Reproducibility with zebrafish models of human health and disease
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1 Reproducibility with zebrafish models of human health and disease Monte Westerfield, University of Oregon, Eugene, USA Sponsored by the Office of the Director National Institutes of Health, the National Human Genome Research Institute, the National Institute of Child Health & Development, the National Institute of Deafness & Other Disorders the National Eye Institute, and the Megan and Vision for a Cure Foundations VisionForACure.com
2 Reproducibility with zebrafish models of human health and disease Why zebrafish? Current use in biomedical research Improving reproducibility
3
4 zebrafish zebrafish fly human
5 Study development directly in transparent embryos (Karlstrom & Kane)
6 Reporter constructs & highly efficient transgenesis Tol2kit Tol2 cell-specific promoter GFP Tol2 Inject into zebrafish Screen for rescue Assays: OKR, ERG, tap test, apoptosis, etc. Models for translational studies
7 Study physiology directly
8 Study cellular interactions (Wilkinson laboratory)
9 Study blood flow directly in live animals (Santoro laboratory)
10 Study activity of every neuron in the brain simultaneously (Keller & Ahrens, 2013)
11 High-throughput optical screening (Pardo-Martin et al., 2010)
12 Reproducibility with zebrafish models of human health and disease Why zebrafish? in vivo analyses at high resolution rapid, inexpensive Current use in biomedical research Improving reproducibility
13 Growth of zebrafish research
14 H. sapiens/130 M. musculus/20 D. melanogaster D. rerio C. elegans
15 Human diseases studied with zebrafish models Acute radiation syndrome Age-related macular degeneration Alcoholism Alcoholic liver disease Alzheimers disease Anophthalmia Anxiety disorders Atherosclerosis Autism Autosomal dominant hearing disease Axenfeld-Rieger syndrome Bardet-Biedl syndrome Barth syndrome Bicuspid aortic valve disease Biliary atresia Cancer Colorectal cancer Head and neck squamous cell carcinoma Glioma Invasive carcinoma LEOPARD syndrome Leukemia Liver cancer Lymphoblastic leukemia Melanoma Myologenous leuekmia Neuroblastoma Pancreatic cancer Rhabdomyosarcoma Centronuclear myopathies Cerebral cavernous malformations Cholera Chronic granulomatous disease Chronic kidney disease Circulatory diseases Colitis Congenital cataract Congenital glycoslyation diseases Core binding factor leukemia Corneal dystrophies Cranial facial diseases Crohns disease Cutis laxa Cystic fibrosis Delayed puberty Diabetes Diamond Blackfan anemia Duchenne muscular dystrophy End stage renal disease Emphysema Epilepsy Extrinsic asthma Eye diseases Facioscapulohumeral muscular dystrophy Fanconi anemia Fatty liver disease Fetal alcohol syndrome Focal segmental glomerulosclerosis Fraser syndrome Ganglioside sialidase deficiency disease GAPO progeroid syndrome Glaucoma Glioblastoma Heart disease Atrial fibrillation Cardiac arrhythmia Cardiac conduction diseases Cardiomegaly Congestive heart failure Hypertrophic cardiomyopathy Sudden cardiac arrest Hepatic steatosis Hereditary retinal degenerations Hirschsprung disease HIVAIDS Holt Oram syndrome Hyperammonemia Hypertension Inflammatory bowel disease Intracranial aneurysm Joubert syndrome Juvenile hydrocephalus Kidney diseases Long QT syndrome Lysosomal disease mucolipidosis II Microphthamia Midfacial cleft disease Multiple sclerosis Mucolipidosis type IV Muscular dystrophy Myotonic dystrophy Nephronophthisis Nonsyndromic cleft lip and palate Noonan syndrome Obesity Oral cleft diseases Osteoporosis Parkinson disease Polycystic kidney disease Pseudoxanthoma elasticum Refractory anemia Schizophrenia Scoliosis Skeletal diseases Sleep disorders Spinal cord regeneration Spinal muscular atrophy Stroke Treacher Collins syndrome Tuberculosis Usher syndrome Uveal coloboma Vascular thrombosis von Willebrand disease (NIH funded grants, 2013)
16 Human diseases studied with zebrafish models Acute radiation syndrome Age-related macular degeneration Alcoholism Alcoholic liver disease Alzheimers disease Anophthalmia Anxiety disorders Atherosclerosis Autism Autosomal dominant hearing disease Axenfeld-Rieger syndrome Bardet-Biedl syndrome Barth syndrome Bicuspid aortic valve disease Biliary atresia Cancer Colorectal cancer Head and neck squamous cell carcinoma Glioma Invasive carcinoma LEOPARD syndrome Leukemia Liver cancer Lymphoblastic leukemia Melanoma Myologenous leuekmia Neuroblastoma Pancreatic cancer Rhabdomyosarcoma Centronuclear myopathies Cerebral cavernous malformations Cholera Chronic granulomatous disease Chronic kidney disease Circulatory diseases Colitis Congenital cataract Congenital glycoslyation diseases Core binding factor leukemia Corneal dystrophies Cranial facial diseases Crohns disease Cutis laxa Cystic fibrosis Delayed puberty Diabetes Diamond Blackfan anemia Duchenne muscular dystrophy End stage renal disease Emphysema Alzheimers Epilepsy Disease Extrinsic asthma Eye diseases Facioscapulohumeral muscular dystrophy Fanconi anemia Fatty liver disease Fetal alcohol syndrome Focal segmental glomerulosclerosis Fraser syndrome Ganglioside sialidase deficiency disease GAPO progeroid syndrome Glaucoma Glioblastoma Heart disease Atrial fibrillation Cardiac arrhythmia Cardiac conduction diseases Cardiomegaly Congestive heart failure Hypertrophic cardiomyopathy Sudden cardiac arrest Hepatic steatosis Hereditary retinal degenerations Hirschsprung disease HIVAIDS Holt Oram syndrome Hyperammonemia Hypertension Inflammatory bowel disease Intracranial aneurysm Joubert syndrome Juvenile hydrocephalus Kidney diseases Long QT syndrome Lysosomal disease mucolipidosis II Microphthamia Midfacial cleft disease Multiple sclerosis Mucolipidosis type IV Muscular dystrophy Myotonic dystrophy Nephronophthisis Nonsyndromic cleft lip and palate Noonan syndrome Obesity Oral cleft diseases Osteoporosis Parkinson disease Polycystic kidney disease Pseudoxanthoma elasticum Refractory anemia Schizophrenia Scoliosis Skeletal diseases Sleep disorders Spinal cord regeneration Spinal muscular atrophy Stroke Treacher Collins syndrome Tuberculosis Usher syndrome Uveal coloboma Vascular thrombosis von Willebrand disease (NIH funded grants, 2013)
17 Human diseases studied with zebrafish models Acute radiation syndrome Age-related macular degeneration Alcoholism Alcoholic liver disease Alzheimers disease Anophthalmia Anxiety disorders Atherosclerosis Autism Autosomal dominant hearing disease Axenfeld-Rieger syndrome Bardet-Biedl syndrome Barth syndrome Bicuspid aortic valve disease Biliary atresia Cancer Colorectal cancer Head and neck squamous cell carcinoma Glioma Invasive carcinoma LEOPARD syndrome Leukemia Liver cancer Lymphoblastic leukemia Melanoma Myologenous leuekmia Neuroblastoma Pancreatic cancer Rhabdomyosarcoma Centronuclear myopathies Cerebral cavernous malformations Cholera Chronic granulomatous disease Chronic kidney disease Circulatory diseases Colitis Congenital cataract Congenital glycoslyation diseases Core binding factor leukemia Corneal dystrophies Cranial facial diseases Crohns disease Cutis laxa Cystic fibrosis Delayed puberty Diabetes Diamond Blackfan anemia Duchenne muscular dystrophy End stage renal disease Emphysema Epilepsy Extrinsic asthma Eye diseases Facioscapulohumeral muscular dystrophy Fanconi Autism anemia Fatty liver disease Fetal alcohol syndrome Focal segmental glomerulosclerosis Fraser syndrome Ganglioside sialidase deficiency disease GAPO progeroid syndrome Glaucoma Glioblastoma Heart disease Atrial fibrillation Cardiac arrhythmia Cardiac conduction diseases Cardiomegaly Congestive heart failure Hypertrophic cardiomyopathy Sudden cardiac arrest Hepatic steatosis Hereditary retinal degenerations Hirschsprung disease HIVAIDS Holt Oram syndrome Hyperammonemia Hypertension Inflammatory bowel disease Intracranial aneurysm Joubert syndrome Juvenile hydrocephalus Kidney diseases Long QT syndrome Lysosomal disease mucolipidosis II Microphthamia Midfacial cleft disease Multiple sclerosis Mucolipidosis type IV Muscular dystrophy Myotonic dystrophy Nephronophthisis Nonsyndromic cleft lip and palate Noonan syndrome Obesity Oral cleft diseases Osteoporosis Parkinson disease Polycystic kidney disease Pseudoxanthoma elasticum Refractory anemia Schizophrenia Scoliosis Skeletal diseases Sleep disorders Spinal cord regeneration Spinal muscular atrophy Stroke Treacher Collins syndrome Tuberculosis Usher syndrome Uveal coloboma Vascular thrombosis von Willebrand disease (NIH funded grants, 2013)
18 Human diseases studied with zebrafish models Acute radiation syndrome Age-related macular degeneration Alcoholism Alcoholic liver disease Alzheimers disease Anophthalmia Anxiety disorders Atherosclerosis Autism Autosomal dominant hearing disease Axenfeld-Rieger syndrome Bardet-Biedl syndrome Barth syndrome Bicuspid aortic valve disease Biliary atresia Cancer Colorectal cancer Head and neck squamous cell carcinoma Glioma Invasive carcinoma LEOPARD syndrome Leukemia Liver cancer Lymphoblastic leukemia Melanoma Myologenous leuekmia Neuroblastoma Pancreatic cancer Rhabdomyosarcoma Centronuclear myopathies Cerebral cavernous malformations Cholera Chronic granulomatous disease Chronic kidney disease Circulatory diseases Colitis Congenital cataract Congenital glycoslyation diseases Core binding factor leukemia Corneal dystrophies Cranial facial diseases Crohns disease Cutis laxa Cystic fibrosis Delayed puberty Diabetes Diamond Blackfan anemia Duchenne muscular dystrophy End stage renal disease Emphysema Epilepsy Extrinsic asthma Eye diseases Facioscapulohumeral muscular dystrophy Fanconi anemia Fatty liver disease Fetal alcohol syndrome Focal segmental glomerulosclerosis Fraser syndrome Obesity Ganglioside sialidase deficiency disease GAPO progeroid syndrome Glaucoma Glioblastoma Heart disease Atrial fibrillation Cardiac arrhythmia Cardiac conduction diseases Cardiomegaly Congestive heart failure Hypertrophic cardiomyopathy Sudden cardiac arrest Hepatic steatosis Hereditary retinal degenerations Hirschsprung disease HIVAIDS Holt Oram syndrome Hyperammonemia Hypertension Inflammatory bowel disease Intracranial aneurysm Joubert syndrome Juvenile hydrocephalus Kidney diseases Long QT syndrome Lysosomal disease mucolipidosis II Microphthamia Midfacial cleft disease Multiple sclerosis Mucolipidosis type IV Muscular dystrophy Myotonic dystrophy Nephronophthisis Nonsyndromic cleft lip and palate Noonan syndrome Obesity Oral cleft diseases Osteoporosis Parkinson disease Polycystic kidney disease Pseudoxanthoma elasticum Refractory anemia Schizophrenia Scoliosis Skeletal diseases Sleep disorders Spinal cord regeneration Spinal muscular atrophy Stroke Treacher Collins syndrome Tuberculosis Usher syndrome Uveal coloboma Vascular thrombosis von Willebrand disease (NIH funded grants, 2013)
19 Human diseases studied with zebrafish models Acute radiation syndrome Age-related macular degeneration Alcoholism Alcoholic liver disease Alzheimers disease Anophthalmia Anxiety disorders Atherosclerosis Autism Autosomal dominant hearing disease Axenfeld-Rieger syndrome Bardet-Biedl syndrome Barth syndrome Bicuspid aortic valve disease Biliary atresia Cancer Colorectal cancer Head and neck squamous cell carcinoma Glioma Invasive carcinoma LEOPARD syndrome Leukemia Liver cancer Lymphoblastic leukemia Melanoma Myologenous leuekmia Neuroblastoma Pancreatic cancer Rhabdomyosarcoma Centronuclear myopathies Cerebral cavernous malformations Cholera Chronic granulomatous disease Chronic kidney disease Circulatory diseases Colitis Congenital cataract Congenital glycoslyation diseases Core binding factor leukemia Corneal dystrophies Cranial facial diseases Crohns disease Cutis laxa Cystic fibrosis Delayed puberty Diabetes Diamond Blackfan anemia Duchenne muscular dystrophy End stage renal disease Emphysema Schizophrenia Epilepsy Extrinsic asthma Eye diseases Facioscapulohumeral muscular dystrophy Fanconi anemia Fatty liver disease Fetal alcohol syndrome Focal segmental glomerulosclerosis Fraser syndrome Ganglioside sialidase deficiency disease GAPO progeroid syndrome Glaucoma Glioblastoma Heart disease Atrial fibrillation Cardiac arrhythmia Cardiac conduction diseases Cardiomegaly Congestive heart failure Hypertrophic cardiomyopathy Sudden cardiac arrest Hepatic steatosis Hereditary retinal degenerations Hirschsprung disease HIVAIDS Holt Oram syndrome Hyperammonemia Hypertension Inflammatory bowel disease Intracranial aneurysm Joubert syndrome Juvenile hydrocephalus Kidney diseases Long QT syndrome Lysosomal disease mucolipidosis II Microphthamia Midfacial cleft disease Multiple sclerosis Mucolipidosis type IV Muscular dystrophy Myotonic dystrophy Nephronophthisis Nonsyndromic cleft lip and palate Noonan syndrome Obesity Oral cleft diseases Osteoporosis Parkinson disease Polycystic kidney disease Pseudoxanthoma elasticum Refractory anemia Schizophrenia Scoliosis Skeletal diseases Sleep disorders Spinal cord regeneration Spinal muscular atrophy Stroke Treacher Collins syndrome Tuberculosis Usher syndrome Uveal coloboma Vascular thrombosis von Willebrand disease (NIH funded grants, 2013)
20 Human diseases studied with zebrafish models Acute radiation syndrome Age-related macular degeneration Alcoholism Alcoholic liver disease Alzheimers disease Anophthalmia Anxiety disorders Atherosclerosis Autism Autosomal dominant hearing disease Axenfeld-Rieger syndrome Bardet-Biedl syndrome Barth syndrome Bicuspid aortic valve disease Biliary atresia Cancer Colorectal cancer Head and neck squamous cell carcinoma Glioma Invasive carcinoma LEOPARD syndrome Leukemia Liver cancer Lymphoblastic leukemia Melanoma Myologenous leuekmia Neuroblastoma Pancreatic cancer Rhabdomyosarcoma Centronuclear myopathies Cerebral cavernous malformations Cholera Chronic granulomatous disease Chronic kidney disease Circulatory diseases Colitis Congenital cataract Congenital glycoslyation diseases Core binding factor leukemia Corneal dystrophies Cranial facial diseases Crohns disease Cutis laxa Cystic fibrosis Delayed puberty Diabetes Diamond Blackfan anemia Duchenne muscular dystrophy End stage renal disease Emphysema Epilepsy Extrinsic asthma Eye diseases Facioscapulohumeral muscular dystrophy Fanconi anemia Fatty liver disease Fetal alcohol syndrome Focal segmental glomerulosclerosis Fraser syndrome Ganglioside sialidase deficiency disease GAPO progeroid syndrome Glaucoma Glioblastoma Heart disease Atrial fibrillation Cardiac arrhythmia Cardiac conduction diseases Sleep disorders Cardiomegaly Congestive heart failure Hypertrophic cardiomyopathy Sudden cardiac arrest Hepatic steatosis Hereditary retinal degenerations Hirschsprung disease HIVAIDS Holt Oram syndrome Hyperammonemia Hypertension Inflammatory bowel disease Intracranial aneurysm Joubert syndrome Juvenile hydrocephalus Kidney diseases Long QT syndrome Lysosomal disease mucolipidosis II Microphthamia Midfacial cleft disease Multiple sclerosis Mucolipidosis type IV Muscular dystrophy Myotonic dystrophy Nephronophthisis Nonsyndromic cleft lip and palate Noonan syndrome Obesity Oral cleft diseases Osteoporosis Parkinson disease Polycystic kidney disease Pseudoxanthoma elasticum Refractory anemia Schizophrenia Scoliosis Skeletal diseases Sleep disorders Spinal cord regeneration Spinal muscular atrophy Stroke Treacher Collins syndrome Tuberculosis Usher syndrome Uveal coloboma Vascular thrombosis von Willebrand disease (NIH funded grants, 2013)
21 Reproducibility with zebrafish models of human health and disease Why zebrafish? in vivo analyses at high resolution rapid, inexpensive Current use in biomedical research widespread and growing Improving reproducibility
22 Forward genetics: thousands of mutants
23 Reverse genetics: morpholinos knockdown gene function mrna Translation blocker Splice blocker
24 Reverse genetics: morpholinos knockdown gene function mrna Translation blocker Splice blocker Problems for reproducibility: toxicity effectiveness diluted over time genomic polymorphisms off-target effects
25 Knock out of all protein coding genes
26 CRISPRs provide precise genome editing
27 Zebrafish models of human patients (Ebermann et al.)
28 Potential genetic interaction between PDZD7 & USH2A (Ebermann et al.)
29 Zebrafish model validates genetic interaction between PDZD7 & USH2A (Ebermann et al.)
30 New tools increase reproducibility complete maps of all SNPs rapid, low cost sequencing isogenic strains
31 Reproducibility with zebrafish models of human health and disease Why zebrafish? in vivo analyses at high resolution rapid, inexpensive Current use in biomedical research widespread Improving reproducibility precise models of human patients improved tools needed
32 Reproducibility with zebrafish models of human health and disease Monte Westerfield, University of Oregon, Eugene, USA Sponsored by the Office of the Director National Institutes of Health, the National Human Genome Research Institute, the National Institute of Child Health & Development, the National Institute of Deafness & Other Disorders the National Eye Institute, and the Megan and Vision for a Cure Foundations VisionForACure.com
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