Analysis And Therapy Of Muscular Dystrophy In Zebrafish
Louis M Kunkel, Professor
Children´s Hospital Boston
300 Longwood Ave
boston, Ma 021155737
Grant 5P50NS040828-070001 from National Institute Of Neurological Disorders And Stroke IRG: ZNS1
Sponsored Links Excellgen http://Excellgen.com
ANALYSIS AND THERAPY OF MUSCULAR DYSTROPHY IN ZEBRAFISH
Louis M Kunkel, Professor Of Genetics And Pediatrics
Children´s Hospital Boston, 300 Longwood Ave, Boston, Ma 02115-5737
Abstract: Over the last 5 years, we have been working to establish the zebrafish as a model for muscular dystrophy. In this capacity, we have published the phenotype of zebrafish lacking dystrophin and 8- sarcoglycan, completed a large genetic screen to isolate additional dystrophic mutants, and identified the mutant gene in the runzelmutant. Using our experiences in muscle research and in establishing the zebrafish as a disease model, we now propose to use the fish to investigate the pathogenesis of muscular dystrophy and evaluate cell therapy as a potential treatment option. The first aim in this project proposes to fully characterize two of our available dystrophic zebrafish models (emz and sof) with the goal of better understanding the pathogenesis of muscular dystrophy. These mutants show a muscle degeneration phenotype very similar to the dystrophin mutant (sapje) suggesting that this phenotype is symptomatic of muscular dystrophy. We propose to identify the genetic mutations in these mutants using a traditional mapping approach and then sequencing candidate genes to identify the specific mutation. If the orthologous human genes are not currently associated with muscular dystrophy, these genes will be considered disease candidate genes and sequenced in human patients for which the cause of muscular dystrophy is unknown. Since mutations in seemingly unrelated proteins can manifest as muscular dystrophy, the identification of additional genes would be helpful for establishing disease pathways. Secondly, we have established methods to transplant cell populations in zebrafish at all developmental stages and now propose using this system to identify the cell population most capable of engrafting into and correcting the diseased muscle. Gene expression profiles of muscle engrafting cell populations will be compared with non-engrafting cells to identify genes expressed predominantly in the engrafting cells. Differentially expressed genes will be considered potential markers and used to purify analogous cell populations in mammals for future experimentation and therapy. Finally, we plan to dissect the lineage relationship of various stem cell populations by assaying the developmental potential of zebrafish muscle progenitor cells. This will be accomplished by transplanting limited populations of labeled cells early in development and then following their fate as the fish matures
Keywords: 21+ years old; ALDH; Adult; Alleles; Allelomorphs; Animal Model; Animal Models and Related Studies; Assay; Bioassay; Biologic Assays; Biological Assay; Brachydanio rerio; Candidate Disease Gene; Candidate Gene; Cell Communication and Signaling; Cell Signaling; Cell Therapy; Cell Transplants; Cell surface; Cells; DNA Alteration; DNA mutation; Danio rerio; Development; Disease; Disease Pathway; Disease model; Disorder; Disorder of muscle, unspecified; Dystrophin; Engraftment; Expression Profiling; Expression Signature; Fishes; Future; Gene Alteration; Gene Expression; Gene Mutation; Genes; Genetic Alteration; Genetic Change; Genetic Screening; Genetic defect; Genetic mutation; Genomics; Goals; Human; Human, Adult; Human, General; In Vitro; Individual; Intracellular Communication and Signaling; Label; Lead; Mammalia; Mammals; Mammals, General; Mammals, Mice; Man (Taxonomy); Man, Modern; Maps; Messenger RNA; Methods; Methods and Techniques; Methods, Other; Mice; Mice, Mutant Strains; Modeling; Molecular Fingerprinting; Molecular Profiling; Mother Cells; Murine; Mus; Muscle; Muscle Disease; Muscle Disorders; Muscle Tissue; Muscle disease or syndrome; Muscular Diseases; Muscular Dystrophies; Mutant Strains Mice; Mutation; Myodystrophica; Myodystrophy; Myopathic Conditions; Myopathic Diseases and Syndromes; Myopathic disease or syndrome; Myopathy; Myopathy, unspecified; Pathogenesis; Pathway interactions; Patients; Pb element; Phenotype; Play; Population; Position; Positioning Attribute; Progenitor Cells; Promoter; Promoters (Genetics); Promotor; Promotor (Genetics); Proteins; Publishing; RNA, Messenger; Research; Rhodamine 123; Role; Sarcoglycans; Sequence Alteration; Side; Signal Transduction; Signal Transduction Systems; Signaling; Sorting - Cell Movement; Staging; Staining method; Stainings; Stains; Stem cells; System; System, LOINC Axis 4; Techniques; Therapy, Cell; Transplantation; Transplants, Cell; Work; Xanthylium, 3, 6-diamino-9-(2-(methoxycarbonyl)phenyl)-, chloride; Zebra Danio; Zebra Fish; Zebrafish; adult human (21+); aldehyde dehydrogenases; biological signal transduction; cell-based therapy; disease/disorder; disorder model; experience; experiment; experimental research; experimental study; gene product; genome mutation; heavy metal Pb; heavy metal lead; in vivo; mRNA; mRNA Expression; model organism; molecuar profile; molecular signature; mouse mutant; muscle degeneration; muscular disorder; mutant; novel; pathway; repair; repaired; research study; social role; sorting; stem cell population; transplant; zebrafish genome
5P50NS040828-09_0001 (2010): $281520
PATHOGENESIS AND TREATMENT OF MUSCULAR DYSTROPHY
Louis M Kunkel, Professor Of Genetics And Pediatrics
Children´s Hospital Boston, 300 Longwood Ave, Boston, Ma 02115-5737
Grant 5P50NS040828-09 from National Institute Of Neurological Disorders And Stroke
Abstract: Over the last 5 years, we have been working to establish the zebrafish as a model for muscular dystrophy. In this capacity, we have published the phenotype of zebrafish lacking dystrophin and 5- sarcoglycan, completed a large genetic screen to isolate additional dystrophic mutants, and identified the mutant gene in the runzel mutant. Using our experiences in muscle research and in establishing the zebrafish as a disease model, we now propose to use the fish to investigate the pathogenesis of muscular dystrophy and evaluate cell therapy as a potential treatment option. The first aim in this project proposes to fully characterize one of our available dystrophic zebrafish models with the goal of better understanding the pathogenesis of muscular dystrophy. We have selected the emz mutant for further analysis since its mutation rough maps to a genomic interval void of any genes orthologous to those currently associated with muscular dystrophy. This mutant shows a phenotype very similar to the dystrophin mutant (sapje) suggesting that the emz phenotype of muscle degeneration is symptomatic of muscular dystrophy. We propose to identify the genetic mutation in this mutant using a traditional mapping approach and then sequencing candidate genes to identify the specific mutation. If the orthologous human gene is not currently associated with muscular dystrophy, the gene will be considered a disease candidate and sequenced in human patients for which the cause of muscular dystrophy is unknown. Since mutations in seemingly unrelated proteins can manifest as muscular dystrophy, the identification of additional genes would be helpful for establishing disease pathways. Secondly, we have established, methods to transplant cell populations in zebrafish at all developmental stages and now propose using this system to identify the cell population most capable of engrafting into and correcting the diseased muscle. Gene expression profiles of muscle engrafting cell populations will be compared with non-engrafting cells to identify genes expressed predominantly in the engrafting cells. Differentially expressed genes will be considered potential markers and used to purify analogous cell populations in mammals for future experimentation and therapy. Finally, we plan to dissect the lineage relationship of various stem cell populations by assaying the developmental potential of zebrafish muscle progenitor cells. This will be accomplished by transplanting limited populations of labeled cells early in development and then following their fate as the fish matures
Keywords: No Project Terms available
Project start date: 2000-12-01
Project end date: 2012-03-31
Budget start date: 1-APR-2010
Budget end date: 31-MAR-2011
5P50NS040828-09 (2010): $1469487
Grants awarded to Louis M Kunkel
GENETIC AND CYTOGENETIC STUDIES OF MENTAL RETARDATION
Louis M Kunkel, Professor
Children s Hospital Boston 300 Longwood Ave Boston, Ma 021155737
Grant 5P01HD018658-10 from National Institute Of Child Health And Human Development IRG: HDMR
Abstract: We propose a program of research on the localization and analysis of genetic determinants on human chromosomes, as well as subchromosomal structural changes underlying diseases, with emphasis on the human X, #15, #21, and other regions of the genome in which abnormalities can be associated with mental retardation and/or developmental defects. The research planned utilizes techniques which allow intense study of subchromosomal regions, such as fluorescence activated chromosome sorting and phenol-enhanced differential hybridization, recombinant DNA methodology, cytogenetics, somatic cell genetics and pedigree analysis. The goal is to obtain, localize, and characterize cloned DNA segments, as well as deleted, duplicated, rearranged, or labile subchromosomal regions associated with specific diseases. Using Xp21 deletions in DMD as a prototype and starting point, we will explore adjacent regions and deletions responsible for glycerol kinase deficiency, the McCloud phenotype, as well as X- linked eye diseases such as retinitis pigmentosa, retinoschisis, and choroideremia, the last associated with a deletion in proximal Xq. Structural analysis of Xp21, as well as distal Xp and Xp will be used to study normal and abnormal meiosis and the role of the latter in deletion-related diseases, insertion of a Yp segment into Xp in 46,XX males, and aneusomy. Recombination based screening will be used to obtain molecular probes near the fragile site in distal Xq to study its structural features and unusual mode of mutation. Existing and newly sought molecular probes will be used to analyze MR-associated diseases in 12p and proximal 15q1, in the case to understand better the nature and causes of structural changes associated with the Prader-Willi syndrome, as well as to refine diagnosis of this disease. Other genomic probes, as well as antibodies, will be used to isolate cDNA sequences mapping to chromosome #21 to investigate abnormalities, related to Down syndrome, of tissue-specific #21 DNA expression, e.g. as regards neuronal organization and cell surface changes underlying developmental defects. Of particular interest will be #21-linked amyloid beta protein cDNA and the relationship of amyloid to premature aging in Down syndrome and to Alzheimer disease. The information sought should both be of basic mechanistic interest, and it should translate readily into practical molecular genetic diagnostic applications.
Keywords: chromosome disorder, cytogenetics, gene expression, mental retardation
Project start date: 1983-01-01
Project end date: 1992-12-31
5P01HD018658-10 (1992): $607844
5P01HD018658-15 (1997): $824365
5P01HD018658-13 (1995): $742760
5P01HD018658-12 (1994): $736734
2P01HD018658-11 (1993): $727932
MOLECULAR GENETICS OF DUCHENNE MUSCULAR DYSTROPHY
Louis M Kunkel, Professor
Children s Hospital Boston 300 Longwood Ave Boston, Ma 021155737
Grant 5R01NS023740-05 from National Institute Of Neurological Disorders And Stroke IRG: MGN
Abstract: The region Xp21 of the human genome will reduced to an ordered array of cloned DNA segments by chromosome "walking" from multiple start points from within the region. The locus responsible for Duchenne muscular dystrophy known to lie within Xp21 will be identified and the product encoded by this locus characterized. The manner in which the product of the DMD locus is altered in patients will be determined by direct analysis of the DNA of the affected individuals. Mutations which disrupt normal function will be used as indicators of where the normal product from the locus functions. Structural characteristics of the Xp21 region which might affect normal expression from the locus will be studied with special emphasis given to possible mutational instablity of the region. The genetic distance of the entire region will be precisely determined and meiotic exchange points identified. Accurate prediction of individuals at risk for the disease will result by either direct analysis of the locus or indirect analysis with tightly linked markers. Ultimately, the pathways in neuromuscular development will be identified which are altered in Duchenne muscular dystrophy patients and possibly patients with other neuromuscular diseases.
Keywords: GENETICS, BIOCHEMICAL GENETICS, GENETICS, BIOCHEMICAL GENETICS, GENETIC CODING, GENETICS, GENETIC REGULATION, TRANSCRIPTION, GENETICS, POPULATION GENETICS HUMAN, METABOLIC DISORDERS INBORN, MUSCULAR DYSTROPHIES, genetic mapping, GENETIC DISORDERS DIAGNOSIS, GENETICS, BIOCHEMICAL GENETICS, MOLECULAR CLONING, NUCLEIC ACIDS BIOSYNTHESIS, RNA BIOSYNTHESIS
Project start date: 1986-07-01
Project end date: 1991-06-30
Sponsored Links Excellgen http://Excellgen.com
Louis M Kunkel, Professor
Children s Hospital Boston 300 Longwood Ave Boston, Ma 021155737
Grant 5P30HD018655-219004 from National Institute Of Child Health And Human Development
Abstract: The Multimedia Core provides conventional photographic support, digital imaging services, and advanced digital multimedia and web-based services. The Core is directed by Dr. Louis Kunkel, who provides almost daily input, and the Director of Operations, Mr. Dixon Yun, who has supervised this facility for 20 years. There is quarterly input from the Internal Advisory Board. The Core operates on a fee for service basis with fees proportional to the amount of time or item (e.g., slide or print) requested.
Keywords: Internet, biomedical facility, digital imaging, information dissemination, mental retardation, photography, computer graphics /printing, publication, videotape /videodisc, behavioral /social science research tag, bioimaging /biomedical imaging, human data
CORE--MOLECULAR GENETICS FACILITY
Louis M Kunkel, Professor
Children s Hospital Boston 300 Longwood Ave Boston, Ma 021155737
Grant 5P30HD018655-219014 from National Institute Of Child Health And Human Development
Abstract: The Molecular Genetics Core provides basic genomic services of DNA sequencing and genotyping to Center investigators, with the new addition of expression profiling using microarray technologies. Economies of scale and consolidation of expertise would be utilized to provide fundamental genome-based information to the Center investigators.
Keywords: biomedical facility, mental retardation, molecular genetics, nucleic acid sequence, computer assisted sequence analysis, gene mutation, genotype, linkage mapping, human genetic material tag, human tissue, microarray technology
Louis M Kunkel, Professor
Children s Hospital Boston 300 Longwood Ave Boston, Ma 021155737
Grant 5P30HD018655-219009 from National Institute Of Child Health And Human Development
Abstract: The Cell Sorter Core offers Center investigators access to high quality flow cytometry and cell sorting. It also provides expertise to investigators concerning the best approaches to sample preparation and staining techniques. In addition, Core personnel can assist investigators with data interpretation and presentation.
Keywords: biomedical facility, cell sorting, mental retardation, biomarker, cell population study, flow cytometry, genetic mapping, karyotype, human tissue
RNA EXPRESSION PATTERNS IN AUTISM
Louis M Kunkel, Professor Of Genetics And Pediatrics
Children´s Hospital Boston, 300 Longwood Ave, Boston, Ma 02115-5737
Grant 5R01MH085143-03 from National Institute Of Mental Health
Abstract: The Autism Spectrum Disorders (ASD) encompass a spectrum of disorders which include autism, Pervasive Developmental Disorder - Not Otherwise Specified (PDD-NOS), Asperger"s disorder and Rett syndrome. The working hypothesis of this proposal is that individuals with ASD experience disturbances in the intricate interplay between genetic predisposition, environmental triggers and experience-mediated neuronal activity during a sensitive period of development, and that this leads to an altered program of gene expression. Thus the neurological disabilities that are characteristic of the disorder result from genetic variability in the processes of synaptic development, maturation, refinement, and connectivity that normally shape the brain, and from an environmental insult that perturbs normal experience-dependant synaptic development. This proposal offers a novel approach aimed at unraveling the genetic mechanisms, improving diagnosis, deciphering the interplay between environment and genetics, and developing rational approaches to therapy
Keywords: Affect; Asperger Syndrome; Asperger`s Disorder; Autism; Autism, Early Infantile; Autism, Infantile; Autism-Dementia-Ataxia-Loss of Purposeful Hand Use Syndrome; Autistic Disorder; Behavior; Bio-Informatics; Biochemical Pathway; Bioinformatics; Brain; Cancers; Candidate Disease Gene; Candidate Gene; Cell Communication and Signaling; Cell Signaling; Cerebroatrophic Hyperammonemia; Characteristics; Chromosomal Disorders; Chromosomal Rearrangement; Chromosome Abnormality Disorders; Chromosome Disorders; Code; Coding System; Congenital chromosomal disease; Defect; Development; Diagnosis; Diagnostic tests; Disease; Disorder; Dystrophia Myotonica; Encephalon; Encephalons; Environment; Environmental Factor; Environmental Risk Factor; Epigenetic; Epigenetic Change; Epigenetic Mechanism; Epigenetic Process; Expression Profiling; Expression Signature; Family; Functional RNA; Gene Expression; Gene Expression Profile; Gene Transcription; Genes; Genetic; Genetic Alteration; Genetic Change; Genetic Predisposition; Genetic Predisposition to Disease; Genetic Susceptibility; Genetic Transcription; Genetic defect; Genome; Heritability; Incidence; Individual; Inherited Predisposition; Inherited Susceptibility; Intracellular Communication and Signaling; Kanner`s Syndrome; Lead; Linkage (Genetics); Malignant Neoplasms; Malignant Tumor; Mediating; Metabolic Networks; Minor; Molecular; Molecular Fingerprinting; Molecular Genetic; Molecular Genetics; Molecular Profiling; Monozygotic Twinning; Monozygotic twins; Mutation; Myotonia Atrophica; Myotonia Dystrophica; Myotonic Dystrophy; Nerve Cells; Nerve Unit; Nervous System, Brain; Neural Cell; Neurocyte; Neurologic; Neurological; Neurons; Non-Coding; Non-Coding RNA; PBL; Parents; Pathogenesis; Patients; Pattern; Pb element; Penetrance; Peripheral Blood Lymphocyte; Pervasive Development Disorder; Pervasive Developmental Disorder; Phenotype; Play; Process; Programs (PT); Programs [Publication Type]; RNA Expression; RNA Splicing; Regulatory Element; RegulatoryElement; Research; Rett Disorder; Rett Syndrome; Rett syndrome (RS, RTS); Role; Shapes; Signal Transduction; Signal Transduction Systems; Signaling; Specific qualifier value; Specified; Splicing; Staging; Steinert Disease; Synapses; Synaptic; Therapeutic Studies; Therapy Research; Transcript; Transcription; Transcription, Genetic; Twins, Identical; Variant; Variation; Work; autism spectrum disorder; base; biological signal transduction; brain shape; cohort; developmental disease/disorder; developmental disorder; disability; disease/disorder; environmental risk; experience; family genetics; gene expression signature; genetic etiology; genetic linkage; genetic mechanism of disease; genetic vulnerability; genome mutation; heavy metal Pb; heavy metal lead; improved; malignancy; molecuar profile; molecular signature; neoplasm/cancer; neuron development; neuronal; new approaches; novel; novel approaches; novel strategies; novel strategy; peripheral blood; programs; social role; success; transcriptome
Project start date: 2008-07-22
Project end date: 2013-02-28
Budget start date: 1-MAR-2010
Budget end date: 28-FEB-2011
5R01MH085143-03 (2010): $706052
Gene Expression In Normal And Diseased Muscle Development
Louis M Kunkel, Professor
Children s Hospital Boston 300 Longwood Ave Boston, Ma 021155737
Grant 5P01NS040828-05 from National Institute Of Neurological Disorders And Stroke IRG: NSD
Abstract: PROGRAM The last decade has witnessed remarkable progress in defining primary defects that cause inherited muscle disorders. The genetic heterogeneity of these diseases is enormous; mutations in more than 40 different genes are implicated. Many critical questions remain concerning the pathogenesis of muscle cell degeneration in these diseases and strategies for their treatment. This Program Project will use classical methods of gene and protein analysis and state-of-the-art gene expression array technology to study these questions. The investigators in this program have contributed importantly to the muscular dystrophy field. The proposed 4 projects have unique features but overlapping concepts and methodologies. Project 1 will study the dystrophin-associated complex of proteins, emphasizing the sarcoglycans and the newly described filamin-C. Project 2 will investigate the biology of dysferlin, its potential protein partners, and how these are altered by dysferlin gene mutations. Project 3 will examine the function of myotubularin in normal muscle development and the mechanisms by which its mutations cause developmental myopathies. Project 4 will study the biological and therapeutic properties of muscle stem cells. Three Cores will provide administrative oversight and services essential to the smooth progression of this program. Core B will coordinate sample acquisition and muscle RNA preparation for each project. Core C will perform the microarray analysis of gene expression and provide expertise in bioinformatics and data interpretation. The aim is to identify patterns of gene expression that are global in all dystrophies or distinct to specific sets of dystrophies and myopathies; this will provide insight into the molecular basis of normal muscle development and its dysfunction in these disease states. The long-term goal is to use this information in conjunction with the insights from studies of stem cell biology to devise new approaches to the treatment of the muscular dystrophies and related myopathies.
Keywords: gene expression, muscle protein, muscular dystrophy, myogenesis, protein structure function
Project start date: 2001-09-25
Project end date: 2007-08-31
5P01NS040828-05 (2005): $1506746
Sponsored Links Excellgen http://Excellgen.com
5P01NS040828-04 (2004): $1464230
5P01NS040828-03 (2003): $1437171
5P01NS040828-02 (2002): $1407253
Gene Expression In Normal & Diseased Muscle Development
Louis M Kunkel, Professor
Children´s Hospital Boston
300 Longwood Ave
boston, Ma 021155737
Grant 1P01NS040828-01A1 from National Institute Of Neurological Disorders And Stroke IRG: NSD
Abstract: PROGRAM The last decade has witnessed remarkable progress in defining primary defects that cause inherited muscle disorders. The genetic heterogeneity of these diseases is enormous; mutations in more than 40 different genes are implicated. Many critical questions remain concerning the pathogenesis of muscle cell degeneration in these diseases and strategies for their treatment. This Program Project will use classical methods of gene and protein analysis and state-of-the-art gene expression array technology to study these questions. The investigators in this program have contributed importantly to the muscular dystrophy field. The proposed 4 projects have unique features but overlapping concepts and methodologies. Project 1 will study the dystrophin-associated complex of proteins, emphasizing the sarcoglycans and the newly described filamin-C. Project 2 will investigate the biology of dysferlin, its potential protein partners, and how these are altered by dysferlin gene mutations. Project 3 will examine the function of myotubularin in normal muscle development and the mechanisms by which its mutations cause developmental myopathies. Project 4 will study the biological and therapeutic properties of muscle stem cells. Three Cores will provide administrative oversight and services essential to the smooth progression of this program. Core B will coordinate sample acquisition and muscle RNA preparation for each project. Core C will perform the microarray analysis of gene expression and provide expertise in bioinformatics and data interpretation. The aim is to identify patterns of gene expression that are global in all dystrophies or distinct to specific sets of dystrophies and myopathies; this will provide insight into the molecular basis of normal muscle development and its dysfunction in these disease states. The long-term goal is to use this information in conjunction with the insights from studies of stem cell biology to devise new approaches to the treatment of the muscular dystrophies and related myopathies
Keywords: gene expression, muscle protein, muscular dystrophy, myogenesis, protein structure function
Project start date: 2001-09-25
Project end date: 2006-08-31
1P01NS040828-01A1 (2001): $1402100
GENETIC STUDIES OF INTERSTITIAL CYSTITIS
Louis M Kunkel
Children´s Hospital Boston, 300 Longwood Ave, Boston, Ma 02115-5737
Grant 5R01DK081647-03 from National Institute Of Diabetes And Digestive And Kidney Diseases
Abstract: Painful bladder syndrome/interstitial cystitis (PBS/IC) is a chronic, debilitating clinical syndrome presenting as severe pelvic pain with extreme urinary urgency and frequency in the absence of any known cause. The etiologic mechanisms underlying PBS/IC are unknown, but recurrence risks to siblings of affected individuals, concordance among monozygotic twins, and our own preliminary studies indicate a strong genetic contribution to the cause of PBS/IC. The overall goal of this proposal is to identify the genes containing mutations that result in PBS/IC and determine how the different encoded proteins of these genes interact with one another in a common biological pathway. Ultimately, understanding how mutations in up to five different genes yield the symptoms of PBS/IC should lead to improved diagnosis and possible therapies. We plan to attain our goals via the following specific aims 1) Accurately characterize PBS/IC patients and their family members and enroll them in our genetic studies. Carefully evaluated cohorts of patients and their families are essential to the discovery of the genetic variation underlying PBS/IC. 2) Map the locations of PBS/IC genes by linkage analysis in families in which PBS/IC is segregating. We have already recruited several families with autosomal dominant inheritance of PBS/IC and identified linkage peaks in the most informative pedigrees. 3) Identify within linked regions of the genome the first genes that control inherited risk of PBS/IC and correlate the types of mutations with clinical symptoms. 4) Determine how proteins encoded by these PBS/IC genes interact with one another. Such knowledge should yield a better understanding of the biochemical or developmental pathways leading to PBS/IC. Painful bladder syndrome/interstitial cystitis (PBS/IC) is a severe, chronic, debilitative clinical syndrome with pelvic pain, extreme urinary urgency and frequency without a known underlying cause. This proposal should lead to a better understanding of the genetic components of the disorder and the underlying biochemical and developmental pathways leading to PBS/IC. Such knowledge could lead to improved diagnosis and rational design of therapeutic interventions
Keywords: Affect; Biochemical; Biological; Chronic; Clinical; Collection; DNA; Data; Deoxyribonucleic Acid; Development; Diagnosis; Disease; Disorder; Enrollment; Epidemiology, Family Medical History; Family; Family Medical History; Family history of; Family member; Frequencies (time pattern); Frequency; Future; Gene Proteins; Gene variant; Genes; Genetic; Genetic Alteration; Genetic Change; Genetic Diversity; Genetic Variation; Genetic defect; Genome; Genotype; Goals; Head and Neck, Saliva; Hereditary; Human Genome; Individual; Inherited; Interstitial Cystitis; Knowledge; Lead; Link; Linkage Analysis; Location; Maps; Molecular Diagnosis; Monozygotic Twinning; Monozygotic twins; Mutation; Nature; Pathway interactions; Patients; Pb element; Pedigree; Pelvic Pain; Phenotype; Protein Gene Products; Proteins; Proteomics; Recruitment Activity; Recurrence; Recurrent; Relative; Relative (related person); Risk; Saliva; Sampling; Siblings; Symptoms; Syndrome; Testing; Therapeutic Intervention; Twins, Identical; Urgency of micturition; Urgency to pass urine; Urgent desire to urinate; Urinary System, Urine; Urine; Variation (Genetics); allelic variant; biomarker; clinical Diagnosis; cohort; design; designing; disease/disorder; enroll; family based linkage study; gene function; gene product; genetic linkage analyses; genetic linkage analysis; genetic pedigree; genome mutation; heavy metal Pb; heavy metal lead; improved; intervention therapy; linkage analyses; micturition urgency; painful bladder syndrome; pathway; pedigree structure; proband; public health relevance; recruit; urinary urgency; urination urgency
Project start date: 2008-08-01
Project end date: 2012-07-31
Budget start date: 1-AUG-2010
Budget end date: 31-JUL-2011
PFA/PA: PA-07-070
5R01DK081647-03 (2010): $588462
5R01DK081647-02 (2009): $573947
Pathogenesis And Treatment Of Muscular Dystrophy
Louis M Kunkel, Professor
Children s Hospital Boston 300 Longwood Ave Boston, Ma 021155737
Grant 2P01NS040828-06A1 from National Institute Of Neurological Disorders And Stroke IRG: ZNS1
Abstract: Over the last 5 years, we have been working to establish the zebrafish as a model for muscular dystrophy. In this capacity, we have published the phenotype of zebrafish lacking dystrophin and 5- sarcoglycan, completed a large genetic screen to isolate additional dystrophic mutants, and identified the mutant gene in the runzel mutant. Using our experiences in muscle research and in establishing the zebrafish as a disease model, we now propose to use the fish to investigate the pathogenesis of muscular dystrophy and evaluate cell therapy as a potential treatment option. The first aim in this project proposes to fully characterize one of our available dystrophic zebrafish models with the goal of better understanding the pathogenesis of muscular dystrophy. We have selected the emz mutant for further analysis since its mutation rough maps to a genomic interval void of any genes orthologous to those currently associated with muscular dystrophy. This mutant shows a phenotype very similar to the dystrophin mutant (sapje) suggesting that the emz phenotype of muscle degeneration is symptomatic of muscular dystrophy. We propose to identify the genetic mutation in this mutant using a traditional mapping approach and then sequencing candidate genes to identify the specific mutation. If the orthologous human gene is not currently associated with muscular dystrophy, the gene will be considered a disease candidate and sequenced in human patients for which the cause of muscular dystrophy is unknown. Since mutations in seemingly unrelated proteins can manifest as muscular dystrophy, the identification of additional genes would be helpful for establishing disease pathways. Secondly, we have established, methods to transplant cell populations in zebrafish at all developmental stages and now propose using this system to identify the cell population most capable of engrafting into and correcting the diseased muscle. Gene expression profiles of muscle engrafting cell populations will be compared with non-engrafting cells to identify genes expressed predominantly in the engrafting cells. Differentially expressed genes will be considered potential markers and used to purify analogous cell populations in mammals for future experimentation and therapy. Finally, we plan to dissect the lineage relationship of various stem cell populations by assaying the developmental potential of zebrafish muscle progenitor cells. This will be accomplished by transplanting limited populations of labeled cells early in development and then following their fate as the fish matures.
Project start date: 2000-12-01
Project end date: 2012-03-31
2P01NS040828-06A1 (2007): $1399630
BIOMARKER DISCOVERY IN MUSCLES FROM FSHD PATIENTS
Louis M Kunkel
Boston Biomedical Research Institute, 64 Grove St, Watertown, Ma 02472
Abstract: FSHD is one of the more common forms of muscular dystrophy in humans with a very unusual and poorly understood biochemical, developmental and molecular underpinning. What is clear is that the D4Z4 repeat deletion in some way influences the expression of genes in muscle in a dominant fashion to cause a variable degree of myofiber degeneration and muscle weakness in different patients. We have established by both proteomic and RNA profiling of muscle biopsies from FSHD patient that both proteins and RNA change expression patterns in diseased tissue. In addition, we have observed in 5 FSHD families from Brazil that some asymptomatic carriers of D4Z4 deletions substantially increase expression of 12 genes including 2 chemokines encoded on chromosome 4 which are only modestly changed in symptomatic patients. We propose to follow up on these findings and use protein, microRNA and mRNA expression profiling as an approach to understanding the differences in disease severity in different individuals with the D4Z4 deletion with the hope that this understanding might lead to the discovery of biomarkers which will be useful in evaluating potential treatments of FSHD. We propose to accomplish this goal according to the following specific aims 1) Continue to profile mRNA from FSHD patients, control muscle and cell lines generated from differentially affected muscles and confirm existing and new array data by RT-PCR. 2) Confirm our observation on the differential expression of certain miRNAs in FSHD muscle and look at the change in expression of the predicted targets of our observed miRNAs in the mRNA expression arrays from aim 1. We will also ablate these candidate miRNAs in myogenic cell lines to see if we can recapitulate in culture the gene expression changes we see in skeletal muscle of FSHD patients. 3) Lastly, we will continue parallel experiments to define the changes in the proteome in FSHD muscles and myogenic cell lines
Budget start date: 1-SEP-2010
Budget end date: 31-AUG-2011
5U54HD060848-03_0002 (2010): $327165
5U54HD060848-02_0002 (2009): $330385
GENE EXPRESSION % FILAMIN / SARCOGLYCAN-RELATED DYSTROPHIES
Louis M Kunkel, Professor
Children s Hospital Boston 300 Longwood Ave Boston, Ma 021155737
Grant 5P01NS040828-020001 from National Institute Of Neurological Disorders And Stroke IRG: NSD
Abstract: For more than 10 years, abnormalities of dystrophin have been known to be the major cause of muscle cell degeneration. Dystrophin is part of a larger complex of proteins, some of which are also involved in distinct forms of limb girdle muscular dystrophy (LGMD). This complex of proteins links the internal cytoskeleton to the extracellular matrix, and alteration in this linkage causes membrane instability, and ultimately, muscle cell degeneration. In LGMD, caused by abnormalities in the sarcoglycans, this linkage is preserved, yet muscles still degenerate. The recent identification of FLNC as a sarcoglycan-interacting protein raises the possibility that there is a signaling role to the sarcoglycans, and it is this signaling that is necessary to maintain the muscle cell integrity. To better understand the normal function of the sarcoglycans and FLNC in muscle fiber stability and the potential for therapy, we propose 5 specific aims 1) To look at FLNC in normal and diseased muscle to determine the role it might play in muscle myofiber stability; 2) To look for novel proteins associated with FLNC and the sarcoglycans via yeast two-hybrid and chemical cross-linking; 3) To use gene chip and gene array technologies to characterize the mRNA expression patterns in dystrophin, sarcoglycan, calpain-3 and FLNC deficient human muscle and compare it to normal; 4) To validate the expression results and develop new hypotheses about the pathogenesis of these diseases and begin the confirmation of these findings by more conventional approaches such as co-IP and co-localization; 5) To determine the existence of stem cells in the muscle of animal models of these diseases and determine whether normal muscle stem cells are corrective for these dystrophies. The latter knowledge is essential if these stem cells are ever to be used in human therapy.
Keywords: actin binding protein, dystrophin, gene expression, muscular dystrophy, protein structure function, biological signal transduction, calpain, disease /disorder model, gene mutation, glycoprotein, integrin, messenger RNA, muscle cell, muscle satellite cell, pathologic process, phosphorylation, protein protein interaction, crosslink, human tissue, laboratory mouse, microarray technology, yeast two hybrid system
Sponsored Links Excellgen http://Excellgen.com
DYSTROPHIN IN THE NERVOUS SYSTEM
Louis M Kunkel
Institution:
Grant 5P01HD018658-140015 from National Institute Of Child Health And Human Development
Abstract: Dystrophin, the protein product of the Duchenne/Becker muscular dystrophy locus, is normally present in muscle tissues and the brain. Approximately one third of patients exhibit a mild degree of mental retardation which is presumably caused by abnormalities of dystrophin. We propose to accurately localize dystrophin within the nervous system and determine what role it might play in normal cognitive function. Attempts will be made to isolate as cDNA clones those proteins which interact with dystrophin in neurons, and experiments will be designed to understand the nature of this interaction. Patients will be sought who have abnormalities of dystrophin expressed in the brain, but normal expression of dystrophin in muscle. Clinical findings in such patients may reveal insights into normal neuronal development and how it is altered in mental retardation. Finally, we propose to characterize the novel shorter dystrophin transcripts which are also expressed in normal brain and may have a role in mental development.
Keywords: dystrophin, gene expression, genetic disorder, mental retardation, muscular dystrophy, brain, gene mutation, phenotype, protein, synaptosome, computer assisted sequence analysis, electron microscopy, genetic library, human genetic material tag, human tissue, immunocytochemistry, laboratory mouse, laboratory rabbit, molecular cloning, nucleic acid sequence, polymerase chain reaction, western blotting
DYSTROPHIN AND RELATED PROTEINS IN NEUROMUSCULAR DISEASE
Louis M Kunkel, Professor
Children s Hospital Boston 300 Longwood Ave Boston, Ma 021155737
Grant 5R01NS023740-10 from National Institute Of Neurological Disorders And Stroke IRG: MGN
Abstract: Dystrophin is a large molecular weight component of the muscle and nerve cell cytoskeleton. Disruption of normal function leads to the genetic disorders Duchenne and Becker muscular dystrophy. Deletion mutations are the most common means to disrupt function but other mutations are known to exist. The proposed characterization of these other mutations will greatly increase the accuracy of diagnosis for at risk individuals and should shed additional light on how normal function can be affected. Dystrophin has been shown to be a member of a family of related proteins with overlapping yet discreet functions. Some of these related proteins may partially compensate for absent dystrophin and hence represent potential candidates for therapeutic intervention to strengthen the compensatory ability. These related proteins are themselves prime candidates to be disrupted in other neuromuscular diseases which, like Duchenne and Becker dystrophy, are slowly progressive. New members of the family will be cloned, mapped to chromosomal locations, and tested for disruption in candidate neuromuscular diseases. Functional analysis of these proteins should shed light on their role in the membrane cytoskeleton and how this role is altered in disease. This should result not only in the identification of other disease causing genes, but also proteins with the potential to mitigate the absence of dystrophin in Duchenne dystrophy.
Project start date: 1986-07-01
Project end date: 1998-06-30
5R01NS023740-10 (1995): $211806
5R01NS023740-09 (1994): $195836
5R01NS023740-08 (1993): $193690
5R01NS023740-07 (1992): $196445
5R01NS023740-12 (1997): $221230
Louis M Kunkel
Children´s Hospital Boston, 300 Longwood Ave, Boston, Ma 02115-5737
Abstract: 7a.2. Overall Objective The overall objective of the Cell Sorter Core is to offer Center investigators access to high quality flow cytometry and cell sorting. An important aim of this Core also is to provide expertise to investigators concerning the best approaches to sample preparation and staining techniques. Core personnel also assist investigators with data interpretation and presentation. Thus, our overall objective is to provide both service and training
Keywords: Cell Cycle; Cell Division Cycle; Cell Isolation; Cell Segregation; Cell Separation; Cell Separation Technology; Cell surface; Cells; Cytofluorometry, Flow; DNA Content; DNA Index; DNA Ploidy; Data; Flow Cytofluorometries; Flow Cytometry; Flow Microfluorimetry; Fluorescence; GFP; Green Fluorescent Proteins; Human Resources; Investigators; Life; Manpower; Measurement; Methods and Techniques; Methods, Other; Microfluorometry, Flow; Ploidies; Preparation; Research Personnel; Researchers; Sampling; Services; Sorting - Cell Movement; Staining method; Stainings; Stains; Techniques; Training; base; cell sorting; chromosome complement; flow cytophotometry; personnel; sorting; stem
Budget start date: 1-JUL-2010
Budget end date: 30-JUN-2011
5P30HD018655-29_9006 (2010): $213447
Louis M Kunkel
Children´s Hospital Boston, 300 Longwood Ave, Boston, Ma 02115-5737
Abstract: 6.a. 1.1. Introductory Overview Sequence databases and technology development that emanated from the Human Genome Project now offer avenues of molecular genetic and functional genomic analyses in biomedical research which were previously impossible as recently as the previous funding cycle for this Center Grant. The rapid pace of technological and instrumentation improvements that facilitate cutting edge molecular genetic and genomic approaches requires that we acquire new technologies, and have a highly skilled support staff that is adept at developing and supporting new instrumentation and methodologies. The Molecular Genetics Core has evolved to advance MRDDRC related research projects in three key areas 1) high-throughput sequence analysis, 2) genomewide genetic marker genotype analysis for mapping and positional cloning, and 3) array-based transcriptional profiling. In providing these services to MRDDRC investigators, the Molecular Genetics Core has developed a cost-effective and efficient mechanism for these technically sophisticated approaches to be centrally managed to maintain quality and speed of data turn-around to MRDDRC investigators. The three functional components of the Molecular Genetics Core - Sequencing, Genotyping, and Expression Arrays - are all heavily utilized by MRDDRC investigators. Automated sequencing was established in the Core in the late 1980´s following the cloning of the dystrophin locus, which ultimately led to the complete sequence analysis of three autosomal recessive forms of dystrophy. In its current form, the Core is being utilized in a high throughput sequencing strategy to identify mutations in all the known genes mutated in muscle diseases. Additionally, numerous MRDDRC investigators submit DMA for sequencing in the Core rather than outsourcing projects to commercial vendors because the in-house service is cost effective and rapid. Genotyping technologies supported by the Core include analysis of dinucleotide repeat (microsatellite) markers and single nucleotide polymorphisms (SNPs). These are performed as custom assays using the ABI 3730 capillary DMA analyzer for microsatellite markers, or on the ABI 7900 HT 384-well Real-time PCR machine for Taqman-based SNP genotyping. For whole genome mapping and genetic marker disease association studies, the Core supports microarray analysis of SNPs using both the Affymetrix 10k and 100k SNP genotyping arrays (which query 10,000 or 100,000 polymorphic human genetic markers respectively). In such studies, genomic DNA from affected and unaffected individuals is labeled and hybridized to the genotyping arrays. Using statistical analysis of the genotypes at each of the thousands of markers examined among the control and patient DNA samples, a probability of linkage between the disease trait and each genetic marker (SNP) can be assigned. The Core has supported MRDDRC investigators in mapping genes involved in eye movement disorders and identifying autosomal mutations involved in recessive disorders of brain development. The Affymetrix 10K and 100K SNP genotyping arrays are also being used to map other complex traits such as autism. The expression array component of the facility is an important new addition that has been used extensively by MRDDRC investigators to compare normal and diseased tissue mRNA expression patterns. In these studies, Affymetrix expression arrays are hybridized to probes generated from mRNA isolated from control and diseased tissues. Genes with statistically significant changes in expression (either up- or down-regulated) are identified, and form the basis for new hypothesis driven experiments to understand the molecular basis of the disease. Establishment of this technology was made possible by a Program Project grant to the Core Director. The array component of the Molecular Genetics Core is overseen by Drs. Isaac Kohane and Alan Beggs, and has extensive support from the Children´s Hospital bioinformatics group led by Dr. Kohane. Close ties with this program are indicated by 14 collaborative publications between MRDDRC investigators and the bioinformatics group. Historically, the facility has tried to keep up with the latest technologic advances in sequencing, genotyping and expression arrays. Gene expression and SNP genotyping capabilities were recently upgraded with the purchase of the Illumina Bead Array system, which provides users access to customized arrays at lower cost than other platforms. The Core has increased sequencing capacity with the purchase of a second Applied Biosystems 48-capillary sequencer and further increased capacity by implementing an automated workstation for high throughput low volume DNA cycle-sequencing. The Core recently relocated to newly renovated and increased space on the 10th floor of the Enders research building. The staff is highly trained in all the methodologies used within the facility and continue to update their training as new techniques become available. The staff has extensive experience working with MRDDRC investigators in planning experiments and in the analysis of their data. In aggregate, this Core has a strong history and is essential to the research programs of MRDDRC investigators. 6.a.2. Overall Objective The overall objective of the Molecular Genetics Core is to provide high quality, low cost DNA sequencing, microsatellite genotyping and a variety of SNP genotyping and gene expression analysis technologies for MRDDRC investigators. The Core strives to complement these technologies with very strong biostatistics support. It also strives to serve as a reliable resource for investigators pursuing a broader understanding of the genetic basis of developmental disabilities
Keywords: Affect; Area; Assay; Autism; Autism, Early Infantile; Autism, Infantile; Autistic Disorder; Bio-Informatics; Bioassay; Bioinformatics; Biologic Assays; Biological Assay; Biomedical Research; Biometrics; Biometry; Biometry and Biostatistics; Biostatistics; Blood capillaries; Body Tissues; Brain Diseases; Brain Disorders; Capillaries; Capillary; Capillary, Unspecified; Child Development Disorders; Chromosome Mapping; Cloning; Complement; Complement Proteins; Complex; Computer Programs; Computer software; Custom; DNA; DNA Resequencing; DNA Sequence; DNA Sequence Analysis; DNA Sequencing Facility; DNA purification; Data; Data Analysis, Statistical; Data Banks; Data Bases; Data Interpretation, Statistical; Databank, Electronic; Databanks; Database, Electronic; Databases; Deoxyribonucleic Acid; Development; Developmental Disabilities; Dinucleotide Repeats; Disease; Disease Association; Disorder; Disorder of muscle, unspecified; Dystrophin; Encephalon Diseases; Eye Motility Disorders; Eye Movement Disorders; Floor; Funding; Gene Expression; Gene Localization; Gene Mapping; Gene Mapping, Total Human and Non-Human; Genes; Genetic; Genetic Alteration; Genetic Change; Genetic Markers; Genetic defect; Genetics, Gene Mapping; Genome Mappings; Genomics; Genotype; Grant; History; Housing; Human Genetics; Human Genome Project; IDDRC; IDDRP; Individual; Instrumentation, Other; Intellectual and Developmental Disabilities Research Centers; Intracranial CNS Disorders; Intracranial Central Nervous System Disorders; Investigators; Kanner`s Syndrome; Label; Linkage (Genetics); Linkage Mapping; MRDD Research Center; MRDDRC; Maps; Mental Retardation and Developmental Disabilities Research Centers; Messenger RNA; Method LOINC Axis 6; Methodology; Methods and Techniques; Methods, Other; Microarray Analysis; Microarray-Based Analysis; Microsatellite Markers; Microsatellite Repeats; Microsatellites; Molecular; Molecular Genetic; Molecular Genetics; Muscle Disease; Muscle Disorders; Muscle disease or syndrome; Muscular Diseases; Mutate; Mutation; Myopathic Conditions; Myopathic Diseases and Syndromes; Myopathic disease or syndrome; Myopathy; Myopathy, unspecified; Ocular Motility Disorders; Outsourcing; P01 Mechanism; P01 Program; Patients; Pattern; Pediatric Hospitals; Polymorphism, Single Base; Probability; Program Project Grant; Program Research Project Grants; Programs (PT); Programs [Publication Type]; Publications; R01 Mechanism; R01 Program; RNA, Messenger; RPG; Recording of previous events; Research; Research Grants; Research Personnel; Research Program Projects; Research Project Grants; Research Projects; Research Projects, R-Series; Research Resources; Researchers; Resequencing; Resources; SEQ-AN; SNP; SNPs; Sampling; Scientific Publication; Sequence Analyses; Sequence Analyses, DNA; Sequence Analysis; Sequence Analysis, DNA; Sequencing Core; Services; Single Nucleotide Polymorphism; Software; Speed; Speed (motion); Statistical Data Analyses; Statistical Data Interpretation; System; System, LOINC Axis 4; Techniques; Technology; Time; Tissues; Training; Update; Vendor; Work; base; capillary; clinical data repository; clinical data warehouse; computer program/software; cost; cost effective; data repository; design; designing; disease/disorder; experience; experiment; experimental research; experimental study; functional genomics; genetic linkage; genetic mapping; genome mutation; genome-wide; genotyping technology; instrumentation; mRNA; mRNA Expression; microarray technology; muscular disorder; new technology; positional cloning; programs; relational database; research study; reverse genetics; statistics/biometry; technology development; trait
Budget start date: 1-JUL-2010
Budget end date: 30-JUN-2011
5P30HD018655-29_9001 (2010): $213446
Louis M Kunkel
Children´s Hospital Boston
Project start date: 2008-07-22
Project end date: 2013-02-28
Sponsored Links Excellgen http://Excellgen.com
RNA Expression Patterns In Autism
Louis M Kunkel, Professor
Children´s Hospital Boston
Grant 5R01MH085143-02 from National Institute Of Mental Health IRG: ZHD1
Project start date: 2008-07-22
Project end date: 2013-02-28
Louis M Kunkel, Professor Of Genetics And Pediatrics
Children´s Hospital Boston, 300 Longwood Ave, Boston, Ma 02115-5737
Keywords: Affect; Area; Assay; Autism; Autism, Early Infantile; Autism, Infantile; Autistic Disorder; Bio-Informatics; Bioassay; Bioinformatics; Biologic Assays; Biological Assay; Biomedical Research; Biometrics; Biometry; Biometry and Biostatistics; Biostatistics; Blood capillaries; Body Tissues; Brain Diseases; Brain Disorders; Capillaries; Capillary; Capillary, Unspecified; Child Development Disorders; Chromosome Mapping; Cloning; Complement; Complement Proteins; Complex; Computer Programs; Computer software; Custom; DNA; DNA Resequencing; DNA Sequence; DNA Sequence Analysis; DNA Sequencing Facility; DNA purification; Data; Data Analysis, Statistical; Data Banks; Data Bases; Data Interpretation, Statistical; Databank, Electronic; Databanks; Database, Electronic; Databases; Deoxyribonucleic Acid; Development; Developmental Disabilities; Dinucleotide Repeats; Disease; Disease Association; Disorder; Disorder of muscle, unspecified; Dystrophin; Encephalon Diseases; Eye Motility Disorders; Eye Movement Disorders; Floor; Funding; Gene Expression; Gene Localization; Gene Mapping; Gene Mapping, Total Human and Non-Human; Genes; Genetic; Genetic Alteration; Genetic Change; Genetic Markers; Genetic defect; Genetics, Gene Mapping; Genetics, Human; Genome Mappings; Genomics; Genotype; Grant; History; Hospitals, Pediatric; Housing; Human Genetics; Human Genome Project; IDDRC; IDDRP; Individual; Instrumentation, Other; Intellectual and Developmental Disabilities Research Centers; Intracranial CNS Disorders; Intracranial Central Nervous System Disorders; Investigators; Kanner`s Syndrome; Label; Linkage (Genetics); Linkage Mapping; MRDD Research Center; MRDDRC; Mappings, Genome; Maps; Mental Retardation and Developmental Disabilities Research Centers; Messenger RNA; Method LOINC Axis 6; Methodology; Methods and Techniques; Methods, Other; Microarray Analysis; Microarray-Based Analysis; Microsatellite Markers; Microsatellite Repeats; Microsatellites; Molecular; Molecular Genetic; Molecular Genetics; Muscle Disease; Muscle Disorders; Muscle disease or syndrome; Muscular Diseases; Mutate; Mutation; Myopathic Conditions; Myopathic Diseases and Syndromes; Myopathic disease or syndrome; Myopathy; Myopathy, unspecified; Ocular Motility Disorders; Outsourcing; P01 Mechanism; P01 Program; Patients; Pattern; Pediatric Hospitals; Polymorphism, Single Base; Probability; Program Project Grant; Program Research Project Grants; Programs (PT); Programs [Publication Type]; Publications; R01 Mechanism; R01 Program; RNA, Messenger; RPG; Recording of previous events; Research; Research Grants; Research Personnel; Research Program Projects; Research Project Grants; Research Projects; Research Projects, R-Series; Research Resources; Researchers; Resequencing; Resources; SEQ-AN; SNP; SNPs; Sampling; Scientific Publication; Sequence Analyses; Sequence Analyses, DNA; Sequence Analysis; Sequence Analysis, DNA; Sequencing Core; Services; Single Nucleotide Polymorphism; Software; Speed; Speed (motion); Statistical Data Analyses; Statistical Data Interpretation; System; System, LOINC Axis 4; Techniques; Technology; Time; Tissues; Training; Update; Vendor; Work; base; capillary; clinical data repository; clinical data warehouse; computer program/software; cost; cost effective; data repository; design; designing; disease/disorder; experience; experiment; experimental research; experimental study; functional genomics; genetic linkage; genetic mapping; genome mutation; genome-wide; genotyping technology; instrumentation; mRNA; mRNA Expression; microarray technology; muscular disorder; new technology; positional cloning; programs; relational database; research study; reverse genetics; statistics/biometry; technology development; trait
Budget start date: 1-JUL-2009
Budget end date: 30-JUN-2010
PFA/PA: RFA-HD-05-030
5P30HD018655-28_9001 (2009): $215602
Louis M Kunkel, Professor Of Genetics And Pediatrics
Children´s Hospital Boston, 300 Longwood Ave, Boston, Ma 02115-5737
Keywords: Cell Cycle; Cell Division Cycle; Cell Isolation; Cell Segregation; Cell Separation; Cell Separation Technology; Cell surface; Cells; Cytofluorometry, Flow; DNA Content; DNA Index; DNA Ploidy; Data; Flow Cytofluorometries; Flow Cytometry; Flow Microfluorimetry; Fluorescence; GFP; Green Fluorescent Proteins; Human Resources; IDDRC; IDDRP; Intellectual and Developmental Disabilities Research Centers; Investigators; Life; MRDD Research Center; MRDDRC; Manpower; Measurement; Mental Retardation and Developmental Disabilities Research Centers; Methods and Techniques; Methods, Other; Microfluorometry, Flow; Ploidies; Preparation; Research Personnel; Researchers; Sampling; Services; Sorting - Cell Movement; Staining method; Stainings; Stains; Techniques; Training; base; cell sorting; chromosome complement; flow cytophotometry; personnel; sorting; stem
Budget start date: 1-JUL-2009
Budget end date: 30-JUN-2010
PFA/PA: RFA-HD-05-030
5P30HD018655-28_9006 (2009): $215603
Louis M Kunkel, Professor
Children s Hospital Boston 300 Longwood Ave Boston, Ma 021155737
Grant 5P30HD018655-199004 from National Institute Of Child Health And Human Development
Keywords: biomedical facility, computer graphics /printing, information dissemination, photography, developmental neurobiology, digital imaging, informatics, information display, mental retardation, publication, videotape /videodisc, behavioral /social science research tag, bioimaging /biomedical imaging, human data
Louis M Kunkel, Professor
Children s Hospital Boston 300 Longwood Ave Boston, Ma 021155737
Grant 5P30HD018655-199014 from National Institute Of Child Health And Human Development
Keywords: biomedical facility, genetic disorder diagnosis, nucleic acid sequence, computer assisted sequence analysis, developmental neurobiology, gene mutation, genotype, linkage mapping, mental retardation, molecular genetics, tissue resource /registry, human genetic material tag, human tissue
Louis M Kunkel, Professor
Children s Hospital Boston 300 Longwood Ave Boston, Ma 021155737
Grant 5P30HD018655-189004 from National Institute Of Child Health And Human Development
Keywords: biomedical facility, computer graphics /printing, information dissemination, photography, developmental neurobiology, digital imaging, informatics, information display, mental retardation, publication, videotape /videodisc, behavioral /social science research tag, bioimaging /biomedical imaging, human data
Louis M Kunkel, Professor
Children s Hospital Boston 300 Longwood Ave Boston, Ma 021155737
Grant 5P30HD018655-189014 from National Institute Of Child Health And Human Development
Keywords: biomedical facility, genetic disorder diagnosis, nucleic acid sequence, computer assisted sequence analysis, developmental neurobiology, gene mutation, genotype, linkage mapping, mental retardation, molecular genetics, tissue resource /registry, human genetic material tag, human tissue
Louis M Kunkel, Professor
Children s Hospital Boston 300 Longwood Ave Boston, Ma 021155737
Grant 5P30HD018655-179004 from National Institute Of Child Health And Human Development
Keywords: biomedical facility, computer graphics /printing, information dissemination, photography, developmental neurobiology, digital imaging, informatics, information display, mental retardation, publication, videotape /videodisc, behavioral /social science research tag, human data
Louis M Kunkel, Professor
Children s Hospital Boston 300 Longwood Ave Boston, Ma 021155737
Grant 5P30HD018655-179014 from National Institute Of Child Health And Human Development
Keywords: biomedical facility, genetic disorder diagnosis, nucleic acid sequence, computer assisted sequence analysis, developmental neurobiology, gene mutation, genotype, linkage mapping, mental retardation, molecular genetics, tissue resource /registry, human genetic material tag, human tissue
CORE--TISSUE CULTURE/DISHWASHING CORE
Louis M Kunkel, Professor
Children s Hospital Boston 300 Longwood Ave Boston, Ma 021155737
Grant 5P01HD018658-159001 from National Institute Of Child Health And Human Development
Keywords: biomedical facility, chromosome disorder, cytogenetics, tissue /cell culture
Sponsored Links Excellgen http://Excellgen.com