![Page 1: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/1.jpg)
Metabolomics: an interpreting tool to understand kidney graft recipients
grouping and their recovery trajectory
Italian BioR DayLodi 30 Novembre 2012
Marco Calderisi, [email protected]
Italian BioR DayLodi 30 Novembre 2012
![Page 2: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/2.jpg)
Kidney
Italian BioR DayLodi 30 Novembre 2012
Kidney removes wastes and fluid from the body
1/36
![Page 3: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/3.jpg)
Italian BioR DayLodi 30 Novembre 2012
Kidney graft recipients monitoring
Aim
immunological rejection ischemia/reperfusion injury
immunosuppressant nephrotoxicity2/36
![Page 4: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/4.jpg)
How
Italian BioR DayLodi 30 Novembre 2012
Urine samples + 1H-NMR + Chemometrics = Metabolomics
3/36
![Page 5: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/5.jpg)
Metabolomics
Italian BioR DayLodi 30 Novembre 2012
Determination of multiple metabolites in biofluids
and tissues and their changes over time
Instrumentation: NMR, MS, GC/MS, HPLC/MS
4/36
![Page 6: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/6.jpg)
Chemometrics
Italian BioR DayLodi 30 Novembre 2012
Chemometrics: chemistry + statistic + informatic
5/36
![Page 7: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/7.jpg)
Exploration + Modelling
Italian BioR DayLodi 30 Novembre 2012
PCA
PLS
6/36
![Page 8: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/8.jpg)
Exploration + Modelling
Italian BioR DayLodi 30 Novembre 2012 6/36
![Page 9: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/9.jpg)
Data Import
Italian BioR DayLodi 30 Novembre 2012
Matlab
save datiperR.mat data asseX '-v6'
require(R.matlab)aa <- readMat('datiperR.mat')
dati <- aa$datadati <- t(dati)
asseX <- aa$asseX
7/36
![Page 10: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/10.jpg)
Data Analysis
Italian BioR DayLodi 30 Novembre 2012
Preprocessing and pretreatment
Exploration
Modelling
8/36
![Page 11: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/11.jpg)
Preprocessing + Pretreatment
Italian BioR DayLodi 30 Novembre 2012
baseline and phase correction,referencing to internal standard,signals alignment
normalizationcentering
scalingother transformations
9/36
![Page 12: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/12.jpg)
Preprocessing + Pretreatment
Italian BioR DayLodi 30 Novembre 2012
ptw, dtw, xcms -> MS or LC spectraPROcess -> protein mass spectrometry
peak peaking
binning, TMS alignment
10/36
![Page 13: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/13.jpg)
Preprocessing + Pretreatment
Italian BioR DayLodi 30 Novembre 2012
TMS peaks before and after alignment...with icoshift...
11/36
![Page 14: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/14.jpg)
Preprocessing + Pretreatment
Italian BioR DayLodi 30 Novembre 2012
# 10 points binnings ynew <- colMeans(matrix(dati[1,1:32760], nrow = 10))xnew <- colMeans(matrix(asseX[1:32760], nrow = 10))
# plotplot(asseX[1:32760], dati[1, 1:32760], type = "l", xlab = "asseX ", ylab = "response", main = "binning", col = "red", ylim=c(0,10^8))
lines(xnew, ynew)
binning
12/36
![Page 15: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/15.jpg)
Preprocessing + Pretreatment
Italian BioR DayLodi 30 Novembre 2012
binning
13/36
![Page 16: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/16.jpg)
Preprocessing + Pretreatment
Italian BioR DayLodi 30 Novembre 2012
require(ptw)# baseline driftybase <- 45600*asseX + 20000*asseX^2 - 2000*asseX^3plot(asseX, ybase[1,], type='l', col='blue')y1mod <- dati[1,]+ybase[1,]# plotplot(asseX, y1mod, type='l', ylim=c(-3000000, 10000000), col='blue')lines(asseX, dati[1,], col='red')# correctionlines(asseX, baseline.corr(y1mod), col='grey')abline(h=0, col='green', lty=2)legend('topright', legend=c('modified', 'original', 'baseline corrected'), lty=1, bty='n', col=c('blue', 'red', 'grey'), cex=0.7)
Baseline correction
14/36
![Page 17: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/17.jpg)
Preprocessing + Pretreatment
Italian BioR DayLodi 30 Novembre 2012
Baseline correction
15/36
![Page 18: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/18.jpg)
Preprocessing + Pretreatment
Italian BioR DayLodi 30 Novembre 2012
δ ppm!
Data reduction
xnew <- c(x[p1:p2,], x[p3:p4,])
p1
p2
p3
p4
16/36
![Page 19: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/19.jpg)
Preprocessing + Pretreatment
Italian BioR DayLodi 30 Novembre 2012
Data reduction
plot(dati[1,], type='l')require (TeachingDemos)plot(dati[1,], type='l')# zooming the plotzoomplot(locator(2))# find the pointsp1 <- locator(1)p2 <- locator(1)plot(dati[1,], type='l')# zooming the plotzoomplot(locator(2))p3 <- locator(1)p4 <- locator(1)# new reduced datasetdati.r <- dati[,c(p1$x:p2$x, p3$x:p4$x)]
17/36
![Page 20: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/20.jpg)
Preprocessing + Pretreatment
Italian BioR DayLodi 30 Novembre 2012
Unit area normalizationdati.un <- sweep(dati.r, MARGIN = 1, apply(dati, 1,function(x) sqrt(sum(x^2))), FUN = '/')
par(mfrow=c(2,1))asseX <- t(asseX.r)matplot(asseX.r, t(dati.r), type='l', lwd=1, col=setcol, lty=1)matplot(asseX.r, t(dati.un), type='l', lwd=1, col=setcol, lty=1)
18/36
![Page 21: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/21.jpg)
Preprocessing + Pretreatment
Italian BioR DayLodi 30 Novembre 2012
Unit area normalization
19/36
![Page 22: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/22.jpg)
Preprocessing + Pretreatment
Italian BioR DayLodi 30 Novembre 2012
ScalingCENTERING
AUTOSCALING
PARETO SCALING
(X-Xmedio)/radq(sd)
20/36
![Page 23: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/23.jpg)
Preprocessing + Pretreatment
Italian BioR DayLodi 30 Novembre 2012
Scalingrequire(pls)
dati.c <- scale(dati.r, center = T, scale =F)dati.sc <- scale(dati.r, center = T, scale =T)
dati.p <- sweep(dati.r, MARGIN = 2, sqrt(apply(dati.r, MARGIN = 2, sd)), FUN = "/")
21/36
![Page 24: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/24.jpg)
Preprocessing + Pretreatment
Italian BioR DayLodi 30 Novembre 2012
Scaling
22/36
![Page 25: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/25.jpg)
Exploration + Modelling
Italian BioR DayLodi 30 Novembre 2012
15 patients were sampled all along the hospital recovery period (from 5 to 40 days) and during the first follow up
systematic sampling (one sample a day)
23/36
![Page 26: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/26.jpg)
Exploration + Modelling
Italian BioR DayLodi 30 Novembre 2012 24/36
![Page 27: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/27.jpg)
Exploration + Modelling
Italian BioR DayLodi 30 Novembre 2012
PCAPareto scaling3 patients
Post-operation
Pre-discharge
Follow-up
25/36
![Page 28: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/28.jpg)
Exploration + Modelling
Italian BioR DayLodi 30 Novembre 2012
PCAPareto scaling15 patients
Pre-discharge
Post-operation
Follow-up
26/36
![Page 29: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/29.jpg)
Exploration + Modelling
Italian BioR DayLodi 30 Novembre 2012
require(ChemometricsWithR)
mod <- PCA(dati.p)
figura1a <- scores(mod)
plot(figura1a[,2], figura1a[,3], xlab ='PC 1 (exp. var. 13.96%)', ylab = 'PC 2 (exp. var. 9.88%)', xaxt="n", yaxt="n", mgp=c(2.5, 1, 0), cex.lab = 1, type = 'n')
axis(1, at=seq(-1,1,0.25), lwd = 0, lwd.ticks = 1, tcl = 0.2, cex.axis=1, mgp=c(1.5, 0.5, 0))axis(2, at=seq(-1,1,0.25), lwd = 0, lwd.ticks = 1, tcl = 0.2, cex.axis=1, las = 2, mgp=c(1.5, 0.5, 0))
points(figura1a[,2], figura1a[,3], pch=as.numeric(figura1a[,1]), cex=0.8)
abline(v=0, h=0, col='grey', lty = 2)
27/36
![Page 30: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/30.jpg)
Exploration + Modelling
Italian BioR DayLodi 30 Novembre 2012
PLS-DA analysisone class vs one class
Modelled class: 1 2 3
Calibration Sensitivity 0.96 0.88 0.91
Specificity 1 0.9 0.96
Class. Err 0.02 0.11 0.07
Cross Validation Sensitivity 0.94 0.86 0.91
Specificity 1 0.83 0.92
Class. Err 0.03 0.15 0.08
28/36
![Page 31: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/31.jpg)
Exploration + Modelling
Italian BioR DayLodi 30 Novembre 2012
Post operation stage.The creatinine and creatine signals are rather low and there is a lipoprotein signal
29/36
![Page 32: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/32.jpg)
Exploration + Modelling
Italian BioR DayLodi 30 Novembre 2012
Pre discharge stage.Creatinine and creatine signals pretty high and lipoprotein signal almost disappeared
30/36
![Page 33: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/33.jpg)
Exploration + Modelling
Italian BioR DayLodi 30 Novembre 2012
Follow up stage.The creatinine and creatine signals are very intense
31/36
![Page 34: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/34.jpg)
Exploration + Modelling
Italian BioR DayLodi 30 Novembre 2012
mod <- do.pls(dati.p, class.p, 2, scale="mean")
VIP.scores.Y1 <- mod$VIP[,2]
plot(-axis.scale,VIP.scores.Y1, type='l', xaxt='n', yaxt='n', xlab='[ppm]', ylab='VIP scores', ylim=c(0,350), lwd = 0.5, cex.lab=0.8, main = 'VIP scores class 1', cex.main=0.9, col='black', mgp=c(2.5, 1, 0))
axis(1, at=seq(0,-10,-1), labels = seq(0,10,1), lwd = 0, lwd.ticks = 1, tcl = 0.2, cex.axis=0.8)
axis(2, at=seq(0, 320, 50), lwd = 0, lwd.ticks = 1, tcl = 0.2, cex.axis=0.8, las= 1)
grid(col = "lightgray", lty = "dotted", lwd =1)
32/36
![Page 35: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/35.jpg)
Exploration + Modelling
Italian BioR DayLodi 30 Novembre 2012
require(pls) // do.pls
short command: mod <- do.pls(x,y)extensive command: mod <- do.pls(x,y, ncomp, scale=c("mean", "autoscaling"), graph=c("line", "points"))
aim: to do a pls regression analysis with just one “click”
http://cran.r-project.org/web/packages/pls/http://mevik.net/work/software/pls.html
33/36
![Page 36: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/36.jpg)
Exploration + Modelling
Italian BioR DayLodi 30 Novembre 2012
Input:
•x is the predictor matrix
•y is the response variable
•ncomp it is the requested number of latent variables (optional). The default is the LV’s number corresponding to the lowest
RMSECV(*)
•scale: choose between mean centering (default) and autoscaling
•graph it is the plot layout: choose between points or line for predictors plot (optional).
(*) only Leave-One-Out CV procedure it is implemented
34/36
![Page 37: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/37.jpg)
Exploration + Modelling
Italian BioR DayLodi 30 Novembre 2012
Output:
it is a list.
•modello, it is the usual mvr output
•performance, explained variance for predictors and response variable, RMSE, RMSECV, number of latent variables selected)
•VIP
•coefficients
35/36
![Page 38: Metabolomics: an interpreting tool to understand kidney graft recipients grouping and their recovery trajectory](https://reader034.vdocument.in/reader034/viewer/2022042816/5599776f1a28ab18528b464a/html5/thumbnails/38.jpg)
Exploration + Modelling
Italian BioR DayLodi 30 Novembre 2012 36/36