Correlation between gene methylation status and clinical features
Brain Lower Grade Glioma (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/C1930SDS
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 20117 genes and 8 clinical features across 515 samples, statistically thresholded by P value < 0.05 and Q value < 0.3, 7 clinical features related to at least one genes.

  • 30 genes correlated to 'YEARS_TO_BIRTH'.

    • TCHH ,  TRIM58 ,  LOC150786 ,  SHISA2 ,  RELN ,  ...

  • 30 genes correlated to 'GENDER'.

    • ALG11__1 ,  UTP14C ,  POLDIP3 ,  RNU12 ,  KIF4B ,  ...

  • 30 genes correlated to 'RADIATION_THERAPY'.

    • SIGLEC9 ,  CCL3 ,  FOXS1 ,  KCNK18 ,  MYCT1 ,  ...

  • 30 genes correlated to 'KARNOFSKY_PERFORMANCE_SCORE'.

    • PHTF1 ,  HS6ST1 ,  UBC ,  PIK3AP1 ,  DEGS1 ,  ...

  • 30 genes correlated to 'HISTOLOGICAL_TYPE'.

    • REST ,  MAPKAP1 ,  C2ORF67 ,  GLIS3 ,  SLC2A4RG ,  ...

  • 12 genes correlated to 'RACE'.

    • C6ORF52__1 ,  PAK1IP1__1 ,  RNF135 ,  LOC253039 ,  PSMD5 ,  ...

  • 2 genes correlated to 'ETHNICITY'.

    • THAP1 ,  ASNSD1

  • No genes correlated to 'DAYS_TO_DEATH_OR_LAST_FUP'

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=26 younger N=4
GENDER Wilcoxon test N=30 male N=30 female N=0
RADIATION_THERAPY Wilcoxon test N=30 yes N=30 no N=0
KARNOFSKY_PERFORMANCE_SCORE Spearman correlation test N=30 higher score N=30 lower score N=0
HISTOLOGICAL_TYPE Kruskal-Wallis test N=30        
RACE Kruskal-Wallis test N=12        
ETHNICITY Wilcoxon test N=2 not hispanic or latino N=2 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) 0-211.2 (median=20.8)
  censored N = 393
  death N = 121
     
  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) 42.93 (13)
  Significant markers N = 30
  pos. correlated 26
  neg. correlated 4
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
TCHH 0.5688 2.172e-45 4.37e-41
TRIM58 0.5602 8.304e-44 8.35e-40
LOC150786 0.5234 1.685e-37 1.13e-33
SHISA2 0.5191 8.264e-37 4.16e-33
RELN 0.5046 1.475e-34 5.93e-31
FOXE3 0.4993 1.215e-33 4.08e-30
ADAMTSL3 0.4917 1.193e-32 3.43e-29
SSTR4 0.4732 4.779e-30 1.2e-26
TFAP2B 0.4719 7.287e-30 1.63e-26
KLRG2 0.4715 8.359e-30 1.68e-26
Clinical variable #3: 'GENDER'

30 genes related to 'GENDER'.

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

GENDER Labels N
  FEMALE 230
  MALE 285
     
  Significant markers N = 30
  Higher in MALE 30
  Higher in FEMALE 0
List of top 10 genes differentially expressed by 'GENDER'

Table S5.  Get Full Table List of top 10 genes differentially expressed by 'GENDER'. 0 significant gene(s) located in sex chromosomes is(are) filtered out.

W(pos if higher in 'MALE') wilcoxontestP Q AUC
ALG11__1 64314 9.943e-79 1e-74 0.9811
UTP14C 64314 9.943e-79 1e-74 0.9811
POLDIP3 5627 8.214e-59 4.13e-55 0.9142
RNU12 5627 8.214e-59 4.13e-55 0.9142
KIF4B 16066 2.468e-23 9.93e-20 0.7549
WBP11P1 48367 1.59e-20 5.33e-17 0.7379
LOC389791__1 47946 1.624e-19 4.08e-16 0.7314
PTGES2__1 47946 1.624e-19 4.08e-16 0.7314
TLE1 17852 6.202e-19 1.39e-15 0.7277
ZC3H14 19608 4.42e-15 8.89e-12 0.7009
Clinical variable #4: 'RADIATION_THERAPY'

30 genes related to 'RADIATION_THERAPY'.

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

RADIATION_THERAPY Labels N
  NO 186
  YES 294
     
  Significant markers N = 30
  Higher in YES 30
  Higher in NO 0
List of top 10 genes differentially expressed by 'RADIATION_THERAPY'

Table S7.  Get Full Table List of top 10 genes differentially expressed by 'RADIATION_THERAPY'

W(pos if higher in 'YES') wilcoxontestP Q AUC
SIGLEC9 16404 1.495e-13 2.87e-09 0.7
CCL3 16532 2.852e-13 2.87e-09 0.6977
FOXS1 16783 9.917e-13 4.21e-09 0.6931
KCNK18 16790 1.026e-12 4.21e-09 0.693
MYCT1 16794 1.047e-12 4.21e-09 0.6929
TPRG1 16832 1.261e-12 4.23e-09 0.6922
SMCP 16906 1.808e-12 5.01e-09 0.6908
HSPG2 16926 1.992e-12 5.01e-09 0.6905
LRRC15 16963 2.382e-12 5.32e-09 0.6898
APOBEC3A 17126 5.201e-12 9.74e-09 0.6868
Clinical variable #5: 'KARNOFSKY_PERFORMANCE_SCORE'

30 genes related to 'KARNOFSKY_PERFORMANCE_SCORE'.

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

KARNOFSKY_PERFORMANCE_SCORE Mean (SD) 86.61 (13)
  Significant markers N = 30
  pos. correlated 30
  neg. correlated 0
List of top 10 genes differentially expressed by 'KARNOFSKY_PERFORMANCE_SCORE'

Table S9.  Get Full Table List of top 10 genes significantly correlated to 'KARNOFSKY_PERFORMANCE_SCORE' by Spearman correlation test

SpearmanCorr corrP Q
PHTF1 0.3072 4.143e-08 0.000833
HS6ST1 0.2921 1.877e-07 0.00148
UBC 0.2902 2.265e-07 0.00148
PIK3AP1 0.2856 3.591e-07 0.00148
DEGS1 0.2814 5.385e-07 0.00148
FLOT1__1 0.2812 5.478e-07 0.00148
CDCP1 0.2805 5.868e-07 0.00148
SIAH2 0.2798 6.284e-07 0.00148
RCAN1 0.2793 6.606e-07 0.00148
TWF2 0.277 8.223e-07 0.0015
Clinical variable #6: 'HISTOLOGICAL_TYPE'

30 genes related to 'HISTOLOGICAL_TYPE'.

Table S10.  Basic characteristics of clinical feature: 'HISTOLOGICAL_TYPE'

HISTOLOGICAL_TYPE Labels N
  ASTROCYTOMA 194
  OLIGOASTROCYTOMA 130
  OLIGODENDROGLIOMA 191
     
  Significant markers N = 30
List of top 10 genes differentially expressed by 'HISTOLOGICAL_TYPE'

Table S11.  Get Full Table List of top 10 genes differentially expressed by 'HISTOLOGICAL_TYPE'

kruskal_wallis_P Q
REST 5.16e-38 1.04e-33
MAPKAP1 2.178e-33 2.19e-29
C2ORF67 6.636e-32 4.45e-28
GLIS3 4.735e-31 2.38e-27
SLC2A4RG 8.814e-31 3.55e-27
BVES 4.255e-30 1.41e-26
ARID1A 4.892e-30 1.41e-26
TMC6 1.223e-29 2.73e-26
TMC8 1.223e-29 2.73e-26
CBX2 2.709e-29 5.45e-26
Clinical variable #7: 'RACE'

12 genes related to 'RACE'.

Table S12.  Basic characteristics of clinical feature: 'RACE'

RACE Labels N
  AMERICAN INDIAN OR ALASKA NATIVE 1
  ASIAN 8
  BLACK OR AFRICAN AMERICAN 21
  WHITE 475
     
  Significant markers N = 12
List of top 10 genes differentially expressed by 'RACE'

Table S13.  Get Full Table List of top 10 genes differentially expressed by 'RACE'

kruskal_wallis_P Q
C6ORF52__1 5.799e-09 5.83e-05
PAK1IP1__1 5.799e-09 5.83e-05
RNF135 3.362e-08 0.000225
LOC253039 2.949e-07 0.00119
PSMD5 2.949e-07 0.00119
ENTPD6 5.862e-06 0.0197
ISCA1 5.645e-05 0.158
ASH1L 7.246e-05 0.158
LOC645676 7.246e-05 0.158
DHRS7 8.268e-05 0.158
Clinical variable #8: 'ETHNICITY'

2 genes related to 'ETHNICITY'.

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

ETHNICITY Labels N
  HISPANIC OR LATINO 32
  NOT HISPANIC OR LATINO 449
     
  Significant markers N = 2
  Higher in NOT HISPANIC OR LATINO 2
  Higher in HISPANIC OR LATINO 0
List of 2 genes differentially expressed by 'ETHNICITY'

Table S15.  Get Full Table List of 2 genes differentially expressed by 'ETHNICITY'

W(pos if higher in 'NOT HISPANIC OR LATINO') wilcoxontestP Q AUC
THAP1 c("3966", "2.282e-05") c("3966", "2.282e-05") 0.263 0.724
ASNSD1 c("3989", "2.61e-05") c("3989", "2.61e-05") 0.263 0.7224
Methods & Data
Input
  • Expresson data file = LGG-TP.meth.by_min_clin_corr.data.txt

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

  • Number of patients = 515

  • Number of genes = 20117

  • Number of clinical features = 8

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)