Clinicopathological features of colorectal cancer patients among KRAS wild type, p.G13D and other mutations: Results from a multicenter, cross-sectional study by the Japan Study Group of KRAS Mutation in Colorectal Cancer Hiroyuki Uetake1, Toshiaki Watanabe2, Takayuki Yoshino3, Kentaro Yamazaki4, Megumi Ishiguro1, Kenichi Sugihara1, Yasuo Ohashi5 １Tokyo Medical and Dental University, Graduate School, Tokyo, Japan, 2Teikyo University School of Medicine, Tokyo, Japan, 3National Cancer Center Hospital East, Chiba, Japan, 4Shizuoka Cancer Center, Shizuoka, Japan, 5Public Health Research Foundation, Tokyo, Japan ASCO 2011 #3605 Background Summary of the previous presentation Results: KRAS WT, KRAS MT (G13D), KRAS MT (others) Conclusions Many studies have reported that 30-40% of CRC patients have KRAS mutations and they lacked in response to anti-epidermal growth factor receptor (EGFR) antibodies.1-3 Recently, improved outcomes for association of KRAS p.G13D mutation (p.G13D MT) in patients with mCRC treated with cetuximab have been reported.4, 5 We have previously reported the correlation between KRAS mutation rate and sample backgrounds;6, 7 however, the clinicopathological features of KRAS p.G13D mutation have not been fully clarified. Study profile Gender Clinicopathological features: WT, MT (G13D), MT (others) KRAS p.G13D mutation was remarkably higher in female (P<0.0001). 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%, P=0.0515). 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. Other KRAS MT  had similar trend with KRAS p.G13D in primary tumor site. This study suggested that clinicopathological features were not similar among 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. 　 Sample registration, n=5,887 　 from 389 facilities From Oct. 2009 to Mar. 2010 Cut-off: Apr. 2010 KRAS WT KRAS MT Mutation rate P<0.0001 n KRAS　WT KRAS　MT (G13D) (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 (30.3%) 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%) 32 17 (53.1%) 4 (12.5%) 11 (34.4%) KRAS WT vs. KRAS MT (G13D) KRAS MT (G13D) vs. KRAS MT (others) Odds-ratio 95%CI P value 1.500 1.224 - 1.838 <0.0001 0.805 0.650 - 0.997 0.0465 0.667 0.544 - 0.817 1.243 1.003 - 1.539 1.098 0.785 - 1.536 0.3549 1.383 0.960 - 1.994 0.1992 1.038 0.813 - 1.327 1.024 0.790 - 1.327 1.127 0.910 - 1.396 0.831 0.663 - 1.040 0.826 0.668 - 1.022 1.059 0.846 - 1.324 0.527 0.114 - 2.448 0.588 0.133 - 2.598 0.0558 0.490 0.397 - 0.604 1.105 0.889 - 1.374 2.325 1.842 - 2.934 0.729 0.571 - 0.932 0.871 0.704 - 1.077 1.231 0.983 - 1.542 - 1.008 0.699 - 1.453 0.9673 0.988 0.672 - 1.454 0.9514 0.992 0.688 - 1.431 1.012 0.688 - 1.489 1.332 0.749 - 2.371 0.5733 0.940 0.510 - 1.734 0.8520 0.803 0.361 - 1.785 1.531 0.639 - 3.666 1.293 0.303 - 5.520 0.634 0.143 - 2.819 1.136 0.345 - 3.746 0.953 0.270 - 3.359 0.662 0.276 - 1.587 0.824 0.341 - 1.992 0.497 0.166 - 1.483 1.507 0.478 - 4.756 100% Excluded, n=97 　 Cancelation 14 　 Ineligible 1 　 Uncollected 82　 90% 80% 　 KRAS test, n=5,790 　　 Direct sequencing 5,479 　　 Luminex 311 70% 1,334 2,243 60% Undetectable samples, n=58 　 Direct sequencing 56 　 Luminex 2　 Mutation 50% 40.9% KRAS test detectable, n=5,732 　　 Direct sequencing 5,423 　　 Luminex 309 35.5% 40% 30% Objective 20% 923 1,232 10% Sample Background This study aimed to examine whether the clinicopathological features of tumors with p.G13D MT is more similar to those of KRAS wild type or other KRAS mutations in large-scale Japanese population (n=5,887). 0% 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 < 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%) < 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%) Male Female n=3,475 n=2,257 Age Methods KRAS WT KRAS MT Mutation rate P=0.0007 100% 90% * not included in the logistic regression Key eligibility criteria Histologically confirmed colorectal adenocarcinoma Adequate tumor samples 80% Gender Primary tumor site Comparison of KRAS mutation results 70% 798 1,289 1,101 389 60% KRAS　MT G13D KRAS　MT (others) G12D G12V G12C G12S G12A Gender Male 205 (52.0%) 424 (56.8%) 270 (59.0%) 102 (67.1%) 61 (54.5%) 55 (54.5%) Female 189 (48.0%) 323 (43.2%) 188 (41.0%) 50 (32.9%) 51 (45.5%) 46 (45.5%) Age < 60 127 (32.2%) 206 (27.6%) 141 (30.8%) 40 (26.3%) 33 (29.5%) 28 (27.7%) 60-69 128 (32.5%) 279 (37.3%) 172 (37.6%) 61 (40.1%) 42 (37.5%) 43 (42.6%) 70 =< 139 (35.3%) 262 (35.1%) 145 (31.7%) 51 (33.6%) 37 (33.0%) 30 (29.7%) Primary tumor site* right-sided colon 158 (40.1%) 316 (42.3%) 155 (33.8%) 76 (50.0%) 32 (28.6%) 32 (31.7%) left-sided colon 98 (24.9%) 215 (28.8%) 154 (33.6%) 39 (25.7%) 48 (42.9%) 36 (35.6%) rectum 138 (35.0%) 216 (28.9%) 149 (32.5%) 37 (24.3%) 33 (32.7%) Site of the sample obtained Primary tumor 362 (91.9%) 683 (91.4%) 427 (93.2%) 136 (89.5%) 106 (94.6%) 94 (93.1%) Metastasis 32 ( 8.1%) 64 ( 8.6%) 31 ( 6.8%) 16 (10.5%) 6 ( 5.4%) 7 ( 6.9%) liver 11 (34.4%) 26 (40.6%) 13 (41.9%) 8 (50.0%) 3 (50.0%) 4 (57.1%) lung 7 (21.9%) 4 ( 6.3%) 5 (16.1%) 2 (12.5%) 1 (16.7%) 2 (28.6%) others 14 (43.8%) 34 (53.1%) 6 (37.5%) 2 (33.3%) 1 (14.3%) KRAS WT KRAS MT (G13D) KRAS MT (others) KRAS WT KRAS MT (G13D) KRAS MT (others) 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) Mutation 50% 39.9% 40% 36.6% 38.1% appendix n=25 36.0% 8.0% 56.0% 30.5% 30% Male n=3,475 6.4 % 64.5% 29.1% 20% 460 792 732 171 right-sided colon n=1,713 10% 51.8% 9.9% 38.4% 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 ( 8.3%) liver 216 (45.6%) lung 74 (15.6%) lymph node 37 ( 7.8%) local 45 ( 9.5%) dissemination 70 (14.8%) 32 ( 6.8%) 0% <50 50-59 60-69 70=< n=560 n=1,258 n=2,081 n=1,833 left-sided colon n=2,160 70.7% 4.7 % 24.5% References Primary tumor site Female n=2,257 59.1% 8.8% 32.1% 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. De Roock W. et al. JAMA. 2010;304:1812-1820. 5. Bando H., Yoshino T. et al. ASCO-GI 2011 #448 6. Yamazaki K. et al. ESMO 2010 #595P 7. Yoshino T. et al. ASCO-GI 2011 #407 KRAS WT KRAS MT Mutation rate P<0.0001 100% rectum n=1,817 62.9% 8.1% 29.1% 90% 80% 887 70% 1,142 1,528 KRAS mutation 60% Age Site of the sample obtained * G13D vs. G12V P=0.0171, vs. G12C P=0.0399, vs. G12S P=0.0009 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 48.2% Mutation 50% No. of sample KRAS　WT KRAS　MT 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 28.7-31.1 7.0-8.4 Summary of the results 37.1% KRAS WT KRAS MT (G13D) KRAS MT (others) KRAS WT KRAS MT (G13D) KRAS MT (others) 40% 29.3% Hospitals Laboratories Data center 1. Sample registration 2. Enrollment confirmation 6. Result 3. Sample 2’. Registration information 5. Result 4. KRAS test KRAS WT KRAS MT (G13D) KRAS MT (Others) Gender Male > Female 64.5% 59.1% Male < Female 6.4% 8.8% 29.1% 32.1% P< 0.0001 P=0.0465 Age Young > Old 69.5%/63.4%/61.9%/60.1%* Young ≒ Old 7.5%/7.2%/6.7%/8.0%* Young < Old 23.0%/29.3%/31.3%/31.9%* P=0.3549 P=0.1992 Primary tumor site Right < Left 51.8% 70.7% Right > Left 9.9% 4.7% 38.4% 24.5% P<0.0001 P=0.0558 Site of the sample obtained Liver ≒ Lung 67.6% 64.9% Liver < Lung 6.0% 9.5% 26.4% 25.7% P=0.5733 P=0.8520 Acknowledgement 30% P=0.0001* P=0.5285* P=0.0002* Primary tumor n=5,258 62.4% 7.3% 30.3% 826 <50 n=560 20% 69.5% 7.5% 23.0% 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. 675 632 Metastasis n=474 10% 62.4% 7.4% 30.2% n % Codon12 1,714 GAT (G12D) GTT (G12V) TGT (G12C) AGT (G12S) GCT (G12A) CGT Others 814 493 162 120 107 15 3 47.5 28.8 9.5 7.0 6.2 0.9 0.1 Codon13 441 GAC (G13D) TGC CGC GAG 420 11 6 1 95.2 2.5 1.4 0.2 0.7 0% others 30.3％ (n=1,735) right-sided colon left-sided colon rectum 50-59 n=1,258 liver n=216 63.4% 7.2% 29.3% 67.6% 6.0% 26.4% n=1,713 n=2,160 n=1,817 lung n=74 64.9% 9.5% 25.7% KRAS MT 37.6% (n=2,155) The frequency of KRAS mutation (37.6%) in Japanese CRC patients is similar to those reported in previous studies from western countries. There are the significant difference of the frequency of KRAS mutation between male (35.5%) and female (40.9%) left-sided colon (29.3%) and right-sided colon (48.2%). 　　 As the age is higher there is more frequent KRAS mutation. 60-69 n=2,081 61.9% 6.7% 31.3% lymph node n=37 59.5% 5.4% 35.1% KRAS WT 62.4% (n=3,577) local n=45 64.4% 6.7% 28.9% G13D 7.3％ (n=420) 70=< n=1,833 60.1% 8.0% 31.9% dissemination n=70 48.6% 8.6% 42.9% *Cochran-Armitage trend test *< 50/50-59/60-69/70=<

Clinicopathological features of colorectal cancer patients among KRAS wild type, p.G13D and other mutations: Results from a multicenter, cross-sectional study by the Japan Study Group of KRAS Mutation in Colorectal Cancer Hiroyuki Uetake1, Toshiaki Watanabe2, Takayuki Yoshino3, Kentaro Yamazaki4, Megumi Ishiguro1, Kenichi Sugihara1, Yasuo Ohashi5 １Tokyo Medical and Dental University, Graduate School, Tokyo, Japan, 2Teikyo University School of Medicine, Tokyo, Japan, 3National Cancer Center Hospital East, Chiba, Japan, 4Shizuoka Cancer Center, Shizuoka, Japan, 5Public Health Research Foundation, Tokyo, Japan ASCO 2011 #3605 Background Summary of the previous presentation Results: KRAS WT, KRAS MT (G13D), KRAS MT (others) Conclusions Many studies have reported that 30-40% of CRC patients have KRAS mutations and they lacked in response to anti-epidermal growth factor receptor (EGFR) antibodies.1-3 Recently, improved outcomes for association of KRAS p.G13D mutation (p.G13D MT) in patients with mCRC treated with cetuximab have been reported.4, 5 We have previously reported the correlation between KRAS mutation rate and sample backgrounds;6, 7 however, the clinicopathological features of KRAS p.G13D mutation have not been fully clarified. Study profile Gender Clinicopathological features: WT, MT (G13D), MT (others) KRAS p.G13D mutation was remarkably higher in female (P<0.0001). 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%, P=0.0515). 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. Other KRAS MT  had similar trend with KRAS p.G13D in primary tumor site. This study suggested that clinicopathological features were not similar among 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. 　 Sample registration, n=5,887 　 from 389 facilities From Oct. 2009 to Mar. 2010 Cut-off: Apr. 2010 KRAS WT KRAS MT Mutation rate P<0.0001 n KRAS　WT KRAS　MT (G13D) (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 (30.3%) 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%) 32 17 (53.1%) 4 (12.5%) 11 (34.4%) KRAS WT vs. KRAS MT (G13D) KRAS MT (G13D) vs. KRAS MT (others) Odds-ratio 95%CI P value 1.500 1.224 - 1.838 <0.0001 0.805 0.650 - 0.997 0.0465 0.667 0.544 - 0.817 1.243 1.003 - 1.539 1.098 0.785 - 1.536 0.3549 1.383 0.960 - 1.994 0.1992 1.038 0.813 - 1.327 1.024 0.790 - 1.327 1.127 0.910 - 1.396 0.831 0.663 - 1.040 0.826 0.668 - 1.022 1.059 0.846 - 1.324 0.527 0.114 - 2.448 0.588 0.133 - 2.598 0.0558 0.490 0.397 - 0.604 1.105 0.889 - 1.374 2.325 1.842 - 2.934 0.729 0.571 - 0.932 0.871 0.704 - 1.077 1.231 0.983 - 1.542 - 1.008 0.699 - 1.453 0.9673 0.988 0.672 - 1.454 0.9514 0.992 0.688 - 1.431 1.012 0.688 - 1.489 1.332 0.749 - 2.371 0.5733 0.940 0.510 - 1.734 0.8520 0.803 0.361 - 1.785 1.531 0.639 - 3.666 1.293 0.303 - 5.520 0.634 0.143 - 2.819 1.136 0.345 - 3.746 0.953 0.270 - 3.359 0.662 0.276 - 1.587 0.824 0.341 - 1.992 0.497 0.166 - 1.483 1.507 0.478 - 4.756 100% Excluded, n=97 　 Cancelation 14 　 Ineligible 1 　 Uncollected 82　 90% 80% 　 KRAS test, n=5,790 　　 Direct sequencing 5,479 　　 Luminex 311 70% 1,334 2,243 60% Undetectable samples, n=58 　 Direct sequencing 56 　 Luminex 2　 Mutation 50% 40.9% KRAS test detectable, n=5,732 　　 Direct sequencing 5,423 　　 Luminex 309 35.5% 40% 30% Objective 20% 923 1,232 10% Sample Background This study aimed to examine whether the clinicopathological features of tumors with p.G13D MT is more similar to those of KRAS wild type or other KRAS mutations in large-scale Japanese population (n=5,887). 0% 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 < 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%) < 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%) Male Female n=3,475 n=2,257 Age Methods KRAS WT KRAS MT Mutation rate P=0.0007 100% 90% * not included in the logistic regression Key eligibility criteria Histologically confirmed colorectal adenocarcinoma Adequate tumor samples 80% Gender Primary tumor site Comparison of KRAS mutation results 70% 798 1,289 1,101 389 60% KRAS　MT G13D KRAS　MT (others) G12D G12V G12C G12S G12A Gender Male 205 (52.0%) 424 (56.8%) 270 (59.0%) 102 (67.1%) 61 (54.5%) 55 (54.5%) Female 189 (48.0%) 323 (43.2%) 188 (41.0%) 50 (32.9%) 51 (45.5%) 46 (45.5%) Age < 60 127 (32.2%) 206 (27.6%) 141 (30.8%) 40 (26.3%) 33 (29.5%) 28 (27.7%) 60-69 128 (32.5%) 279 (37.3%) 172 (37.6%) 61 (40.1%) 42 (37.5%) 43 (42.6%) 70 =< 139 (35.3%) 262 (35.1%) 145 (31.7%) 51 (33.6%) 37 (33.0%) 30 (29.7%) Primary tumor site* right-sided colon 158 (40.1%) 316 (42.3%) 155 (33.8%) 76 (50.0%) 32 (28.6%) 32 (31.7%) left-sided colon 98 (24.9%) 215 (28.8%) 154 (33.6%) 39 (25.7%) 48 (42.9%) 36 (35.6%) rectum 138 (35.0%) 216 (28.9%) 149 (32.5%) 37 (24.3%) 33 (32.7%) Site of the sample obtained Primary tumor 362 (91.9%) 683 (91.4%) 427 (93.2%) 136 (89.5%) 106 (94.6%) 94 (93.1%) Metastasis 32 ( 8.1%) 64 ( 8.6%) 31 ( 6.8%) 16 (10.5%) 6 ( 5.4%) 7 ( 6.9%) liver 11 (34.4%) 26 (40.6%) 13 (41.9%) 8 (50.0%) 3 (50.0%) 4 (57.1%) lung 7 (21.9%) 4 ( 6.3%) 5 (16.1%) 2 (12.5%) 1 (16.7%) 2 (28.6%) others 14 (43.8%) 34 (53.1%) 6 (37.5%) 2 (33.3%) 1 (14.3%) KRAS WT KRAS MT (G13D) KRAS MT (others) KRAS WT KRAS MT (G13D) KRAS MT (others) 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) Mutation 50% 39.9% 40% 36.6% 38.1% appendix n=25 36.0% 8.0% 56.0% 30.5% 30% Male n=3,475 6.4 % 64.5% 29.1% 20% 460 792 732 171 right-sided colon n=1,713 10% 51.8% 9.9% 38.4% 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 ( 8.3%) liver 216 (45.6%) lung 74 (15.6%) lymph node 37 ( 7.8%) local 45 ( 9.5%) dissemination 70 (14.8%) 32 ( 6.8%) 0% <50 50-59 60-69 70=< n=560 n=1,258 n=2,081 n=1,833 left-sided colon n=2,160 70.7% 4.7 % 24.5% References Primary tumor site Female n=2,257 59.1% 8.8% 32.1% 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. De Roock W. et al. JAMA. 2010;304:1812-1820. 5. Bando H., Yoshino T. et al. ASCO-GI 2011 #448 6. Yamazaki K. et al. ESMO 2010 #595P 7. Yoshino T. et al. ASCO-GI 2011 #407 KRAS WT KRAS MT Mutation rate P<0.0001 100% rectum n=1,817 62.9% 8.1% 29.1% 90% 80% 887 70% 1,142 1,528 KRAS mutation 60% Age Site of the sample obtained * G13D vs. G12V P=0.0171, vs. G12C P=0.0399, vs. G12S P=0.0009 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 48.2% Mutation 50% No. of sample KRAS　WT KRAS　MT 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 28.7-31.1 7.0-8.4 Summary of the results KRAS WT KRAS MT (G13D) KRAS MT (others) KRAS WT KRAS MT (G13D) KRAS MT (others) 40% 37.1% 29.3% Hospitals Laboratories Data center 1. Sample registration 2. Enrollment confirmation 6. Result 3. Sample 2’. Registration information 5. Result 4. KRAS test KRAS WT KRAS MT (G13D) KRAS MT (Others) Gender Male > Female 64.5% 59.1% Male < Female 6.4% 8.8% 29.1% 32.1% P< 0.0001 P=0.0465 Age Young > Old 69.5%/63.4%/61.9%/60.1%* Young ≒ Old 7.5%/7.2%/6.7%/8.0%* Young < Old 23.0%/29.3%/31.3%/31.9%* P=0.3549 P=0.1992 Primary tumor site Right < Left 51.8% 70.7% Right > Left 9.9% 4.7% 38.4% 24.5% P<0.0001 P=0.0558 Site of the sample obtained Liver ≒ Lung 67.6% 64.9% Liver < Lung 6.0% 9.5% 26.4% 25.7% P=0.5733 P=0.8520 Acknowledgement 30% P=0.0001* P=0.5285* P=0.0002* Primary tumor n=5,258 62.4% 7.3% 30.3% 826 <50 n=560 20% 69.5% 7.5% 23.0% 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. 675 632 Metastasis n=474 10% 62.4% 7.4% 30.2% n % Codon12 1,714 GAT (G12D) GTT (G12V) TGT (G12C) AGT (G12S) GCT (G12A) CGT Others 814 493 162 120 107 15 3 47.5 28.8 9.5 7.0 6.2 0.9 0.1 Codon13 441 GAC (G13D) TGC CGC GAG 420 11 6 1 95.2 2.5 1.4 0.2 0.7 0% others 30.3％ (n=1,735) right-sided colon left-sided colon rectum 50-59 n=1,258 liver n=216 63.4% 7.2% 29.3% 67.6% 6.0% 26.4% n=1,713 n=2,160 n=1,817 lung n=74 64.9% 9.5% 25.7% KRAS MT 37.6% (n=2,155) The frequency of KRAS mutation (37.6%) in Japanese CRC patients is similar to those reported in previous studies from western countries. There are the significant difference of the frequency of KRAS mutation between male (35.5%) and female (40.9%) left-sided colon (29.3%) and right-sided colon (48.2%). 　　 As the age is higher there is more frequent KRAS mutation. 60-69 n=2,081 61.9% 6.7% 31.3% lymph node n=37 59.5% 5.4% 35.1% KRAS WT 62.4% (n=3,577) local n=45 64.4% 6.7% 28.9% G13D 7.3％ (n=420) 70=< n=1,833 60.1% 8.0% 31.9% dissemination n=70 48.6% 8.6% 42.9% *Cochran-Armitage trend test *< 50/50-59/60-69/70=<