Monte Westerfield, University of Oregon, Eugene, USASponsored 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
Reproducibility with zebrafish modelsof human health and disease
VisionForACure.com
• Why zebrafish?
• Current use in biomedical research
• Improving reproducibility
Reproducibility with zebrafish modelsof human health and disease
zebrafish fly
human
zebrafish
Study development directly in transparent embryos
(Karlstrom & Kane)
cell-specific promoter GFPTol2 Tol2Tol2kit
Inject into zebrafish
Reporter constructs & highly efficient transgenesis
Screen for rescue Assays: OKR, ERG, tap test, apoptosis, etc.
Models for translational studies
Study physiology directly
(Wilkinson laboratory)
Study cellular interactions
Study blood flow directly in live animals
(Santoro laboratory)
(Keller & Ahrens, 2013)
Study activity of every neuron in the brain simultaneously
High-throughput optical screening
(Pardo-Martin et al., 2010)
• Why zebrafish?• in vivo analyses at high resolution
• rapid, inexpensive
• Current use in biomedical research
• Improving reproducibility
Reproducibility with zebrafish modelsof human health and disease
Growth of zebrafish research
H. sapiens/130
M. musculus/20
D. melanogaster
D. rerioC. elegans
Acute radiation syndromeAge-related macular degenerationAlcoholismAlcoholic liver diseaseAlzheimers diseaseAnophthalmiaAnxiety disordersAtherosclerosisAutismAutosomal dominant hearing diseaseAxenfeld-Rieger syndromeBardet-Biedl syndromeBarth syndromeBicuspid aortic valve diseaseBiliary atresiaCancer
Colorectal cancerHead and neck squamous cell carcinomaGliomaInvasive carcinomaLEOPARD syndromeLeukemiaLiver cancerLymphoblastic leukemiaMelanomaMyologenous leuekmiaNeuroblastomaPancreatic cancerRhabdomyosarcoma
Centronuclear myopathiesCerebral cavernous malformationsCholeraChronic granulomatous diseaseChronic kidney diseaseCirculatory diseasesColitisCongenital cataractCongenital glycoslyation diseasesCore binding factor leukemia
Corneal dystrophiesCranial facial diseasesCrohns diseaseCutis laxa Cystic fibrosisDelayed pubertyDiabetes Diamond Blackfan anemiaDuchenne muscular dystrophyEnd stage renal diseaseEmphysemaEpilepsy Extrinsic asthmaEye diseases
Facioscapulohumeral muscular dystrophyFanconi anemiaFatty liver diseaseFetal alcohol syndromeFocal segmental glomerulosclerosisFraser syndromeGanglioside sialidase deficiency diseaseGAPO progeroid syndromeGlaucomaGlioblastomaHeart disease
Atrial fibrillationCardiac arrhythmiaCardiac conduction diseasesCardiomegalyCongestive heart failureHypertrophic cardiomyopathySudden cardiac arrest
Hepatic steatosisHereditary retinal degenerationsHirschsprung diseaseHIVAIDSHolt Oram syndromeHyperammonemia
Hypertension Inflammatory bowel diseaseIntracranial aneurysm Joubert syndromeJuvenile hydrocephalusKidney diseasesLong QT syndromeLysosomal disease mucolipidosis IIMicrophthamiaMidfacial cleft diseaseMultiple sclerosisMucolipidosis type IVMuscular dystrophyMyotonic dystrophyNephronophthisisNonsyndromic cleft lip and palateNoonan syndromeObesityOral cleft diseasesOsteoporosisParkinson diseasePolycystic kidney diseasePseudoxanthoma elasticumRefractory anemiaSchizophreniaScoliosisSkeletal diseasesSleep disordersSpinal cord regenerationSpinal muscular atrophyStroke Treacher Collins syndromeTuberculosisUsher syndromeUveal colobomaVascular thrombosisvon Willebrand disease
Human diseases studied with zebrafish models
(NIH funded grants, 2013)
Acute radiation syndromeAge-related macular degenerationAlcoholismAlcoholic liver diseaseAlzheimers diseaseAnophthalmiaAnxiety disordersAtherosclerosisAutismAutosomal dominant hearing diseaseAxenfeld-Rieger syndromeBardet-Biedl syndromeBarth syndromeBicuspid aortic valve diseaseBiliary atresiaCancer
Colorectal cancerHead and neck squamous cell carcinomaGliomaInvasive carcinomaLEOPARD syndromeLeukemiaLiver cancerLymphoblastic leukemiaMelanomaMyologenous leuekmiaNeuroblastomaPancreatic cancerRhabdomyosarcoma
Centronuclear myopathiesCerebral cavernous malformationsCholeraChronic granulomatous diseaseChronic kidney diseaseCirculatory diseasesColitisCongenital cataractCongenital glycoslyation diseasesCore binding factor leukemia
Corneal dystrophiesCranial facial diseasesCrohns diseaseCutis laxa Cystic fibrosisDelayed pubertyDiabetes Diamond Blackfan anemiaDuchenne muscular dystrophyEnd stage renal diseaseEmphysemaEpilepsy Extrinsic asthmaEye diseases
Facioscapulohumeral muscular dystrophyFanconi anemiaFatty liver diseaseFetal alcohol syndromeFocal segmental glomerulosclerosisFraser syndromeGanglioside sialidase deficiency diseaseGAPO progeroid syndromeGlaucomaGlioblastomaHeart disease
Atrial fibrillationCardiac arrhythmiaCardiac conduction diseasesCardiomegalyCongestive heart failureHypertrophic cardiomyopathySudden cardiac arrest
Hepatic steatosisHereditary retinal degenerationsHirschsprung diseaseHIVAIDSHolt Oram syndromeHyperammonemia
Hypertension Inflammatory bowel diseaseIntracranial aneurysm Joubert syndromeJuvenile hydrocephalusKidney diseasesLong QT syndromeLysosomal disease mucolipidosis IIMicrophthamiaMidfacial cleft diseaseMultiple sclerosisMucolipidosis type IVMuscular dystrophyMyotonic dystrophyNephronophthisisNonsyndromic cleft lip and palateNoonan syndromeObesityOral cleft diseasesOsteoporosisParkinson diseasePolycystic kidney diseasePseudoxanthoma elasticumRefractory anemiaSchizophreniaScoliosisSkeletal diseasesSleep disordersSpinal cord regenerationSpinal muscular atrophyStroke Treacher Collins syndromeTuberculosisUsher syndromeUveal colobomaVascular thrombosisvon Willebrand disease
Human diseases studied with zebrafish models
(NIH funded grants, 2013)
Alzheimers Disease
Acute radiation syndromeAge-related macular degenerationAlcoholismAlcoholic liver diseaseAlzheimers diseaseAnophthalmiaAnxiety disordersAtherosclerosisAutismAutosomal dominant hearing diseaseAxenfeld-Rieger syndromeBardet-Biedl syndromeBarth syndromeBicuspid aortic valve diseaseBiliary atresiaCancer
Colorectal cancerHead and neck squamous cell carcinomaGliomaInvasive carcinomaLEOPARD syndromeLeukemiaLiver cancerLymphoblastic leukemiaMelanomaMyologenous leuekmiaNeuroblastomaPancreatic cancerRhabdomyosarcoma
Centronuclear myopathiesCerebral cavernous malformationsCholeraChronic granulomatous diseaseChronic kidney diseaseCirculatory diseasesColitisCongenital cataractCongenital glycoslyation diseasesCore binding factor leukemia
Corneal dystrophiesCranial facial diseasesCrohns diseaseCutis laxa Cystic fibrosisDelayed pubertyDiabetes Diamond Blackfan anemiaDuchenne muscular dystrophyEnd stage renal diseaseEmphysemaEpilepsy Extrinsic asthmaEye diseases
Facioscapulohumeral muscular dystrophyFanconi anemiaFatty liver diseaseFetal alcohol syndromeFocal segmental glomerulosclerosisFraser syndromeGanglioside sialidase deficiency diseaseGAPO progeroid syndromeGlaucomaGlioblastomaHeart disease
Atrial fibrillationCardiac arrhythmiaCardiac conduction diseasesCardiomegalyCongestive heart failureHypertrophic cardiomyopathySudden cardiac arrest
Hepatic steatosisHereditary retinal degenerationsHirschsprung diseaseHIVAIDSHolt Oram syndromeHyperammonemia
Hypertension Inflammatory bowel diseaseIntracranial aneurysm Joubert syndromeJuvenile hydrocephalusKidney diseasesLong QT syndromeLysosomal disease mucolipidosis IIMicrophthamiaMidfacial cleft diseaseMultiple sclerosisMucolipidosis type IVMuscular dystrophyMyotonic dystrophyNephronophthisisNonsyndromic cleft lip and palateNoonan syndromeObesityOral cleft diseasesOsteoporosisParkinson diseasePolycystic kidney diseasePseudoxanthoma elasticumRefractory anemiaSchizophreniaScoliosisSkeletal diseasesSleep disordersSpinal cord regenerationSpinal muscular atrophyStroke Treacher Collins syndromeTuberculosisUsher syndromeUveal colobomaVascular thrombosisvon Willebrand disease
Human diseases studied with zebrafish models
(NIH funded grants, 2013)
Autism
Acute radiation syndromeAge-related macular degenerationAlcoholismAlcoholic liver diseaseAlzheimers diseaseAnophthalmiaAnxiety disordersAtherosclerosisAutismAutosomal dominant hearing diseaseAxenfeld-Rieger syndromeBardet-Biedl syndromeBarth syndromeBicuspid aortic valve diseaseBiliary atresiaCancer
Colorectal cancerHead and neck squamous cell carcinomaGliomaInvasive carcinomaLEOPARD syndromeLeukemiaLiver cancerLymphoblastic leukemiaMelanomaMyologenous leuekmiaNeuroblastomaPancreatic cancerRhabdomyosarcoma
Centronuclear myopathiesCerebral cavernous malformationsCholeraChronic granulomatous diseaseChronic kidney diseaseCirculatory diseasesColitisCongenital cataractCongenital glycoslyation diseasesCore binding factor leukemia
Corneal dystrophiesCranial facial diseasesCrohns diseaseCutis laxa Cystic fibrosisDelayed pubertyDiabetes Diamond Blackfan anemiaDuchenne muscular dystrophyEnd stage renal diseaseEmphysemaEpilepsy Extrinsic asthmaEye diseases
Facioscapulohumeral muscular dystrophyFanconi anemiaFatty liver diseaseFetal alcohol syndromeFocal segmental glomerulosclerosisFraser syndromeGanglioside sialidase deficiency diseaseGAPO progeroid syndromeGlaucomaGlioblastomaHeart disease
Atrial fibrillationCardiac arrhythmiaCardiac conduction diseasesCardiomegalyCongestive heart failureHypertrophic cardiomyopathySudden cardiac arrest
Hepatic steatosisHereditary retinal degenerationsHirschsprung diseaseHIVAIDSHolt Oram syndromeHyperammonemia
Hypertension Inflammatory bowel diseaseIntracranial aneurysm Joubert syndromeJuvenile hydrocephalusKidney diseasesLong QT syndromeLysosomal disease mucolipidosis IIMicrophthamiaMidfacial cleft diseaseMultiple sclerosisMucolipidosis type IVMuscular dystrophyMyotonic dystrophyNephronophthisisNonsyndromic cleft lip and palateNoonan syndromeObesityOral cleft diseasesOsteoporosisParkinson diseasePolycystic kidney diseasePseudoxanthoma elasticumRefractory anemiaSchizophreniaScoliosisSkeletal diseasesSleep disordersSpinal cord regenerationSpinal muscular atrophyStroke Treacher Collins syndromeTuberculosisUsher syndromeUveal colobomaVascular thrombosisvon Willebrand disease
Human diseases studied with zebrafish models
(NIH funded grants, 2013)
Obesity
Acute radiation syndromeAge-related macular degenerationAlcoholismAlcoholic liver diseaseAlzheimers diseaseAnophthalmiaAnxiety disordersAtherosclerosisAutismAutosomal dominant hearing diseaseAxenfeld-Rieger syndromeBardet-Biedl syndromeBarth syndromeBicuspid aortic valve diseaseBiliary atresiaCancer
Colorectal cancerHead and neck squamous cell carcinomaGliomaInvasive carcinomaLEOPARD syndromeLeukemiaLiver cancerLymphoblastic leukemiaMelanomaMyologenous leuekmiaNeuroblastomaPancreatic cancerRhabdomyosarcoma
Centronuclear myopathiesCerebral cavernous malformationsCholeraChronic granulomatous diseaseChronic kidney diseaseCirculatory diseasesColitisCongenital cataractCongenital glycoslyation diseasesCore binding factor leukemia
Corneal dystrophiesCranial facial diseasesCrohns diseaseCutis laxa Cystic fibrosisDelayed pubertyDiabetes Diamond Blackfan anemiaDuchenne muscular dystrophyEnd stage renal diseaseEmphysemaEpilepsy Extrinsic asthmaEye diseases
Facioscapulohumeral muscular dystrophyFanconi anemiaFatty liver diseaseFetal alcohol syndromeFocal segmental glomerulosclerosisFraser syndromeGanglioside sialidase deficiency diseaseGAPO progeroid syndromeGlaucomaGlioblastomaHeart disease
Atrial fibrillationCardiac arrhythmiaCardiac conduction diseasesCardiomegalyCongestive heart failureHypertrophic cardiomyopathySudden cardiac arrest
Hepatic steatosisHereditary retinal degenerationsHirschsprung diseaseHIVAIDSHolt Oram syndromeHyperammonemia
Hypertension Inflammatory bowel diseaseIntracranial aneurysm Joubert syndromeJuvenile hydrocephalusKidney diseasesLong QT syndromeLysosomal disease mucolipidosis IIMicrophthamiaMidfacial cleft diseaseMultiple sclerosisMucolipidosis type IVMuscular dystrophyMyotonic dystrophyNephronophthisisNonsyndromic cleft lip and palateNoonan syndromeObesityOral cleft diseasesOsteoporosisParkinson diseasePolycystic kidney diseasePseudoxanthoma elasticumRefractory anemiaSchizophreniaScoliosisSkeletal diseasesSleep disordersSpinal cord regenerationSpinal muscular atrophyStroke Treacher Collins syndromeTuberculosisUsher syndromeUveal colobomaVascular thrombosisvon Willebrand disease
Human diseases studied with zebrafish models
(NIH funded grants, 2013)
Schizophrenia
Acute radiation syndromeAge-related macular degenerationAlcoholismAlcoholic liver diseaseAlzheimers diseaseAnophthalmiaAnxiety disordersAtherosclerosisAutismAutosomal dominant hearing diseaseAxenfeld-Rieger syndromeBardet-Biedl syndromeBarth syndromeBicuspid aortic valve diseaseBiliary atresiaCancer
Colorectal cancerHead and neck squamous cell carcinomaGliomaInvasive carcinomaLEOPARD syndromeLeukemiaLiver cancerLymphoblastic leukemiaMelanomaMyologenous leuekmiaNeuroblastomaPancreatic cancerRhabdomyosarcoma
Centronuclear myopathiesCerebral cavernous malformationsCholeraChronic granulomatous diseaseChronic kidney diseaseCirculatory diseasesColitisCongenital cataractCongenital glycoslyation diseasesCore binding factor leukemia
Corneal dystrophiesCranial facial diseasesCrohns diseaseCutis laxa Cystic fibrosisDelayed pubertyDiabetes Diamond Blackfan anemiaDuchenne muscular dystrophyEnd stage renal diseaseEmphysemaEpilepsy Extrinsic asthmaEye diseases
Facioscapulohumeral muscular dystrophyFanconi anemiaFatty liver diseaseFetal alcohol syndromeFocal segmental glomerulosclerosisFraser syndromeGanglioside sialidase deficiency diseaseGAPO progeroid syndromeGlaucomaGlioblastomaHeart disease
Atrial fibrillationCardiac arrhythmiaCardiac conduction diseasesCardiomegalyCongestive heart failureHypertrophic cardiomyopathySudden cardiac arrest
Hepatic steatosisHereditary retinal degenerationsHirschsprung diseaseHIVAIDSHolt Oram syndromeHyperammonemia
Hypertension Inflammatory bowel diseaseIntracranial aneurysm Joubert syndromeJuvenile hydrocephalusKidney diseasesLong QT syndromeLysosomal disease mucolipidosis IIMicrophthamiaMidfacial cleft diseaseMultiple sclerosisMucolipidosis type IVMuscular dystrophyMyotonic dystrophyNephronophthisisNonsyndromic cleft lip and palateNoonan syndromeObesityOral cleft diseasesOsteoporosisParkinson diseasePolycystic kidney diseasePseudoxanthoma elasticumRefractory anemiaSchizophreniaScoliosisSkeletal diseasesSleep disordersSpinal cord regenerationSpinal muscular atrophyStroke Treacher Collins syndromeTuberculosisUsher syndromeUveal colobomaVascular thrombosisvon Willebrand disease
Human diseases studied with zebrafish models
(NIH funded grants, 2013)
Sleep disorders
• Why zebrafish?• in vivo analyses at high resolution
• rapid, inexpensive
• Current use in biomedical research• widespread and growing
• Improving reproducibility
Reproducibility with zebrafish modelsof human health and disease
Forward genetics: thousands of mutants
Reverse genetics: morpholinos knockdown gene function
Translationblocker
Spliceblocker
mRNA
Problems for reproducibility:• toxicity• effectiveness diluted over time• genomic polymorphisms• off-target effects
Translationblocker
Spliceblocker
mRNA
Reverse genetics: morpholinos knockdown gene function
Knock out of all protein coding genes
CRISPRs provide precise genome editing
(Ebermann et al.)
Zebrafish models of human patients
(Ebermann et al.)
Potential genetic interaction between PDZD7 & USH2A
(Ebermann et al.)
Zebrafish model validates genetic interaction between PDZD7 & USH2A
New tools increase reproducibility
• complete maps of all SNPs
• rapid, low cost sequencing
• isogenic strains
• 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
Reproducibility with zebrafish modelsof human health and disease
Monte Westerfield, University of Oregon, Eugene, USASponsored 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
Reproducibility with zebrafish modelsof human health and disease
VisionForACure.com
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