n KRAS WTKRAS MT (p.G13D)
KRAS MT (others)
Gender Male 3,475 2,243 (64.5%) 222 ( 6.4%) 1,010 (29.1%)Female 2,257 1,334 (59.1%) 198 ( 8.8%) 725 (32.1%)
Age < 50 560 389 (69.5%) 42 ( 7.5%) 129 (23.0%)50-59 1,258 798 (63.4%) 91 ( 7.2%) 369 (29.3%)60-69 2,081 1,289 (61.9%) 140 ( 6.7%) 652 (31.3%)70 =< 1,833 1,101 (60.1%) 147 ( 8.0%) 585 (31.9%)
Primary tumor site
appendix 25 9 (36.0%) 2 ( 8.0%) 14 (56.0%)right-sided colon 1,713 887 (51.8%) 169 ( 9.9%) 657 (38.4%)left-sided colon 2,160 1,528 (70.7%) 102 ( 4.7%) 530 (24.5%)rectum 1,817 1,142 (62.9%) 147 ( 8.1%) 528 (29.1%)others* 17 11 (64.7%) 0 ( 0.0%) 6 (35.3%)
Site of the sample obtained
Primary tumor 5,258 3,281 (62.4%) 385 ( 7.3%) 1,592 (30.0%)Metastasis 474 296 (62.4%) 35 ( 7.4%) 143 (30.2%)
liver 216 146 (67.6%) 13 ( 6.0%) 57 (26.4%)lung 74 48 (64.9%) 7 ( 9.5%) 19 (25.7%)lymph node 37 22 (59.5%) 2 ( 5.4%) 13 (35.1%)local 45 29 (64.4%) 3 ( 6.7%) 13 (28.9%)dissemination 70 34 (48.6%) 6 ( 8.6%) 30 (42.9%)others* 32 17 (53.1%) 4 (12.5%) 11 (34.4%)
n KRAS MT P value
Roth A.D. et al. 20109right
left
516 783
207 (40.1%) 274 (35.0%)
0.070
Abubaker J. et al. 200910right colon
left colon
44 241
18 (40.9%) 62 (25.7%)
0.0454
Bennani B. et al. 201011proximal colon
distal colon
rectum
14 20 28
4 (28.6%) 4 (20.0%) 10 (35.7%)
NS
Barault L. et al 200812proximal colon
distal colon
244 331
89 (36.5%) 109 (32.9%)
0.377
Tie J. et al 201013 right-sided colon
left-sided colon
rectum
185 153 172
70 (38%) 43 (28%) 54 (31%)
0.07
The present study
appendix
right-sided colon
left-sided colon
rectum
others
251,7132,1601,817 17
16 (64.0%) 826 (48.2%) 632 (29.3%) 675 (37.1%) 6 (35.3%)
<0.0001
Clinicopathological features in metastatic colorectal cancer patients with KRAS wild type versus codon 12 and codon 13 mutant: Results from a multicenter, cross-sectional study by the Japan Study Group of KRAS Mutation in Colorectal Cancer
Clinicopathological features in metastatic colorectal cancer patients with KRAS wild type versus codon 12 and codon 13 mutant: Results from a multicenter, cross-sectional study by the Japan Study Group of KRAS Mutation in Colorectal Cancer
Takayuki Yoshino,1 Toshiaki Watanabe,2 Kentaro Yamazaki,3 Hiroyuki Uetake,4 Megumi Ishiguro,4 Kenichi Sugihara,4 Yasuo Ohashi5Takayuki Yoshino,1 Toshiaki Watanabe,2 Kentaro Yamazaki,3 Hiroyuki Uetake,4 Megumi Ishiguro,4 Kenichi Sugihara,4 Yasuo Ohashi5
1National Cancer Center Hospital East, Chiba, Japan, 2Teikyo University School of Medicine, Tokyo, Japan, 3Shizuoka Cancer Center, Shizuoka, Japan, 4Tokyo Medical and Dental University, Graduate School, Tokyo, Japan, 5Public Health Research Foundation, Tokyo, Japan
1National Cancer Center Hospital East, Chiba, Japan, 2Teikyo University School of Medicine, Tokyo, Japan, 3Shizuoka Cancer Center, Shizuoka, Japan, 4Tokyo Medical and Dental University, Graduate School, Tokyo, Japan, 5Public Health Research Foundation, Tokyo, Japan
The KRAS mutation mainly located in the codon 12 and 13 indicates patients with metastatic colorectal cancer (CRC) lack of response to anti-epidermal growth factor receptor (EGFR) antibodies.
Many studies have reported that approximately 30-40% of CRC patients have KRAS mutations.1-3
We have previously reported the correlation between KRAS mutation rate and sample backgrounds;4 however, the clinicopathological features of KRAS mutant CRC have not been fully clarified.
Recently, some analysis showed improved outcomes for association of KRAS p.G13D mutation in patients with chemotherapy-refractory metastatic colorectal cancer treated with cetuximab.5-6
This study aimed to clarify the clinicopathological features of KRAS mutant CRC in comparison with KRAS wild type in large-scale Japanese population.
To clarify the clinicopathological features between KRAS codon 12 and 13 mutant CRC.
To examine whether the clinicopathological features of KRAS p.G13D mutation is more similar to those of KRAS wild type or other KRAS mutations.
BackgroundBackground
ObjectivesObjectives
MethodsMethods
Sample size: 5,000
Data collection:
Sample for KRAS test– Surgically resected specimen or biopsy from primary tumor or
metastases
– Paraffin-embedded tumor blocks or thinly sliced tumor sections
Patients and sample backgrounds– Gender, age – Primary tumor site – Type of sample (surgically resected / biopsy) – Date of the sample obtained– Site of the sample obtained (primary tumor / metastases) – Stage (l / ll / lll / IV / recurrence / unknown)– Duration of formalin fixation (<24h / 24-48h / 48h< / unknown)– Formalin concentration (10% / 20% / unknown)
Key eligibility criteriaHistologically confirmed colorectal adenocarcinomaAdequate tumor samples
Send formalin-fixed paraffin-embedded tumor blocks or thinly sliced tumor sections to commercial laboratories ( 10µm 5 slices and 3µm 1 slice for HE staining )
Investigate KRAS point mutations in the codon 12 and 13 by following laboratories’ SOP
Hospitals
LaboratoriesData center
1. Sample registration
2. Enrollment confirmation
6. Result
3. Sample
2’. Registration information
5. Result
4. KRAS test
Study profileStudy profileFrom Oct. 2009 to Mar. 2010Cut-off: Apr. 2010 Sample registration, n=5,887
from 389 facilitiesExcluded, n=97 Cancelation 14 Ineligible 1 Uncollected 82
Undetectable samples, n=58 Direct sequencing 56 Luminex 2
KRAS test detectable, n=5,732 Direct sequencing 5,423 Luminex 309
KRAS test, n=5,790 Direct sequencing 5,479 Luminex 311
ConclusionsConclusions
This study was funded by Comprehensive Support Project for Oncology Research (CSPOR) of Public Health Research Foundation.
The research institutes are Tokyo Medical and Dental University, Teikyo University School, and National Cancer Center Hospital East.
We would like to thank the 389 sample providing hospitals.
AcknowledgementAcknowledgement
ReferencesReferences 1. Karapetis CS. et al. N Engl J Med. 2008;359:1757-1765. 2. Amado RG. et al. J Clin Oncol. 2008;26:1626-1634. 3. Van Cutsem E. et al. N Engl J Med. 2009;360:1408-1417. 4. Yamazaki K. et al. ESMO 2010 Abst.595P 5. De Roock W. et al. JAMA. 2010;304:1812-1820. 6. Bando H., Yoshino T. et al. ASCO-GI 2011 #448 7. Ferreira C.G. et al. J Clin Oncol 2010;28,suppl:3614. 8. Andreyev HJN. et al. J Natl Cancer Inst 1998;90:675-684. 9. Roth A.D. et al. J clin oncol 2010;28:466-474.10. Abubaker J. et al. J Pathology 2009;219:435-445.11. Bennani B. et al. Int J Biol Markers 2010;25:179-84.12. Barault L. et al. Int J Cancer 2008;122:2255-9.13. Tie J. et al. Int J Cancer. 2010 Jul 15. [online ahead of print]14. Andreyev HJN. Br J Cancer 2001;85:692-696.15. Winawer S.J. et al. N Engl J Med. 1993;328:901-906.16. Rashid A. et al. Gut. 1999;44:826-33.
As far as we know, this is one of the largest observational study of KRAS mutational status in CRC.
The frequency of KRAS mutation (37.6%) in Japanese CRC patients is similar to those reported in previous studies from other countries.
Independent factors correlated with KRAS mutation were gender, age, date of sample obtained and primary tumor site.
Clinicopathological features were similar between KRAS codon 12 and 13 mutant CRC patients.
KRAS p.G13D mutation was remarkably higher in female and right-sided colon; however, it was not influenced by age. Another possible finding was that this mutation may be higher in lung metastasis.
However, this study suggested that clinicopathological features were not similar between KRAS wild type, KRAS p.G13D and other mutations.
It suggests a possibility that KRAS p.G13D mutation might be a different tumor than other KRAS mutations, since some different trends were seen between those mutations.
Presently, any KRAS mutation is regarded to be the same; however, this study suggests that it might be an aggregation from different tumors.
We found the independent factors correlated with KRAS mutation were gender, age, date of sample obtained and primary tumor site. It suggests that some correlations exist between KRAS mutation and gender or primary tumor site since these factors were consistent with previous reports
Although there is no consistent trend regarding age in the previous reports, this large cohort showed significantly more frequent KRAS mutation as the age was higher.
Some reports showed “Adenomatous polyps increase with age”;15 while, another report showed “adenomatous polyps with KRAS mutation decreased with age”.16 Thus, it remains unknown why more frequent KRAS mutation occurred with age.
There is no clear reason why KRAS mutation rate was lower in older samples. Since the concordance rate was very high between primary and metastases in this study, although these were not obtained from the same patients, it would be better to obtain new specimen if possible.
DiscussionsDiscussions
KRAS mutation rate in codon 12 and 13n Codon 12 Codon 13
Andreyev HJN. et al. 200114 3,439 900 (26.2%) 297 (8.6%)
Ferreira C.G. et al. 20107 3,129 933 (29.8%) 171 (5.5%)
Andreyev HJN. et al. 19988 2,721 755 (27.7%)* 146 (6.6%)
Roth A.D. et al. 20109 1,299 372 (28.6%) 102 (7.9%)
The present study 5,732 1,714 (29.9%) 441 (7.7%)
* 755/2,214
Gender n KRAS MT P value
Ferreira C.G. et al. 20107MaleFemale
1,5661,563
515 (32.9%) 589 (37.7%)
0.005
Andreyev HJN. et al. 19988MaleFemale
1,1871,009
430 (36.2%) 399 (39.5%)
0.11
Roth A.D. et al. 20109MaleFemale
749 550
280 (37.4%) 201 (36.5%)
0.80
Abubaker J. et al. 200910MaleFemale
135 150
37 (27.4%) 43 (28.7%)
0.8132
Bennani B. et al. 201011MaleFemale
31 31
7 (22.6%) 11 (35.5%)
0.263
The present studyMaleFemale
3,4752,257
1,232 (35.5%) 923 (40.9%)
<0.0001
Age
Primary tumor site
KRAS p.G13D mutation was remarkably higher in female.
KRAS p.G13D mutation occurred at a constant rate regardless of age (P=0.5285); however, KRAS other mutations increased with age (P=0.0002). In contrast, KRAS wild type decreased with age (P=0.0001).
KRAS p.G13D mutation seemed remarkably higher in right-sided colon than left-sided colon. (right : 9.9%, left : 4.7%)
KRAS p.G13D mutation was tend to be higher in lung metastasis (9.5%) compared to other metastatic sites; however, this is uncertain because the sample size has varied through the metastatic sites.
ResultsResultsSample Background
n
Gender Male 3,475 (60.6%)Female 2,257 (39.4%)
Age < 50 560 ( 9.8%)
Median:65 ( 12-92 )
50-59 1,258 (21.9%)60-69 2,081 (36.3%)70 =< 1,833 (32.0%)
Type of sample Surgically resected 5,364 (93.6%)Biopsy 368 ( 6.4%)
Year Surgically resected
< 2006 748 (13.0%)2006 445 ( 7.8%)2007 761 (13.3%)2008 1,255 (21.9%)2009 1,843 (32.2%)2010 312 ( 5.4%)
Biopsy < 2009 110 ( 1.9%)2009 =< 258 ( 4.5%)
Stage I 166 ( 2.9%) II 814 (14.2%)III 1,765 (30.8%)IV 2,805 (48.9%)recurrence 152 ( 2.7%)unknown 30 ( 0.5%)
Primary tumor site appendix 25 ( 0.4%)right-sided colon 1,713 (29.9%)left-sided colon 2,160 (37.7%)rectum 1,817 (31.7%)others 17 ( 0.3%)
Site of the sample obtained
Primary tumor 5,258 (91.7%)Metastasis 474 ( 8.2%)
liver 216 (45.6%)lung 74 (15.6%)lymph node 37 ( 7.8%)local 45 ( 9.5%)dissemination 70 (14.8%)others 32 ( 6.8%)
ResultsResults
KRAS mutation rate
n %
Codon12 1,714
AGTCGTGATGCTGTTTGTothers
120158141074931623
7.00.947.56.228.89.50.1
Codon13 441
CGCGACGAGTGCothers
64201113
1.495.20.22.50.7
No. of sample
KRAS WT KRAS MT
n % 95%CI n % 95%CI
All 5,732 3,577 62.4 61.1-63.7 2,155 37.6 36.3-38.9
Codon12 Codon13
1,714 441
29.9 7.7
28.7-31.17.0-8.4
KRAS WT62.4%
(n=3,577)
KRAS MT37.6%
(n=2,155)
p.G13D7.3 %(n=420)
others30.3 %(n=1,735)
KRAS WT, KRAS MT
n KRAS WT KRAS MT P value
Gender Male 3,475 (60.6%) 2,243 (64.6%) 1,232 (35.5%)<0.0001
Female 2,257 (39.4%) 1,334 (59.1%) 923 (40.9%)
Age < 50 560 ( 9.8%) 389 (69.5%) 171 (30.5%)
0.000750-59 1,258 (21.9%) 798 (63.4%) 460 (36.6%)
60-69 2,081 (36.3%) 1,289 (61.9%) 792 (38.1%)
70 =< 1,833 (32.0%) 1,101 (60.1%) 732 (39.9%)
Year Surgically resected
< 2006 748 (13.0%) 497 (66.4%) 251 (33.6%)
0.02852006 445 ( 7.8%) 282 (63.4%) 163 (36.6%)
2007 =< 4,171 (72.8%) 2,561 (61.4%) 1,610 (38.6%)
Biopsy < 2009 110 ( 1.9%) 79 (71.8%) 31 (28.2%)0.0524
2009 =< 258 ( 4.5%) 158 (61.2%) 100 (38.8%)
Primary tumor site
appendix 25 ( 0.4%) 9 (36.0%) 16 (64.0%)
<0.0001
right-sided colon 1,713 (29.9%) 887 (51.8%) 826 (48.2%)
left-sided colon 2,160 (37.7%) 1,528 (70.7%) 632 (29.3%)
rectum 1,817 (31.7%) 1,142 (62.9%) 675 (37.1%)
others* 17 ( 0.3%) 11 (64.7%) 6 (35.3%)
* not included in the logistic regression
Logistic regression: WT vs. MT
reference odds-ratio 95% CI
Gender Female vs. Male 1.210 1.082 - 1.354
Age 50 – 59 vs. < 50 1.308 1.053 - 1.624
60 – 69 vs. < 50 1.365 1.113 - 1.674
70 =< vs. < 50 1.399 1.137 - 1.720
Year Surgically resected (2006)
vs.Surgically resected
(<2006)1.150 0.896 - 1.474
Surgically resected (2007=<)
vs.Surgically resected
(<2006)1.188 1.005 - 1.403
Biopsy(<2009)
vs.Surgically resected
(<2006)1.003 0.751 - 1.341
Biopsy(2009=<) vs. Biopsy (<2009) 1.482 0.907 - 2.423
Primary tumor site
right-sided colon vs. left-sided colon 2.272 1.972 - 2.625
right-sided colon vs. rectum 1.570 1.379 - 1.789
[Odds ratio of biopsy ( 2009=< ) against surgery specimen ( <2006 ) is estimated as (1.003×1.482)=1.486]
There were the significant difference of the frequency of KRAS mutation
Female (40.9%) and male (35.5%) (Odds-ratio 1.210)As the age was higher, more frequent KRAS mutation (Odds-ratio 1.308-1.399)Date of the sample obtained : after 2007 (38.6 % ), 2006 (36.6 % ), before 2005 (33.6 % ) (Odds-ratio 1.118) Right-sided colon (48.2%) and left-sided colon (29.3%) (Odd ratio 2.272)
KRAS MT(codon 12), KRAS MT(codon 13)
reference odds-ratio 95% CI
Gender Female vs. Male 0.823 0.665 - 1.018
Age 50-59 vs. <50 1.243 0.820 - 1.885
60-69 vs. <50 1.422 0.959 - 2.108
70=< vs. <50 1.252 0.844 - 1.857
Primary tumor site
right-sided colon vs. left-sided colon 0.806 0.619 - 1.050
right-sided colon vs. appendix 0.473 0.106 - 2.114
right-sided colon vs. rectum 1.081 0.841 - 1.389
Clinicopathological features of KRAS codon 13 mutation were similar to those of KRAS codon 12 mutation
Logistic regression: MT(codon 12) vs. MT(codon 13)
KRAS WT, KRAS MT(p.G13D), KRAS MT(others)
* not included in the logistic regression
KRAS WT vs. KRAS MT(p.G13D) KRAS MT(p.G13D) vs. KRAS MT(others)
Odds-ratio 95%CI P value Odds-ratio 95%CI P value
1.500 1.224 - 1.838 <0.0001 0.805 0.650 - 0.997 0.04650.667 0.544 - 0.817 1.243 1.003 - 1.5391.098 0.785 - 1.536 0.3549 1.383 0.960 - 1.994 0.19921.038 0.813 - 1.327 1.024 0.790 - 1.3271.127 0.910 - 1.396 0.831 0.663 - 1.0400.826 0.668 - 1.022 1.059 0.846 - 1.3240.527 0.114 - 2.448 <0.0001 0.588 0.133 - 2.598 0.05580.490 0.397 - 0.604 1.105 0.889 - 1.3742.325 1.842 - 2.934 0.729 0.571 - 0.9320.871 0.704 - 1.077 1.231 0.983 - 1.542- - - - - -
1.008 0.699 - 1.453 0.9673 0.988 0.672 - 1.454 0.95140.992 0.688 - 1.431 1.012 0.688 - 1.4891.332 0.749 - 2.371 0.5733 0.940 0.510 - 1.734 0.85200.803 0.361 - 1.785 1.531 0.639 - 3.6661.293 0.303 - 5.520 0.634 0.143 - 2.8191.136 0.345 - 3.746 0.953 0.270 - 3.3590.662 0.276 - 1.587 0.824 0.341 - 1.9920.497 0.166 - 1.483 1.507 0.478 - 4.756
reference odds-ratio 95% CI
Gender Female vs. Male 0.710 0.576 - 0.876
Age 50 – 59 vs. < 50 0.919 0.620 - 1.361
60 – 69 vs. < 50 0.980 0.676 - 1.421
70 =< vs. < 50 0.837 0.577 - 1.214
Primary tumor site
right-sided colon vs. left-sided colon 0.365 0.281 - 0.475
right-sided colon vs. rectum 0.727 0.569 - 0.930
reference odds-ratio 95% CI
Gender Female vs. Male 0.799 0.643 - 0.955
Age 50 – 59 vs. < 50 1.338 0.875 - 2.046
60 – 69 vs. < 50 1.549 1.037 - 2.313
70 =< vs. < 50 1.288 0.862 - 1.924
Primary tumor site
right-sided colon vs. left-sided colon 0.750 0.570 - 0.986
right-sided colon vs. rectum 1.104 0.855 - 1.425
Logistic regression: WT vs. MT(p.G13D)
Gender Age
Primary tumor site Site of the sample obtained
n KRAS MT P value
Roth A.D. et al. 20109=< 6060 <
644 655
227 (35.2%) 254 (38.8%)
0.21
Abubaker J. et al. 200910=< 5050 <
87 198
29 (33.3%) 51 (25.8%)
0.1940
Bennani B. et al. 201011=< 4041-6060 <
16 30 16
5 (31.3%) 13 (43.3%) 0 ( 0.0%)
NS
Barault L. et al. 200812=< 6566-7475 =<
149 160 267
54 (36.2%) 55 (34.4%) 89 (33.3%)
0.836
The present study
< 5050-5960-6970 =<
5601,2582,0811,833
171 (30.5%) 460 (36.6%) 792 (38.1%) 732 (39.9%)
0.0007
KRAS WT KRAS MT(p.G13D) KRAS MT(others)
female(n=2,257)
male(n=3,475)
59.1%
64.5%
8.8%
6.4%
32.1%
29.1%
*Cochran-Armitage trend test
70=<(n=1,833)
60-69(n=2,081)
50-59(n=1,258)
<50(n=560)
60.1%
61.9%
63.4%
69.5%
8.0%
6.7%
7.2%
7.5%
31.9%
31.3%
29.3%
23.0%
P=0.0001* P=0.5285* P=0.0002*
62.9%
70.7%
51.8%
36.0%
8.1%
4.7%
9.9%
8.0%
29.1%
24.5%
38.4%
56.0%
rectum(n=1,817)
left-sided colon
(n=2,160)
right-sided colon
(n=1,713)
appendix(n=25)
48.6%
64.4%
59.5%
64.9%
67.6%
62.4%
62.4%
8.6%
6.7%
5.4%
9.5%
6.0%
7.4%
7.3%
42.9%
28.9%
35.1%
25.7%
26.4%
30.2%
30.3%
dissemination(n=70)
local(n=45)
lymph node(n=37)
lung(n=74)
liver(n=216)
Metastasis(n=474)
Primary tumor(n=5,258)
ASCO GI 2011 #407ASCO GI 2011 #407
Clinicopathological features: WT vs. MT
* not included in the logistic regression
n KRAS MT (codon 12)
KRAS MT (codon 13) P value
Gender Male 3,475 996 (28.7%) 236 ( 6.8%) 0.0820
Female 2,257 718 (31.8%) 205 ( 9.1%)
Age < 50 560 129 (23.0%) 42 ( 7.5%) 0.3496
50-59 1,258 363 (28.9%) 97 ( 7.7%)
60-69 2,081 643 (30.9%) 149 ( 7.2%)
70 =< 1,833 579 (31.6%) 153 ( 8.3%)
Primary tumor site
appendix 25 14 (56.0%) 2 ( 8.0%) 0.1710
right-sided colon 1,713 650 (37.9%) 176 (10.3%)
left-sided colon 2,160 519 (24.0%) 113 ( 5.2%)
rectum 1,817 525 (28.9%) 150 ( 8.3%)
others* 17 6 (35.3%) 0 ( 0.0%)
Clinicopathological features: MT(codon 12) vs. MT(codon 13)
Clinicopathological features: WT, MT(p.G13D), MT(others)
Logistic regression: MT(p.G13D) vs. MT(others)
KRAS WT KRAS MT(p.G13D) KRAS MT(others)
KRAS WT KRAS MT(p.G13D) KRAS MT(others)
KRAS WT KRAS MT(p.G13D) KRAS MT(others)