genetics of gene expression primer
DESCRIPTION
Broad Institute/MPG primer lecture given 2008-12-11. Thanks to Alkes Price and Steve McCarroll for contributing slides.TRANSCRIPT
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Outline
Background Significance Experimental design Definitions
Basic findings Tissue specificity Population differences Biological insight (?) Last thoughts
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In the beginning...
King and Wilson, Science 1975 Chimp – human coding sequence differences
don't match phenotypic distance Gene regulation responsible for rest? Basis for evo-devo, GGE
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Genetics of gene expression
Find genetic variation that (partially) controls levels of RNA
If RNA levels are different, maybe protein levels are also different
If protein concentration is different, maybe there is a phenotype?
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Experimental design
Protein measurements 2D gels; quantitation hard; resolution?
RNA measurements Expression chips ca. 1995 (Schena et al, Science)
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Experimental design - methods
Allelic discrimination SBE Pyrosequencing SNP arrays
Whole-genome expression assays Genotypes?
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Experimental design - samples
Pedigrees – multiplex/trios HapMap
Population samples Inbred strain panels
genotypes New crosses Inter-specific hybrids
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Definitions
Cis
Trans
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Outline
Background Basic findings Tissue specificity Population differences Biological insight (?) Last thoughts
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First forays
Damerval et al 1994 Protein levels are different in 42/72 maize gene
products 2D electrophoresis, eyeball spot quantitation Problems:
genome coverage quantitation post-translational modifications
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Cowles et al 2002
F1 mice 69 genes in three tissues 4 variant; 2 only in liver
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Yeast WG approaches
Brem et al Science 2002 Linkage in 40 offspring of lab x wild strain cross 1528/6215 DE between parents 570 map in cross
multiple QTLs 32% of 570 have cis linkage
262 not DE in parents also map
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Trans hotspots
Brem et al Science 2002
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Yvert et al Nat Genet 2003
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Mammals I
F2 mice on atherogenic diet Expression arrays; WG linkage
Schadt et alNature 2003
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Mammals II
Chesler et al Nat Genet 2005
10% !!
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Mammals III
No major trans loci in humans Cheung et al Nature 2003 Monks et al AJHG 2004 Stranger et al PLoS Genet 2005, Science 2007 Further scans (WT, Cheung, Schadt etc)
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What's the deal with trans?
Artefacts Normalization robustness Replication? Clusters of correlated genes
Power trans often weak effects Small sample sizes
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Outline
Background Basic findings Tissue specificity Population differences Biological insight (?) Last thoughts
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trans-acting factors may be tissue specific TFs, signalling molecules, etc etc
cis regulatory sequences may be used by TS factors
So, shouldn't genetic effects on GE be tissue specific?
Logic
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Examples
Chesler vs Bystrykh (Nat Genet 2005) 39/101 (39%) forebrain cis in HSC 297/1218 (24%) HSC cis in forebrain
Hubner (Nat Genet 2005) 311/4297 (7%) shared between fat and kidney
Campbell et al (Gen Res 2008) 12% of ~100 genes vary in at least one of three
tissues
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Correlation networks persist across tissues
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Outline
Background Basic findings Tissue specificity Population differences Biological insight (?) Last thoughts
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Slide courtesy Alkes Price
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Population differences could have non-genetic basis
• Differences due to environment? (Idaghdour et al. 2008)
• Differences in cell line preparation? (Stranger et al. 2007)
• Differences due to batch effects? (Akey et al. 2007)
(Reviewed in Gilad et al. 2008)
Slide courtesy Alkes Price
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Gene expression experiment
Does gene expression in 60 CEU + 60 YRI vary with ancestry?
Does gene expression in 89 AA vary with % Eur ancestry?
60 CEU + 60 YRI from HapMap, 89 AA from Coriell HD100AAGene expression measurements at 4,197 genes obtained using Affymetrix Focus array
c
Slide courtesy Alkes Price
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Gene expression differences in African Americans validate CEU-YRI differences
c = 0.43 (± 0.02)(P-value < 10-25)
12% ± 3%in cis
Slide courtesy Alkes Price
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Outline
Background Basic findings Tissue specificity Population differences Biological insight Last thoughts
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Does GGE matter?
Where is the phenotype? Case/control differences?
Does it translate to protein? Can we reconstruct networks/pathways?
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Insight
Mechanism? trans – coregulated groups?
pathways Model QTs Molecular variation snapshots
What does it mean? Ancillary information
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IRGM harbors a 20-kilobase deletion polymorphism immediately upstream
CEU 0.06 1.0
J CH 0.41 1.0
YRI 0.53 1.0
Deletion allele
frequency
LD (deletion,
rs13361189)
IRGM
Ind
ivid
ua
ls
Position on chr5
rs13361189(strongest Crohn’s-associated SNP)
Slide courtesy Steve McCarrollMcCarroll et al., Nature Genetics, Sept. 2008
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Slide adapted, courtesy Steve McCarroll
IRGM structural haplotypes have altered expression
McCarroll et al., Nature Genetics, Sept. 2008
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IRGM expression levels influence the efficacy of autophagy
McCarroll et al., Nature Genetics, Sept. 2008 Slide adapted, courtesy Steve McCarroll
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Outline
Background Basic findings Tissue specificity Population differences Biological insight (?) Last thoughts
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Success?
Baseline variation - tricky Perturbations and phenotypes
mouse obesity crosses (Schadt et al) Drug screening (Choi, Yelensky et al) Crohn's disease (McCarroll et al)
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Stuff we haven't covered
Coregulated sets Disease context Annotation/pathway analysis Modelling QTLs Regression WGAS Multiple data sets/types Post-transcriptional event variation
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