This is a test version of Biostars. For the public version, visit https://www.biostars.org.
Somatic mutation calling without matched normal

Dear everyone,

I have been asked to comment on an experiment design that involves WES or WGS of cancer cell lines which lack matched normal.

I know that this design is far from ideal, but I was wondering if there are people who have already stream-lined it (to the extent possible of course). I can think of a couple of common variation filters to keep out the germline variants, but I am not sure how to go beyond.

I am sorry if this is a duplicate question; I couldn't find a related hit.

Thank you,

Noushin

mutation calling somatic
• 7,562 views • link
updated by Ram Ram • written by Noushin N Noushin N

2 answers

The bottom line is that without a matched normal, you're just not going to be able to call the somatic status for the vast majority of sites. That said, you can winnow down a list to those you *suspect* are somatic. Some ideas:

  • Weed out sites with high frequency in the population
  • If your tumor is very impure, you can take advantage of the fact that the frequencies of somatic variants will be shifted away from 50%/100%
• 0 views • link
written by Chris Miller Chris Miller

Note that the variant allele frequencies will be shifted from 50%/100% even in a pure tumor such as a cell line if you have copy number alterations. Depending on the cancer type, cancer cell lines can easily be triploid or tetraploid, with many copy number aberrations.

• 0 views • link
updated by Ram Ram • written by Christof Winter Christof Winter

That's a good point, but one is probably safe in assuming relatively high purity for cell lines (please correct me if I am wrong) compared to tissue biopsies. The issue about aberrant karyotype distorting allelic fractions is absolutely true, and needs to be accounted for.

• 0 views • link
written by Noushin N Noushin N

I think unless there are contaminating cells in the culture, your cell line DNA extracts should be close to 100% pure.

• 0 views • link
written by Christof Winter Christof Winter

Since you are asking about cell lines, maybe these have already been sequenced. There are bigger studies I'm aware of:

  1. Cancer Cell Line Encyclopedia (CCLE), see http://www.ncbi.nlm.nih.gov/pubmed/22460905
    Browse and download the data: http://www.broadinstitute.org/ccle/home
  2. NCI-60 cell line, see e.g. here: http://www.cbioportal.org/public-portal/study.do?cancer_study_id=cellline_nci60

Then I would check against COSMIC for known somatic mutations. And maybe the cell line in question even has data in COSMIC, see By Sample at http://cancer.sanger.ac.uk/cancergenome/projects/cosmic/

• 0 views • link
updated by Ram Ram • written by Christof Winter Christof Winter

Thanks for pointing out these useful resources Christof. The cell lines in question are not previously characterized. Using COSMIC to identify known somatic mutations sounds good. But it seems to me it will be hard to call novel somatic mutations as they will be almost indistinguishable from private/rare germline variants, unless for those which are present at smaller cellular frequencies. Does this seem right?

• 0 views • link
written by Noushin N Noushin N

Yes, I would also say it's close to impossible to distinguish between novel somatic and private or rare germline variants that are not dbSNP/1000genomes/ESP6500. Looking at both mutation allele frequency and copy number at that position might help for cases where a somatic mutation occurred after a copy number change, but it's still rather guessing than identifying.

• 0 views • link
updated by Ram Ram • written by Christof Winter Christof Winter

Log in to answer this question.

More posts like this