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Nitric oxide synthase gene therapy: A potent therapeutic in metastatic breast cancer
Abstracts/Nitric Oxide 42 (2014) 99–153
inflammatory or anticancer agents. We have synthesized the first HNO-donating NSAIDs, which like structurally related NO donors show similar anti-inflammatory properties but significantly lower gastrointestinal ulceration than the NSAID alone. Both the HNO and NO donor prodrugs also exhibit higher toxicity to breast cancer cells and significantly reduce tumor growth of breast cancer xenografts in nude mice than the parent NSAID. Detailed analysis has indicated that the cytotoxic and antiproliferative effects of the NO and HNO donor adducts are induced by distinct pathways, which will be described. Overall, HNO-NSAIDs appear to be a unique and promising new class of chemotherapeutic agent. Keywords: Nitric oxide; Nitroxyl; NSAID; Anticancer.
Oral 1922-3. Nitric oxide synthase gene therapy: A potent therapeutic in metastatic breast cancer http://dx.doi.org/10.1016/j.niox.2014.09.039 Cian McCrudden, John McBride, Jonathan Coulter, Victoria Kett, Tracy Robson, Helen McCarthy Queen’s University, Belfast
Primary breast cancer, the most common malignancy in women, is a disease with many treatment options, and impressive response rates. However, treatment options are limited once tumors metastasize. Docetaxel is the front line therapy for metastatic breast cancer, but resistance eventually develops, limiting its potency. Nitric oxide (NO•) exerts anticancer effects by inducing DNA damage, modifying DNA repair proteins, and inhibiting anti-apoptotic factor NF-κB. We have previously reported the therapeutic benefit of inducible NO• synthase (iNOS) gene therapy in several xenograft models of human cancer. The purpose of the current study was to investigate the antimetastatic potential of NO• using a transcriptionally targeted iNOS gene construct and our novel delivery technology. We have developed a peptide-based gene therapy delivery vehicle that produces cationic nanoparticles (NPs) when incubated with plasmid iNOS (piNOS). The NPs overcome the biological barriers to gene therapy, efficiently delivering piNOS to the nucleus of breast cancer cells. piNOS-loaded NPs (constitutively active CMV or transcriptionally-regulated human osteocalcin [hOC] promoters) evoked iNOS gene expression in MDA-MB-231-Luc-D3H1 breast cancer cells in vitro, which manifested impressive nitrite accumulation in culture medium (indicative of NO• generation). C57/BL6 mice tolerated multiple administrations of pDNA-loaded NPs, and their sera lacked NP-neutralizing antibodies. These results indicated that iNOSloaded NPs have therapeutic potential in in vivo models of breast cancer metastasis. Metastatic breast cancer was established in female BALB/c SCID mice by inoculation with 2 × 10 5 MDA-MB-231-LucD3H1 via the left ventricle, and disease progression was assessed via bioluminescence imaging and body weight determination. Treated mice received 50 μg CMV-iNOS/hOC-iNOS in five intravenous biweekly doses. Serum nitrite levels were elevated in mice that received either iNOS gene therapy. Mice that received CMV-iNOS (median survival 40 days; p < 0.001) or hOCiNOS (38.5 days; p = 0.001) therapy had significantly improved survival compared with control mice (31.5 days). Moreover, combining docetaxel therapy with either CMV-iNOS (51 days) or hOC-iNOS (46 days) improved median survival of mice beyond that observed with docetaxel alone (44 days). Targeted NO• gene therapy has shown excellent therapeutic potential for metastatic breast cancer, and studies are ongoing to develop our NP formulation toward a potential clinical product.
111
Keywords: Gene therapy; Breast cancer; Metastasis; Nitric oxide. Oral 2011-1. The therapeutic effects of a nitric oxide generating lozenge that utilizes natural product chemistry http://dx.doi.org/10.1016/j.niox.2014.09.040 Nathan BryanPhD The University of Texas Health Science Center
The production and availability of nitric oxide (NO) is critical for cardiovascular health. The complex production of NO from L-arginine by nitric oxide synthase becomes compromised with age and certain elements of lifestyle. As this primary pathway for NO becomes dysfunctional, it leads to conditions of NO deficiency with subsequent clinical sequel including hypertension and onset and progression of cardiovascular disease. A complementary endogenous pathway for NO production emerged wherein inorganic nitrate and nitrite, produced from both the oxidation of NO and dietary nitrate-rich functional foods such as leafy green vegetables and beets, can be reduced back to NO through enterosalivary cycling. Utilizing intellectual property developed out of the University of Texas Health Science Center in Houston (US patents 8,298,589, 8,303,995 & 8,435,570), Neogenis Laboratories has licensed this technology and developed a GMP certified, over the counter, all natural formulation that provides a system for generating NO in an endothelium-dependent and independent manner. This technology has been clinically tested in placebo controlled designed studies. A single administration of an oral, active, naturebased NO supplement significantly lowers blood pressure, improves vascular compliance and restores endothelial function in subjects with hypertension. Thirty day administration also leads to normalization of blood pressure in pre-hypertensive patients. Pharmaceutical approaches for NO restoration present long-term safety issues; however, functional, nutraceutical approaches may offer select advantages for restoring NO availability. This treatment may be beneficial as routine supplementation for cardiovascular protection. Keywords: Nitrite; Nutraceutical; Diet. Oral 2011-2. Ammonia-oxidizing bacteria accelerate wound closure in diabetic mice http://dx.doi.org/10.1016/j.niox.2014.09.041 Ioannis Gryllos a, Shomir Ghosh a, Neeraja Vajrala b, David Whitlock a, Luis Sayavedra-Soto b, Spiros Jamas a a AOBIOME LLC, Cambridge, MA, USA b Department of Botany and Plant Pathology, Oregon State University, Corvallis, OR, USA
Diabetic foot ulcers (DFUs) are among the most common and serious complications of diabetes. They present in approximately 15% of diabetic patients, leading to ~80,000 amputations per year in the United States alone. Effective strategies for the treatment of DFUs are currently limited, thus highlighting an urgent unmet medical need. Topical delivery of nitric oxide (NO) at the wound site via NO-generating compounds, or by using acidified nitrite as a NO source, has been shown to promote healing in murine models of diabetes by reversing dysregulated expression of inflammatory cytokines, growth factors and matrix metalloproteinases among others. In the present study, we have evaluated the use of ammonia-oxidizing bacteria (AOB) of the genus Nitrosomonas as natural self-regulating biological delivery systems of NO/NOx in vivo. Nitrosomonas are ubiquitous Gram-negative obligate chemolithoautotrophic bacteria that exclusively oxidize ammonia as an energy source to generate nitrite and NOx species. As proof-of-concept, we proceeded to examine
inflammatory or anticancer agents. We have synthesized the first HNO-donating NSAIDs, which like structurally related NO donors show similar anti-inflammatory properties but significantly lower gastrointestinal ulceration than the NSAID alone. Both the HNO and NO donor prodrugs also exhibit higher toxicity to breast cancer cells and significantly reduce tumor growth of breast cancer xenografts in nude mice than the parent NSAID. Detailed analysis has indicated that the cytotoxic and antiproliferative effects of the NO and HNO donor adducts are induced by distinct pathways, which will be described. Overall, HNO-NSAIDs appear to be a unique and promising new class of chemotherapeutic agent. Keywords: Nitric oxide; Nitroxyl; NSAID; Anticancer.
Oral 1922-3. Nitric oxide synthase gene therapy: A potent therapeutic in metastatic breast cancer http://dx.doi.org/10.1016/j.niox.2014.09.039 Cian McCrudden, John McBride, Jonathan Coulter, Victoria Kett, Tracy Robson, Helen McCarthy Queen’s University, Belfast
Primary breast cancer, the most common malignancy in women, is a disease with many treatment options, and impressive response rates. However, treatment options are limited once tumors metastasize. Docetaxel is the front line therapy for metastatic breast cancer, but resistance eventually develops, limiting its potency. Nitric oxide (NO•) exerts anticancer effects by inducing DNA damage, modifying DNA repair proteins, and inhibiting anti-apoptotic factor NF-κB. We have previously reported the therapeutic benefit of inducible NO• synthase (iNOS) gene therapy in several xenograft models of human cancer. The purpose of the current study was to investigate the antimetastatic potential of NO• using a transcriptionally targeted iNOS gene construct and our novel delivery technology. We have developed a peptide-based gene therapy delivery vehicle that produces cationic nanoparticles (NPs) when incubated with plasmid iNOS (piNOS). The NPs overcome the biological barriers to gene therapy, efficiently delivering piNOS to the nucleus of breast cancer cells. piNOS-loaded NPs (constitutively active CMV or transcriptionally-regulated human osteocalcin [hOC] promoters) evoked iNOS gene expression in MDA-MB-231-Luc-D3H1 breast cancer cells in vitro, which manifested impressive nitrite accumulation in culture medium (indicative of NO• generation). C57/BL6 mice tolerated multiple administrations of pDNA-loaded NPs, and their sera lacked NP-neutralizing antibodies. These results indicated that iNOSloaded NPs have therapeutic potential in in vivo models of breast cancer metastasis. Metastatic breast cancer was established in female BALB/c SCID mice by inoculation with 2 × 10 5 MDA-MB-231-LucD3H1 via the left ventricle, and disease progression was assessed via bioluminescence imaging and body weight determination. Treated mice received 50 μg CMV-iNOS/hOC-iNOS in five intravenous biweekly doses. Serum nitrite levels were elevated in mice that received either iNOS gene therapy. Mice that received CMV-iNOS (median survival 40 days; p < 0.001) or hOCiNOS (38.5 days; p = 0.001) therapy had significantly improved survival compared with control mice (31.5 days). Moreover, combining docetaxel therapy with either CMV-iNOS (51 days) or hOC-iNOS (46 days) improved median survival of mice beyond that observed with docetaxel alone (44 days). Targeted NO• gene therapy has shown excellent therapeutic potential for metastatic breast cancer, and studies are ongoing to develop our NP formulation toward a potential clinical product.
111
Keywords: Gene therapy; Breast cancer; Metastasis; Nitric oxide. Oral 2011-1. The therapeutic effects of a nitric oxide generating lozenge that utilizes natural product chemistry http://dx.doi.org/10.1016/j.niox.2014.09.040 Nathan BryanPhD The University of Texas Health Science Center
The production and availability of nitric oxide (NO) is critical for cardiovascular health. The complex production of NO from L-arginine by nitric oxide synthase becomes compromised with age and certain elements of lifestyle. As this primary pathway for NO becomes dysfunctional, it leads to conditions of NO deficiency with subsequent clinical sequel including hypertension and onset and progression of cardiovascular disease. A complementary endogenous pathway for NO production emerged wherein inorganic nitrate and nitrite, produced from both the oxidation of NO and dietary nitrate-rich functional foods such as leafy green vegetables and beets, can be reduced back to NO through enterosalivary cycling. Utilizing intellectual property developed out of the University of Texas Health Science Center in Houston (US patents 8,298,589, 8,303,995 & 8,435,570), Neogenis Laboratories has licensed this technology and developed a GMP certified, over the counter, all natural formulation that provides a system for generating NO in an endothelium-dependent and independent manner. This technology has been clinically tested in placebo controlled designed studies. A single administration of an oral, active, naturebased NO supplement significantly lowers blood pressure, improves vascular compliance and restores endothelial function in subjects with hypertension. Thirty day administration also leads to normalization of blood pressure in pre-hypertensive patients. Pharmaceutical approaches for NO restoration present long-term safety issues; however, functional, nutraceutical approaches may offer select advantages for restoring NO availability. This treatment may be beneficial as routine supplementation for cardiovascular protection. Keywords: Nitrite; Nutraceutical; Diet. Oral 2011-2. Ammonia-oxidizing bacteria accelerate wound closure in diabetic mice http://dx.doi.org/10.1016/j.niox.2014.09.041 Ioannis Gryllos a, Shomir Ghosh a, Neeraja Vajrala b, David Whitlock a, Luis Sayavedra-Soto b, Spiros Jamas a a AOBIOME LLC, Cambridge, MA, USA b Department of Botany and Plant Pathology, Oregon State University, Corvallis, OR, USA
Diabetic foot ulcers (DFUs) are among the most common and serious complications of diabetes. They present in approximately 15% of diabetic patients, leading to ~80,000 amputations per year in the United States alone. Effective strategies for the treatment of DFUs are currently limited, thus highlighting an urgent unmet medical need. Topical delivery of nitric oxide (NO) at the wound site via NO-generating compounds, or by using acidified nitrite as a NO source, has been shown to promote healing in murine models of diabetes by reversing dysregulated expression of inflammatory cytokines, growth factors and matrix metalloproteinases among others. In the present study, we have evaluated the use of ammonia-oxidizing bacteria (AOB) of the genus Nitrosomonas as natural self-regulating biological delivery systems of NO/NOx in vivo. Nitrosomonas are ubiquitous Gram-negative obligate chemolithoautotrophic bacteria that exclusively oxidize ammonia as an energy source to generate nitrite and NOx species. As proof-of-concept, we proceeded to examine