1985 — 1987 |
Curiel, David T |
F32Activity Code Description: To provide postdoctoral research training to individuals to broaden their scientific background and extend their potential for research in specified health-related areas. |
Alpha 1 Antitrypsin Gene Sequence Polymorphism @ U.S. National Institutes of Health |
0.909 |
1992 — 1993 |
Curiel, David T |
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. |
Conjugate-Mediated Gene Transfer to Airway Epithelium @ University of Alabama At Birmingham
Genetic correction of CF will require the development of vectors capable of achieving direct, in vivo gene transfer to the cells of the airway epithelia. Molecular conjugate vectors offer may potential advantages for this application. The present proposal will evaluate the feasibility of gen delivery to CF respiratory epithelia employing molecular conjugate vectors towards the ultimate goal of achieving gene therapy for CF. One Specific Aim of this proposal is thus to determine if gene transfer can be accomplished to CF airway epithelial cells by the receptor-mediated endocytosis pathway via conjugate vectors utilizing a targetable cell surface receptor. These studies will utilize primary cultures of human airway epithelial cells. To achieve genetic correction by direct in vivo delivery, conjugates must accomplish efficient gene transfer to target cells. Since it is known that gene transfer mediated by molecular conjugates can be dramatically augmented by agents that cause target cell endosomolysis, the second Specific Aim of this proposal is to devise conjugate vectors of higher efficiency by the incorporation of a specific endosomolysis mechanism into the conjugate design. The strategy employed will focus on utilization of adenoviral-mediated endosomolysis by identification of the adenovirus endosomolysis principle residing within the capsid structure. The selective incorporation of this agent would allow employment of viral entry functions without the biosafety limitations inherent in recombinant viral vectors. As it may be a physiologic constraint to selectively express CFTR within a defined airway epithelial cell subset, the third Specific Aim of this proposal is the design of molecular conjugate vectors with the capacity to target these cells. This strategy will capitalize on the cell-specific tropism of influenza virus and Mycoplasma pneumoniae for ciliated respiratory epithelial cells. The cell binding domains of these agents will be prepared by recombinant techniques and incorporated into conjugate design to yield a molecular conjugate vector with a corresponding specificity. In summary, the present proposal seeks to: 1) demonstrate the feasibility of conjugate-mediated gene delivery to airway epithelial cells; 20 confer upon such a conjugate the higher efficiency gene transfer capacity required for in vivo delivery; and 3) target these conjugates to achieve cell-specific delivery to a physiologically relevant airway epithelial cells subset. Achievement of these Aims will potentially yield a vector system with the features required to achieve gene therapy for CF.
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1 |
1994 — 1996 |
Curiel, David T |
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. |
Conjugate Mediated Gene Transfer to Airway Epithelium @ University of Alabama At Birmingham |
1 |
1996 — 1999 |
Curiel, David T |
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. |
Gene Therapy For Ovarian Cancer @ University of Alabama At Birmingham
DESCRIPTION: (Applicant's Abstract) A variety of strategies have been developed to accomplish gene therapy for cancer. The strategy of mutation compensation attempts to achieve genetic correction of cancer cells by specifically targeting the genetic events etiologic of neoplastic progression. This has been previously attempted employing antisense methods of gene inhibition. Whereas phenotypic reversion has been demonstrated in several contexts using this method, problems related to antisense instability and delivery have limited clinical application of this approach. As an alternative, the applicant has developed a novel method to achieve specific ablation of relevant oncogenes. In this regard, the applicant has developed methods to accomplish intracellular expression of a single-chain antibody (sFv) directed against the oncoprotein. In this schema, the newly synthesized oncoprotein would be entrapped during synthesis, and thus unable to exert its transforming effect. As an initial target, the applicant has developed methods to achieve selective ablation of the oncoprotein erbB-2, which has been shown to be important in the pathogenesis of ovarian and other carcinomas. For the approach that the applicant proposes, anti-erbB-2 single-chain immunoglobulin (sFv) genes were constructed to direct expression of intracellular anti-erbB-2 antibodies. Preliminary studies showed that expression of an endoplasmic reticulum (ER) form of the anti-erbB-2 sFv resulted in a profound down-regulation of cell surface erbB-2 in an erbB-2 over-expressing ovarian carcinoma cell line. In addition, expression of the intracellular antibody resulted in a direct cytocidal effect in erbB-2 overexpressing tumor cells. Further analysis demonstrated that this cytocidal effect was on the basis of induction of apoptosis. Thus, mislocalization of the erbB-2 oncoprotein in the ER of tumor cells accomplished a novel means of inducing cellular apoptosis. Importantly, expression of the ER anti-erbB-2 sFv in non-overexpressing cells did not induce cell death or apoptosis. Thus, this form of genetic intervention is capable of achieving a targeted killing of erbB-2 overexpressing tumor cells. This finding has been translated into a gene therapy approach for ovarian carcinoma whereby a recombinant adenovirus encoding the anti-erbB-2 sFv could be shown to accomplish in vivo tumor cell killings in a mouse xenograft model of human ovarian carcinoma. Based upon these results, the applicant will further explore the mechanisms and utility of the anti-oncogene sFv technique. In this regard, studies will be undertaken to evaluate the molecular determinants of sFv-mediated apoptosis after oncoprotein mislocalization. In addition, other ovarian cancer knock-out targets will be addressed to determine their role in neoplastic progression and the therapeutic utility of achieving their functional knock-out in the context of ovarian carcinoma. Finally, in vivo studies will be carried out to develop the methods for translation of these strategies into a gene therapy approach for ovarian carcinoma.
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1 |
1997 — 2000 |
Curiel, David T |
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. |
Targeted Vectors For Cystic Fibrosis Gene Therapy @ University of Alabama At Birmingham
A variety of vector strategies have been proposed to accomplish gene therapy for cystic fibrosis (CF). In this regard, a great deal of interest has focused on the utility of replication-defective adenoviral vectors to achieve direct in situ CFTR gene delivery to affected airway epithelium. Towards this goal, this vector has demonstrated the capacity to achieve efficient gene transfer in vitro to human airway epithelial cells and in vivo to intact airway epithelium in animal models. This latter capacity to accomplish relatively efficient in vivo gene delivery to pulmonary epithelia has distinguished this vector, and thus recommended its employ for human clinical gene therapy trials for CF. Despite these considerations, a number of factors have now been appreciated which have limited the utility of this vector agent for CF gene therapy, at least in its present form. In this context, vector administration has been associated with significant inflammatory and immunologic consequences. In addition, the actual efficacy of transduction of mature airway epithelium is not as efficient as initially reported. Thus, to render these vehicles useful for CF gene therapy, strategies must be developed to address the issues of vector immunogenicity as well as efficacy of target cell transduction. In this latter regard. The basis of vector transductional inefficiency for airway epithelium is understood to derive from a relative paucity of receptors for adenoviral binding and internalization. Thus, methods to redirect the virus into receptor pathways relevant to pulmonary cellular targets would be predicted to augment overall gene transfer efficacy. This approach, however, is undermined by the relative paucity of characterized pulmonary epithelial-specific receptors. Thus, the definition of such cellular markers for airway epithelium would be adjunctive to re-targeting strategies to augment adenoviral vector efficacy. It is thus our hypothesis that we can modify adenoviral tropism as a means to augment transductional efficiency of mature airway epithelium. Further, we propose that the definition of airway cell-specific markers will facilitate this approach by providing selective cellular pathways for vector targeting.
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1 |
1997 — 2001 |
Curiel, David T |
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. |
Adenoviral Mediated Targeted Gene Delivery @ University of Alabama At Birmingham
DESCRIPTION: It is the purpose of the present proposal to develop a gene transfer vector capable of specific and selective transduction of disseminated neoplastic cells. The development of a vector with this capacity would allow the targeting of malignant cells in situ after systemic delivery. Capitalizing on the unique capacity of recombinant adenoviral vectors to accomplish high efficiency in vivo gene delivery, it is the hypothesis of the investigators that further modifications of this vector may be achieved to alter parent virus tropism such that selective transduction of malignant cells may be accomplished. The specific aim of this proposal is thus to modify the tropism of recombinant adenoviral vectors employing genetic and immunologic methods to modify the adenoviral fiber protein. The capacity of the tropism-modified adenoviral vectors to accomplish targeted tumor cell-specific transduction in vivo in the context of animal models of disseminated neoplastic disease will be determined. The derivation of this 'targetable injectable' vector system would represent a potentially powerful technical advance allowing the implementation of a variety of strategies to accomplish gene therapy for cancer.
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1 |
1997 — 1999 |
Curiel, David T |
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. |
Sfv Mediated Oncogene Knockout For Chemosensitization @ University of Alabama At Birmingham
The development of drug resistance represents a major limitation of presently employed chemotherapeutic agents. In this regard, diverse mechanisms have been shown to be etiologic of the drug resistant phenotype. In many instances, the specific molecular basis of drug resistance has been defined. Mutational changes that lead to the drug resistant phenotype include modifications in the structure or level of topoisemerase, increased detoxification reactions, interference with delivery of cytotoxic drugs to intracellular targets,or overexpression of selected growth factor receptors. In addition, alterations affecting the regulation of the cell cycle and apoptosis are highly associated with drug resistance. Thus, based upon an understanding of the molecular basis ofdrug resistance, strategies may be proposed to correct the genetic lesions etiologic of the drug resistant phenotype. To this end, we have developed a novel method to achieve targeted oncoprotein ablation based on intracellular expression of a single-chain antibody. We have shown that this method can accomplish phenotypic alterations in tumor cells based upon downregulationof the target oncoprotein. In addition, we have demonstrated that the technique of anti-oncogene sFv-mediated knockout can enhance tumor cell chemosensitivity. It is thus our hypothesis that the sFv-mediated knockout technique can be developed as a means to achieve enhanced chemosensitization as a therapeutic modality for ovarian neoplasms of epithelial origin. In this proposal, we will thus determine the parameters that modulate chemosensitivity induced by intracellular expression of an anti-erbB-2 sFv. In addition, we will evaluate single chain antibodies targeting other gene products relevant to the pathogenesis of ovarian carcinoma and to explore their utility in accomplishing tumor cell chemosensitization in animal models relevant to human carcinoma of the ovary. It is evident that novel therapeutic modalities are needed for the successful treatment of epithelial ovarian carcinoma. We have demonstrated that sFv-mediated modulation of target oncogene products may allow enhanced sensitivity to chemotherapeutic agents. It is anticipated that the experiments described herein will allow the definition of th eoptimal biologic parameters relevant to implementation of this strategy. These studies would thus establish the basis for the development of a human clinicalprotocol for epithelial ovarian carcinoma based upon this novel approach.
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1 |
1998 — 2008 |
Curiel, David Terry |
T32Activity Code Description: To enable institutions to make National Research Service Awards to individuals selected by them for predoctoral and postdoctoral research training in specified shortage areas. |
Cancer Gene Therapy Training Program @ University of Alabama At Birmingham
ESCRIPTION (provided by applicant): Purpose and Program Characteristics: The primary goal of this training grant is to provide intensive training to predoctoral and postdoctoral fellows in the interdisciplinary field of Cancer Gene Therapy. This program will support physician scientists and Ph.D. scientists at the interface of basic and clinical science. Research training will be provided by faculty members representing basic science and translational research. Recognizing the current lack of individuals trained in cancer gene therapy, the overall goal of the program is to generate young academic investigators with a broad background and an excellent understanding of the integrated approach necessary to translate basic research into a clinical setting. The Gene Therapy Center at the University of Alabama at Birmingham offers a unique training resource for the development of young scientists who are likely to play an important role in bringing new technologies from the lab into a clinical setting in the coming years. The interdisciplinary and translational nature of this program provides a unique resource to expand the pool of young investigators who are qualified to carry out this research. Trainees: We request funding for three postdoctoral fellows and one predoctoral fellow. The research training period for postdoctoral fellows will have a duration of at least two years, while support for the predoctoral fellow will be for three years. We anticipate supporting a mix of Ph.D. and physician scientists at the postdoctoral level. The predoctoral trainee will be selected from those students pursuing the gene therapy subspecialty of the Molecular and Cellular Pathology graduate program. Selection of fellows for these training positions will be based on our assessment of the trainee's likelihood of establishing an independent research career in an area relevant to cancer gene therapy. Facilities: The Gene Therapy Center of the University of Alabama at Birmingham will be the major site of the training program. The training faculty occupies more than 20,000 feet of excellent quality, well-equipped laboratory space. As all of the faculty members acting as preceptors on this training grant are members of the Comprehensive Cancer Center, the trainees will have full access to the extensive resources of the Center. The university also has modern animal facilities supervised by veterinarians and run by trained animal care personnel working under NIH guidelines.
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1 |
1999 — 2002 |
Curiel, David T |
U19Activity Code Description: To support a research program of multiple projects directed toward a specific major objective, basic theme or program goal, requiring a broadly based, multidisciplinary and often long-term approach. A cooperative agreement research program generally involves the organized efforts of large groups, members of which are conducting research projects designed to elucidate the various aspects of a specific objective. Substantial Federal programmatic staff involvement is intended to assist investigators during performance of the research activities, as defined in the terms and conditions of award. The investigators have primary authorities and responsibilities to define research objectives and approaches, and to plan, conduct, analyze, and publish results, interpretations and conclusions of their studies. Each research project is usually under the leadership of an established investigator in an area representing his/her special interest and competencies. Each project supported through this mechanism should contribute to or be directly related to the common theme of the total research effort. The award can provide support for certain basic shared resources, including clinical components, which facilitate the total research effort. These scientifically meritorious projects should demonstrate an essential element of unity and interdependence. |
Genetic Modification of Dendritic Cells For Tolerance @ University of Alabama At Birmingham
The long-range goal of this project is to develop a novel and effective strategy for allograft tolerance induction. To accomplish this goal, we will create an "engineered veto cell" and apply it in a preclinical non-human primate model of kidney transplant to induce durable tolerance with non immunosuppressive treatment beyond the peritransplant tolerance induction period. This study is unique in that it uses novel gene transfer technology to exploit the veto effect to kill or inactivate recipient alloreactive cells. Our approach to tolerance induction combines genetic modification of donor dendritic cells (DC) with brief immunosuppressive treatment, a combination that builds upon our previous studies of tolerance induction with donor bone marrow in the rhesus model. The research plan will develop and utilize customized gene transfer methods to constitutively express TGF-beta1 in donor dendritic cells (dDC) in order to deliver within the microenvironment of T cell-dDC cross talk, a lethal signal to the responding donor-specific T cells. We propose that when used in conjunction with a short treatment with anti-thymocyte globulin, TGF-beta1 engineered dDC will effectively function as "ersatz veto cells" to induce clonal deletion and specific tolerance. A requisite condition for achieving our goals is the genetic modification of dDC with high efficacy and in a manner that does not impact unfavorably in their functionality as antigen presenting cells (APC). A variety of vector approaches have been explored to achieve effective transduction of DC to express heterologous genes relevant to this strategy and others. Available gene transfer methods, however, have represented a major limitation, as current generation vectors are generally of low efficiency and/or may be associated with significant toxicity for DC. For successful transplantation to clinical practice in transplantation and also for other immunological applications (e.g., autoimmune disease, cancer, and vaccines), there is a need to develop improved vector systems for genetically modifying DC. In this regard, we have developed methods to alter the tropism of adenoviral (Ad) vectors as a means to enhance their efficacy profile. We have demonstrated that an immunologic re-target approach allows re-routing of Ad vectors via the CD40 pathway of human DC with dramatic enhancements of efficiency and with beneficial effects in immune presentation function. In the proposed studies, the Ad TGF-beta1 DC cellular construct will be first optimized for in vitro reduction of allospecific cytotoxic T cell (CTL) responses in the monkey model. The final goal is proof of principal for the employment of TGF-beta1 dDC in a preclinical setting using the well-established kidney transplant model in rhesus macaques. Although we expect that these studies will yield novel approaches for induction of tolerance in the context of kidney transplantation, our general strategy might also be extended to the transplantation of other organs and tissues. In addition, the propose tolerance strategy will also be applicable to those recipients, who by virtue of high diphtheria sensitization or other causes, are not candidates for treatment with the potent diphtheria based anti-CD3 immunotoxin described in Projects 1 and 2. Finally, vector developments and analyses proposed herein may allow the use of tropism-modifier adenovirus as a gene delivery method for genetic modification of DC, and will elucidate key aspects of the immune response generated in vivo against Ad vectors.
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1 |
1999 — 2002 |
Curiel, David T |
P50Activity Code Description: To support any part of the full range of research and development from very basic to clinical; may involve ancillary supportive activities such as protracted patient care necessary to the primary research or R&D effort. The spectrum of activities comprises a multidisciplinary attack on a specific disease entity or biomedical problem area. These grants differ from program project grants in that they are usually developed in response to an announcement of the programmatic needs of an Institute or Division and subsequently receive continuous attention from its staff. Centers may also serve as regional or national resources for special research purposes. |
Gene Therapy Program @ University of Alabama At Birmingham
DESCRIPTION: (Applicant's Description) Defining new therapeutic paradigms for patients with ovarian cancer is of critical importance, given that this is a uniformly fatal disease for the vast majority of women afflicted by it. Many human cancers, including ovarian cancer, are now recognized to be the result of accumulated genetic events, which culminate in the transformed malignant phenotype. Therefore, it is rational to investigate the utility of various gene therapy strategies as a means to improve clinical outcome. Of note, distinct gene therapeutic initiatives have been rapidly translated into clinical trials for ovarian cancer, several of which have been conducted by the investigators of this proposal. The implementation of these trials, however, has revealed limitations that relate primarily to the inadequacy of current vector systems to achieve efficient and specific tumor cell targeting and to the logistics and morbidity associated with invasive assessment of in vivo gene transfer. It is our hypothesis that the therapeutic utility of cancer gene therapy approaches for ovarian cancer will be substantially improved by the employment of a new generation of vector systems capable of efficient and tumor cell-specific transduction and by the application of novel non-invasive gene transfer imaging techniques. To address this hypothesis, we propose: (1) to develop and validate a genetically modified adenovirus that augments tumor targeting and achieves an enhanced anti-tumor effect in the context of ovarian cancer, (2) to develop and validate, in the context of ovarian cancer, a gene transfer imaging methodology based on radionuclide imaging of genetically transferred receptors, (3) to prepare a Good Manufacturing Practice (GMP) adenoviral vector for a human clinical gene therapy trial in the context of ovary cancer that is genetically modified to augment tumor tropism and that encodes both a therapeutic gene and a receptor for gene transfer imaging, and (4) to conduct a phase I/II trial in the context of ovary cancer utilizing a GMP adenoviral vector that is genetically modified to augment tumor tropism and that encodes both a therapeutic gene and a receptor for gene transfer imaging. Success in these endeavors will overcome the obstacles that are currently preventing gene therapy from reaching its potential as a clinically relevant option for ovarian cancer. Pursuant to this SPORE application, completion of these objectives will allow for establishment of gene therapy as a novel and effective treatment paradigm for ovarian cancer.
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1 |
1999 — 2000 |
Curiel, David T |
N01Activity Code Description: Undocumented code - click on the grant title for more information. |
Novel Technologies For Noninvasive Detection, Diagnosis @ University of Alabama At Birmingham
neoplasm /cancer diagnosis; neoplasm /cancer therapy; technology /technique development; transfection /expression vector
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1 |
2000 — 2004 |
Curiel, David T |
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. |
Dendritic Cell Specific Vector @ University of Alabama At Birmingham
DESCRIPTION: (derived from the applicant's abstract) This proposal explores the utilization of adenoviral vectors targeted dendritic cells (DC) for the expression of a tumor specific antigen as a means to induce maturation and antigen presentation of these cells to elicit a primary antitumor response. The key element of this study is the CD40 targeting and simultaneous stimulation of the DCs using a modified adenoviral vector possessing altered tropism. The efficacy of the vector transduced DC-based vaccination will be assessed. The validity of this proposal is based on numerous recent publications advancing the progress towards vaccination-based adjuvant therapies using antigen primed DCs and the development and understanding of the role that DCs play in the presentation of antigen to elicit cellular and humor immune responses. Ultimately, advances made during this study will help in translating the current knowledge into clinical practice for the treatment of cancer by inducing a tumor-specific immune response.
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1 |
2000 — 2008 |
Curiel, David Terry |
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. |
Replicative Adenoviruses With Enhanced Infectivity @ University of Alabama At Birmingham
DESCRIPTION (provided by applicant): Conditionally replicative adenoviruses (CRAds) represent novel therapeutic agents which have been recently applied in the context of cancer therapy. Indeed, recent reports investigating CRAds in the context of squamous cell carcinomas of the head and neck represent the first true clinical efficacy results achieved in human clinical cancer gene therapy to date. Clearly, the application of CRAds to cancer of the ovary thus represents a highly desirable endeavor whereby these promising agents may be evaluated in this desperate disease context. Of note, an initial clinical trial has demonstrated the feasibility of utilizing CRAds in an ovarian cancer clinical context. We postulate that the limited clinical effect noted in this initial trial may be related to deficiencies in two key biologic principles that mitigate CRAd efficacy. These important characteristics include (1) the ability of the virus to infect target tumor cells and to infect laterally post-replication and (2) the ability of the virus to replicate specifically within the tumor target. Addressing deficiencies in these areas will likely enhance the therapeutic index of CRAds in the context of ovarian cancer and is the basis of this proposal. The foregoing considerations have established several key principles with respect to our overall strategy to develop the optimal CRAd agent for carcinoma of the ovary. To reiterate, CRAd efficacy is clearly predicated upon the achievement of effective primary and lateral tumor cell infection and upon the achievement of tumor-specific replication. We hypothesize that the incorporation of infectivity enhancement and replication specific properties that address deficiencies in these key areas in current CRAd systems will facilitate the development of a CRAd agent with an enhanced therapeutic index for treatment of patients with ovarian carcinoma. Support via this competitive renewal will allow construction and validation of advanced generation CRAds that will combine new approaches to achieve enhanced infectivity and tissue specific replication. As well, proof-of principle studies in model systems will establish the rationale for an advanced human clinical trial for cancer of the ovary with derived advanced generation CRAd agents.
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1 |
2001 — 2004 |
Curiel, David T |
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. |
Endothelial Gene Delivery For Pulmonary Hypertension @ University of Alabama At Birmingham
Pulmonary vascular disease constitutes a major cause of mortality, morbidity and healthcare expenditure. Although the underlying causes are mutifactorial, this condition is generally characterized by increased pulmonary vascular resistance, which develops on the basis of both vasoconstriction and arterial vascular remodeling. Within this context, pathobiologic changes in the pulmonary endothelium are understood to playa central role in the development of both of these characteristic disease features. Studies of pathologic material from patients have demonstrated dysregulated expression of factors relevant to vasomotor tone and vascular wall remodeling. It is clear. however, that a fuller understanding of these pulmonary endothelial abnormalities will be required to allow rational interventional strategies. In this regard, methods allowing specific and directed perturbations in gene expression profiles within pulmonary endothelium would potentially provide unique insight into disease pathophysiology. On this basis, it would be desirable to develop technologies to achieve the goal of transient alterations in gene expression patterns selectively within pulmonary endothelium. Such gene-based methods would allow rational analysis of candidate genes with respect to their role in the pathobiology of this condition. Of note, these same methods would potentially allow directed therapeutic interventions. We have recently developed methods to alter the tropism of Ad vectors. These modifications have allowed the achievement of cell-specific gene delivery to pulmonary vascular endothelium in vivo after systemic vascular administration of the vector. We will use these modified Ad vectors to deliver relevant genes to the pulmonary endothelium in rat models of pulmonary hypertension and thrombosis, as a means to discover new insights into the pathophysiology of pulmonary hypertension. In these analyses we will asses the impact of gene delivery on pulmonary vascular pressures, vascular remodeling and thrombosis. In addition, it is our hypothesis that we can further improve the targeting properties of adenoviral vectors by genetic means, thereby improving the efficiency and specificity for I pulmonary endothelial markers and generating agents of potential clinical utility.
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1 |
2002 — 2006 |
Curiel, David T |
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. |
Enhanced Crad For Pancreatic Cancer @ University of Alabama At Birmingham
Overall survival among patients affected with pancreatic carcinoma has remained essentially static despite the availability of contemporary surgery and chemotherapy modalities. This fact has argued strongly for the development of novel therapeutic approaches for pancreatic cancers. In this field, conditionally replicative adenoviruses (CRAds) agents offer a novel and potent modality to approach cancer of the pancreas. However, pancreatic cancer cells are especially resistant to infection by adenovirus, thus limiting the capability of CRAd agents to achieve effective lateralization within the tumor. In addition, the absence of promoter elements that demonstrate selective inducibility in pancreatic cancer cells has hampered the construction of CRAd agents with desired replicating specificity, the key to the achievement of acceptable therapeutic index. In this proposal, we construct enhanced CRAds to overcome these CRAd limitations. In the first instance, we have developed methods to alter the tropism of the adenovirus, thereby achieving infectivity enhancement for tumor cells. These maneuvers have dramatically improved the infectious potency of adenovirus onto pancreatic cancer tumor targets. As well, we have identified and characterized two novel promoter elements, which exhibit selective activity in pancreatic tumor targets. Here, we have shown that the cyclooxygenase-2 (cox-2) and midkine (MK) promoters exhibit the "tumor ON / liver OFF" expression profile, which is critical for their utility in an adenoviral context. These findings directly address the limits of those agents for their application for pancreatic cancers but also offer the potential to derive CRAd agents for this target disease. Based on those considerations, it is clear that the combination of infectivity enhancement and cox-2 and/or MK controlled replication will offer the derivation of a CRAd agent with characteristics predicting its utility for cancer of the pancreas. The utility of these vectors will be established from the toxicological and anti-tumor effect standpoints.
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1 |
2005 — 2009 |
Curiel, David Terry |
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. |
Core--Viral Vector @ Columbia University Health Sciences
The Vector Development Core (Core B) is a shared facility that will provide vector reagents to facilitate the hypothesis-driven studies proposed in Project 1 (Fisher) relating to prostate cancer, Project 2 (Dent) investigating malignant glioma and Project 3: (Curiel) studying ovarian cancer. In this regard, all of these projects employ recombinant adenovirus vector systems for in vitro and in vivo studies. The core will construct vectors for each of these projects. Additional adenoviral vectors will be prepared which utilize progression elevated gene-3 promoter to drive virus replication and concomitantly express mda-7/IL-24 for Project 1 ;(Fisher) and after validation for use in Project 2 (Dent) and Project 3 (Curiel), will also be produced. Specific requirements for non-replicative adenoviruses, including those expressing bcl-2 and bcl-xL Project 1 (Fisher) and Project 2 (Dent), will be provided by Core B. Project 3 (Curiel) will produce and characterize "complex mosaic" adenoviruses with enhanced cancer infectivity tropisms for enhanced cancer-specific expression of mda-7/IL-24. Once these viruses are characterized, this Core will provide these vectors for use in prostate cancer (Project 1: Fisher) and in malignant gliomas (Project 2: Dent). Core B will also serve to validate the identity, function, and quality of the various viral vector reagents to provide uniform quality controlled materials for the various projects. The Vector Production Core will provide preparative amounts of validate vectors and additional vectors as required by specific projects. The concentration of these activities in a shared resource context will enable these reagents to be derived in an efficient and cost-effective manner and will capitalize on programmatic expertise in this area.
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0.939 |
2005 — 2009 |
Curiel, David Terry |
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. |
Capsid-Labeled Adenovirus For Virotherapy Monitoring @ University of Alabama At Birmingham
[unreadable] DESCRIPTION (provided by applicant): Conditionally replicative adenoviruses (CRAds) represent an important novel approach for cancer therapy. CRAd agents are designed to specifically replicate in and kill tumor cells to yield an effective yet safe therapeutic outcome. Although their great potential have awarded them rapid translation into human clinical trials where the safety of CRAds have been clearly highlighted, little data have been obtained with regards to the critical functions of CRAds efficient infection of tumor cells, tumor-specific replication, and lateral spread. Without deeper understanding of the nature of these replicative agents especially in a patient context, further development of CRAds would be greatly hindered. The crux of a general dilemma in the virotherapy field is lack of a monitoring system compatible with replicative agents. Unfortunately, the bulk of gene therapy vector detection schemes have been specifically designed for the assessment of gene expression. These modalities are not suitable for the monitoring of CRAds because of the very nature of their program to kill infected tumor cells, a concept at odds with the notion of viable transgene expression. Furthermore, reporters by themselves cannot accurately depict the underlying level of replication as well as true physical distribution of viral progeny, two crucial functions of replicative agents. Clinical trials to date have had to rely on traditional histological analysis of biopsy specimens which are error-prone and cannot portray the multiplicative nature of CRAds. The ideal monitoring system for CRAds should embody the following features: (1) report the level of viral replication, (2) allow direct detection of viral spread, (3) permit dynamic detection of viral activity, (4) minimally perturb replication and spread efficiency of the virus, and (5) possess the power for noninvasive detection. To address this issue, we hypothesized that a genetic adenovirus labeling system using a structural reporter fusion protein would dynamically represent viral replication and spread. We propose adenovirus capsid labeling with IX-EGFP and core labeling with mu-EGFP, V-EGFP, and VII-EGFP. It is evident that a monitoring system for CRAds is needed for advancing the field. A genetic structural labeling system for adenovirus would offer noninvasive dynamic detection of replication and spread. Not only would this system be indispensable in developing advanced CRAds, it would also be applicable for monitoring CRAd therapy in patients. [unreadable] [unreadable]
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1 |
2005 — 2009 |
Curiel, David Terry |
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. |
Mda-7/Il-24: Therapy of Ovarian Cancer @ Columbia University Health Sciences
Conventional therapies, including surgery coupled with chemotherapy, have not resulted in significant improvements in overall survival rates for ovarian cancer. In this context, novel therapeutic modalities are required for this disease. Gene therapy represents a rational and potentially effective approach for ovarian carcinoma. To this end, a number of distinct cancer gene therapy approaches have been developed for ovarian carcinomas, which are predicated upon direct gene delivery to tumor cells. To achieve effective gene delivery, recombinant adenoviral vectors have been employed based on their superior in vivo efficacy characteristics. Whereas adenoviral vectors are understood to exhibit superior levels of in vivo gene transfer compared to available alternative vector systems, present levels may nonetheless be sub optimal for ovarian cancer gene therapy applications. On this basis, we have endeavored to modify the tropism of adenoviral vectors to achieve enhanced gene delivery to tumor cells as a means to improve the overall feasibility of these cancer gene therapy strategies. We have achieved adenovirus retargeting using genetic strategies to alter tropism. In the present proposal, we seek to further develop these approaches to allow the development of adenoviral vectors, which allow maximal enhancement of gene delivery to ovarian cancer cells. With this objective in mind, we have developed a highly novel strategy of Ad tropism alteration whereby multiple, and distinct, retargeting ligands are genetically incorporated into the viral capsid "complex mosaic". The ability to achieve biological access to multiple cellular entry pathways potentially allows for maximized gene delivery to tumor cells deficient in the primary Ad receptor. We hypothesize that these "complex mosaic" adenoviral vectors will allow the achievement of an improved therapeutic index in the context of mda-7/IL-24 based gene therapy approaches for ovarian carcinoma. In principle, these "complex mosaic" adenoviral vectors should also prove beneficial for treatingother cancers, including prostate cancer (Project 1: Fisher) and malignant glioma (Project 2: Dent). This strategy may also be amenable to increase the infectivity of adenoviral vectrs in which replication is restricted to cancer cells by the progression elevated gene-3 promoter and mda-7/IL-24 is expressed in a different region of the adenoviral genome (proposed in Project 1:Fisher). This vector would provide a direct means of enforcing replication in cancer cells resulting in their cytolysis with concomitant production of mda-7/IL-24. Such a "complex mosaic" adenoviral vector would offer profound cancer specific activity, not only in ovarian, but also in other cancers and be of direct relevance to the theme of this program project, to increase the translational aspects of mda- 7/IL-24 for cancer therapy.
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0.939 |
2007 — 2011 |
Curiel, David Terry |
R21Activity Code Description: To encourage the development of new research activities in categorical program areas. (Support generally is restricted in level of support and in time.) R33Activity Code Description: The R33 award is to provide a second phase for the support for innovative exploratory and development research activities initiated under the R21 mechanism. Although only R21 awardees are generally eligible to apply for R33 support, specific program initiatives may establish eligibility criteria under which applications could be accepted from applicants demonstrating progress equivalent to that expected under R33. |
Capsid-Incorporation of Hiv Antigens as a Novel Adenovirus Hiv Vaccine Approach
DESCRIPTION (provided by applicant): Despite the many potential advantages of Ad vectors for vaccine application, full utility of current Ad vaccines may be limited by the host anti-vector immune response. Specifically, the anti-Ad humoral immunity abrogates the effectiveness of subsequent administrations of the Ad vector, confounding expression of the encoded transgene, and thus practically restricting the gains that might be accrued via booster effect. In order to exploit the inherent antigenicity of the Ad vector we have developed a vaccination approach based on incorporation of the immunizing antigen epitope directly into the Ad capsid. This novel paradigm is based upon Ad presenting the antigen as a component of the capsid rather than an encoded transgene. Incorporation of immunogenic peptides into the Ad capsid offers potential advantages. Most noteworthy, the processing of the capsid incorporated antigen via the exogenous pathway should result in a strong humoral response akin to the response provoked by native Ad capsid proteins. In addition, since anti-Ad capsid responses are augmented by repeated vector administration, immune responses against antigenic epitopes that are part of the Ad capsid should be augmented by repeated administration as well, thus allowing boosting. These considerations suggest that this novel capsid-incorporated antigen approach may offer exciting potentials to realize Ad-based vaccine strategies that circumvent the major limitations associated with Ad vectors. Critical to the realization of this approach is to define the optimal configuration of antigen in the adenoviral capsid context. To this end, we have established several key technologies that will enable us to reach our goal. In particular, we have developed the means to incorporate heterologous peptide epitopes within the surface-exposed domains of the major Ad capsid protein hexon. We have begun to determine the size and structural factors that predicate functional utility of these domains in the hexon. In addition, we have developed the means to apply cryoelectron microscopy (cryoEM) single particle reconstruction methods to allow us to explore the capsid-incorporated peptide localization with unprecedented, subnanometer resolution. Based on these technologies, we will be able to establish the critical correlates between antigen locale/accessibility within the capsid context and vaccine efficacy. On the basis of these established feasibilities, we hypothesize that Ad vectors can be created with novel capsid-incorporated antigens that can serve as vaccine agents against HIV in animal models. CryoEM-guided capsid design will be applied to develop an optimized vector with optimal anti-HIV immunization. We envision that our proposed structural studies will provide complementary information to in vitro assays and biological readouts and thereby will enable us to understand the functional determinants of incorporated HIV epitopes. This project will design new and innovative methodologies to create HIV vaccines, in hopes of preventing the spread of HIV disease.
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1 |
2007 — 2011 |
Curiel, David Terry |
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. |
Monitoring of Advanced Virotherapy For Ovarian Cancer @ University of Alabama At Birmingham
DESCRIPTION (provided by applicant): The mandate to realize surrogate endpoints in the context of human clinical trials with conditionally replicative adenovirus (CRAd) agents has thus suggested the utility of imaging analysis. In theory, this type of assay could provide critical information with respect to CRAd replication, amplification, and localization. These types of studies could thereby provide key insight into CRAd function in a human clinical context. To address this key issue we have developed a novel labeling system based upon structural incorporation of imaging reporter motifs into the adenovirus capsid. We hypothesize that this approach will provide the basis of deriving useful surrogate endpoint read-outs allowing valid imaging analysis of a human CRAd intervention for carcinoma of the ovary. In this regard, we have recently demonstrated our ability to incorporate complex heterologous fusion proteins at adenovirus minor capsid sites. Proof-of-principle studies with direct incorporation of fluorescent or SPECT/PET-linked imaging motifs validated that functional reporters could be incorporated within the adenovirus capsid and provide imaging signal. These demonstrated capacities provide the framework for understanding if such imaging detection systems can be employed in combination via capsid incorporation of dual modality reporter fusions which embody the capacity for SPECT/PET and fluorescent monitoring. Furthermore, the validation of this approach in relevant animal model systems will provide the rationale for clinical translation of this strategy as a means to derive useful surrogate endpoints of CRAd function in the context of human clinical employ. On the basis of these considerations, the Specific Aims of our proposal are: Specific Aim #1: To develop candidate ovarian cancer CRAd agents which embody capsid incorporated dual modality structural reporter fusions allowing dynamic fluorographic and SPECT/PET-based imaging analysis. Specific Aim #2: To employ the derived dual imaging modality CRAds in orthotopic murine models of carcinoma of the ovary to validate SPECT/PET and fluorographic imaging analysis of CRAd functionally. Specific Aim #3: To employ the optimal imaging modality for CRAd-based imaging analysis in the context of our next generation CRAd agent to be employed in a Phase I human clinical trial for carcinoma of the ovary to validate the analytical value of the structural reporter fusion method for CRAd monitoring.
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1 |
2008 — 2009 |
Curiel, David Terry |
P41Activity Code Description: Undocumented code - click on the grant title for more information. |
Visualizing Gold-Labeled Adenoviral Vectors Inside Cancer Cells @ University of California San Diego
Ablation; Adenoviral Vector; Adenovirus Vector; Animal Experiments; Au element; CRISP; Cancer Treatment; Cancers; Capsid; Computer Retrieval of Information on Scientific Projects Database; Coupled; Coupling; Deep; Depth; Disease; Disorder; Drug Delivery; Drug Delivery Systems; Drug Targeting; Drug Targetings; Dysfunction; Electromagnetic, Laser; Electromagnetic, Microwave; Experiments, Animal; Fever; Focused Ultrasound; Focused Ultrasound Therapy; Functional disorder; Funding; Gene Therapy Vectors; Gene Transduction Agent; Gene Transduction Vectors; Gold; Grant; Heating; High Power Focused Ultrasound; Hyperthermia; Image; Institution; Investigators; Label; Lasers; Light; Location; Malignant Cell; Malignant Neoplasm Therapy; Malignant Neoplasm Treatment; Malignant Neoplasms; Malignant Tumor; Methods; Microwaves; NIH; Nanoscale Science; Nanotechnology; National Institutes of Health; National Institutes of Health (U.S.); Normal Cell; Numbers; Permeability; Photoradiation; Physiopathology; Predisposition; Pyrexia; Radiation, Laser; Research; Research Personnel; Research Resources; Researchers; Resources; Source; Susceptibility; System; System, LOINC Axis 4; Therapeutic; Therapy (Focused US); Tumor Burden; Tumor Cell; Tumor Load; United States National Institutes of Health; Virion; Virus Particle; adeno vector; adenovector; anticancer therapy; base; cancer cell; cancer therapy; clinical applicability; clinical application; clinical hyperthermia; coat (nonenveloped virus); combination gene therapy; combinatorial; design; designing; disease/disorder; febrile; febris; hyperthermia treatment; imaging; malignancy; microwave electromagnetic radiation; microwave radiation; nano particle; nano scale Science; nano tech; nano technology; nanoparticle; nanotech; neoplasm/cancer; neoplastic cell; novel; pathophysiology; tumor
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0.946 |
2009 — 2010 |
Curiel, David Terry |
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. |
Development of Double-Targeted Vectors For Long-Term Vascular Expression in Vivo
DESCRIPTION (provided by applicant): To create a clinically relevant vector for vascular-based diseases, we must advance three pivotal efforts. First, it is critical to formally correct an established disease phenotype via systemic administration of a vector that de-targets the liver and thus reduces adenovirus (Ad) associated liver toxicity. Second, development of a safer, less immunogenic Ad vector capable of long-term gene expression will be required to realize human clinical translation of a gene therapy for diseases involving endothelium. To this end, we will combine our current targeting technology with "gutless" Ad vectors, known to provide long-term gene expression in vivo as a result of their reduced immunogenicity. Finally, to further increase the clinical utility of our vector, we propose the development of a single component targeted Ad vector compatible with large scale production and FDA approval. In this regard, we have recently developed the methodology to incorporate functional single chain antibody (scFv) targeting moieties into the Ad capsid resulting in highly selective gene delivery to specific cell surface markers. This key technological development will allow us to extend our established targeting paradigm to a single component vector with genetically incorporated targeting. We hypothesize that the combination of these technologies will allow the realization of a clinically relevant gene therapy vector for diseases requiring systemic injection and significant pulmonary endothelium involvement. Our specific aims are thus: Specific Aim 1: To evaluate the ability of double-targeted Ad vectors to correct disease involving the pulmonary endothelium. Specific Aim 2: To create targeted gutless Ad vectors capable of long term gene expression in the pulmonary endothelium. Specific Aim 3: To create a single-component, targeted gutless Ad employing genetically incorporated recombinant antibodies to target the pulmonary endothelium. The successful completion of these specific aims will provide important and timely data regarding the efficacy of endothelial targeted gene therapy and lay the groundwork for other endothelial based diseases. Further, our studies will clearly establish therapeutic gain associated with decreased vector immunogenicity and long term gene expression. Finally, the studies proposed herein will provide a critical step toward a safer, single component vector compatible with large-scale production and clinical translation. PUBLIC HEALTH RELEVANCE: Adenovirus (Ad) based vectors are widely used in clinical trials due to their well characterized biology and positive safety profile. However, the majority of clinical applications employing Ad based vectors utilize local administration due to limitations associated with first generation Ad vectors. In this proposal, we seek to engineer a safer, less immunogenic Ad vector capable of long-term gene expression.
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1 |
2010 — 2014 |
Curiel, David Terry |
P50Activity Code Description: To support any part of the full range of research and development from very basic to clinical; may involve ancillary supportive activities such as protracted patient care necessary to the primary research or R&D effort. The spectrum of activities comprises a multidisciplinary attack on a specific disease entity or biomedical problem area. These grants differ from program project grants in that they are usually developed in response to an announcement of the programmatic needs of an Institute or Division and subsequently receive continuous attention from its staff. Centers may also serve as regional or national resources for special research purposes. |
Development of Oncolytic Adenovirus Targeting Pancreatic Cancer Stem Cells @ University of Alabama At Birmingham
The poor response of existing treatnnent options for unresectable pancreatic cancer is the result of a subset of cancer cells exhibiting chemoresistance. Evidence has shown that the capacity of a tumor to grow and propagate is dependent on this small subset of cells, identified as pancreatic cancer stem (or stem-like) cells (CSCs). Due to their vital role in tumor maintenance and the formation of metastases, CSCs are integral targets to treat pancreatic cancer, as current therapeutic regimens cannot effectively treat this cell population because of its intrinsically resistant nature. Adenovirus (Ad)-based oncolytic viruses have unique advantages for targeting and eradicating pancreatic CSCs along with inclusively targeting the general pancreatic cancer cell population and thereby represent a promising novel therapeutic strategy. Ad replication specificity can be achieved by placing viral replication genes under the control of tumor-specific promoters, thereby generating conditionally replicative adenoviruses (CRAds). The keys to developing a pancreatic cancer CRAd that inclusively targets CSCs are: 1) effective transduction and 2) replication specificity of pancreatic cancer cells and CSCs. We have developed innovative strategies and breakthrough technologies which can be rapidly applied for designing such CRAds. Our advanced infectivity enhancement strategies allow efficient transduction of pancreatic cancer cells and CSCs. To confer replication specificity, we have developed the CXCR4 promoter as a CRAd replication control element. CXCR4 expression Is a signature of pancreatic cancer and CSCs. SA 1: To construct infectivity-enhanced, CXCR4 promoter controlled CRAds for pancreatic cancer, inclusively targeting pancreatic CSCs. SA 2: To validate the efficacy of CRAds with in vitro models of chemoresistant pancreatic CSCs. SA 3: To demonstrate efficacious targeting and replication of CXCR4 CRAds with in vivo models of pancreatic cancer. SA 4: To validate the efficacy and specificity of pancreatic CSC-targeted CRAds with primary CSCs derived from human patient tumors. Development of an effective therapy for the advanced pancreatic cancer Is predicated on efficiently targeting and eradicating pancreatic CSCs, which represent the chemoresistant nature of this disease. The strategy proposed herein, of exploiting CRAd agents based on the CXCR4 axis to inclusively target pancreatic CSCs offers a promising treatment approach. On the basis of these studies, we will thus understand the direct utility of our novel system for clinical application and will facilitate a rapid clinical translation. We will submit an IND application concurrently with this proposal to rapidly translate the agents herein for clinical trial.
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1 |
2011 — 2015 |
Curiel, David Terry Mathis, J. Michael |
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. |
Targeted-and Image-Based Adenovirus Cancer Therapeutic Vectors
DESCRIPTION (provided by applicant): We hypothesize that a novel capsid incorporation approach will allow comprehensive in vivo monitoring of a cancer retargeted vector, and permit the assessment of our novel vector engineering strategy to enable targeted therapy of disseminated neoplastic disease. By investigating adenovirus (Ad) vector design, we anticipate the new information gained from this initiative will demonstrate the utility of our highly novel imaging approach, and validate the specificity of retargeted tumor transduction. The goal of this application is to develop a multi-functional Ad vector that combines both imaging and targeted therapy. To fulfill this goal, we will combine three unique technologies to advance therapeutic targeting of disseminated neoplastic disease: cancer-specific retargeting, liver-detargeting, and non-invasive imaging. We have developed Ad vectors capable of cell-specific targeting, by incorporating targeting single chain antibodies (scFv) into the viral capsid in combination with hexon modification for liver detargeting and evasion of pre-existing immunity. Our innovative Ad targeting approach provides a novel way to circumvent the problem of structural and biosynthetic incompatibility between Ad and complex targeting ligands such as scFv, and could facilitate Ad targeting to a wide variety of clinically important cell populations using novel targeting ligands including recombinant antibodies and growth factors The goal of this application is to develop a multi-functional Ad vector that combines both imaging and targeted therapy. To fulfill this goal, we will combine three unique technologies to advance therapeutic targeting of disseminated neoplastic disease: cancer-specific retargeting, liver-detargeting, and non-invasive imaging. We have developed Ad vectors capable of cell-specific targeting, by incorporating targeting single chain antibodies (scFv) into the viral capsid in combination with hexon modification for liver detargeting and evasion of pre-existing immunity. Our innovative Ad targeting approach provides a novel way to circumvent the problem of structural and biosynthetic incompatibility between Ad and complex targeting ligands such as scFv, and could facilitate Ad targeting to a wide variety of clinically important cell populations Our innovative use of a structural fusion protein incorporating metallothionein into adenovirus pIX gene provides the non-invasive imaging advantages of detecting physical biodistribution and spread of Ad vectors after administration that is not possible employing a reporter gene. Further, the ability to noninvasively observe Ad function on a whole-body level allows the possibility of detecting virus dissemination outside the tumor site(s) for monitoring clinical safety. By combining these modifications in Ad vector design, we anticipate the new information gained from this initiative will demonstrate the utility of our highly novel imaging approach, and validate the specificity of targeted tumor transduction. This work addresses the significant unmet need of new therapies for advanced stage metastatic cancer. The novel design of the proposed imaging approach is distinct from previously described vector imaging modalities, which have been based exclusively on monitoring the expression of reporter genes. In addition, our combined technologies have broad impact on the diverse field of gene therapy, in which the ability to achieve both cell- specific transduction and monitoring is universally required. To accomplish these goals, we have assembled an outstanding team of investigators with expertise in Ad vector design and construction, adenovirus retargeting approaches, preclinical models using Ad vectors, non-invasive PET and SPECT imaging, medical physics, colon cancer oncology and clinical trials, clinical pathology, biostatistics, and preclinical toxicology.
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1 |
2012 — 2013 |
Curiel, David Terry Shoemaker, Charles Bix |
R21Activity Code Description: To encourage the development of new research activities in categorical program areas. (Support generally is restricted in level of support and in time.) |
Motor Neuron-Targeted Adenovirus Antidotes For Botulism @ Tufts University Boston
DESCRIPTION (provided by applicant): Botulism is caused by Clostridium botulinum neurotoxin (BoNT), a CDC Category A biodefense threat agent for which no antidote exists. BoNT is the most potent toxin known and can be produced with relative ease. If large numbers of people were exposed to even a small dose of this toxin, they would become paralyzed and require assisted breathing which could easily overwhelm the limited supplies of respirators. We have developed small protein agents that, when expressed within intoxicated neurons, promote rapid degradation of BoNT proteases and improve rates of recovery from intoxication. We call these biomolecule agents targeted F-boxes (TFBs) because they consist of a 15 kDa F-box domain fused to a 14 kDa camelid VHH domain with binding specificity for a BoNT protease. Here we propose to develop adenovirus-based botulism antidotes that will specifically target motor neurons and lead to cytosolic expression of a TFB agent within intoxicated neurons. Adenoviruses (Ads) are ideal vehicles for delivery of TFBs to motor neurons for several reasons. First, Ad does not integrate their genome into infected cells and Ad vectors have been produced that are non-replicating and safe for therapeutic use. Second, Ad can be modified to dramatically reduce the normal tropism (liver) while increasing infection of a selected cell population. Thirdly, Ad vectors can be modified to produce a transient burst of transgene expression lasting only a few weeks after which the infected cells revert to normal. Finally, Ads have been found to infect motor neurons leading to transgene expression and, surprisingly, BoNT intoxicated motor neurons are much more efficiently infected than normal neurons. In the R21 phase of this proposed project, we will develop an adapter protein that will block normal Ad tropism while imparting tropism for motor neurons by employing a neuron-specific receptor binding domain (RBD) that derives from BoNT. Recombinant Ad will be engineered to express a TFB agent that targets the protease from BoNT serotype A (BoNT/A) for degradation and will be pre-treated with the neuron-targeting adapter. This modified Ad will then be tested as a botulism antidote by local injection into BoNT/A intoxicated muscles in mice. If successful, in the R33 phase we will make additional modifications to the Ad vector to further improve specificity for motor neurons and obviate the need for an adapter protein. We will also engineer an Ad antidote for another BoNT serotype, BoNT/B. These Ad vectors will be tested for the ability to accelerate recovery from botulism paralysis in mice following a systemic administration of the therapeutic agent. If successful, similar agents can be readily developed for all seven known BoNT serotypes by simply replacing the VHH domain of the TFBs with others having specificity for different BoNT proteases. In addition, the neuron-targeted Ad vehicles we develop may have applications in other important neuronal pathologies.
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0.942 |
2012 — 2013 |
Curiel, David Terry |
R21Activity Code Description: To encourage the development of new research activities in categorical program areas. (Support generally is restricted in level of support and in time.) |
A Gene-Therapy Based Functional Restoration of Salivary Glands
DESCRIPTION (provided by applicant): Xerostomia is a frequent side effect of external beam radiation therapy (XRT) for head and neck cancer and is not amenable to palliative therapies. This medical and quality of life issue thus remains an unmet healthcare need for the increasing number of individuals who will receive XRT treatment for this disease of increasing incidence. A recent approach for this disorder is based upon gene therapy restoration of salivary gland function. In this strategy, a recombinant adenoviral vector (Ad) is employed to deliver the aquaporin gene to epithelial cells of the salivary glands via direct intra-ductal instillation. Preliminary phase I human clinical trials at the NIH/National Institute of Dental Research have clearly shown that this approach can restore salivary flow with an amelioration of xerostomia symptoms. Based on these promising findings, it is logical to hypothesize that vector design advancements to improve further aquaporin gene delivery could enhance functional restoration of the salivary gland. In addition, the achievement of long term expression of the aquaporin gene would extend the symptomatic benefits of this gene therapy approach for xerostomia. Herein we will seek to address these vector aspects of xerostomia gene therapy to enable its full patient benefit value to be realized as a translational therapeutic. In the first regard, we hve developed tropism modified Ad which embody dramatically enhanced target cell infectivity. We hypothesize that these infectivity enhanced Ad can accomplish improved aquaporin gene transfer for ductal epithelial cells of the salivary gland thereby augmenting functional restoratio. In addition, we have developed high capacity gutless adenoviral vectors (HCAd) that allow long term gene expression. We hypothesize that this vector capacity will allow us to maintain long term aquaporin expression levels to allow for sustained functional reconstitution of the salivary gland. The studies we propose herein will establish the full rationale for the ultimate development of a novel gene therapy approach for XRT-induced xerostomia based on aquaporin delivery to the ductal epithelium of the salivary gland. We also note that our approach is clearly generalizable for the diverse pathobiologies etiologic of xerostermia (Sjorgen's syndrome, chronic sialadentitis, etc.). Data acquired herein thru this R21 Pilot Award can thus be leveraged for the acquisition of NCI translational mechanisms (NExT, etc.) to fully realize our bench-to-bed translational goals.
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0.948 |
2017 — 2018 |
Curiel, David Terry |
R21Activity Code Description: To encourage the development of new research activities in categorical program areas. (Support generally is restricted in level of support and in time.) |
Gorilla Adenovirus Zika Vaccine For Humans
Zika virus (ZIKV) is an arthropod-borne flavivirus that has spread rapidly across the Americas with 35 countries and territories reporting active viral transmission over the past year; this epidemic has prompted the WHO to declare ZIKV a worldwide public health emergency. An effective vaccine for ZIKV would be a cost-effective intervention that limits clinical disease and controls the spread of the infection since there is no antiviral treatment available. The goal of this proposal is to design novel ZIKV vaccine candidates and conduct initial preclinical testing in newly-generated mouse models of infection and disease. In this regard, vector-based vaccine systems offer key advantages including their robust efficiency to elicit humoral and cellular immunity by mimicking infection while producing ZIKV antigens de novo. Adenovirus (Ad) vectors have emerged as one of the most promising vaccine platforms that stimulate both innate and adaptive immune responses, and have been evaluated extensively in clinical trials in the cancer vaccine and infectious disease fields. The use of simian Ad species isolated from chimpanzee and gorilla offers a unique ability to bypass pre-existing immunity to human Ad. Here, we will test whether gorilla-based Ad (GAd) vectors expressing soluble envelope (E) protein or intact subviral particles (SVPs) can elicit neutralizing humoral and cellular immune responses and protect against ZIKV infection and disease. Furthermore, we hypothesize that targeting GAd vectors specifically to dendritic cells (DCs) will facilitate direct presentation of ZIKV structural antigens and improve vaccination outcome compared to conventional Ad-based vaccine. To test our hypothesis we have developed methods to ablate native Ad tropism via genetic modifications of the viral capsid while allowing recognition of the target cell receptor and the possibility of cell-specific delivery in vivo. We showed the utility of incorporating single domain antibody (sdAb) species into viral particles for Ad targeting to cancer cell types and immature myeloid murine DCs. We propose to use GAd developed by our commercial partner GenVec, Inc., as this vector platform has greater potential for clinical translation. In support of this study, we generated a panel of sdAb candidates that bind to the DC-specific receptor Clec9A that can be exploited to induce robust T- and B-cell immune responses following vaccination. The Diamond laboratory has developed new murine models of ZIKV pathogenesis including a model of maternal-fetal transmission. These models will allow us to assess rapidly the efficacy of our different candidate vaccines. In summary, our collaborative group has the necessary reagents, technology, and expertise to develop and evaluate novel GAd-vectored ZIKV vaccine candidates using relevant animal models. We believe the translation of these results could have a significant impact on reducing ZIKV disease and spread, a beneficial effect on human health.
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0.948 |
2017 — 2021 |
Curiel, David Terry |
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. |
Novel Targeted Adenovirus
ABSTRACT A central mandate to realize effective gene therapy is the ability to accomplish cell specific delivery. Capitalizing on the in vivo delivery efficacy of adenoviral vectors (Ad), recent studies have highlighted the capacity of targeted Ad to accomplish cell selectivity in this stringent delivery context. Systemic employment of Ad for gene delivery, however, is currently limited by vector particle sequestration in the liver. Recent work in several laboratories, however, has identified the biologic dictates of vector hepatotropism. Based on this understanding, it has now been possible to un-target the liver thereby facilitating strategies designed to achieve cell specific gene delivery in the context of systemic vector administration. Of note, we have recently shown that such liver un-targeting strategies can synergize with described vector targeting methods such as those based upon restricting delivered transgene expression to target cells with a tissue/tumor selective promoter (transcriptional targeting). The dramatic synergistic specificity gains noted with combination of these two approaches logically suggests that further gains may accrue additional targeting methods exploited in combination. In this regard, strategies have been proposed to target Ad based upon re-directing vector binding to target cell specific cell surface markers. Such transductional targeting methods would offer potential synergies with the targeting methods we note above. Such an endeavor has been limited to this point by the inability of current vector engineering to achieve capsid incorporation of antibody targeting species. Herein we seek to address this key limit. First, we have developed a method to replace the native adenovirus fiber with a substitute chimera devoid of the native fiber's knob binding domain. This maneuver eliminates native tropism and allows for the incorporation of a wider range of large/complex candidate targeting ligands. Second, we have demonstrated that the single domain antibody species derived from camelids (sdAb) possess the unique attributes allowing biologic compatibility with adenovirus capsid synthesis and assembly. In the aggregate, these two technologies now allow for the functional incorporation of antibody targeting species into the Ad capsid for the achievement of cell-specific targeting. This additional level of targeting provides for potential synergies with the defined liver un-targeting and transcriptional vector targeting methods we have explored.
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1 |
2017 — 2021 |
Curiel, David Terry George, Steven Carl [⬀] Rentschler, Stacey Lynn (co-PI) [⬀] |
UG3Activity Code Description: As part of a bi-phasic approach to funding exploratory and/or developmental research, the UG3 provides support for the first phase of the award. This activity code is used in lieu of the UH2 activity code when larger budgets and/or project periods are required to establish feasibility for the project. UH3Activity Code Description: The UH3 award is to provide a second phase for the support for innovative exploratory and development research activities initiated under the UH2 mechanism. Although only UH2 awardees are generally eligible to apply for UH3 support, specific program initiatives may establish eligibility criteria under which applications could be accepted from applicants demonstrating progress equivalent to that expected under UH2. |
A 3d in Vitro Disease Model of Atrial Conduction @ University of California At Davis
PROJECT SUMMARY Nearly 1 in 10 adults over the age of 65 in the U.S. suffer from atrial fibrillation (AF) leading to approximately $6 billion annually in healthcare costs. Because advanced age is a primary risk factor for developing AF, the overall incidence is expected to rise steadily over the coming decades as our population ages. Current therapeutic interventions have remarkably poor efficacy and/or untoward side effects due in large part to our inability to target the root cause of the disease and provide specificity for the atria and the patient. The central objective of this proposal is to create and validate a robust 3D microphysiological model of abnormal human atrial conduction using induced pluripotent stem cells from the patient. The model will simulate important elements of AF, such as conduction velocity, and develop novel therapeutic strategies that employ adenoviral delivery of gene interference (CRISPRi) that target altered gene regulatory pathways as the source of AF. We will accomplish this objective by completing the following specific aims: 1) characterize the transcriptome, epigenome, and electrophysiology of adult human atrial cardiomyocytes (normal and AF); 2) create a 3D in vitro disease model of human atrial conduction leveraging human iPS cell-derived atrial cardiomyocytes (iPS- aCM) and atrial regulatory gene expression; 3) design and test an adenoviral gene delivery strategy to specifically target atrial (not ventricular or nodal) cardiomyocytes; 4) demonstrate atrial specific adenoviral delivery (SA3) of CRISPRi and gene interference of PITX2 in ex vivo human atrial tissue; and 5) create iPS- aCM and the corresponding in vitro model of atrial conduction from a cohort of normal subjects and patients with AF; characterize drug efficacy and gene delivery in the in vitro models using a panel of existing drugs and our atrial specific adenoviral construct for CRISPR gene interference of PITX2 (SA3). Completing the specific aims will provide a model of human atrial conduction that can be used as a broad platform to understand drug efficacy and safety for diseases such as AF.!
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0.943 |
2019 — 2021 |
Curiel, David Terry |
UG3Activity Code Description: As part of a bi-phasic approach to funding exploratory and/or developmental research, the UG3 provides support for the first phase of the award. This activity code is used in lieu of the UH2 activity code when larger budgets and/or project periods are required to establish feasibility for the project. |
Endothelial-Targeted Adenovirus For Organ-Selective Gene Editing in Vivo
PROJECT SUMMARY A wide range of biological applications have derived from the CRISPR/Cas9 site-specific nuclease system in recent years. Of note, the capacity to accomplish gene editing in a targeted manner has also impacted the design of gene therapy strategies for an expanding repertoire of disorders. Critical to realizing the gene editing functions of the CRISPR/Cas9 system in a gene therapeutic context is the requirement to accomplish effective co-delivery in vivo of the constituent components. This delivery issue has been approached applying both non-viral and viral vector systems. In selected instances, successful gene-editing facilitated gene therapies have been accomplished in model systems of inherited genetic disease. Despite these elegant proof-of-principle studies, limits in available vector technology have greatly restricted the application of CRISPR/Cas9-facilitated gene therapy. In this regard, effective in vivo co-delivery of CRISPR/Cas9 to target somatic cells is required for many of these applications. Such delivery should be restricted exclusively to the key cellular targets in vivo to minimize off-target effects. In addition, the mandated co-delivery must be accomplished in the potential presence of pre-formed anti-vector immunity. Finally, methods to limit Cas9 expression must be endeavored to limit the potential of off-target editing. Of note, these functionalities should ideally be configured into the context of a single vector particle context to facilitate practical upscaling and human clinical translation. To this end, we have exploited the molecular promiscuities of adenovirus (Ad) to address the requirements of CRISPR/Cas9-facilitated gene therapy. In this regard, we have endeavored capsid engineering of adenovirus to achieve targeted modifications of vector tropism. In addition to allowing for re-directed tropism, capsid engineering provides the means to allow Ad to circumvent pre-formed vector immunity. We have also applied a strategy of capsid engineering to accomplish transient expression of heterologous proteins. On this basis, during the UG3 Phase (3 years) we will establish proof-of-principle with respect to delivery of genome editing machinery into disease relevant cells and tissues in vivo. The follow-on UH3 Phase (1 year) will address scale up and testing of our novel approach in a large animal model. This will be accomplished in collaboration with the SCGE Large Animal Testing Centers.
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2019 — 2021 |
Curiel, David Terry |
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. |
Novel Vector Platform For Gene Therapy
ABSTRACT Inherited serum deficiency disorders, such as hemophilia and ?1-antitrypsin (AAT) deficiency, have been considered ideal candidates for corrective gene therapy. In this regard, viral vector-mediated transduction of the liver has been proposed for a number of such disorders wherein hepatocytes represent the normal physiologic source of the deficient serum factor. Specifically, adeno-associated virus (AAV)-based vectors have demonstrated utility in hemophilia in animal models and this approach is presently being translated to the context of human clinical trials. Despite these findings, a number of considerations have led to the recognition that alternative vector approaches may be required. In the first instance, vector-related liver toxicities make consideration of non-hepatic sourcing of serum factors desirable. In addition, for some inherited deficiency disorders, such as AAT deficiency, AAV-mediated in vivo transduction of the liver has not achieved adequate levels of the deficient serum factor to achieve effective gene therapy. We propose here to test our highly original approach to accomplish genetic correction of a prototype serum deficiency disorders ? AAT deficiency lung disease. Our technology platform will clearly have broad field impact. In the first regard, we provide the technical basis for a new translational approaches for the full range of inherited serum deficiency disorders. In the second regard, we will engineer Ad as a vehicle capable of long term gene expression. This fundamental change in this important vector's functionalities will dramatically expand the range of utilities whereby it can be employed.
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