My Blog List

Tuesday, August 2, 2011

K=6 STRUCTURE run: results for participants V157-V222

I've recently finished STRUCTURE analysis for the participants V157-V222. The fixing of STRUCTURE problems for the whole set of genomic SNP-markers was somewhat problematic, therefore i decided to run series of complimentary STRUCTURE on each chromosome separately (after splitting the initial PLINK dataset into chromosomal groups). Since in BGA analysis it is important not to overlook effects of  SNP-genotyping and linkage disequilibrium (LD), the dataset was pruned for low-genotyping rate SNPs and SNP markers with high LD with each other. The question of how far back one has to look to find unadmixed ancestors is relevant for determining ancestry admixture because it relates to whether or not, and to what extent, markers are in linkage disequilibrium (LD) with one another. Assuming that the markers are unlinked often produces admixture estimates that show subtle differences from those obtained when assuming the markers may be linked. Not having a knowledge of an individual’s genealogy  before testing them, we cannot predict the extent to which markers will be in LD beforehand, so we must be able to accommodate uncertainty in the linkage between markers if we hope to improve the precision of admixture estimates. The best way to accommodate uncertainty is to perform LD-based prunning in PLINK.

STRUCTURE method (Pritchard et al.2000) is called a model-based clustering method because it assumes a model bywhich it classifies samples into groups (either discretely or proportionally). However, there is  the very important problem with discrete population models. To measure the bias of the method and make real sense of it, one should first to define  absolute 100% ancestry—but how do we define absolute ancestry? The design of  "amateurish" BGA project (like our one) is usually involve the selection of arbitrary reference populations that are impossible to justify. But merely selecting the "reference" is not  sufficient to guarantee the success of BGA project. However, some (Dodecad and Harappa project, most notably) tried to circumvent this problem by simulating allele frequencies of  "ancestral" populations (Dodecad's "zombies") that represented a perfect surrogate/proxy for "real" ancestry, then as long as definition does not bias toward, say, one type of Eurasian over another (which could occur if we sampled one part of the Eurasian continent but not others), the definition would be reasonable for our purposes. Had the "simulating" experiment with "zombies"succeeded, it would have been a perfect surrogate of absolute ancestry.Once the definition of absolute ancestry had been established, it would be possible to then define the error of an admixture method relative to this reference point (as it is in the suprevised model of ADMIXTURE). However, one should keep in minds that this method has its own limitations. For example, given that we can estimate the parental allele frequency considering that of its modern-day descendents, we are still left with the problem of how to define which modern-day groups are bona-fide descendents of the parental groups and which are not.
Perhaps a  skeptic could admit that it is more safe to use assumption-free, objective approaches, such the clustering method, implemented by  Pritchard et al. (2000). The difference, the sceptic's claim would go, lies in methodology: running STRUCTURE with a given set of markers and a given set of samples, we define the best way to subdivide the sample given the genotype data and so by definition, if our sample is a good one, we establish the most appropriate population model for the markers. In that this particular method does not require a definition of structure a priori to infer population structure and assign individual samples to groups, its use obviates the need to define who belongs to a parental group beforehand, or even how many parental groups there are.

Another obvious reason for running STRUCTURE on each chromosome separately is the distribution of SNPs. Clearly, given that mixed ancestry can be unequally distributed throughout the genome, the more dense the marker spacing the more precise the estimates of ancestry and the less likely low levels of recent admixture (which might be focused on a small number of chromosomes) would be missed. 

Some final considerations:  the results (coeffecients of admixture) presented below are clustered using CLUMPP software ( CLUMPP was used to resolve the label switching and compute the average admixture coefficients for 22 subsequent runs) in 1000 permutations. Since the permutations of clusters was involved,  it is more appropriate to think of  results in terms of combined ancestry (i.e N-European + East-European, mixed (Balto-)Slavs, West-European+East-European etc.). 

You can check individual coefficients of ancestry in spreadsheet. 



Monday, July 11, 2011

STRUCTURE results: to be published soon

The co-admin of  the MDL project, Leon (of HIRSearch) has managed to get STRUCTURE to work with the whole MDL dataset on one of his servers. I expect the STRUCTURE job to be completed in a couple of days.

After that i will use either CLUMPP or DISTRUCT  from Noah Rosenberg's lab for sorting the cluster labels and producing nice graphical displays of STRUCTURE results.

Friday, July 1, 2011

A close-up on "the core" of the MDL project

The PCA and MDS plots fleshed out very stable clusters of what appears to be the core of the MDLP projects. I would label these clusters after major population groups constituning a core set of the project's populations.

1) "Russian" cluster: V164, V163, V207
2) The intermediate cluster of admixed eastern Slavs (russians+belarussians, ukrainians+russians etc.,): V219, V165, V186, V187, V161, V189, V185, V201, V202, V205, V215, V196
3) "Belarussian" cluster: V188, V158,V157,V174
4)  The intermediate cluster of  Belarussian-admixed Lithuanians and vice versa: V162, V214, V210, V220
5) "Lithuanian" cluster: V191, V192,V184, V211, V216, V183, V190, V218, V170
6) "Polish" cluster:  V169, V205, V160, V208, V180, V187, V181, V176, V177


ADMIXTURE results for all participants up to V222

Dienekes was kind enough to share his C++ code for converting the 1000 genomes dataset  to PLINK format. Since many participants of our project have Central European and British ancestry i have added the converted dataset of GBR (“British from England and Scotland”),  along with Orcadian sample from HGDP project and CEU panel from HapMap project.  These samples proved to be useful in the delineation of NWE ancestry in the participants of our project.

I have also added Finns from the 1000genomes project to access the possible North-Western Asian component in the reference set of Russian population.

Here are results of  unsupervised and supervised runs of ADMIXTURE on the pruned MDLP datasets. I haven't intentionally labeled the inferred components, since there is still considerable disagreement among genomic bloggers as to what these components might be. 

Unsupervised ADMIXTURE run:




Supervised  ADMIXTURE run


PS. I have also tried to run STRUCTURE software on the whole MDLP dataset (in STRUCTURE input format circa 145 Mb file), but this trial run have ended in disaster and suspension of one from my LINUX accounts :/  It means that i won't be able to run STRUCTURE analysis for the whole dataset in a couple of months. Instead, i would try to analyse each chromosome separately.

PS.PS. Pat Berge has requested me to publish ADMIXTURE/PCA/MDS data in spreadsheet
ADMIXTURE results
PCA/MDS plot data

Sunday, June 26, 2011

SNP Map application: a new genomic toy

Smallaxe of DNA-forums.com has created a platform for developing tools to help exploring RAW data files.
Basically it is useful for identifying possible areas of your genome that align with different regions and populations. The user interface at this point is fairly rudimentary, but it makes doing the SPSMart sort of thing quite a bit more easy. It can accept 23andMe and FTDNA FamilyFinder data files.
Here are guidelines for using SNPMap tool:
The general sequence when using the tool is:
1. Load your genetic data using the File menu. The program can accept either 23andMe or FTDNA Family Finder unzipped data files. Note that the program is running locally on your computer, so your data stays with you. No need to send your data to me.

2. Select the populations you're interested in. There are two lists on the left. The top list shows the world regions. The bottom list shows the population data sets within the selected region. You can check/uncheck the regions and populations you want used in the analysis. If you've checked more than one region, then the analysis will compare those regions against each other. If you've checked only a single region, then the analysis will compare the individual populations within that region against each other. You may have to click twice on an item to toggle the check box.

3. Choose the chromosome. For speed, and to make the display of large amounts of info easier, the analysis works with a single chromosome at a time. There is a dropdown box for selecting the chromosome. If you are using FTDNA data, remember that you will have two files - one with just the X chromosome data, and one with chromosomes 1-22.

4. Click on the Recalculate button. This will perform the analysis of your data against the reference population data.
The symbol that looks like an equals sign with a slash through it means the person's genetic data is very unlike that region/population for that SNP, and other regions/populations are more likely. If the person is homozygous, then two of those symbols show up. One symbol means the person is at most 50% ancestry of that region/population at that location of their genome. Two symbols means the person may be 0% ancestry of that region/population at that location. The circle symbol (it's actually a happy face) means the person is very like that region/population, and the SNP is a good marker for that region/population vs. all the others being compared. One happy face means probably at least 50% that region/population, and two means probably 100% that region/population.

As you get more familiar with the program, you can adjust the Reliable/Noisy slider to change the threshold the program uses to distinguish SNPs of interest.

By default, only SNPs of interest are displayed. If you check the All SNPs checkbox and then Recalculate, all the SNPs in your data for which there is any population data will be displayed.

If you select a row Populations list and then right click in the list, a menu will pop up with some options, including opening the Yale ALFRED website with more detailed information about the selected population.

You can select one or more rows in the SNP results list and then right click in the list to see a menu of options. If you select a single row, the menu will have items for opening the Yale ALFRED website with detailed population information about the selected SNP, and an item for opening the NIH database website containing a variety of detailed SNP information. If you've selected one or more rows, the menu will let you copy the list of selected SNP ids to the clipboard. This is useful for pasting into the SNP list entry on the SPSMart website for further SNP exploration. You can also copy all the selected data to the clipboard in comma delimited format suitable for pasting into a spreadsheet program such as Excel or OpenOffice.

Tuesday, June 14, 2011

Interpretome -personal genome analysis webservice

David of Eurogenes blog and Zack of Harappa DNA project have posted a link to a  very promising  personal genome analysis webservice Interpretome.  The webservice has an extended set of online utilities useful for different types of analysis (imputation of missing SNPS in 23andme raw data file; clinical assesment of genetic risks; different types of the ancestry analysis, such as IBS, PCA and chromosome painting). However,  the coolest feature of this webservice is that webservice leverages data from various genetic diversity panels, such as HGDP, POPRES, and other data sources (below), which have generated genotype data from individuals around the world. For the individuals of European origin, the most informative genetic diversity  panel is The Population Reference Sample POPRES, whose controlled-access data can otherwise only be obtained if a user has been authorized by the appropriate Data Access Committee (DAC).
I, therefore, encourage all participants of the MDL project to analyze their ancestry using Intrepretome's ancestry online utilities. To get started, choose a reference panel onto which to project yourself and the number of SNPs to use for the projection. First, start with "HGDP: World" or "Hapmap: World" to explore your ancestry on a global population scale. For more detail in a specific region, choose the reference panel from the relevant population. If applicable, choose a level of detail for the results to be plotted.

For the POPRES dataset use 43,000 SNPs (23andme Raw data v2) or 74,000 SNPs (23andme Raw data v3). If you want to recapitulate the results of  paper Novembre J. Genes mirror geography within Europe. Nat 456:98 (2008), plot PC1 vs. PC4.


Thursday, June 2, 2011

Graphoanalytical approach to the analysis of genetic relationships

It goes without saying that the admixture analysis and MDA&PCA (statistical) calculations are most typical methods for inferring the underlying genetic structures in the human populations. But sometimes it is very useful to visualize the shared genetic components (or, rather, segments) as graphs. Graph visualization can be used to better understand underlying patterns of genetic effects (such as the genetic drift, the founder's effect etc). 
Earlier i attempted to  re-formulate the basic aspects of the graphoanalytical  approach  , proposing graph visualization techniques, which can be easily applied to the analysis of IBD (identical by descent) segments. I even succeeded in  producing a neat graphical output for IBD files, but the method itself required more amending and refining efforts. 

As I was willing to invest more time and efforts to make of my preliminary considerations of graph visualization something meaningful,  then this moning i decided to repeat my previous experiment. Prior to running PLINK routine commands for detecting IBD  segments, i removed almost all  individuals with 3 or more standard deviations from the mean Z-score. Another subset of individuals (c.9% of all dataset)  was removed due to the suprisingly significant values of inbreeding coefficient F (in PLINK, the calculations of inbreeding coeffecient F are based on the observed versus expected number of homozygous genotypes). The champions of inbreeding coefficient F are Orcadians, and some Lithuanians/Belarusians, whose F-score is significantly higher than European average value of F. As some of project's participants might be interested in the re-evaluation of Z-score and F-inbreeding coeffecient, i've uploaded the corresponding NEAREST and HET files. 

Then i reduced the dataset by performing linkage disequilibrium based SNP pruning  and removing low quality SNPs. Then i created the genetic map (in cM unit) for the remainig set of  circa 72000 SNPs (this time i sorted&joined the interpolated data from HapMap)  and ran the segmental sharing analysis in PLINK (it took c.70 minutes to complete this task, since the scope of the analysis was restricted to the low sharing 1cM/1Mb smps).The resulting file (available for donwloading here)  has been converted into delimited table format and then read into R tool for network analysis with Igraph.

I used values of NSEG (number of pairwise shared segments -see INDIV.SUMMARY file) to create a weighted undirected graph. The created graph was then analyzed using custom Igraph's methods of community detection by extended modularity and the calculation of weighted betweenness  in igraph. The vertex and edge betweenness are  defined by the number of geodesics (shortest paths) going through a vertex or an edge. With all preprocessing of graph done in R, i converted Igraph object into GraphXML and pulled it into GEPHI  , to apply some minor visual and aesthetic tweaks.







You can see the final graph in its full glory in PDF format (with a loseless invariant scaling), although the better representation of graph is achieved in GEPHI (compare the pictures uploaded above).