexes was reduced by altering RNA 3-D structure and providing relocation of the 5V/3V-ends. The use of such protein-free nanoparticles, which are by composition poor immunogens, as therapeutic reagents would allow for long-term administration to avoid the immune response due to repeated treatment for chronic diseases.
studying virus-cell surface interactions and how these contribute to pathogenesis and disease caused by viruses. There she also initiated work in the development of viruses as nanoparticles for therapeutics and vaccines. In 2002 she moved to the Center for Integrative Molecular Biosciences (CIMBio) in the Department of Cell Biology at Scripps. Currently she is an Associate Professor of Cell Biology, and she is the Director of two interdisciplinary research programs in nanotechnology studying virus-based nanoparticles for cancer therapeutics and diagnostics, and developing nanoparticle-based therapeutics for biodefense.
Dr. Peixuan Guo is a Professor of Molecular Virology and Biomedical Engineering at Purdue University where he serves as the Director of Purdue Bionanotechnology Interdisciplinary Graduate Program. He constructed an in-vitro phi29 DNA packaging motor (the most powerful biomotor constructed to date) (PNAS, 1986); discovered the motor pRNA (the viral RNA that binds ATP) (Science, 1987) and the formation of hexameric pRNA (Molecular Cell, 1998). He also demonstrated that RNA can serve as building blocks for nano-devices (Nano Letters, 2003, 2005). Dr. Guo is an editor or editorial board member for six journals, including four in nanotechnology and bionanotechnology. http://www.vet.purdue.edu/peixuanguo/
Diagnostic and therapeutic applications of bacteriophage and adenoassociated virus technologies in pulmonary emphysema Petrache I, Arap W, Pasqualini R, Flotte T, Rubin M. Tuder, Indiana University School of Medicine, Indianapolis, Indiana, USA
doi:10.1016/j.nano.2006.10.080 70
Sunday, September 10th (11:05) Concurrent Symposium XIV: Nanotechnology in Gene Delivery and Therapy for Cancer or Viral Diseases
Targeted therapy using virus-based nanoparticles (VNPs) Manchester M, Department of Cell Biology, Center for Integrative Molecular Biosciences, The Scripps Research Institute, La Jolla, California, USA The ability to specifically target therapies to the site of a developing tumor, while avoiding healthy tissue, is an important goal for cancer research. Similarly, there is a tremendous need to identify, image, and monitor tumors particularly at early stages and during treatment. There are many different classes of nanoparticles currently in development that combine targeted imaging and treatment functions. We focus on virus-based nanoparticles (VNPs) as platforms for the development of tissue-specific targeting and imaging agents in vivo. Two of the viruses we study are canine parvovirus (CPV), and cowpea mosaic virus (CPMV). CPV is a small icosahedral virus that has a natural affinity for tumor cells, by virtue of its interaction with human transferrin receptors on the cell surface. CPMV, while a plant virus, also has a natural affinity for vertebrate endothelial cells and we have shown that it may be targeted specifically to tumors in vitro and in vivo. The ability to harness the natural surface properties of viruses to effectively target tumor cells provides a novel mechanism for drug delivery and imaging of tumors in vivo. Marianne Manchester received her training in virology at the Lineberger Comprehensive Cancer Center at the University of North Carolina, Chapel Hill, where she was a Lineberger Fellow and received her Ph.D. in Genetics in 1992. From 1993 to 1998 she was a postdoctoral fellow at The Scripps Research Institute and a fellow of the World Health Organization’s Programme for Vaccines and Immunization. In 1998 she joined the faculty of the Department of Neuropharmacology at Scripps,
doi:10.1016/j.nano.2006.10.081
71
Sunday, September 10th (11:30) Concurrent Symposium XIV: Nanotechnology in Gene Delivery and Therapy for Cancer or Viral Diseases
Emphysema is a chronic lung disease induced primarily by cigarette smoke and several genetic defects, such as alpha-1 antitrypsin deficiency. Emphysema is a common manifestation of chronic obstructive pulmonary diseases (COPD), manifesting as permanent disappearance of structural elements of the lung. The alveolus (the gas-exchange unit of the lung) is composed of a functional syncitial network composed of alveolar epithelial and endothelial cells, and fibroblasts, suggesting the existence of an intercellular crosstalk of molecular signaling pathways. The disruption of this complex interaction by apoptosis, oxidative stress, and/or proteaseantiprotease imbalance may underlie the destruction of alveolar septae by cigarette smoke. As no early diagnostic or prognostic markers have been developed thus far, nanotechnology may provide significant improvement of our abilities to model the disease, assess severity, and permit lung specific drug delivery. To address the specific role of alveolar endothelial cell apoptosis in the development of emphysema, we utilized the bacteriophage display methodology to assign bzipQ addresses to endothelial cells in vivo. We identified a lung endothelial cell unique peptide (called lung homing peptide) using cultured endothelial cells for multiple rounds of enrichment and positive selection of peptide binding motifs. The peptide bound specifically to lung endothelial cells and when co-delivered to mice with a pro-apoptotic peptide, induced alveolar cell apoptosis and emphysema in vivo. The phages can be used to assemble viable nanoplatforms, which can specifically target the lung cell compartments. Therapeutically, these nanophage devices can target lung specific delivery of protective molecules such as alpha-1 antitrypsin (A1AT) as its deficiency allows for unopposed action of elastases and as we have recently described, of proapoptotic active caspase-3. A1AT prevents emphysema by inhibiting apoptosis when delivered with an adeno-associated viral-mediated augmentation therapy, thus suggesting that increase of A1AT at the lung cellular levels can be used therapeutically. Irina Petrache, M.D. is an Associate professor of Medicine at the Indiana University. Her lab is investigating mechanisms of lung injury pertinent to cigarette smoke-induced chronic lung disease, employing a translational approach, from basic science to animal models and human samples. A major focus is the role of sphingolipid homeostasis in the maintenance of the alveolar-capillary structures and the role of endothelial cell apoptosis in the destruction of alveolar architecture in emphysema. Her laboratory is examining the protective effects of alpha-1 antitrypsin on the pulmonary microvascular endothelium as it relates to prevention of emphysema.
doi:10.1016/j.nano.2006.10.082
Abstracts / Nanomedicine: Nanotechnology, Biology, and Medicine 2 (2006) 269–312
We recently showed that RNA can be used as a building block for bottom-up assembly in nanotechnology (Nano Letters, 2004, 4:1717). Here we report the assembly of 30–40 nm trimeric RNA particles that can simultaneously codeliver three therapeutic RNAs to specific cancer cells. The RNA building block harboring the therapeutic molecule was subsequently fabricated into a trimer by utilizing engineered right and left interlocking RNA loops. Incubation of such nanoscale devices containing receptor-binding aptamer or other ligands resulted in binding and co-entry of the trivalent therapeutic particles into cells, subsequently modulating the apoptosis of cancer or viral infected cells. The specificity and efficiency were further confirmed in animal trials. Stable RNA nanoparticles were constructed via the use of chemical modification. Toxicity of the deliverable complexes was reduced by altering RNA 3-D structure and providing relocation of the 5V/3V-ends. The use of such protein-free nanoparticles, which are by composition poor immunogens, as therapeutic reagents would allow for long-term administration to avoid the immune response due to repeated treatment for chronic diseases.
studying virus-cell surface interactions and how these contribute to pathogenesis and disease caused by viruses. There she also initiated work in the development of viruses as nanoparticles for therapeutics and vaccines. In 2002 she moved to the Center for Integrative Molecular Biosciences (CIMBio) in the Department of Cell Biology at Scripps. Currently she is an Associate Professor of Cell Biology, and she is the Director of two interdisciplinary research programs in nanotechnology studying virus-based nanoparticles for cancer therapeutics and diagnostics, and developing nanoparticle-based therapeutics for biodefense.
Dr. Peixuan Guo is a Professor of Molecular Virology and Biomedical Engineering at Purdue University where he serves as the Director of Purdue Bionanotechnology Interdisciplinary Graduate Program. He constructed an in-vitro phi29 DNA packaging motor (the most powerful biomotor constructed to date) (PNAS, 1986); discovered the motor pRNA (the viral RNA that binds ATP) (Science, 1987) and the formation of hexameric pRNA (Molecular Cell, 1998). He also demonstrated that RNA can serve as building blocks for nano-devices (Nano Letters, 2003, 2005). Dr. Guo is an editor or editorial board member for six journals, including four in nanotechnology and bionanotechnology. http://www.vet.purdue.edu/peixuanguo/
Diagnostic and therapeutic applications of bacteriophage and adenoassociated virus technologies in pulmonary emphysema Petrache I, Arap W, Pasqualini R, Flotte T, Rubin M. Tuder, Indiana University School of Medicine, Indianapolis, Indiana, USA
doi:10.1016/j.nano.2006.10.080 70
Sunday, September 10th (11:05) Concurrent Symposium XIV: Nanotechnology in Gene Delivery and Therapy for Cancer or Viral Diseases
Targeted therapy using virus-based nanoparticles (VNPs) Manchester M, Department of Cell Biology, Center for Integrative Molecular Biosciences, The Scripps Research Institute, La Jolla, California, USA The ability to specifically target therapies to the site of a developing tumor, while avoiding healthy tissue, is an important goal for cancer research. Similarly, there is a tremendous need to identify, image, and monitor tumors particularly at early stages and during treatment. There are many different classes of nanoparticles currently in development that combine targeted imaging and treatment functions. We focus on virus-based nanoparticles (VNPs) as platforms for the development of tissue-specific targeting and imaging agents in vivo. Two of the viruses we study are canine parvovirus (CPV), and cowpea mosaic virus (CPMV). CPV is a small icosahedral virus that has a natural affinity for tumor cells, by virtue of its interaction with human transferrin receptors on the cell surface. CPMV, while a plant virus, also has a natural affinity for vertebrate endothelial cells and we have shown that it may be targeted specifically to tumors in vitro and in vivo. The ability to harness the natural surface properties of viruses to effectively target tumor cells provides a novel mechanism for drug delivery and imaging of tumors in vivo. Marianne Manchester received her training in virology at the Lineberger Comprehensive Cancer Center at the University of North Carolina, Chapel Hill, where she was a Lineberger Fellow and received her Ph.D. in Genetics in 1992. From 1993 to 1998 she was a postdoctoral fellow at The Scripps Research Institute and a fellow of the World Health Organization’s Programme for Vaccines and Immunization. In 1998 she joined the faculty of the Department of Neuropharmacology at Scripps,
doi:10.1016/j.nano.2006.10.081
71
Sunday, September 10th (11:30) Concurrent Symposium XIV: Nanotechnology in Gene Delivery and Therapy for Cancer or Viral Diseases
Emphysema is a chronic lung disease induced primarily by cigarette smoke and several genetic defects, such as alpha-1 antitrypsin deficiency. Emphysema is a common manifestation of chronic obstructive pulmonary diseases (COPD), manifesting as permanent disappearance of structural elements of the lung. The alveolus (the gas-exchange unit of the lung) is composed of a functional syncitial network composed of alveolar epithelial and endothelial cells, and fibroblasts, suggesting the existence of an intercellular crosstalk of molecular signaling pathways. The disruption of this complex interaction by apoptosis, oxidative stress, and/or proteaseantiprotease imbalance may underlie the destruction of alveolar septae by cigarette smoke. As no early diagnostic or prognostic markers have been developed thus far, nanotechnology may provide significant improvement of our abilities to model the disease, assess severity, and permit lung specific drug delivery. To address the specific role of alveolar endothelial cell apoptosis in the development of emphysema, we utilized the bacteriophage display methodology to assign bzipQ addresses to endothelial cells in vivo. We identified a lung endothelial cell unique peptide (called lung homing peptide) using cultured endothelial cells for multiple rounds of enrichment and positive selection of peptide binding motifs. The peptide bound specifically to lung endothelial cells and when co-delivered to mice with a pro-apoptotic peptide, induced alveolar cell apoptosis and emphysema in vivo. The phages can be used to assemble viable nanoplatforms, which can specifically target the lung cell compartments. Therapeutically, these nanophage devices can target lung specific delivery of protective molecules such as alpha-1 antitrypsin (A1AT) as its deficiency allows for unopposed action of elastases and as we have recently described, of proapoptotic active caspase-3. A1AT prevents emphysema by inhibiting apoptosis when delivered with an adeno-associated viral-mediated augmentation therapy, thus suggesting that increase of A1AT at the lung cellular levels can be used therapeutically. Irina Petrache, M.D. is an Associate professor of Medicine at the Indiana University. Her lab is investigating mechanisms of lung injury pertinent to cigarette smoke-induced chronic lung disease, employing a translational approach, from basic science to animal models and human samples. A major focus is the role of sphingolipid homeostasis in the maintenance of the alveolar-capillary structures and the role of endothelial cell apoptosis in the destruction of alveolar architecture in emphysema. Her laboratory is examining the protective effects of alpha-1 antitrypsin on the pulmonary microvascular endothelium as it relates to prevention of emphysema.
doi:10.1016/j.nano.2006.10.082