A web resource of theBeta Cell Biology Consortium

EPConDB

http://www.cbil.upenn.edu/EPConDB

Introduction

• The EPConDB website provides an integrated view of pancreas- and islet-related gene annotation, expression and regulation data.

1. Data in EPConDB

2. Browsing Individual Genes

3. Working with Gene Lists

The Data in EPConDB• Science-driven bioinformatics resource• Composed of heterogeneous information• Maximal cross-talk between types of information

GeneralInformation

Research ResourcesE

PC

onD

B

MicroarraysNews

Downloads

MethodsHelp

Data Tools

Expression experiments

ChIP experiments

DoTS transcripts

T1DBase gene pages

BLAST

TFBS search

Predicted TFBS

T1DBase genome browser

Expression profilesUp/down regulated genes

Reportmaker

Query History

EPConDB Current Data ContentDoTS Transcripts/Genes940,837 human DoTS transcripts 57,382 are associated with 22,268 unique Entrez genes 22,630 GeneCard genes 746,668 mouse DoTS transcripts 60,931 are associated with 33,276 unique Entrez genes 26,580 MGI genes

Expression Data Sets 32 pancreas-related expression data sets contributed by 24 different groups 5 private studies (BCBC viewable) 5 tissue expression surveys (include pancreas and/or islet) 3 expression studies in progress11 lists of differentially expressed genes from studies30 studies have gene expression profiles

Chromatin IP Data Sets 1 ChIP-on-Chip study 2 lists of TF target genes Predicted Human binding sites - 70,100 Predicted Mouse binding sites - 72,921 2 ChIP-on-Chip studies in progress

Home Page• BCBC links

• Main navigation bar

• Find Gene • By Name• Expression profiles• On arrays

• Find gene by sequence

EPConDBBetaCell.org T1DBase.org

Three Integrated WebsitesPancreas

Functional GenomicsType 1 Diabetes

Genetics & GenomicsBeta Cell

Research Informatics

• Reciprocal links to both web sites

• Reagents, e.g. Antibodies • Gene page & Genome Browser

1. Enter gene symbol or name and click Search.

2. Finds three transcripts from mouse or human genes.

3. Click on DoTS ID to see summary view for that transcript.

Finding Individual Genes

Queries to quickly find transcripts for genes

FoxA2 Transcripts

Transcript Information

• EPConDB contains many pancreas- or islet-related studies.

• Very detailed information is easily visualized for each study.

Expression Profiles

Working with Gene Lists

Browse lists of genes differentially regulated in mRNA studies.

1. Follow Experiments link in nav bar.

• Lists 32 mRNA expression experiments loaded into EPConDB.

• Categorized and selectable by experimental design

• Links to

• Study details

• 11 lists of differentially expressed genes

(Analysis Result Link)

• 1 ChIP-on-Chip experiment with gene lists

• RSS link can provide a nice searchable, sortable view.

• Display browser specific

• Use to update studies displayed at betacell.org

Experiments Page

Analyzed with PaGE.

May be multiple analyses for a study.

Sortable by confidence or fold change.

Links to analysis details.

Red is up

Green is down

Grant G.R., Liu J., Stoeckert C.J.Jr. (2005) A practical false discovery rate approach to identifying patterns of differential expression in microarray data, Bioinformatics 11, 2684-2690.

Differentially Expressed Genes

To make the data more useful, EPConDB provides the ability to work with gene lists using queries and query history.

These tools provide you the ability to generate lists of genes matching your criteria to identify candidate genes for your research.

Browsing genes is nice but you can do more

Querying for Gene Lists

Which human genes have promoters bound by HNF1alpha in the islet and have a predicted HNF1alpha site in their promoter?

EPConDB does not have this query …

but

… it does have two simpler queries we

can combine.

Create a Gene List

Genes with promoters bound by HNF1alpha

in the islet

Genes with predicted HNF1alpha sites

in promoter

1. Go to the Query Transcripts Page via Navbar.

2. Click on the Query TF Target Genes identified by genome-wide location analysis (ChIP-Chip)

3. Choose a TF from the list, ‘Binding Sites for HNF1alpha’

4. Adjust fold-change and p-value if desired

5. Click Submit

Which human genes have promoters bound by HNF1alpha in the islet?Gene List #1

Which human genes have a predicted HNF1alpha site in their promoter?Gene List #2

1. Go to the Query Transcripts Page using the Navbar.

2. Click on the Predicted Transcription Factor Target Genes query.

3. Select HNF1 (Tcf1) and human genome from the menus

4. Click Submit

5. TFBS predicted on conserved regions of promoter in both Hs and Mm.

6. See methods page to see details and logos of TFBS PWM motifs.

7. Connected DTs to all TFBS within 1KB of start site.

Click on the ‘My transcripts’ button to go to the history page.

ClickIntersect

button

IntersectionResult

Checkboxes With the query

history page you can create unions, intersections, and subtract gene lists from each other.

You can also delete unwanted gene lists.

Click on view to see Result Page

or Download

Intersect Gene Lists with History Page

Which human genes have promoters bound by HNF1alpha in the islet and have a predictedHNF1alpha site in their promoter?

Two simple queries we combined.

Result page with 27 transcripts and their corresponding genes.

Combined Gene Lists

Genes with promoters bound by HNF1alpha

in the islet

Genes with predicted HNF1alpha sites

in promoter

2912 (transcripts)

218 (transcripts)

27

Result Page Gene List Links to DT, Gene, Expression profiles, and genome browser.

Summary for this part of the talk

We have covered:

• What data EPConDB contains.

• How to browse info for individual genes.

• How to browse lists of differentially expressed genes.

• Gene list queries. How to combine lists to create gene lists that meet your criteria, e.g, genes with promoters bound by HNF1alpha in islet

• Visit us at our poster

Future EPConDB Site Plans• New web site design in progress

• User input will guide development of new site to make site easier to use and navigate

Let us know what you think at poster, please use meeting as opportunity to talk with us

What web site features/views should be maintained?

• Sharing logins, BCBC web site and EPConDB

Home

Query Transcripts

Mock ups

Planned Functional Genomics Study Pages on betacell.orgProposed to contain key information frominformation already existing in EPConDB and supplemented with manual annotations (e.g., goal, experimental paradigms, genes of interest, take home message)

• Title, authors, date, etc…• Summary, with biological relevance• Lists of up/down-regulated genes• Downloads of raw/analysis data• Links to more detail at EPConDB/RAD/PAGE

We ask for your feedback. What do you wanton these pages?

J.-P. Cartailler

Abstract from Hardy et al. Mol. Endocrinol. 2006: Recent advances in functional genomics afford the opportunity to interrogate the expression profiles of thousands of genes simultaneously and examine the function ofthese genes in a high throughput manner. In this study, we describe a rational and efficient approach to identifying novel regulators of insulin secretion by the pancreatic beta cell. Computational analysis of expression profiles of several mouse and cellular modelsof impaired insulin secretion identified 373 candidate genes involved in regulation of insulin secretion. Using RNA interference, we assessed the requirements of ten of these candidates and identified four genes (40%) as being essential for normal insulinsecretion. Among the genes identified was short-chain L-3-hydroxyacyl-CoAdehydrogenase (Hadhsc), which encodes SCHAD, an enzyme of mitochondrial oxidation of fatty acids whose mutation results in congenital hyperinsulinism. RNAi mediated gene suppression of Hadhsc in insulinoma cells and primary rodent islets revealed enhanced basal but normal glucose stimulated insulin secretion. This increase in basal insulin secretion was not attenuated by opening of the KATP channel with diazoxide,suggesting that SCHAD regulates insulin secretion through a KATP channel independent mechanism. Our results suggest a molecular explanation for the hyperinsulinemiahypoglycemic seen in patients with SCHAD deficiency.

Data Sets and EPConDB Experiments in future EPConDB releases:

• Three public microarray data sets (published)• Petri, A. et al. “The effect of neurogenin3 deficiency on pancreatic gene expression in embryonic mice.” J Mol Endocrinol. 2006 37:301-16. • Liu,Y. et al. “Activation of the Reg family genes by pancreatic-specific IGF-I gene deficiency and after streptozotocin-induced diabetes in mouse pancreas.” Am J Physiol Endocrinol Metab. 2006 291:E50-8.• Sherwood, R.I. et al. “Prospective isolation and global gene expression analysis of definitive

and visceral endoderm.” Dev Biol. 2007 304:541-55.

• BCBC experiments that become public• Expression data, Chromatin IP binding data

• What data sets would you like to see included in EPConDB or are missing?

• What can you contribute to EPConDB to increase the visibility and thus the value of your data?

Microarray analysis on E8.25 definitive and visceral endoderm

Melton GroupData analysis/spreadsheet available

Go to http://www.betacell.org/Click on Resources > Data Resources >

Research Data

Raw data is also being provided and willbe annotated/loaded into EPConDB

J.-P. Cartailler

• Easily submitted to ArrayExpress for you

• Included on the EPConDB Experiments page

• Used for Gene Expression Profiles

• Used to create Gene Lists

Experiments in EPConDB are:

Having many experiments in EPConDB will allow for comparisons among data sets.

Extend the analysis to meet your criteria and refine your list of genes

Data Set Submissions

• Simple process of submitting your data

• Contact us about your data set• use email link or contact us directly

• Provide initial experiment description

• We create a spread sheet for you

• Review your completed spread sheet

• Place your data files on our download site

A simple spreadsheet-based, MIAME-supportive format for microarray data: MAGE-TAB. Tim F. Rayner et al. (2006) BMC Bioinformatics, 7, 489

EPConDB Data Submission

Step 1: The Sample Submission form provides basic information about your experiment. You fill this out. (4 sections)

Step 2: We complete the form and send it back to you for review.

Also, review the automatically generated graph of your experiment. (You may be asked to answer some final questions or provide missing details at this step.)

Step 3: Upload your data files (.gpr, .cel, .txt) onto our secure ftp site.

Step 4: (Optional): Login and view your finalized experiment prior to an EPConDB release (this requires a temporary password).

CBIL Stoeckert Lab

• John Brestelli• Junmin Liu• Jonathan Schug• Elisabetta Manduchi• Joan Mazzarelli

Nat Goodman & Luc Smink

• Barry Healy• Erin Hulbert• Burak Kutlu• James Allen• Victor Cassen

BCBC - Mark Magnuson• J.-P. Cartailler• Christian Shaffer• Robert McClellan• Lisa Rouse

Functional Genomics Core• Klaus Kaestner• Peter White

We thank all of the labs who have already contributed data.

Acknowledgements

Funding• NIDDK• JDRF