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Boolean Analysis of Large Gene-expression Datasets

Debashis SahooPhD Candidate, Electrical Engineering

Joint work with David Dill, Andrew Gentles, Rob Tibshirani, Sylvia Plevritis

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Outline

Standard microarray work flowData collection and preprocessingBoolean analysisBiological insightsConclusion and future work

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Microarray Work Flow

mRNA Hybridization Scanning

Image processingNormalizationData analysis

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Data Collection

There are thousands of microarray freely available

GEOArrayExpressSMDCelsius

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Preprocessing

Get original RAW CEL files for one platform together.

Typical number of CEL files : 2,000-11,000

Use RMA to normalize the CEL filesNeed a memory efficient algorithmGenerates expression values for each probeset

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Existing Methods

Correlation analysisConditional probabilityMutual information

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Boolean Analysis

Get RAW Data Normalize

Determine thresholds

Discover Boolean relationshipsNew Biology

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Example

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Determine threshold

Sort the gene expressionsUse StepMiner to determine the threshold

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Determine threshold

Its hard to determine a threshold for this gene.StepMiner usually puts a threshold in the middle for this case.

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Discover Boolean Relationships

Analyze scatter plots between two genes.Divide the space into four different regions using the thresholds (quadrants).Determine sparse quadrants.Determine the Boolean relationships.

WNT5A high PAX5 low

0

1 3

2

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Statistical Tests

Compute the expected number of points under the independence model

Compute maximum likelihood estimate of the error rate

statistic =(expected – observed)

expected√

a00

(a00+ a01)

a00

(a00+ a10)+( )1

2error rate =

a00

a01 a11

a10

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Boolean Relationships

Tightly co-regulated genes forms two sparse quadrants.There are six possible Boolean relationships

EquivalentOppositeA low B lowA low B highA high B lowA high B high

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Boolean Relationships

Equivalent

Opposite

PTPRC low CD19 low XIST high RPS4Y1 low

COL3A1 high COL1A1 highFAM60A low NUAK1 high

Symmetric Asymmetric

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Boolean Implication Network

Directed graphNodes:

For each gene AA highA low

Edges:A high to B low

A high B low

A high

B low

A low

B high

C high

C low

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New Biology

This slide is under construction!!

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Biological Insights

Gender Organ Tissue

Development Differentiation Co-expression

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Example Application

ImmunologyB Cell differentiationGoal:

Discover genes that mark unique B Cell precursors

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Differentiation Tree

Hematopoietic stem cell differentiation is a tree

Root: HSCLeaf

LymphocytesB Cell, T Cell, NK cell, Dendritic cell

ErythrocytesGranulocytes: Basophil, Neutrophil, EosinophilMonocytes: Dendritic cellThrombocytes

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KIT high A high B low B220 low CD19 low

KIT

A

B

B22

0C

D19

A high

B low

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Conclusion

Boolean analysisDirectly visible on the scatter plot.Enables discovery of asymmetric relationship.Follow biology.Potential application to Immunology

Future workCancer progression New biology

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Acknowledgements

The Felsher Lab:Natalie WuCathy ShachafDean Felsher

Funding: ICBP Program (NIH grant: 5U56CA112973-02)

Leonore A HerzenbergJames Brooks Joe LipsickGavin SherlockHoward ChangStuart Kim

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The END