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TUTORIAL - CANCER GENE PRIORITIZATION

                Prioritize a list of
                candidate genes based
                on CGPrio probabilities
                and oncogenomic
                alterations
STEP 1


Go to www.intogen.org
& Click Browser
STEP 2

Click on Upload filter.
STEP 3
                                         Select the type of
                                         gene IDs for the list
                                         that you are
                                         providing




                                 STEP 4
                         Paste the list of genes
                         to prioritize.                          NOTE
                         You can also upload a         For this tutorial we are
                         file with the list of         going to load a list of 37
                         genes.                        candidate cancer genes
                                                       identified after a mutation
                                                       screening of 20661 human
                                                       genes in 22 human tumor
                                                       samples of Glioblastoma
       STEP 5
                                                       multiforme.
Click on Create filter
                                                       (Parson et al Science 2008)
STEP 6
                                                      Select the type of cancer in
                                                      which you are interested. In
                                                      this case we will select brain
                                                      tumors since we are studying a
                                                      list of candidate cancer genes
                                                      for glioblastoma.


                                                      These values are based on
                                                      predictions by CGPrio method.
                                                      Genes with a higher probability
                                                      rank are more likely to be
                                                      involved in cancer.

                                                      The CGPrio method is based on
This indicates that the                               computational classifiers that
filter is applied. You can                            use different combinations of
click the red x to                                    sequence and functional data
eliminate the filter and                              including sequence
return to the list of all                             conservation, protein domains
genes.                                                and interactions, and regulatory
                                                      data. For more information see
                                                      Furney et al., NAR 2008
          This table informs if the
          gene is in the Cancer
          Gene Census of the          With these links you
          Sanger Center and           can go to the next
          which type of mutations     page to see the rest
          are annotated there.        of genes. In total
                                      there are 37.
Note that now we are on
                                                     the page for brain and
                                                     related tumor types

                                                This table gives a
                                                summary of alterations
                                                found for each gene in
                                                the selected tumor type.

                                                                               The colors
                                                                               indicate
                                                                               significance.
                                                                               Color toward
                                                                               red means
                                                                               that this gene
                                                                               is significantly
                                                                               altered in this
                                                                               tumor type.


Color scale of corrected p-values:
Gray = no significantly altered
Red/Yellow = significantly altered



                                     Gray means that this        White means that
                                     gene is not significantly   there is no data
                                     altered (CNA gain in this   for this gene in
                                     case) in this tumor type    this tumor type
STEP 7
                    STEP 5
You can sort the table by clicking the
title of any column, to have the genes
with lowest or highest values on top.
Click Onco twice to sort by those genes
with the highest oncogene prediction
rank.




                         This column gives
                         information on mutations in
                         these genes found and
                         studied in this tumor type.
                         This information is imported
                         from COSMIC.
STEP 8
                            Now in the top of the table   You can sort the table by
                            we have the genes that        clicking to the title of any
                            are more significantly up-    column, to have the genes
                            regulated in brain and        more significantly altered in
                            related tumors                the top. Click Up to sort by
                                                          up-regulation.




                   STEP 9
            For any of the genes you
            can get detailed
            information. For example
            click on the Ensembl ID for
            EGFR .

Note that you can
retrieve the data in
a tabulated file by
clicking on ‘CSV file’
under ‘Export'.
Note that now we are on
                                                              the page for EGFR for
                                                              brain and related tumor
                                                              types.

                                                 STEP 10
                                           Click on Tumor types
                                           Tab.

This box shows details of
mutations of EGFR in brain cancer.
The number of samples with
mutations (103), the number of
samples analyzed (1643) and the
link to COSMIC for this gene.




                                                                         This box shows details
                                                                         of prediction ranks for
                                                                         EGFR gene obtained
                                                  This box shows
                                                                         with CGPrio for the
                                                  some gene details
                                                                         different sets of
            This box indicate that this gene      and link to ensembl
                                                                         properties analyzed.
            is in the Cancer Gene Census,
            and it give details about the type
            of mutations identified.
STEP 11
                                  Now click on
                                  Experiments Tab to
                                  see details




This table gives information on the
alterations found in EGFR in
different morphologies of brain
cancer. Note that this gene is
amplified and up-regulated in
many morphologies.
This is the list of experiments
                            that have analyzed EGFR in
                            brain cancer and related
                            tissues.




The color indicates                             N = number
that the region of                              of samples
this gene is                                    analyzed
amplified in a
significant number of
the 171 samples
analyzed.

   White cells means no
   data. In this case, it
   means that this
   experiment has not
   analyzed
   transcriptomic
   alterations
STEP 12
                      Click Up to have the
                      experiments on top in which
                      this gene is significantly up-
                      regulated.




     STEP 13
Click on the
experiment by Sun L
et al to see the
details.
Note that now we
                                are on the page of
                                EGFR for a particular
                                experiment of brain
                                cancer.

This box gives
details of the
experiment,
including authors,
title and link to the
publication or
original source of
data.

This box gives details of up-
regulation of EGFR in this
experiment. 152 samples have
been analyzed, of which of
which 80 show over-expression
of this gene. The expected
number of samples with
alteration by chance is about
4.6. The over-expression is
therefore highly significant.
STEP 13
Click “all” in
Experiments and “all” in
Tumor type to see the
information of known
cancer alterations in
EGFR in other tumor
types. Go to the tumor
type tab.
Note that now we on the
page for EGFR for all tumor
types

                        STEP 14
                    Click all in Genes.
                    Then go to the genes
                    tab.




                              This table gives
                              information of known
                              cancer alterations in
                              different tumor types
                              found in the EGFR gene.
                              In this case we can see
                              that the region of this
                              gene is significantly
                              amplified (gained) in
                              several tumor types, it is
                              transcriptionally altered
                              in some and a number of
                              mutations have been
                              found in different tumor
                              types.
Now we see the page
                               showing all the tumor
                               types for all genes
                               included in the filter and all
                               tumor types.




The filter is still applied,
so we can continue
navigating over the
evidences for
involvement in cancer of
our group of genes of
interest.
THANKS FOR USING INTOGEN


       You will find more tutorials
       and documentation in
       www.intogen.org

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Cancer Gene Prioritization

  • 1. TUTORIAL - CANCER GENE PRIORITIZATION Prioritize a list of candidate genes based on CGPrio probabilities and oncogenomic alterations
  • 2. STEP 1 Go to www.intogen.org & Click Browser
  • 3. STEP 2 Click on Upload filter.
  • 4. STEP 3 Select the type of gene IDs for the list that you are providing STEP 4 Paste the list of genes to prioritize. NOTE You can also upload a For this tutorial we are file with the list of going to load a list of 37 genes. candidate cancer genes identified after a mutation screening of 20661 human genes in 22 human tumor samples of Glioblastoma STEP 5 multiforme. Click on Create filter (Parson et al Science 2008)
  • 5. STEP 6 Select the type of cancer in which you are interested. In this case we will select brain tumors since we are studying a list of candidate cancer genes for glioblastoma. These values are based on predictions by CGPrio method. Genes with a higher probability rank are more likely to be involved in cancer. The CGPrio method is based on This indicates that the computational classifiers that filter is applied. You can use different combinations of click the red x to sequence and functional data eliminate the filter and including sequence return to the list of all conservation, protein domains genes. and interactions, and regulatory data. For more information see Furney et al., NAR 2008 This table informs if the gene is in the Cancer Gene Census of the With these links you Sanger Center and can go to the next which type of mutations page to see the rest are annotated there. of genes. In total there are 37.
  • 6. Note that now we are on the page for brain and related tumor types This table gives a summary of alterations found for each gene in the selected tumor type. The colors indicate significance. Color toward red means that this gene is significantly altered in this tumor type. Color scale of corrected p-values: Gray = no significantly altered Red/Yellow = significantly altered Gray means that this White means that gene is not significantly there is no data altered (CNA gain in this for this gene in case) in this tumor type this tumor type
  • 7. STEP 7 STEP 5 You can sort the table by clicking the title of any column, to have the genes with lowest or highest values on top. Click Onco twice to sort by those genes with the highest oncogene prediction rank. This column gives information on mutations in these genes found and studied in this tumor type. This information is imported from COSMIC.
  • 8. STEP 8 Now in the top of the table You can sort the table by we have the genes that clicking to the title of any are more significantly up- column, to have the genes regulated in brain and more significantly altered in related tumors the top. Click Up to sort by up-regulation. STEP 9 For any of the genes you can get detailed information. For example click on the Ensembl ID for EGFR . Note that you can retrieve the data in a tabulated file by clicking on ‘CSV file’ under ‘Export'.
  • 9. Note that now we are on the page for EGFR for brain and related tumor types. STEP 10 Click on Tumor types Tab. This box shows details of mutations of EGFR in brain cancer. The number of samples with mutations (103), the number of samples analyzed (1643) and the link to COSMIC for this gene. This box shows details of prediction ranks for EGFR gene obtained This box shows with CGPrio for the some gene details different sets of This box indicate that this gene and link to ensembl properties analyzed. is in the Cancer Gene Census, and it give details about the type of mutations identified.
  • 10. STEP 11 Now click on Experiments Tab to see details This table gives information on the alterations found in EGFR in different morphologies of brain cancer. Note that this gene is amplified and up-regulated in many morphologies.
  • 11. This is the list of experiments that have analyzed EGFR in brain cancer and related tissues. The color indicates N = number that the region of of samples this gene is analyzed amplified in a significant number of the 171 samples analyzed. White cells means no data. In this case, it means that this experiment has not analyzed transcriptomic alterations
  • 12. STEP 12 Click Up to have the experiments on top in which this gene is significantly up- regulated. STEP 13 Click on the experiment by Sun L et al to see the details.
  • 13. Note that now we are on the page of EGFR for a particular experiment of brain cancer. This box gives details of the experiment, including authors, title and link to the publication or original source of data. This box gives details of up- regulation of EGFR in this experiment. 152 samples have been analyzed, of which of which 80 show over-expression of this gene. The expected number of samples with alteration by chance is about 4.6. The over-expression is therefore highly significant.
  • 14. STEP 13 Click “all” in Experiments and “all” in Tumor type to see the information of known cancer alterations in EGFR in other tumor types. Go to the tumor type tab.
  • 15. Note that now we on the page for EGFR for all tumor types STEP 14 Click all in Genes. Then go to the genes tab. This table gives information of known cancer alterations in different tumor types found in the EGFR gene. In this case we can see that the region of this gene is significantly amplified (gained) in several tumor types, it is transcriptionally altered in some and a number of mutations have been found in different tumor types.
  • 16. Now we see the page showing all the tumor types for all genes included in the filter and all tumor types. The filter is still applied, so we can continue navigating over the evidences for involvement in cancer of our group of genes of interest.
  • 17. THANKS FOR USING INTOGEN You will find more tutorials and documentation in www.intogen.org