1985 — 2000 |
Klintworth, Gordon Kenneth |
P30Activity Code Description: To support shared resources and facilities for categorical research by a number of investigators from different disciplines who provide a multidisciplinary approach to a joint research effort or from the same discipline who focus on a common research problem. The core grant is integrated with the center's component projects or program projects, though funded independently from them. This support, by providing more accessible resources, is expected to assure a greater productivity than from the separate projects and program projects. |
Core Grant For Vision Research
The primary objectives of this Core Grant for Vision Research are to strengthen ongoing collaborative research activities and stimulate new interdisciplinary and interdepartmental research by providing core support for several modules to be made available as common research facilities to investigators of Vision and its Disorders. These modules include: an Animal Surgery Module, a Morphology Module, a Biochemistry/Molecular Biology Module, and an Administrative Module. These facilities provide services and shared resources for collaborative studies on eye related research projects that conform with the NEI national research plan. The various modules will enable us to strengthen our collaborative efforts among scientists of various disciplines in the Departments of Biochemistry, Cell Biology, Immunology, Medicine, Neurobiology, Ophthalmology, Pathology, Psychology, and Radiology at Duke University and the Departments of Cell Biology, Anatomy and Physiology at the University of North Carolina and the Departments of Anatomy/Physiology at North Carolina State University. The Animal Surgery Module will foster collaborative research by making it possible for investigators from diverse disciplines to carry out surgery and experiments together in a facility specifically designed and equipped for ophthalmic procedures in a Eye Center in which basic scientists and clinical ophthalmologists work in harmony. The Ocular Morphology Module with its components in the Departments of Cell Biology and Ophthalmology will provide services enabling co-investigators from different disciplines to correlate histologic, autoradiographic and ultrastructural observations with biochemical, physiological or clinical alterations in ocular disease. The Biochemistry/Molecular Biology Module will provide equipment and facilities that would not otherwise be available to individual investigators. The Administrative Module will coordinate the various eye related research activities and provide the necessary resources for the administration of the Core Grant.
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1985 — 1992 |
Klintworth, Gordon Kenneth |
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. |
Experimental Approach to Diseases of the Cornea
The proposed research is concerned with an experimental study of corneal diseases. The main emphasis is: a) To investigate the pathogenesis of corneal vascularization. These studies will be based upon a systematic investigation of the biological properties of the cornea and those factors inducing vascularity, with particular attention to those of clinical significance. An attempt will be made to quantitate corneal vascularizaton in rodents in reproducible experimental models by digitizing images obtained from flat preparations of corneas with colloidal carbon filled blood vessels and by using radioactive microspheres that are known to lodge in the microvasculature in direct proportion to blood flow. An attempt will also be made to quantitate corneal neovascularization in living animals by using an image processor to digitize images created by blood vessels. Part of this research will evaluate the relative roles of the angiogenic effect of mediators of the inflammatory response as well as of cellular elements of the blood (polymorphonuclear leukocytes, lymphocytes, monocytes and platelets) and plasma constituents. To minimize individual host responses many experiments on corneal vascularization will be performed on inbred mice with the same genome. b) To study the ultrastructural, and biochemical characteristics and pathophysiological reactions of corneal diseases, especially those in which abnormal materials deposit in the cornea. It is hoped to integrate the ultrastructural, biochemical, and metabolic aspects of certain corneal diseases. Particular attention will be paid to glycosaminoglycans, proteoglycans, collagen, and other proteins in the corneal stroma. Special attention will be devoted to keratoconus and the corneal dystrophies, especially macular corneal dystrophy. The studies will involve state-of-the-art energy dispersive x-ray microanalytical and image processing equipment, cell and organ cultures, monoclonal antibodies, immunochemistry and sensitive biochemical analytical methods. A registry of known cases of macular corneal dystrophy will be maintained and a genealogic and clinicopathologic investigation will be performed on these cases. Selected communities with a high gene frequency for macular corneal dystrophy will be screened for early cases with a sensitive ELISA technique that can detect keratan sulfate in serum and other body fluids.
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1989 — 1993 |
Klintworth, Gordon Kenneth |
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. |
Genetic Linkage Study Macular Corneal Dystrophy
Macular corneal dystrophy (MCD) is a rare, autosomal recessive ocular stora e disease which leads to progressive bilateral visual loss. Clinical progression of the disease usually causes visual impairment during the seco d decade of life and generally worsens until penetrating keratoplasty (cornea transplantation) is required to restore vision usually by the fourth decade of life. A keratan sulfate like glycosaminoglycan accumulates in the corne . The location of the gene for MCD is unknown and no presymptomatic diagnosti test is available to individuals who are at risk. Nor is there a reliable form of carrier detection. The objective of the proposed research is to define the genomic location of the MCD gene(s) while concurrently attempting to characterize the storage substance. Participants will be selected from a registry of affected individuals and their unaffected relatives maintained by the Principal Investigator. Blood samples will be obtained from affected individuals, their parents, and siblings to establish lymphoblastoid cell lines as permanent sources of DNA. In addition, serum will be tested for a specific epitope of sulfated keratan sulfate which is known to be markedly reduced i many affected individuals. Linkage analysis of MCD with restriction fragme t length polymorphisms randomly located throughout the genome as well as candidate genes that become available will be performed using state-of-the- art statistical methodology. The establishment of a linkage relationship and identification of the chromosomal location are the first steps in identifying the gene. Fine gen mapping, gene expression, the nature of the abnormal accumulates, and its mechanism of generation may then be possible. In addition establishment of linkage relationships may clarify the possibility of genetic heterogeneity in this disorder. These developments will lead to a method of presymptomat c diagnosis of MCD and detection of carriers of the MCD gene. Eventually the work may lead to an alternative means of treating MCD when further techniqu s of genetic engineering become available.
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1993 — 1995 |
Klintworth, Gordon Kenneth |
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. |
Experimental Approach to Deseases of the Cornea
DESCRIPTION (applicant's abstract): The proposed research is part of an ongoing long-term experimental study of corneal diseases. The current main emphasis is on an investigation of corneal vascularization, corneal proteins in health and disease, and certain corneal dystrophies. Special attention would be devoted especially to macular, lattice and granular corneal dystrophies. The pathogenesis of corneal vascularization is being studied in vivo by evaluating variables that influence its severity, with particular attention to those of clinical significance. Corneal vascularization is being quantified in rodents in reproducible experimental models by using digitized images obtained from flat preparations of corneas with colloidal carbon filled blood vessels. To minimize individual host responses many experiments on corneal vascularization would be performed on inbred mice with the same genome. An attempt is also being made to quantify corneal neovascularization in living animals by using an image processor to digitize video images created by blood vessels. Part of this research would evaluate the relative roles of mediators of the inflammatory response as well as of cellular elements of the blood (polymorphonuclear leukocytes, lymphocytes, monocytes and platelets) and plasma constituents in the angiogenic response. An attempt would be made to detect putative angiogenic cytokines and growth factors in the cornea at the protein and mRNA levels prior to and during angiogenesis using immunochemical methods and the recently described techniques for quantification mRNA by the polymerase chain reaction. Other studies would focus on the major corneal soluble protein, which has an apparent molecular weight of 54,000 daltons and on diseases in which proteins accumulate within the cornea in pathologic states. The studies would involve state-of-the-art energy dispersive x-ray microanalytical and image processing equipment, cell and organ cultures, monoclonal antibodies, immunochemistry and sensitive biochemical and molecular biological analytical methods. A registry of known cases of muscular corneal dystrophy would be maintained and a genealogic and clinicopathologic investigation would be performed on these cases. Selected communities with a high gene frequency for macular corneal dystrophy would be screened for early cases with a sensitive ELISA technique that can detect keratan sulfate in serum and other body fluids. A registry of cases of granular and lattice corneal dystrophy would be expanded.
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1994 — 2001 |
Klintworth, Gordon Kenneth |
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. |
Genetic Linkage Study--Macular Corneal Dystrophy
Macular corneal dystrophy (MCD) is an autosomal recessive corneal dystrophy which leads to progressive bilateral visual loss. The disease may begin during infancy or as late as the seventh decade of life, but vision usually becomes impaired during the second decade of life and worsens until penetrating keratoplasty is required to restore vision. A keratan sulfate like glycosaminoglycan accumulates in the cornea. Two types of MCD have been characterized and the location of the gene for both MCD type I and II is unknown and no presymptomatic diagnostic test is available to individuals who are at risk. Nor is there a reliable form of carrier detection. The objective of the proposed research is to define the genomic location of the MCD gene(s) while concurrently attempting to characterize the storage substance. Blood has been collected from 205 individuals belonging to sixteen families of MCD. Over 110 markers have been screened for genomic localization and additional markers are being used in an ongoing attempt to localize the MCD gene. Once localization occurs, we will use selected YAC's and band specific microdissection libraries to develop highly polymorphic microsatellites that will closely flank these genes. We will use a variety of techniques including meiotic mapping and linkage disequilibrium to narrow the region containing this gene. The YAC megabase library will be used to construct a physical map spanning the disease locus. Coding regions will be identified through direct selection and if needed, exon amplification. As they become available candidate genes will be isolated and tested for mutations using PFGE, heteroduplex and SSCP. Once the defect is known we can begin to evaluate the basic physiologic mechanisms of the MCD gene.
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1994 — 1995 |
Klintworth, Gordon Kenneth |
P30Activity Code Description: To support shared resources and facilities for categorical research by a number of investigators from different disciplines who provide a multidisciplinary approach to a joint research effort or from the same discipline who focus on a common research problem. The core grant is integrated with the center's component projects or program projects, though funded independently from them. This support, by providing more accessible resources, is expected to assure a greater productivity than from the separate projects and program projects. |
Vision Research
The primary objectives of this Core Grant for Vision Research are to strengthen ongoing collaborative research activities and stimulate new interdisciplinary and interdepartmental research by providing core support for several modules to be made available as common research facilities to investigators of Vision and its Disorders. These modules include: an Animal Surgery Module, a Morphology Module, a Biochemistry/Molecular Biology Module, and an Administrative Module. These facilities provide services and shared resources for collaborative studies on eye related research projects that conform with the NEI national research plan. The various modules will enable us to strengthen our collaborative efforts among scientists of various disciplines in the Departments of Biochemistry, Cell Biology, Immunology, Medicine, Neurobiology, Ophthalmology, Pathology, Psychology, and Radiology at Duke University and the Departments of Cell Biology, Anatomy and Physiology at the University of North Carolina and the Departments of Anatomy/Physiology at North Carolina State University. The Animal Surgery Module will foster collaborative research by making it possible for investigators from diverse disciplines to carry out surgery and experiments together in a facility specifically designed and equipped for ophthalmic procedures in a Eye Center in which basic scientists and clinical ophthalmologists work in harmony. The Ocular Morphology Module with its components in the Departments of Cell Biology and Ophthalmology will provide services enabling co-investigators from different disciplines to correlate histologic, autoradiographic and ultrastructural observations with biochemical, physiological or clinical alterations in ocular disease. The Biochemistry/Molecular Biology Module will provide equipment and facilities that would not otherwise be available to individual investigators. The Administrative Module will coordinate the various eye related research activities and provide the necessary resources for the administration of the Core Grant.
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2000 — 2003 |
Klintworth, Gordon Kenneth |
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. |
Bigh3 Wild Type and Mutant Proteins
Several corneal dystrophies [granular corneal dystrophy (GCD), lattice corneal dystrophy (LCD) types I and IIIA, combined UCD-LCD type I, Thiel-Behnke dystrophy and Reis-Bucklers dystrophy] have been found to result from mutations in BIGH3. These conditions are characterized by the deposition within the cornea of proteins that react with antibodies to beta-ig-h3 (the protein product of the BIGH3 gene). For each dystrophy the accumulations have a characteristic ultrastructure. This proposal is to purify and characterize beta-ig-h3 from plasma of persons with and without different BIGH3 corneal dystrophies and from cell cultures of COS- 7, CHO-K1 and baculovirus infected insect cells transfected with vectors containing the mutations that produce BIGH3 corneal dystrophies. Wild-type and mutant beta-ig-h3 proteins will be compared to test the hypothesis that the structure of the proteins that accumulate within the cornea in the BIGH3 corneal dystrophies are related to the mutated protein and that a relationship exists between different phenotypes and the BIGH3 genotypes and between the amount of the mutant protein that is expressed. The nature of the amyloid that accumulates in certain BIGH3 corneal dystrophies and the reason for its production will be investigated. We will determine whether BIGH3 related amyloid is a specific fragment of mutant beta-ig-h3. Because beta-ig-h3 interacts with components of the extracellular matrix we will attempt to identify protein- protein interactions that may account for the accumulations that characterize the BIGH3 corneal dystrophies. The biosynthesis of beta-ig-h3 will be characterized in rabbit corneal epithelial cells.
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2002 — 2005 |
Klintworth, Gordon Kenneth |
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. |
Macular Corneal Dystrophy-Molecular Pathobiology
DESCRIPTION (provided by applicant): The proposed research is part of an ongoing study of macular corneal dystrophy (MCD) and the CHST6 gene. We will maintain and expand a computerized registry and genealogical database of individuals with MCD for relevant studies. Mutation analyses of the CHST6 gene are being determined in families with and without MCD from different populations using DNA extracted from the peripheral blood or from pathologic archival corneal tissue. This is being done with the intent of differentiating disease-producing mutations in the CHST6 gene from inconsequential single nucleotide polymorphisms (SNPs). Genotype-phenotype correlations are also being performed from this information. The molecular basis for the observed immunophenotypes in MCD is being sought in relationship to specific mutations in CHST6. To determine the distribution of the manifestations of MCD we will continue to examine various tissues in affected persons when available. We are mining nucleotide and protein computerized databases to further knowledge about CHST6 and the encoded carbohydrate sulfotransferase. The recombinant protein synthesized by mammalian and insect cell lines transfected with vectors containing wild-type CHST6 is being purified and characterized biochemically and enzymatically. The role of CHST6 in the sulfation of specific carbohydrate moieties is being determined. We are studying the subcellular localization of the expressed CHST6 gene product and are developing model systems to study the pathobiology of MCD in cell culture systems. An attempt is being made to develop an intracellular storage disorder comparable to MCD in cultured keratocytes (corneal fibroblasts) using antisense oligonucleotides targeted against specific sequences in CHST6 mRNA.
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2004 — 2008 |
Klintworth, Gordon Kenneth |
K12Activity Code Description: For support to a newly trained clinician appointed by an institution for development of independent research skills and experience in a fundamental science within the framework of an interdisciplinary research and development program. |
Nei Institute Clinical Scientist Development Program
DESCRIPTION (provided by applicant): This is an institutional training grant proposal to educate the next generation of clinician-scientists for academic positions in Ophthalmology. For over 38 years, the Duke University Eye Center (DUEC) has been a national leader in training academic Ophthalmologists. The earliest generations of these trainees now serve in numerous leadership positions in academic medical centers across the country and abroad. They have roles as Deans, Department Chairs, and faculty members and serve as mentors for academicians. We propose to continue this tradition through an intensive program designed to provide the guidance, resources, and didactic teaching necessary to produce clinician-scientists for academic positions in Ophthalmology. We plan to select two trainees each year after their completion of a residency in Ophthalmology or Pathology for an intense 3 year educational program that will lead to independent peer-reviewed grant funding and a faculty position in an academic institution. The trainees will undergo an intensive didactic curriculum combined with a close mentorship by senior faculty at Duke University. Each trainee will receive mandatory training in areas of clinical research, genetics, statistics, scientific writing, grant preparation, ethics, and the responsible conduct of research. Each trainee will participate in significant original research and learn the essential skills to develop a successful and independent academic career. Additional aspects of the curriculum of each trainee will be tailored by the program director and lead mentor based upon the selected primary research track and his/her skills, background and career goals. We propose five primary research tracks: 1) Ophthalmic Genomics and Proteomics, 2) Ophthalmic Pathology, 3) Translational Medicine and Clinical Trials, 4) Clinic to Bench, 5) Outcomes and Health Policy. Because of an international shortage of Ophthalmic Pathologists involved in research, we will attempt to recruit Pathologists and train them for a research career to further knowledge about ocular diseases using tissue derived from patients and experimental models.
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2007 — 2011 |
Klintworth, Gordon Kenneth |
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. |
Study of Genetic Basis of Fuchs Corneal Dystrophy
DESCRIPTION (provided by applicant): Fuchs corneal dystrophy (FCD) is a common bilateral late onset disorder of the corneal endothelium chracterized by a slowly progressive dysfunction of corneal endothelial cells associated with the loss of these cells and the formation of excrescences (corneal guttae) and excessive thickening of Descemet's membrane. This age-related disorder has a striking female:male preponderance. Evidence for Mendelian forms of FCD comes from large multigenerational FCD families. However, the majority of FCD are found in small families, which exhibit a complex form of FCD. FCD has also been mapped to loci on several chromosomes. The objective of this application by an experienced multidisciplinary team of investigators is to expand family recruitment and further our understanding of FCD with genetic studies. This study has five specific aims: (1) To continue recruiting families of FCD with at least one affected individual and up to two unaffected siblings as well as multigenerational FCD families, (2) To sequence and genotype the COL8A2 gene in each affected individual, (3) To perform a whole genome linkage screen using Illumina's fourth-generation SNP-based linkage panel, (4) To perform association mapping for up to four linkage regions to identify candidate genes associated with FCD and to follow up of these candidate genes, and (5) To perform positional cloning for large multigenerational families. To achieve these goals, an ocular examination will be performed on all participants to determine whether they are affected or not. Families with COL8A2 mutations will be removed from the linkage and follow-up analyses. Linkage analysis will be performed on various datasets of families of different size. This strategy will allow chromosonal regions that are responsible for Mendelian and complex forms of genetics in FCD to be identified. The long-term objective of this study is to understand the basic pathobiology of FCD and to identify genetic components of the disorder that will be valuable from the standpoint of developing molecular diagnostic tests for early diagnosis and for developing novel therapeutic procedures for this debilitating disease. Statement of Relevance: Knowledge about the genes that place individuals at risk for FCD will lead to a better understanding of this important disease, early diagnostic tests and novel non surgical methods of treatment.
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2008 — 2011 |
Klintworth, Gordon Kenneth |
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. |
Bigh3 Wild-Type and Mutant Proteins
[unreadable] DESCRIPTION (provided by applicant): The cornea contains an abundant poorly understood clinically relevant protein important for the preservation of corneal transparency. Mutations in the TGFBI (BIGH3) gene are responsible for several phenotypically different inherited corneal diseases that have no apparent non-ocular manifestations. These disorders include several varieties of lattice corneal dystrophy and corneal amyloidoses, granular corneal dystrophy, Reis- B|cklers corneal dystrophy, Thiel-Behnke dystrophy, and several atypical corneal disorders. The particular clinical and histopathologic phenotypes are dependent upon the precise mutation in TGFBI, but a molecular explanation for the different phenotypes remains to be determined. The mutated extracellular transforming growth factor beta induced protein (TGFBIp) encoded by TGFBI accumulates in the corneal stroma in these disorders which are apparently limited to the cornea. The long-term objectives are to understand the properties of this unique protein and to investigate the molecular mechanisms responsible for the specific deposits that accumulate within the cornea in patients with mutated TGFBI. The Specific Aims of this proposal are: (1) to screen subjects with inherited corneal diseases for TGFBI mutations, (2) to analyze abnormal deposits isolated from surgically excised corneal tissue by laser capture micro-dissection and liquid chromatography/tandem mass spectrometry (LC MS/MS) to determine whether part or all of the mutated TGFBIp accumulates in the cornea and what other protein(s) are closely linked to it, (3) to determine the biochemical and biophysical properties of the FAS4 domain of purified recombinant wild-type and disease producing TGFBIp and (4) to solve the three-dimensional structure at atomic resolution of recombinant wild- type TGFBIp and the FAS4 domains of recombinant wild-type and disease producing mutants using X-ray crystallography and nuclear magnetic resonance spectroscopy (NMR). PUBLIC HEALTH RELEVANCE. This is a study of an important poorly understood protein (TGFBIp). Mutations in the gene (TGFBI) encoding for this protein cause several corneal diseases (dystrophies) and a better understanding of TGFBIp will lead to better methods of treating the resulting impaired vision and debilitating symptoms. [unreadable] [unreadable] [unreadable]
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2010 — 2012 |
Klintworth, Gordon Kenneth |
K12Activity Code Description: For support to a newly trained clinician appointed by an institution for development of independent research skills and experience in a fundamental science within the framework of an interdisciplinary research and development program. |
Nei Mentored Clincial Scientist Development Program Award (K12)
This is an institutional training grant proposal to educate the next generation of clinician-scientists for academic positions in Ophthalmology. For over 38 years, the Duke University Eye Center (DUEC) has been a national leader in training academic Ophthalmologists. The earliest generations of these trainees now serve in numerous leadership positions in academic medical centers across the country and abroad. They have roles as Deans, Department Chairs, and faculty members and serve as mentors for academicians. We propose to continue this tradition through an intensive program designed to provide the guidance, resources, and didactic teaching necessary to produce clinician-scientists for academic positions in Ophthalmology. We plan to select two trainees each year after their completion of a residency in Ophthalmology or Pathology for an intense 3 year educational program that will lead to independent peer-reviewed grant funding and a faculty position in an academic institution. The trainees will undergo an intensive didactic curriculum combined with a close mentorship by senior faculty at Duke University. Each trainee will receive mandatory training in areas of clinical research, genetics, statistics, scientific writing, grant preparation, ethics, and the responsible conduct of research. Each trainee will participate in significant original research and learn the essential skills to develop a successful and independent academic career. Additional aspects of the curriculum of each trainee will be tailored by the program director and lead mentor based upon the selected primary research track and his/her skills, background and career goals. We propose five primary research tracks: (1) translational medicine and clinical trials, (2) clinics and bedside to the bench, (3) Ophthalmic genomics and proteomics, (4) ophthalmic pathology, (5) outcomes and health policy. Because of an international shortage of Ophthalmic Pathologists we will attempt to recruit and appropriate individuals for a research career involving the use of ocular tissues derived from patients and experimental models.
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