Transcript Linkage Disequilibrium, HapMap and Chip Design

Understanding GWAS Chip
Design – Linkage Disequilibrium
and HapMap
Peter Castaldi
January 29, 2013
Objectives
• Introduce the concept of linkage disequilibrium
(LD)
• Describe how the HapMap project provides
publically available information on genetic variation
and LD structure
• Review how LD enables genome-wide screens with
only a subset of genome-wide SNP markers
• Describe the design of chip-based genotype assays
Human Genome
• 3 billion base pairs, 23
paired chromosomes
• 99.9% sequence
similarity between
individuals
• ~12 million variant sites
What are the Different Types
of Genetic Variation?
• Single base pair change (ACGT  ATGT), aka Single
Nucleotide Polymorphism
• ~12 million across the genome
• Insertions/Deletions (TGGTTTCTA  TGGT---TA)
• Can be of variable size
• Trinucelotide repeats (microsatellites)
• Highly polymorphic, less common than SNPs
• Responsible for certain clinic disorders (Huntington’s, Fragile
X, myotonic dystrophy)
SNPs in detail
• SNPs can have up to four possible alleles (A,C,G,T),
most have only two alleles present in human populations
• Each person has two SNP alleles (one for each copy of
the chromosome)
• when both copies are the same, you’re homozygous (i.e. AA,
CC, GG, TT). When they’re different (AT), your
heterozygous.
• Each allele has a frequency in which it appears in a given
population
• major allele (more common), minor allele (less common)
• they sum to 1 (or 100%)
SNPs are Used as Genetic
Markers for GWAS Chips
• Properties of SNPs that make them good markers for
GWAS
• densely spaced across the genome
• usually bi-allelic (only 2 alleles in the population, simplifies
statistical tests)
• GWAS chips can effectively represent most common
variation with just a subset of SNPs
• with ~500,000 SNPs, most common variation can be
captured
• this is because there is significant correlation between
neighboring SNPs
Linkage Disequilibrium Causes
Correlation Between Neighboring SNPs
• Mendel’s laws state that genes (alleles) are
independently transferred across generations
(random assortment – linkage equilibrium).
• This is not the case when two genetic loci are
physically close to each other.
• When two physically close genetic loci are not
randomly assorted, this is called linkage
disequilibrium.
Linkage Equilibrium Arises Because of
Meiotic Recombination
http://kenpitts.net/hbio/8cell_repro/meiosis_pics.htm
Linkage and Recombination
Paternal DNA
Gametogenesis
Maternal DNA
X
Y
x
y
X
y
X
y
Z
z
z
z
From Paternal
grandfather
From Paternal
grandmother
X
Y
X
y
z
z
Recombination Breaks Up
Chromosomal Segments
Over Generations
•
recombination is not uniform
across the genome
(recombination hotspots).
•
SNPs within the yellow region
are correlated with each other
and form haplotypes.
•
Because of this correlation, one
can often use a single SNP from
a haplotype to represent all the
SNP variation within a
haplotype.
Haplotype Structure Reflects
Evolutionary History
•
The structure of haplotype
blocks varies across racial
groups
•
African populations have
short LD blocks, reflecting
the longer evolutionary
history of those populations
~500,000 SNP Markers Can Reasonably Represent Most
of the Common Genetic Variation in European Genomes
• GWAS relies upon linkage disequilibrium and the
ubiquitous nature of SNP markers to enable
genome-wide surveys of the impact of common
variation on disease susceptibility
Pe’er et al. Nat Gen. 2006
The HapMap Project is a catalog of
human variation across populations
• The Human Genome project provided the complete
human sequence for a small number of individuals
• To get an accurate sense of variable sites, data from many
individuals is needed
• HapMap has three iterations (http://hapmap.ncbi.nlm.nih.gov/)
• dense genotype data from multiple populations groups
• CEU – individuals of Northern and Western European
ancestry from Utah
• YRI – Yorubans from Nigeria
• JPT – Japanese from Tokyo
• CHB – Han Chinese from Beijing
Data from the HapMap Project Enabled
GWAS Chip Design
• Information from HapMap Used in chip design
• panel of potential SNPs to use in a genotype chip
• population specific LD structure to allow the
identification of tag SNPs that effectively tag
haplotypes
Using Linkage Disequilibrium
to find Genes
• Linkage disequilibrium (LD) means that sites of
genetic variation can serve as “markers” for larger
chromosomal segments.
• Correlation between markers is quantified with rsquared and D’.
GWAS identify novel disease loci, but
additional localization is often necessary
Genotype Chip Technology
http://scienceeducation.nih.gov/newsnapshots/TOC_Chips/Chips_RITN/How_Chips_Wo
rk_1/how_chips_work_1.html
Kang et al. The American Journal of Human Genetics Volume 74, Issue 3 2004 495 - 510
Summary
• Genetic material is transmitted across generations in
blocks called haplotypes.
• Linkage disequilibrium and haplotype blocks allow
for SNP tagging approaches that enable GWAS
chips to capture common genetic variation with a
subset of genetic markers.
• Haplotype structure varies across ancestral groups.
• The HapMap project catalogs human genetic
variation and LD structure across populations.