I would like to use R to perform hierarchical clustering with two groups of variables describing the same samples. One group is microarray gene expression data (for specific genes) that have been normalized and batch effect corrected. The other group also has some quantitative clinical parameters that describe the same samples. However, these clinical variables have not been normalized or subjected to any kind of transformation(i.e. raw continuous values).
For example, one variable of these could have range of values from 2 to 35, whereas another from 0.1 to 0.9, etc.
Thus, as my ultimate goal in to implement hierarchical clustering and use both groups simultaneously (merged in a matrix/dataframe), in order to inspect which of these clinical variables cluster with specific genes, etc:
1) Is an initial transformation in the group of the clinical variables necessary before merging with the genes and perform the clustering ? For example: log2 transformation, which has also been done to part of my gene expression data !!
2) Or, a row scaling (that is the total features in the input data) would take into account this discrepancy ?
3) For a similar analysis/approach, like constructing a correlation plot of the above total variables, would a simple scaling be sufficient?
Without having seen your gene expression data, I can only provide you some general suggestions based on your description, in the context of the 3 questions you asked:
1) You should definitely check the distribution of each group. In R, you may use one or more of the following function to visualize the distribution:
hist(expression_data) ##histogram
plot(density(expression_data)) ##density plot; alternative to histogram
qqnorm(expression_data); qqline(expression_data) #QQ plot
Since my understanding is that one of your expression data group is log2 transformed, that particular group should have a normal distribution (i.e. a bell curve shape in the histogram and a straight line in the QQ plot). Whether to transform the group that has not yet been transformed will depend on what you want to do with the data. For instance, if you want to use a t-test to compare the two groups, then you definitely need a transformation, as there is a normality assumption associated with a t-test. With regard to hierarchical clustering, if you decide to use both groups in a single clustering analysis, then why would you ever keep one transformed and the other not?
2) Scaling by features is a reasonable approach. Here is a clustering lecture from a Utah State Univ. stats course, with an example. scale=TRUE is an option for you if you decide to use heatmap function in R.
3) I don't think there is a definitive answer to your third question. It has to depend on how many available features you have and what analyses you will be doing downstream. Similar to question 1, I would argue that simple scaling may be sufficient for visualizing your data by hierarchical clustering. However, do keep in mind that, say you decide to perform a linear model (which is very common with microarray gene expression data), you might want to consider more sophisticated data scaling.
Related
I'm performing hierarchical cluster analysis using Ward's method on a dataset containing 1000 observations and 37 variables (all are 5-point likert-scales).
First, I ran the analysis in SPSS via
CLUSTER Var01 to Var37
/METHOD WARD
/MEASURE=SEUCLID
/ID=ID
/PRINT CLUSTER(2,10) SCHEDULE
/PLOT DENDROGRAM
/SAVE CLUSTER(2,10).
FREQUENCIES CLU2_1.
I additionaly performed the analysis in R:
datA <- subset(dat, select = Var01:Var37)
dist <- dist(datA, method = "euclidean")
hc <- hclust(d = dist, method = "ward.D2")
table(cutree(hc, k = 2))
The resulting cluster sizes are:
1 2
SPSS 712 288
R 610 390
These results are obviously confusing to me, as they differ substentially (which becomes highly visible when observing the dendrograms; also applies for the 3-10 clusters solutions). "ward.D2" takes into account the squared distance, if I'm not mistaken, so I included the simple distance matrix here. However, I tried several (combinations) of distance and clustering methods, e.g. EUCLID instead of SEUCLID, squaring the distance matrix in R, applying "ward.D" method,.... I also looked at the distance matrices generated by SPSS and R, which are identical (when applying the same method). Ultimately, I excluded duplicate cases (N=29) from my data, guessing that those might have caused differences when being allocated (randomly) at a certain point. All this did not result in matching outputs in R and SPSS.
I tried running the analysis with the agnes() function from the cluster package, which resulted in - again - different results compared to SPSS and even hclust() (But that's a topic for another post, I guess).
Are the underlying clustering procedures that different between the programs/packages? Or did I overlook a crucial detail? Is there a "correct" procedure that replicates the results yielded in SPSS?
If the distance matrices are identical and the merging methods are identical, the only thing that should create different outcomes is having tied distances handled differently in two algorithms. Tied distances might be present with the original full distance matrix, or might occur during the joining process. If one program searches the matrix and finds two or more distances tied at the minimum value at that step, and it selects the first one, while another program selects the last one, or one or both select one at random from among the ties, different results could occur.
I'd suggest starting with a small example with some data with randomness added to values to make tied distances unlikely and see if the two programs produce matching results on those data. If not, there's a deeper problem. If so, then tie handling might be the issue.
I am trying to carry out hierarchical cluster analysis (based on Ward's method) on a large dataset (thousands of records and 13 variables) representing multi-species observations of marine predators, to identify possible significant clusters in species composition.
Each record has date, time etc and presence/absence data (0 / 1) for each species.
I attempted hierarchical clustering with the function pvclust. I transposed the data (pvclust works on transposed tables), then I ran pvclust on the data selecting Jacquard distances (“binary” in R) as a distance measure (suitable for species pres/abs data) and Ward’s method (“ward.D2”). I used “parallel = TRUE” to reduce computation time. However, using a default of nboots= 1000, my computer was not able to finish the computation in hours and finally I got ann error, so I tried with lower nboots (100).
I cannot provide my dataset here, and I do not think it makes sense to provide a small test dataset, as one of the main issues here seems to be the size itself of the dataset. However, I am providing the lines of code I used for the transposition, clustering and plotting:
tdata <- t(data)
cluster <- pvclust(tdata, method.hclust="ward.D2", method.dist="binary",
nboot=100, parallel=TRUE)
plot(cluster, labels=FALSE)
This is the dendrogram I obtained (never mind the confusion at the lower levels due to overlap of branches).
As you can see, the p-values for the higher ramifications of the dendrogram all seem to be 0.
Now, I understand that my data may not be perfect, but I still think there is something wrong with the method I am using, as I would not expect all these values to be zero even with very low significance in the clusters.
So my questions would be
is there anything I got wrong in the pvclust function itself?
may my low nboots (due to “weak” computer) be a reason for the non-significance of my results?
are there other functions in R I could try for hierarchical clustering that also deliver p-values?
Thanks in advance!
.............
I have tried to run the same code on a subset of 500 records with nboots = 1000. This worked in a reasonable computation time, but the output is still not very satisfying - see dendrogram2 .dendrogram obtained for a SUBSET of 500 records and nboots=1000
I have two groups. The treatment group is exposure to media; the control group is no media. They are distinguished by a categorial variable in the data frame. (exposure to media = 1, no media = 0)
Now, I want to examine whether there are any clear differences between these two groups. To do this, apply the k-means algorithm with two clusters to four variables (proportion of black population, proportion of male population, proportion of hispanic population, median income on the logarithmic scale).
How to do this in R? Could anyone give some hints? Thanks!
Try this:
km <-kmeans(your data, 2, nstart=10)
your data here as a data.frame (your whole data or you can select the variables that you are interesting about them). You need to select the number of clusters (here is 2). A good practice to understand your data is to apply different number of cluster and then see which one fit your data better (use for example any criteria methods such as AIC or BIC).
k-means is an approach applied to cluster data. Where this data come from different distribution and we would like to know from where each observation come from (from which distribution).
You can also have a look at many tutorials about kmeans in R. For example,
https://onlinecourses.science.psu.edu/stat857/node/125
https://www.r-statistics.com/2013/08/k-means-clustering-from-r-in-action/
http://www.statmethods.net/advstats/cluster.html
my aim is to cluster 126 time-series concerning 26 weeks (so each time-series has 26 observation). I used pam{cluster} = partitioning around medoids to cluster these time-series.
Before clustering I wanted to compare which distance measure is the most appropriate: euclidean, manhattan or dynamic time warping. I used each distance to cluster and compare by silhouette plot. Is there any way I can compare different distance measure?
For example I know that procedure clValid {clValid} to validate cluster results, however I cannot implement dtw to calculate indexes.
So how can I compare different distance metrics (not only by silhouette)?
Additional question: is GAP statistic enough to decide how many clusters choose? Or should I evaluate number of clusters with different methods or compare two or three ways how to do it?
I would be grateful for any suggestions.
I have just read the book "cluster analysis, fifth edition" by Brian S. Everitt, etc. And currently, I adopt the following strategy to select method to calculate distance matrix, clustering and validation:
for distance: using cmdscale{stats} function to calculate multidimentional scaling, and plot the scatterplot of the two scaling dimensions with density information. As expected, if there is distinct clusters or nested clusters, the scatterplot will give some hints.
for clustering: for every clustering method, calculate cophenetic correlation between clustering results and the distance, this can be calculated using cophenetic{stats} function. The best clustering method will give higher correlation. However, this is only working for hierarchical clustering. I haven't idea for other clustering methods, like pam, or kmeans.
for partition evaluation: package {clusterSim} give several function to calculate the index to evaluate the clustering quality. Another package {NbClust} also calculate so many as 30 index to evaluate the combination of "distance", "clustering" and "number of clusters". However, this package partition the hierarchical tree using {cutree}, which is not suitable for nested clustering structure. Another method provided by {dynamicTreeCut} give reasonable results.
for cluster number determination: will added later.
Cluster data for which you have class labels, and use the RAND index to measure cluster quality.
50 such datasets are at the UCR time series archive
This paper does something similar
http://www.cs.ucr.edu/~eamonn/ClusteringTimeSeriesUsingUnsupervised-Shapelets.pdf
I want to do clustering of my data (kmeans or hclust) in R language (coding). My data is ordinal, which means that the data is Likert scale to measure the causes of cost escalation (I have 41 causes "variables") that scaled from 1 to 5, which 1 is no effect to 5 major effect (I have about 160 observations "who rank the causes")... any help of how to cluster the 41 cause based on the observations ... do I have to convert the scale to percentage or z score before clustering or any thing that help ...... I really need your help!! here is the data to play with https://docs.google.com/spreadsheet/ccc?key=0AlrR2eXjV8nXdGtLdlYzVk01cE96Rzg2NzRpbEZjUFE&usp=sharing
I want to cluster the variables (the columns) in terms of similarity of occurrence in observations... I follow the code in statmethods.net/advstats/cluster.html; but I couldn't cluster the variables (the columns) in terms of similarity of occurrence in observations and also I follow the work at mattpeeples.net/kmeans.html#help; but I don't know why he convert the data to percentage and then to Z-score standardize.
It isn't clear to me if you want to cluster the rows (the observations) in terms of similarity in the variables, or cluster the variables (the columns) in terms of similarity of occurrence in observations?
Anyway, see package cluster. This is a recommended package that comes with all R installations.
Read ?daisy for details of what is done with ordinal data. This metric can be used in functions such as agnes (for hierarchical clustering) or pam (for partitioning about medoids, a more robust version of k-means).
By default, these will cluster the rows/observations. Simply transpose the data object using t() if you want to cluster the columns (variables). Although that may well mess up the data depending on how you have stored them.
Converting the data to percentage is called normalization of data so all the variables are in the range of 0 - 1.
If data is not normalized you run the risk of bias towards dimensions with large values