Correlation between gene methylation status and clinical features
Testicular Germ Cell Tumors (Primary solid tumor)
21 August 2015  |  analyses__2015_08_21
Maintainer Information
Citation Information
Maintained by Juok Cho (Broad Institute)
Cite as Broad Institute TCGA Genome Data Analysis Center (2015): Correlation between gene methylation status and clinical features. Broad Institute of MIT and Harvard. doi:10.7908/C13F4NXZ
Overview
Introduction

This pipeline uses various statistical tests to identify genes whose promoter methylation levels correlated to selected clinical features.

Summary

Testing the association between 20645 genes and 10 clinical features across 134 samples, statistically thresholded by P value < 0.05 and Q value < 0.3, 6 clinical features related to at least one genes.

  • 30 genes correlated to 'YEARS_TO_BIRTH'.

    • PTPRD ,  MOCS1 ,  SCG3 ,  DCLK1 ,  HS6ST3 ,  ...

  • 30 genes correlated to 'PATHOLOGIC_STAGE'.

    • HAS1 ,  NLGN1 ,  ELMO1 ,  PLXDC2 ,  PDE3A ,  ...

  • 2 genes correlated to 'PATHOLOGY_T_STAGE'.

    • LOC286016 ,  TNPO3

  • 30 genes correlated to 'PATHOLOGY_M_STAGE'.

    • CLDN20__1 ,  SNRK ,  TFB1M__1 ,  LRP2BP ,  ATXN7L3 ,  ...

  • 30 genes correlated to 'RADIATION_THERAPY'.

    • BANK1 ,  GPR18 ,  UBAC2__2 ,  STK10 ,  LYN ,  ...

  • 4 genes correlated to 'ETHNICITY'.

    • BLOC1S3 ,  TRAPPC6A__1 ,  KIAA1704 ,  NUFIP1

  • No genes correlated to 'DAYS_TO_DEATH_OR_LAST_FUP', 'PATHOLOGY_N_STAGE', 'KARNOFSKY_PERFORMANCE_SCORE', and 'RACE'.

Results
Overview of the results

Complete statistical result table is provided in Supplement Table 1

Table 1.  Get Full Table This table shows the clinical features, statistical methods used, and the number of genes that are significantly associated with each clinical feature at P value < 0.05 and Q value < 0.3.

Clinical feature Statistical test Significant genes Associated with                 Associated with
DAYS_TO_DEATH_OR_LAST_FUP Cox regression test   N=0        
YEARS_TO_BIRTH Spearman correlation test N=30 older N=30 younger N=0
PATHOLOGIC_STAGE Kruskal-Wallis test N=30        
PATHOLOGY_T_STAGE Spearman correlation test N=2 higher stage N=0 lower stage N=2
PATHOLOGY_N_STAGE Spearman correlation test   N=0        
PATHOLOGY_M_STAGE Wilcoxon test N=30 class1 N=30 class0 N=0
RADIATION_THERAPY Wilcoxon test N=30 yes N=30 no N=0
KARNOFSKY_PERFORMANCE_SCORE Spearman correlation test   N=0        
RACE Kruskal-Wallis test   N=0        
ETHNICITY Wilcoxon test N=4 not hispanic or latino N=4 hispanic or latino N=0
Clinical variable #1: 'DAYS_TO_DEATH_OR_LAST_FUP'

No gene related to 'DAYS_TO_DEATH_OR_LAST_FUP'.

Table S1.  Basic characteristics of clinical feature: 'DAYS_TO_DEATH_OR_LAST_FUP'

DAYS_TO_DEATH_OR_LAST_FUP Duration (Months) 0.1-244.5 (median=41.5)
  censored N = 130
  death N = 3
     
  Significant markers N = 0
Clinical variable #2: 'YEARS_TO_BIRTH'

30 genes related to 'YEARS_TO_BIRTH'.

Table S2.  Basic characteristics of clinical feature: 'YEARS_TO_BIRTH'

YEARS_TO_BIRTH Mean (SD) 31.99 (9.3)
  Significant markers N = 30
  pos. correlated 30
  neg. correlated 0
List of top 10 genes differentially expressed by 'YEARS_TO_BIRTH'

Table S3.  Get Full Table List of top 10 genes significantly correlated to 'YEARS_TO_BIRTH' by Spearman correlation test

SpearmanCorr corrP Q
PTPRD 0.3959 2.191e-06 0.00744
MOCS1 0.3878 3.67e-06 0.00744
SCG3 0.3808 5.668e-06 0.00744
DCLK1 0.3789 6.342e-06 0.00744
HS6ST3 0.3781 6.678e-06 0.00744
GATA6 0.377 7.139e-06 0.00744
PCDHA1__5 0.3741 8.498e-06 0.00744
PCDHA10__5 0.3741 8.498e-06 0.00744
PCDHA11__5 0.3741 8.498e-06 0.00744
PCDHA12__5 0.3741 8.498e-06 0.00744
Clinical variable #3: 'PATHOLOGIC_STAGE'

30 genes related to 'PATHOLOGIC_STAGE'.

Table S4.  Basic characteristics of clinical feature: 'PATHOLOGIC_STAGE'

PATHOLOGIC_STAGE Labels N
  STAGE I 19
  STAGE IA 26
  STAGE IB 11
  STAGE II 5
  STAGE IIA 6
  STAGE IIB 1
  STAGE IIC 1
  STAGE III 2
  STAGE IIIA 1
  STAGE IIIB 6
  STAGE IIIC 5
  STAGE IS 46
     
  Significant markers N = 30
List of top 10 genes differentially expressed by 'PATHOLOGIC_STAGE'

Table S5.  Get Full Table List of top 10 genes differentially expressed by 'PATHOLOGIC_STAGE'

kruskal_wallis_P Q
HAS1 3.168e-05 0.065
NLGN1 4.106e-05 0.065
ELMO1 4.146e-05 0.065
PLXDC2 5.561e-05 0.065
PDE3A 6.749e-05 0.065
ADAMTS20 7.262e-05 0.065
NEDD4L 8.55e-05 0.065
BASP1__1 8.651e-05 0.065
LOC285696__1 8.651e-05 0.065
SMAD6 8.834e-05 0.065
Clinical variable #4: 'PATHOLOGY_T_STAGE'

2 genes related to 'PATHOLOGY_T_STAGE'.

Table S6.  Basic characteristics of clinical feature: 'PATHOLOGY_T_STAGE'

PATHOLOGY_T_STAGE Mean (SD) 1.47 (0.58)
  N
  T1 76
  T2 51
  T3 6
     
  Significant markers N = 2
  pos. correlated 0
  neg. correlated 2
List of 2 genes differentially expressed by 'PATHOLOGY_T_STAGE'

Table S7.  Get Full Table List of 2 genes significantly correlated to 'PATHOLOGY_T_STAGE' by Spearman correlation test

SpearmanCorr corrP Q
LOC286016 -0.3688 1.252e-05 0.129
TNPO3 -0.3688 1.252e-05 0.129
Clinical variable #5: 'PATHOLOGY_N_STAGE'

No gene related to 'PATHOLOGY_N_STAGE'.

Table S8.  Basic characteristics of clinical feature: 'PATHOLOGY_N_STAGE'

PATHOLOGY_N_STAGE Mean (SD) 0.25 (0.51)
  N
  N0 46
  N1 11
  N2 2
     
  Significant markers N = 0
Clinical variable #6: 'PATHOLOGY_M_STAGE'

30 genes related to 'PATHOLOGY_M_STAGE'.

Table S9.  Basic characteristics of clinical feature: 'PATHOLOGY_M_STAGE'

PATHOLOGY_M_STAGE Labels N
  class0 115
  class1 4
     
  Significant markers N = 30
  Higher in class1 30
  Higher in class0 0
List of top 10 genes differentially expressed by 'PATHOLOGY_M_STAGE'

Table S10.  Get Full Table List of top 10 genes differentially expressed by 'PATHOLOGY_M_STAGE'

W(pos if higher in 'class1') wilcoxontestP Q AUC
CLDN20__1 450 0.001211 0.0915 0.9783
SNRK 450 0.001211 0.0915 0.9783
TFB1M__1 450 0.001211 0.0915 0.9783
LRP2BP 448 0.001342 0.0915 0.9739
ATXN7L3 447 0.001412 0.0915 0.9717
ATP7B__1 445 0.001564 0.0915 0.9674
GLDC 445 0.001564 0.0915 0.9674
MCPH1 445 0.001564 0.0915 0.9674
NCRNA00174 445 0.001564 0.0915 0.9674
ZNF543 445 0.001564 0.0915 0.9674
Clinical variable #7: 'RADIATION_THERAPY'

30 genes related to 'RADIATION_THERAPY'.

Table S11.  Basic characteristics of clinical feature: 'RADIATION_THERAPY'

RADIATION_THERAPY Labels N
  NO 110
  YES 19
     
  Significant markers N = 30
  Higher in YES 30
  Higher in NO 0
List of top 10 genes differentially expressed by 'RADIATION_THERAPY'

Table S12.  Get Full Table List of top 10 genes differentially expressed by 'RADIATION_THERAPY'

W(pos if higher in 'YES') wilcoxontestP Q AUC
BANK1 275 3.155e-07 0.000826 0.8684
GPR18 286 4.635e-07 0.000826 0.8632
UBAC2__2 286 4.635e-07 0.000826 0.8632
STK10 291 5.511e-07 0.000826 0.8608
LYN 303 8.314e-07 0.000826 0.855
C12ORF27 309 1.019e-06 0.000826 0.8522
C3ORF62 313 1.166e-06 0.000826 0.8502
NUDT3 315 1.247e-06 0.000826 0.8493
LAT 317 1.333e-06 0.000826 0.8483
NAGK 317 1.333e-06 0.000826 0.8483
Clinical variable #8: 'KARNOFSKY_PERFORMANCE_SCORE'

No gene related to 'KARNOFSKY_PERFORMANCE_SCORE'.

Table S13.  Basic characteristics of clinical feature: 'KARNOFSKY_PERFORMANCE_SCORE'

KARNOFSKY_PERFORMANCE_SCORE Mean (SD) 94.9 (6)
  Score N
  80 5
  90 40
  100 53
     
  Significant markers N = 0
Clinical variable #9: 'RACE'

No gene related to 'RACE'.

Table S14.  Basic characteristics of clinical feature: 'RACE'

RACE Labels N
  ASIAN 4
  BLACK OR AFRICAN AMERICAN 6
  WHITE 119
     
  Significant markers N = 0
Clinical variable #10: 'ETHNICITY'

4 genes related to 'ETHNICITY'.

Table S15.  Basic characteristics of clinical feature: 'ETHNICITY'

ETHNICITY Labels N
  HISPANIC OR LATINO 12
  NOT HISPANIC OR LATINO 111
     
  Significant markers N = 4
  Higher in NOT HISPANIC OR LATINO 4
  Higher in HISPANIC OR LATINO 0
List of 4 genes differentially expressed by 'ETHNICITY'

Table S16.  Get Full Table List of 4 genes differentially expressed by 'ETHNICITY'

W(pos if higher in 'NOT HISPANIC OR LATINO') wilcoxontestP Q AUC
BLOC1S3 c("1165", "2.147e-05") c("1165", "2.147e-05") 0.222 0.8746
TRAPPC6A__1 c("1165", "2.147e-05") c("1165", "2.147e-05") 0.222 0.8746
KIAA1704 c("186", "4.368e-05") c("186", "4.368e-05") 0.225 0.8604
NUFIP1 c("186", "4.368e-05") c("186", "4.368e-05") 0.225 0.8604
Methods & Data
Input
  • Expresson data file = TGCT-TP.meth.by_min_clin_corr.data.txt

  • Clinical data file = TGCT-TP.merged_data.txt

  • Number of patients = 134

  • Number of genes = 20645

  • Number of clinical features = 10

Selected clinical features
  • Further details on clinical features selected for this analysis, please find a documentation on selected CDEs (Clinical Data Elements). The first column of the file is a formula to convert values and the second column is a clinical parameter name.

  • Survival time data

    • Survival time data is a combined value of days_to_death and days_to_last_followup. For each patient, it creates a combined value 'days_to_death_or_last_fup' using conversion process below.

      • if 'vital_status'==1(dead), 'days_to_last_followup' is always NA. Thus, uses 'days_to_death' value for 'days_to_death_or_fup'

      • if 'vital_status'==0(alive),

        • if 'days_to_death'==NA & 'days_to_last_followup'!=NA, uses 'days_to_last_followup' value for 'days_to_death_or_fup'

        • if 'days_to_death'!=NA, excludes this case in survival analysis and report the case.

      • if 'vital_status'==NA,excludes this case in survival analysis and report the case.

    • cf. In certain diesase types such as SKCM, days_to_death parameter is replaced with time_from_specimen_dx or time_from_specimen_procurement_to_death .

  • This analysis excluded clinical variables that has only NA values.

Survival analysis

For survival clinical features, Wald's test in univariate Cox regression analysis with proportional hazards model (Andersen and Gill 1982) was used to estimate the P values using the 'coxph' function in R. Kaplan-Meier survival curves were plot using the four quartile subgroups of patients based on expression levels

Correlation analysis

For continuous numerical clinical features, Spearman's rank correlation coefficients (Spearman 1904) and two-tailed P values were estimated using 'cor.test' function in R

Wilcoxon rank sum test (Mann-Whitney U test)

For two groups (mutant or wild-type) of continuous type of clinical data, wilcoxon rank sum test (Mann and Whitney, 1947) was applied to compare their mean difference using 'wilcox.test(continuous.clinical ~ as.factor(group), exact=FALSE)' function in R. This test is equivalent to the Mann-Whitney test.

Q value calculation

For multiple hypothesis correction, Q value is the False Discovery Rate (FDR) analogue of the P value (Benjamini and Hochberg 1995), defined as the minimum FDR at which the test may be called significant. We used the 'Benjamini and Hochberg' method of 'p.adjust' function in R to convert P values into Q values.

Download Results

In addition to the links below, the full results of the analysis summarized in this report can also be downloaded programmatically using firehose_get, or interactively from either the Broad GDAC website or TCGA Data Coordination Center Portal.

References
[1] Andersen and Gill, Cox's regression model for counting processes, a large sample study, Annals of Statistics 10(4):1100-1120 (1982)
[2] Spearman, C, The proof and measurement of association between two things, Amer. J. Psychol 15:72-101 (1904)
[3] Mann and Whitney, On a Test of Whether one of Two Random Variables is Stochastically Larger than the Other, Annals of Mathematical Statistics 18 (1), 50-60 (1947)
[4] Benjamini and Hochberg, Controlling the false discovery rate: a practical and powerful approach to multiple testing, Journal of the Royal Statistical Society Series B 59:289-300 (1995)