1997 |
Amalfitano, Andrea Na |
R55Activity Code Description: Undocumented code - click on the grant title for more information. |
Modified Adenovirus Vectors For the Use in Gene Therapy
Adenovirus (Ad) based vectors have demonstrated great promise for the treatment of many human diseases via gene therapy. Ad vectors have the ability to deliver transgenes to a variety of cell types, in vitro and in vivo, and unlike retrovirus based vectors, Ad vectors can also efficiently transduce mitotically quiescent cells. Therefore, the potential treatment of many different diseases both genetic and non-genetic can be envisioned with the use of Ad vectors. For example, Ad vectors have been demonstrated to be capable of delivering genes to 1) liver cells for the potential treatment of many metabolic disorders, 2) muscle cells (skeletal and cardiac) for the potential treatment of myopathies and storage disorders, 3) brain and nervous system tissues for the potential treatment of neurologic diseases like Parkinson disease, and 4) respiratory epithelium for the treatment of pulmonary disorders like cystic fibrosis. in addition, many other common diseases like AIDS and various forms of cancer have all been demonstrated to be potentially treated by Ad mediated gene transfer strategies. While there is an enormous potential for the treatment of many human diseases, there are several problems with current Ad vectors that must be addressed before Ad mediated gene therapy becomes a clinical reality. The most serious problem with current Ad vectors is the transient duration of transgene expression after successful gene delivery into the tissues of immunocompetent animals. Other problems include the generation o replication competent Ad (RCA), and the inability of Ad vectors to carry larger genes. This grant proposal outlines a series of experiments that will address each of the limitations of current Ad vectors. In so doing, we will isolate modified Ad vectors that are predicted to allow for longer durations of transgene expression in vivo, decrease the incidence of RCA generation, and significantly increase Ad vector caring capacity. initially, the modified Ad vectors will be analyzed in mouse models of liver and muscle (cardiac and skeletal) cell gene therapy. The result will be the isolation of new Ad vectors capable of efficacious use in animal models of human disease, as well as for eventual use in the therapy of a great number of human conditions.
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0.928 |
1998 — 2001 |
Amalfitano, Andrea Na |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Modified Adenovirus Vectors For Use in Gene Therapy
Ad vectors have the ability to deliver transgenes to a variety of cell types, in vitro and in vivo, and unlike retrovirus based vectors, Ad vectors can also efficiently transduce mitotically quiescent cells. Therefore, the potential treatment of many different diseases, both genetic and non-genetic can be envisioned with the use of Ad vectors. For example, Ad vectors have been demonstrated to be capable of delivering genes to 1) liver cells for the potential treatment of many metabolic disorders, 2) muscle cells (skeletal and cardiac) for the potential treatment of myopathies and storage disorders, 3) brain and nervous system tissues for the potential treatment of neurologic diseases like Parkinson disease, and 4) respiratory epithelium for the treatment of pulmonary disorders like cystic fibrosis. In addition, many other common diseases like AIDS and various forms of cancer have all been demonstrated to be potentially treated by Ad mediated gene transfer strategies. While there is an enormous potential for the treatment of many human diseases, there are several problems with current Ad vectors that must be addressed before Ad mediated gene therapy becomes a clinical reality. The most serious problem with current Ad vectors is the transient duration of transgene expression after successful gene delivery into the tissues of immunocompetent animals. Other problems include the generation of replication competent Ad (RCA), and the inability of Ad vectors to carry larger genes or tissue-specific promoter/enhancer elements. This grant proposal outlines a series of experiments that will address each of the limitations of current Ad vectors. In so doing, we will isolate modified Ad vectors that are predicted to allow for longer durations of transgene expression in vivo, decrease the incidence of RCA generation, and significantly increase Ad vector carrying capacity. Initially, the modified Ad vectors will be analyzed in mouse models of liver and muscle cell gene therapy. The result will be the isolation of new Ad vectors capable of efficacious use in animal models of human disease, as well as for eventual use in the therapy of a great number of human conditions.
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0.928 |
2004 — 2006 |
Amalfitano, Andrea Na |
P01Activity Code Description: For the support of a broadly based, multidisciplinary, often long-term research program which has a specific major objective or a basic theme. A program project generally involves the organized efforts of relatively large groups, members of which are conducting research projects designed to elucidate the various aspects or components of this objective. Each research project is usually under the leadership of an established investigator. The grant can provide support for certain basic resources used by these groups in the program, including clinical components, the sharing of which facilitates the total research effort. A program project is directed toward a range of problems having a central research focus, in contrast to the usually narrower thrust of the traditional research project. Each project supported through this mechanism should contribute or be directly related to the common theme of the total research effort. These scientifically meritorious projects should demonstrate an essential element of unity and interdependence, i.e., a system of research activities and projects directed toward a well-defined research program goal. |
Adoptive Immunotherapy With Recombinant Adenvirus Vector
Several different vaccine strategies have been evaluated and combined in an attempt to amplify T-cell responses toward induction of anti-tumor immunity. The model tumor antigen used in many of these studies was carcinoembryonic antigen (CEA). While initial T-cell activation studies were conducted in conventional mice and then tested, results from the clinical trials suggested immune and clinical responses less dramatic than in the murine models. One strategy to improve the clinical outcome has been the use of recombinant viral vectors encoding CEA modified dendritic cells. Based upon several lines of observation, this strategy appears to be capable of further improvement when using a heterologous prime-boost vaccination strategy, using alternative means of introducing the tumor antigen. Therefore, we propose pre-clinical and clinical studies of combined vaccine strategy studies, in this instance capitalizing upon the known efficacy of fowlpox CEA virus based constructs, but now combining this expertise with use of adenovirus based vectors also encoding CEA. Exciting data from the HIV vaccine literature suggest that heterologous prime-boost vaccine strategies have significantly benefited from the utilization of first generation Ad based vectors, showing dramatically improved evidence of inducing immune critical T-cell responses in human subjects. Uniquely, our group has previously constructed several new generations of Ad vector that will allow us to investigate and optimize the use of Ad vectors as vaccines for a variety of antigens. Once the most optimized Ad encoding CEA is delineated, we will determine the efficacy of the vector alone, or in heterologous prime-boost vaccine strategies utilizing rigorous animal models. A key innovation will be our ability to synergize with the other projects and cores in this program project, for example we will evaluate the anti-tumor efficacy of heterologous prime-boost strategies utilizing the optimal Ad-CEA vector vaccine, combined with either the aforementioned fowlpox-CEA vector vaccine, or an alphavirus based CEA vector vaccine (the latter being developed in Project #2 of this overall proposal). These studies are intended to demonstrate that the use of heterologous prime-boost regimens (via the use of two different recombinant vectors) can further amplify T-cell responses toward tumor associated antigens such as CEA. Finally, we will initiate pre-clinical studies and a pilot project of active immunotherapy using the most optimized adenovirus+CEA vector based vaccine, a prelude to a combined pox/Ad or alphavirus/Ad heterologous prime-boost clinical trial.
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0.928 |
2005 — 2007 |
Amalfitano, Andrea Na |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Adenovirus Vectors and Complement System @ Michigan State University
DESCRIPTION (provided by applicant): Ongoing studies make it clear that the intravenous injection of high doses of Adenovirus (Ad) based vectors offer a tremendous potential for treatment of disseminated cancers, inherited diseases, vascular diseases, as well for gene therapy of the liver for a variety of hepatic/genetic diseases. However, the systemic administration of Ad based vectors is currently viewed as a highly risky maneuver, as significant acute toxicities can be incurred, including chemokine/cytokine release, endothelial cell damage, Kupffer cell activation, hypotension, thrombocytopenia (low platelet counts), and as noted in the Gelsinger tragedy, the systemic inflammatory response syndrome and death. Precious little is known about the innate systems that are immediately triggered by injection of an Ad vector at high doses. Activation of these systems is predicted to be pivotal in initiating and/or modulating subsequent host immune responses, both innate and adaptive. One of these first line innate host defense systems is the complement system. Inappropriate complement activation can result in a number of dire consequences, including anaphylactoid reactions, ARDS (adult respiratory distress syndrome), hypotensive shock, DIC (disseminated intravascular coagulation), cytokine release and systemic inflammatory response syndromes. More specifically, complement activation products released after complement activation (i.e: C3a, C5a) are potent inflammatory mediators, which are known to activate Kupffer cells, as well activate vascular endothelium, leading to neutrophil recruitment, and platelet aggregation. Intriguingly, many of these consequences are also noted after high dose Ad injections. Based upon these considerations, we forward the following Central Hypothesis: "Ad vector interactions with the complement system results in many of the limitations and toxicities typically attributed to the use of Ad vectors". The aims of this proposal will directly test the central hypothesis. The results of this line of investigation will move the field of gene transfer using systemically administered Ad vectors forward.
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0.958 |
2007 — 2008 |
Amalfitano, Andrea Na |
P01Activity Code Description: For the support of a broadly based, multidisciplinary, often long-term research program which has a specific major objective or a basic theme. A program project generally involves the organized efforts of relatively large groups, members of which are conducting research projects designed to elucidate the various aspects or components of this objective. Each research project is usually under the leadership of an established investigator. The grant can provide support for certain basic resources used by these groups in the program, including clinical components, the sharing of which facilitates the total research effort. A program project is directed toward a range of problems having a central research focus, in contrast to the usually narrower thrust of the traditional research project. Each project supported through this mechanism should contribute or be directly related to the common theme of the total research effort. These scientifically meritorious projects should demonstrate an essential element of unity and interdependence, i.e., a system of research activities and projects directed toward a well-defined research program goal. |
Adoptive Immunotherapy With Recombinant Adenovirus Based Vectors
Several different vaccine strategies have been evaluated and combined in an attempt to amplify T-cell responses toward induction of anti-tumor immunity. The model tumor antigen used in many of these studies was carcinoembryonic antigen (CEA). While initial T-cell activation studies were conducted in conventional mice and then tested, results from the clinical trials suggested immune and clinical responses less dramatic than in the murine models. One strategy to improve the clinical outcome has been the use of recombinant viral vectors encoding CEA modified dendritic cells. Based upon several lines of observation, this strategy appears to be capable of further improvement when using a heterologous prime-boost vaccination strategy, using alternative means of introducing the tumor antigen. Therefore, we propose pre-clinical and clinical studies of combined vaccine strategy studies, in this instance capitalizing upon the known efficacy of fowlpox CEA virus based constructs, but now combining this expertise with use of adenovirus based vectors also encoding CEA. Exciting data from the HIV vaccine literature suggest that heterologous prime-boost vaccine strategies have significantly benefited from the utilization of first generation Ad based vectors, showing dramatically improved evidence of inducing immune critical T-cell responses in human subjects. Uniquely, our group has previously constructed several new generations of Ad vector that will allow us to investigate and optimize the use of Ad vectors as vaccines for a variety of antigens. Once the most optimized Ad encoding CEA is delineated, we will determine the efficacy of the vector alone, or in heterologous prime-boost vaccine strategies utilizing rigorous animal models. A key innovation will be our ability to synergize with the other projects and cores in this program project, for example we will evaluate the anti-tumor efficacy of heterologous prime-boost strategies utilizing the optimal Ad-CEA vector vaccine, combined with either the aforementioned fowlpox-CEA vector vaccine, or an alphavirus based CEA vector vaccine (the latter being developed in Project #2 of this overall proposal). These studies are intended to demonstrate that the use of heterologous prime-boost regimens (via the use of two different recombinant vectors) can further amplify T-cell responses toward tumor associated antigens such as CEA. Finally, we will initiate pre-clinical studies and a pilot project of active immunotherapy using the most optimized adenovirus+CEA vector based vaccine, a prelude to a combined pox/Ad or alphavirus/Ad heterologous prime-boost clinical trial.
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0.928 |
2010 — 2014 |
Amalfitano, Andrea Na |
R01Activity Code Description: To support a discrete, specified, circumscribed project to be performed by the named investigator(s) in an area representing his or her specific interest and competencies. |
Er-Localized Aminopeptidases in Ankylosing Spondylitis @ Michigan State University
DESCRIPTION (provided by applicant): Ankylosing spondylitis (AS) is a painful, incurable autoimmune disease that affects millions of people worldwide. Although it is clear that both environmental and genetic factors are involved in the disease, the cause and pathogenesis of AS remain unclear. A strong link with the MHC class I allele HLA-B27 has implicated antigen processing as an underlying factor, and this hypothesis was strengthened by the recent demonstration that polymorphisms in ERAP1 (an aminopeptidase that we and others have shown is associated with antigen processing) affect the risk of developing AS. We hypothesize that ERAP1 polymorphisms either directly or indirectly affect substrate selection and trimming, so that different ERAP1 alleles alter the levels of arthritogenic or protective peptides presented on HLA-B27. We will test this hypothesis using biochemical assays for substrate specificity, cultured cells to measure effects on antigen processing, and transgenic mice to test effects on disease in vivo. We will also test the effects of ERAP1 polymorphisms on other pathways that have been proposed to affect AS pathogenesis, including the assembly and stability of HLA- B27 and shedding of cytokine receptors. The Aims of this project are to determine the functional effects of ERAP1 polymorphisms and to understand the molecular pathogenesis of AS. The long-term goals of this project are to predict the risk of AS in individuals, and to develop methods to prevent and treat AS. We will also apply the understanding of AS pathogenesis to other autoimmune diseases that are linked to MHC class I alleles.
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0.958 |