Colon/Rectal Adenocarcinoma: Correlation between miRseq expression and clinical features
Maintained by Juok Cho (Broad Institute)
Overview
Introduction

This pipeline uses various statistical tests to identify miRs whose expression levels correlated to selected clinical features.

Summary

Testing the association between 420 genes and 11 clinical features across 521 samples, statistically thresholded by Q value < 0.05, 9 clinical features related to at least one genes.

- 7 genes correlated to 'AGE'.

- HSA-MIR-432 , HSA-MIR-153-2 , HSA-MIR-141 , HSA-MIR-26A-1 , HSA-MIR-410 , ...

- 35 genes correlated to 'PRIMARY.SITE.OF.DISEASE'.

- HSA-MIR-10B , HSA-MIR-1201 , HSA-MIR-30C-2 , HSA-MIR-1259 , HSA-MIR-20A , ...

- 1 gene correlated to 'GENDER'.

- HSA-MIR-651

- 45 genes correlated to 'HISTOLOGICAL.TYPE'.

- HSA-MIR-10B , HSA-MIR-92A-1 , HSA-MIR-1201 , HSA-MIR-20A , HSA-MIR-30C-2 , ...

- 4 genes correlated to 'PATHOLOGY.T'.

- HSA-MIR-144 , HSA-MIR-206 , HSA-MIR-502 , HSA-MIR-501

- 2 genes correlated to 'PATHOLOGY.N'.

- HSA-MIR-146A , HSA-MIR-625

- 31 genes correlated to 'PATHOLOGICSPREAD(M)'.

- HSA-LET-7F-2 , HSA-MIR-106A , HSA-MIR-628 , HSA-MIR-142 , HSA-MIR-616 , ...

- 2 genes correlated to 'TUMOR.STAGE'.

- HSA-MIR-146A , HSA-MIR-625

- 41 genes correlated to 'NEOADJUVANT.THERAPY'.

- HSA-MIR-106A , HSA-MIR-103-2 , HSA-MIR-26A-1 , HSA-LET-7F-2 , HSA-MIR-331 , ...

- No genes correlated to 'Time to Death', and 'RADIATIONS.RADIATION.REGIMENINDICATION'.

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 Q value < 0.05.

Clinical feature | Statistical test | Significant genes | Associated with |  | Associated with |
--- | --- | --- | --- | --- | --- | ---
Time to Death | Cox regression test | N=0 |  |  |  |
AGE | Spearman correlation test | N=7 | older | N=5 | younger | N=2
PRIMARY SITE OF DISEASE | t test | N=35 | rectum | N=29 | colon | N=6
GENDER | t test | N=1 | male | N=0 | female | N=1
HISTOLOGICAL TYPE | ANOVA test | N=45 |  |  |  |
PATHOLOGY T | Spearman correlation test | N=4 | higher pT | N=0 | lower pT | N=4
PATHOLOGY N | Spearman correlation test | N=2 | higher pN | N=0 | lower pN | N=2
PATHOLOGICSPREAD(M) | ANOVA test | N=31 |  |  |  |
TUMOR STAGE | Spearman correlation test | N=2 | higher stage | N=0 | lower stage | N=2
RADIATIONS RADIATION REGIMENINDICATION | t test | N=0 |  |  |  |
NEOADJUVANT THERAPY | t test | N=41 | yes | N=32 | no | N=9

Clinical variable #1: 'Time to Death'

No gene related to 'Time to Death'.

Table S1. Basic characteristics of clinical feature: 'Time to Death'

Time to Death | Duration (Months) | 0.1-135.5 (median=8)
| censored | N = 327
| death | N = 59
| Significant markers | N = 0

Clinical variable #2: 'AGE'

7 genes related to 'AGE'.

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

AGE | Mean (SD) | 66.82 (13)
| Significant markers | N = 7
| pos. correlated | 5
| neg. correlated | 2

List of 7 genes significantly correlated to 'AGE' by Spearman correlation test

Table S3. Get Full Table List of 7 genes significantly correlated to 'AGE' by Spearman correlation test

| SpearmanCorr | corrP | Q
--- | --- | --- | ---
HSA-MIR-432 | -0.2249 | 2.511e-07 | 0.000105
HSA-MIR-153-2 | 0.2109 | 1.19e-06 | 0.000499
HSA-MIR-141 | 0.2021 | 3.318e-06 | 0.00139
HSA-MIR-26A-1 | 0.1904 | 1.216e-05 | 0.00507
HSA-MIR-410 | -0.1756 | 6.142e-05 | 0.0256
HSA-MIR-616 | 0.1743 | 7.021e-05 | 0.0291
HSA-MIR-33B | 0.1706 | 9.244e-05 | 0.0383

Figure S1. Get High-res Image As an example, this figure shows the association of HSA-MIR-432 to 'AGE'. P value = 2.51e-07 with Spearman correlation analysis. The straight line presents the best linear regression.

Clinical variable #3: 'PRIMARY.SITE.OF.DISEASE'

35 genes related to 'PRIMARY.SITE.OF.DISEASE'.

Table S4. Basic characteristics of clinical feature: 'PRIMARY.SITE.OF.DISEASE'

PRIMARY.SITE.OF.DISEASE | Labels | N
| COLON | 377
| RECTUM | 140
| Significant markers | N = 35
| Higher in RECTUM | 29
| Higher in COLON | 6

List of top 10 genes differentially expressed by 'PRIMARY.SITE.OF.DISEASE'

Table S5. Get Full Table List of top 10 genes differentially expressed by 'PRIMARY.SITE.OF.DISEASE'

| T(pos if higher in 'RECTUM') | ttestP | Q | AUC
--- | --- | --- | --- | ---
HSA-MIR-10B | -9.25 | 1.427e-17 | 6e-15 | 0.7571
HSA-MIR-1201 | 8.31 | 3.75e-15 | 1.57e-12 | 0.7157
HSA-MIR-30C-2 | 7.78 | 1.331e-13 | 5.56e-11 | 0.6956
HSA-MIR-1259 | 7.6 | 3.24e-13 | 1.35e-10 | 0.6798
HSA-MIR-20A | 6.73 | 9.408e-11 | 3.91e-08 | 0.6793
HSA-MIR-1977 | 6.45 | 4.377e-10 | 1.82e-07 | 0.6582
HSA-MIR-425 | 6.46 | 5.171e-10 | 2.14e-07 | 0.6754
HSA-MIR-191 | 6.03 | 4.834e-09 | 2e-06 | 0.6589
HSA-MIR-224 | 5.98 | 6.626e-09 | 2.73e-06 | 0.6517
HSA-LET-7G | 5.86 | 1.513e-08 | 6.22e-06 | 0.6572

Figure S2. Get High-res Image As an example, this figure shows the association of HSA-MIR-10B to 'PRIMARY.SITE.OF.DISEASE'. P value = 1.43e-17 with T-test analysis.

Clinical variable #4: 'GENDER'

One gene related to 'GENDER'.

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

GENDER | Labels | N
| FEMALE | 249
| MALE | 272
| Significant markers | N = 1
| Higher in MALE | 0
| Higher in FEMALE | 1

List of one gene differentially expressed by 'GENDER'

Table S7. Get Full Table List of one gene differentially expressed by 'GENDER'

| T(pos if higher in 'MALE') | ttestP | Q | AUC
--- | --- | --- | --- | ---
HSA-MIR-651 | -3.99 | 7.466e-05 | 0.0314 | 0.6032

Figure S3. Get High-res Image As an example, this figure shows the association of HSA-MIR-651 to 'GENDER'. P value = 7.47e-05 with T-test analysis.

Clinical variable #5: 'HISTOLOGICAL.TYPE'

45 genes related to 'HISTOLOGICAL.TYPE'.

Table S8. Basic characteristics of clinical feature: 'HISTOLOGICAL.TYPE'

HISTOLOGICAL.TYPE | Labels | N
| COLON ADENOCARCINOMA | 327
| COLON MUCINOUS ADENOCARCINOMA | 48
| RECTAL ADENOCARCINOMA | 127
| RECTAL MUCINOUS ADENOCARCINOMA | 10
| Significant markers | N = 45

List of top 10 genes differentially expressed by 'HISTOLOGICAL.TYPE'

Table S9. Get Full Table List of top 10 genes differentially expressed by 'HISTOLOGICAL.TYPE'

| ANOVA_P | Q
--- | --- | ---
HSA-MIR-10B | 1.408e-18 | 5.91e-16
HSA-MIR-92A-1 | 1.435e-12 | 6.01e-10
HSA-MIR-1201 | 3.035e-12 | 1.27e-09
HSA-MIR-20A | 4.542e-12 | 1.89e-09
HSA-MIR-30C-2 | 2.873e-11 | 1.2e-08
HSA-MIR-1259 | 3.785e-10 | 1.57e-07
HSA-MIR-224 | 6.751e-10 | 2.79e-07
HSA-MIR-181D | 1.014e-09 | 4.19e-07
HSA-LET-7G | 1.215e-09 | 5.01e-07
HSA-MIR-425 | 1.609e-09 | 6.61e-07

Figure S4. Get High-res Image As an example, this figure shows the association of HSA-MIR-10B to 'HISTOLOGICAL.TYPE'. P value = 1.41e-18 with ANOVA analysis.

Clinical variable #6: 'PATHOLOGY.T'

4 genes related to 'PATHOLOGY.T'.

Table S10. Basic characteristics of clinical feature: 'PATHOLOGY.T'

PATHOLOGY.T | Mean (SD) | 2.84 (0.63)
|  | N
| T0 | 1
| T1 | 18
| T2 | 90
| T3 | 361
| T4 | 47
| Significant markers | N = 4
| pos. correlated | 0
| neg. correlated | 4

List of 4 genes significantly correlated to 'PATHOLOGY.T' by Spearman correlation test

Table S11. Get Full Table List of 4 genes significantly correlated to 'PATHOLOGY.T' by Spearman correlation test

| SpearmanCorr | corrP | Q
--- | --- | --- | ---
HSA-MIR-144 | -0.1775 | 4.933e-05 | 0.0207
HSA-MIR-206 | -0.2221 | 7.771e-05 | 0.0326
HSA-MIR-502 | -0.1697 | 0.0001053 | 0.044
HSA-MIR-501 | -0.1692 | 0.0001104 | 0.046

Figure S5. Get High-res Image As an example, this figure shows the association of HSA-MIR-144 to 'PATHOLOGY.T'. P value = 4.93e-05 with Spearman correlation analysis.

Clinical variable #7: 'PATHOLOGY.N'

2 genes related to 'PATHOLOGY.N'.

Table S12. Basic characteristics of clinical feature: 'PATHOLOGY.N'

PATHOLOGY.N | Mean (SD) | 0.61 (0.78)
|  | N
| N0 | 296
| N1 | 127
| N2 | 94
| Significant markers | N = 2
| pos. correlated | 0
| neg. correlated | 2

List of 2 genes significantly correlated to 'PATHOLOGY.N' by Spearman correlation test

Table S13. Get Full Table List of 2 genes significantly correlated to 'PATHOLOGY.N' by Spearman correlation test

| SpearmanCorr | corrP | Q
--- | --- | --- | ---
HSA-MIR-146A | -0.1814 | 3.327e-05 | 0.014
HSA-MIR-625 | -0.1777 | 4.832e-05 | 0.0202

Figure S6. Get High-res Image As an example, this figure shows the association of HSA-MIR-146A to 'PATHOLOGY.N'. P value = 3.33e-05 with Spearman correlation analysis.

Clinical variable #8: 'PATHOLOGICSPREAD(M)'

31 genes related to 'PATHOLOGICSPREAD(M)'.

Table S14. Basic characteristics of clinical feature: 'PATHOLOGICSPREAD(M)'

PATHOLOGICSPREAD(M) | Labels | N
| M0 | 393
| M1 | 67
| M1A | 9
| M1B | 1
| MX | 42
| Significant markers | N = 31

List of top 10 genes differentially expressed by 'PATHOLOGICSPREAD(M)'

Table S15. Get Full Table List of top 10 genes differentially expressed by 'PATHOLOGICSPREAD(M)'

| ANOVA_P | Q
--- | --- | ---
HSA-LET-7F-2 | 2.56e-08 | 1.08e-05
HSA-MIR-106A | 4.642e-08 | 1.95e-05
HSA-MIR-628 | 9.245e-08 | 3.86e-05
HSA-MIR-142 | 4.735e-07 | 0.000197
HSA-MIR-616 | 1.158e-06 | 0.000482
HSA-MIR-199A-2 | 1.516e-06 | 0.000629
HSA-MIR-1271 | 1.535e-06 | 0.000636
HSA-MIR-199B | 1.564e-06 | 0.000646
HSA-MIR-140 | 2.194e-06 | 0.000904
HSA-MIR-199A-1 | 2.266e-06 | 0.000931

Figure S7. Get High-res Image As an example, this figure shows the association of HSA-LET-7F-2 to 'PATHOLOGICSPREAD(M)'. P value = 2.56e-08 with ANOVA analysis.

Clinical variable #9: 'TUMOR.STAGE'

2 genes related to 'TUMOR.STAGE'.

Table S16. Basic characteristics of clinical feature: 'TUMOR.STAGE'

TUMOR.STAGE | Mean (SD) | 2.42 (0.95)
|  | N
| Stage 1 | 90
| Stage 2 | 188
| Stage 3 | 149
| Stage 4 | 75
| Significant markers | N = 2
| pos. correlated | 0
| neg. correlated | 2

List of 2 genes significantly correlated to 'TUMOR.STAGE' by Spearman correlation test

Table S17. Get Full Table List of 2 genes significantly correlated to 'TUMOR.STAGE' by Spearman correlation test

| SpearmanCorr | corrP | Q
--- | --- | --- | ---
HSA-MIR-146A | -0.2106 | 1.934e-06 | 0.000812
HSA-MIR-625 | -0.2004 | 6.05e-06 | 0.00253

Figure S8. Get High-res Image As an example, this figure shows the association of HSA-MIR-146A to 'TUMOR.STAGE'. P value = 1.93e-06 with Spearman correlation analysis.

Clinical variable #10: 'RADIATIONS.RADIATION.REGIMENINDICATION'

No gene related to 'RADIATIONS.RADIATION.REGIMENINDICATION'.

Table S18. Basic characteristics of clinical feature: 'RADIATIONS.RADIATION.REGIMENINDICATION'

RADIATIONS.RADIATION.REGIMENINDICATION | Labels | N
| NO | 8
| YES | 513
| Significant markers | N = 0

Clinical variable #11: 'NEOADJUVANT.THERAPY'

41 genes related to 'NEOADJUVANT.THERAPY'.

Table S19. Basic characteristics of clinical feature: 'NEOADJUVANT.THERAPY'

NEOADJUVANT.THERAPY | Labels | N
| NO | 71
| YES | 450
| Significant markers | N = 41
| Higher in YES | 32
| Higher in NO | 9

List of top 10 genes differentially expressed by 'NEOADJUVANT.THERAPY'

Table S20. Get Full Table List of top 10 genes differentially expressed by 'NEOADJUVANT.THERAPY'

| T(pos if higher in 'YES') | ttestP | Q | AUC
--- | --- | --- | --- | ---
HSA-MIR-106A | 7.08 | 1.012e-10 | 4.25e-08 | 0.699
HSA-MIR-103-2 | 6.58 | 1.506e-09 | 6.31e-07 | 0.7083
HSA-MIR-26A-1 | 6.36 | 5.445e-09 | 2.28e-06 | 0.7011
HSA-LET-7F-2 | -6.06 | 1.962e-08 | 8.18e-06 | 0.6702
HSA-MIR-331 | 5.91 | 3.805e-08 | 1.58e-05 | 0.685
HSA-MIR-1826 | 5.95 | 3.95e-08 | 1.64e-05 | 0.7086
HSA-MIR-23A | 5.43 | 3.616e-07 | 0.00015 | 0.6815
HSA-MIR-23B | 5.26 | 7.385e-07 | 0.000305 | 0.6632
HSA-MIR-7-1 | 5.2 | 9.846e-07 | 0.000406 | 0.6741
HSA-LET-7D | 5.19 | 1.01e-06 | 0.000415 | 0.6584

Figure S9. Get High-res Image As an example, this figure shows the association of HSA-MIR-106A to 'NEOADJUVANT.THERAPY'. P value = 1.01e-10 with T-test analysis.

Methods & Data
Input

- Expresson data file = COADREAD.miRseq_RPKM_log2.txt

- Clinical data file = COADREAD.clin.merged.picked.txt

- Number of patients = 521

- Number of genes = 420

- Number of clinical features = 11

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

Student's t-test analysis

For two-class clinical features, two-tailed Student's t test with unequal variance (Lehmann and Romano 2005) was applied to compare the log2-expression levels between the two clinical classes using 't.test' function in R

ANOVA analysis

For multi-class clinical features (ordinal or nominal), one-way analysis of variance (Howell 2002) was applied to compare the log2-expression levels between different clinical classes using 'anova' function in R

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

This is an experimental feature. The full results of the analysis summarized in this report can be downloaded from the TCGA Data Coordination Center.

- Analysis Results (MD5 checksum)

- Auxiliary Data (MD5 checksum)

- MAGE-TAB File (MD5 checksum)

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] Lehmann and Romano, Testing Statistical Hypotheses (3E ed.), New York: Springer. ISBN 0387988645 (2005)
[4] Howell, D, Statistical Methods for Psychology. (5th ed.), Duxbury Press:324-5 (2002)
[5] 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)
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