Nature Medicine doi: /nm.2457

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Transcript:

ature Medicine doi:10.1038/nm.2457

ature Medicine doi:10.1038/nm.2457

ature Medicine doi:10.1038/nm.2457

ature Medicine doi:10.1038/nm.2457

ature Medicine doi:10.1038/nm.2457

Table S1 - Candidate genes for investigation of aberrant splicing events due to MSI in GEE ITROIC POSITIO CADIDATE EXO SYMBOL REPEAT EXO SIZE RTPCR RESULTS IGF1R A(10) -40 18 467 ORMAL KRT20 A(11) -3 7 370 ORMAL OXR1 A(14) 0 5 518 ALTERATIVE EXO RIF1 A(23) -27 15 594 ALTERATIVE EXO MUC13 A(9) -38 5 506 ORMAL ZFYVE9 C(15) -3 5 620 ALTERATIVE EXO ERBB2 C(8) -3 15 541 ORMAL PDCD4 T(10) -12 7 356 ORMAL MRE11 T(11) -4 5 497 EXO SKIPPIG (MSI) PKIB T(11) -10 3 284 ALTERATIVE EXO EK1 T(11) -14 7 458 ALTERATIVE EXO BMPR2 T(11) -37 11 444 ORMAL RIF1 T(11) -39 3 655 ORMAL KIAA1033 T(12) -6 22 399 ORMAL BMPR2 T(12) -22 7 412 ORMAL SLC22A3 T(12) -38 6 416 ALTERATIVE EXO MSH2 T(13) -2 2 533 ORMAL MSH6 T(13) -2 7 521 ORMAL UBR5 T(13) -2 47 591 ORMAL PBRM1 T(13) -3 9 697 ALTERATIVE EXO ATP9A T(13) -16 5 592 ALTERATIVE EXO SUZ12 T(14) -1 5 458 ALTERATIVE EXO FBXW7 T(14) -3 3 594 ORMAL STAG2 T(14) -7 23 581 ORMAL PTE T(15) -1 8 892 ORMAL RB1CC1 T(15) -1 21 575 ORMAL ATM T(15) -2 6 642 ALTERATIVE EXO ATM T(15) -4 8 511 ORMAL APAF1 T(16) -2 23 525 ORMAL TP53BP1 T(16) -2 13 535 ORMAL ATM T(16) -3 12 671 ORMAL CUL2 T(16) -3 17 375 ORMAL HSP110 T(17) -3 9 433 EXO SKIPPIG (MSI) TGFB2 T(17) -4 6 615 ORMAL LAPTM4B T(18) -11 2 331 ORMAL PRKCI T(18) -24 8 484 ORMAL STAG2 T(25) -3 15 295 ORMAL STAG2 T(27) -5 20 389 ORMAL SRI T(8) -6 3 283 ORMAL ETK1 T(8) -10 6 416 ORMAL PSD3 T(8) -17 5 389 ORMAL SX13 T(8) -18 5 566 ORMAL CCG2 T(9) -4 7 371 ORMAL TRPM6 T(9) -7 31 410 ORMAL ROCK2 T(9) -17 32 597 ORMAL A: on Amplified. Position: relatively to the 3' intron-exon junction ature Medicine doi:10.1038/nm.2457

Table S2. HSP110 status in cell lines, control LBLs and primary tumors SAMPLE TYPE SAMPLE AME MSI/MSS STATUS T17 GEOTYPE DELETIO SIZE (in bp)* -CELL LIE CO115 MSI 139/140 7 29,4 -CELL LIE LS174T MSI 140/142 6 193,9 -CELL LIE LOVO MSI 141/141 5 25,7 -CELL LIE TC71 MSI 142/146 4 48,1 -CELL LIE HCT15 MSI 142/143 4 17,1 -CELL LIE HCT116 MSI 143/144 3 14,3 -CELL LIE TC7 MSI 141/143 5 90,2 -CELL LIE SW48 MSI 141/144 5 14,1 -CELL LIE RKO MSI 139/142 6 14,4 -CELL LIE HCT8 MSI 143/145 3 7,3 -CELL LIE KM12 MSI 140/141 6 88,4 -CELL LIE LIM1215 MSI 142/143 4 12,6 -CELL LIE LIM2405 MSI 141/143 5 12,9 -CELL LIE ALA MSS 146 0 8,2 -CELL LIE COLO320 MSS 146 0 6,8 -CELL LIE EB MSS 145 0 5,0 -CELL LIE FET MSS 146 0 6,0 -CELL LIE FRI MSS 145 0 4,6 -CELL LIE GLY MSS 145/146 0 9,1 -CELL LIE SW480 MSS 146 0 8,4 LBL-CELL LIE LBL-1 MSS 145 0 17,9 LBL-CELL LIE LBL-2 MSS 145/146 0 18,0 LBL-CELL LIE LBL-3 MSS 146 0 14,0 LBL-CELL LIE LBL-4 MSS 145 0 10,6 LBL-CELL LIE LBL-5 MSS 146 0 22,6 LBL-CELL LIE LBL-6 MSS 145/146 0 26,0 LBL-CELL LIE LBL-7 MSS 145/146 0 19,3 LBL-CELL LIE LBL-8 MSS 145 0 19,7 LBL-CELL LIE LBL-9 MSS 145/146 0 24,0 LBL-CELL LIE LBL-10 MSS 145 0 18,9 LBL-CELL LIE LBL-11 MSS 146 0 19,9 LBL-CELL LIE LBL-12 MSS 145/146 0 15,4 LBL-CELL LIE LBL-13 MSS 146 0 16,7 LBL-CELL LIE LBL-14 MSS 145 0 26,8 LBL-CELL LIE LBL-15 MSS 145/146 0 18,3 LBL-CELL LIE LBL-16 MSS 145/146 0 23,7 LBL-CELL LIE LBL-17 MSS 145/146 0 20,8 LBL-CELL LIE LBL-18 MSS 145/146 0 24,3 LBL-CELL LIE LBL-19 MSS 145 0 25,3 LBL-CELL LIE LBL-20 MSS 146 0 26,5 PRIMARY TMSI1 MSI 141/145 5 46,8 PRIMARY TMSI10 MSI 143/146 3 11,7 PRIMARY TMSI11 MSI 141/144 5 18,6 PRIMARY TMSI12 MSI 140/142/144 6 44,9 PRIMARY TMSI13 MSI 140/144 6 38,9 PRIMARY TMSI14 MSI 141/142/145/146 5 40,8 PRIMARY TMSI15 MSI 141/144/145 5 24,1 PRIMARY TMSI16 MSI 140/144 6 24,2 PRIMARY TMSI17 MSI 140/143/145 6 26,8 PRIMARY TMSI18 MSI 139/144 7 25,9 PRIMARY TMSI19 MSI 138/146 8 143,3 PRIMARY TMSI2 MSI 142/144 4 18,7 PRIMARY TMSI20 MSI 141/144/146 5 31,2 PRIMARY TMSI21 MSI 143/145/146 3 15,9 PRIMARY TMSI22 MSI 140/142/144 6 39,9 PRIMARY TMSI23 MSI 140/142/145 6 46,2 PRIMARY TMSI24 MSI 141/143/145 5 49,8 HSP110ΔE9 mra EXPRESSIO** (in %) ature Medicine doi:10.1038/nm.2457

SAMPLE TYPE SAMPLE AME MSI/MSS STATUS T17 GEOTYPE DELETIO SIZE (in bp)* PRIMARY TMSI25 MSI 142/146 4 45,5 PRIMARY TMSI26 MSI 140/145 6 106,3 PRIMARY TMSI27 MSI 139/140/144/145 7 252,7 PRIMARY TMSI28 MSI 139/143/146 7 26,7 PRIMARY TMSI29 MSI 140/143/146 6 206,4 PRIMARY TMSI3 MSI 141/142/145 5 28,5 PRIMARY TMSI30 MSI 142/145 4 18,9 PRIMARY TMSI31 MSI 142/143/145 4 22,4 PRIMARY TMSI32 MSI 142/145 4 55,2 PRIMARY TMSI33 MSI 140/141/145 6 288,2 PRIMARY TMSI34 MSI 140/146 6 314,0 PRIMARY TMSI35 MSI 143/145 3 20,8 PRIMARY TMSI36 MSI 141/146 5 58,5 PRIMARY TMSI37 MSI 138/142/145 8 29,7 PRIMARY TMSI38 MSI 141/145/146 5 479,2 PRIMARY TMSI39 MSI 141/145 5 319,5 PRIMARY TMSI4 MSI 141/142/145 5 76,8 PRIMARY TMSI40 MSI 141/144 5 26,2 PRIMARY TMSI41 MSI 140/145 6 27,2 PRIMARY TMSI42 MSI 143/145 3 21,3 PRIMARY TMSI43 MSI 141/144 5 57,5 PRIMARY TMSI5 MSI 140/144 6 69,3 PRIMARY TMSI6 MSI 141/145 5 73,3 PRIMARY TMSI7 MSI 142/143/146 4 25,4 PRIMARY TMSI8 MSI 142/146 4 19,1 PRIMARY TMSI9 MSI 141/145/146 5 38,8 PRIMARY TMSS1 MSS 145 0 12,7 PRIMARY TMSS10 MSS 146 0 11,1 PRIMARY TMSS11 MSS 146/145 0 6,3 PRIMARY TMSS12 MSS 146 0 6,8 PRIMARY TMSS13 MSS 145 0 6,9 PRIMARY TMSS14 MSS 146/145 0 7,4 PRIMARY TMSS15 MSS 146/145 0 6,6 PRIMARY TMSS16 MSS 146/145 0 7,8 PRIMARY TMSS17 MSS 145 0 7,6 PRIMARY TMSS18 MSS 146/145 0 8,7 PRIMARY TMSS19 MSS 146 0 6,0 PRIMARY TMSS2 MSS 146/145 0 10,8 PRIMARY TMSS20 MSS 146 0 8,4 PRIMARY TMSS3 MSS 146 0 18,4 PRIMARY TMSS4 MSS 146/145 0 16,3 PRIMARY TMSS5 MSS 146/145 0 16,4 PRIMARY TMSS6 MSS 146 0 18,7 PRIMARY TMSS7 MSS 145 0 18,4 PRIMARY TMSS8 MSS 146/145 0 16,0 PRIMARY TMSS9 MSS 146 0 16,2 * calculated by substracting the size of the shortest HSP110 allele to 146 in each case ** HSP110ΔE9 relative to HSP110wt expression (Quantitative RTPCR) HSP110ΔE9 mra EXPRESSIO** (in %) ature Medicine doi:10.1038/nm.2457

Table S3. Information on MSI Samples from MSI patients included in survival analyses Series Sample ID TM HSP110ΔE9/HSP110 Stage (% ) Class of HSP110ΔE9/HSP110 expression Age at Gender diagnosis (in years) Tumor Location Chemotherapy Performed Chemotherapy Type DFS DFS delay (in event months) S1 TMSI10 2 11.7 LOW M 61 left colon na 0 17 S1 TMSI17 2 26.8 LOW M 46 right colon na 0 73 S1 TMSI22 2 39.9 LOW F 75 right colon na 0 65 S1 TMSI2 2 18.7 LOW F 75 right colon na 0 65 S1 TMSI23 2 46.2 LOW M 48 right colon na 0 35 S1 TMSI25 2 45.6 LOW M 76 right colon na 1 3 S1 TMSI27 2 252.7 HIGH F 88 right colon na 0 51 S1 TMSI30 2 18.9 LOW F 79 right colon na 0 46 S1 TMSI3 2 28.5 LOW M 88 right colon na 0 2 S1 TMSI34 2 314.0 HIGH M 65 right colon na 0 51 S1 TMSI36 2 58.5 LOW M 71 left colon na 0 2 S1 TMSI37 2 29.7 LOW M 24 right colon na 0 37 S1 TMSI38 2 479.2 HIGH M 77 right colon na 0 43 S1 TMSI39 2 319.5 HIGH M 67 na na 0 35 S1 TMSI41 2 27.2 LOW M 71 right colon na 0 31 S1 TMSI4 2 76.8 HIGH F 69 right colon na 0 57 S1 TMSI5 2 69.3 LOW M 78 right colon na 0 33 S1 TMSI7 2 25.4 LOW M 57 right colon na 0 20 S1 TMSI8 2 19.1 LOW F 81 left colon na 0 0 S1 TMSI9 2 38.8 LOW M 49 left colon na 0 48 S1 TMSI13 3 38.9 LOW F 58 right colon Y FOLFOX 0 32 S1 TMSI14 3 40.8 LOW M 73 right colon Y LV5FU 1 5 S1 TMSI15 3 24.1 LOW M 32 left colon Y LV5FU 0 76 S1 TMSI19 3 143.4 HIGH F 78 right colon Y LV5FU 0 60 S1 TMSI26 3 106.3 HIGH M 59 right colon Y LV5FU 0 43 S1 TMSI29 3 206.4 HIGH F 67 right colon Y LV5FU 0 52 S1 TMSI31 3 22.4 LOW F 59 right colon Y LV5FU 1 24 S1 TMSI35 3 20.8 LOW M 61 na Y LV5FU 0 35 S1 TMSI43 3 57.5 LOW M 46 left colon Y FOLFOX 1 10 S1 TMSI6 3 73.3 LOW M 74 right colon Y LV5FU 1 20 S2 ntmsi44 2 133.3 HIGH F 83 right colon na 0 9 S2 ntmsi45 2 97.7 HIGH M 31 left colon Y FOLFOX 0 28 S2 ntmsi46 2 23.3 LOW F 66 right colon Y FOLFOX 0 20 S2 ntmsi48 2 136.2 HIGH M 74 right colon na 0 36 S2 ntmsi53 2 0.9 LOW F 28 right colon na 0 42 S2 ntmsi54 2 2.2 LOW M 65 right colon Y FOLFOX 1 13 S2 ntmsi55 2 8.9 LOW F 73 right colon na 0 49 S2 ntmsi56 2 91.2 HIGH M 60 right colon na 0 72 S2 ntmsi57 2 0.8 LOW F 80 right colon na 0 42 S2 ntmsi60 2 97.7 HIGH M 67 right colon na 0 60 S2 ntmsi62 2 1.2 LOW F 85 right colon na 0 48 S2 ntmsi63 2 2.1 LOW F 82 right colon na 0 48 S2 ntmsi64 2 9.9 LOW M 83 na na 1 40 S2 ntmsi65 2 73.2 HIGH F 60 left colon na 0 42 S2 ntmsi66 2 0.7 LOW M 26 right colon na 0 60 S2 ntmsi67 2 14.0 LOW M 69 right colon na 0 21 S2 ntmsi68 2 151.4 HIGH M 86 right colon na 0 17 S2 ntmsi69 2 37.7 LOW F 69 right colon na 0 72 S2 ntmsi70 2 0.9 LOW M 50 right colon na 0 42 S2 ntmsi71 2 12.1 LOW F 90 right colon na 0 1 S2 ntmsi72 2 84.6 HIGH F 51 right colon na 0 60 S2 ntmsi73 2 110.5 HIGH M 82 right colon na 0 1 S2 ntmsi74 2 19.8 LOW M 48 right colon na 0 72 S2 ntmsi75 2 69.9 HIGH M 59 right colon na 0 24 S2 ntmsi76 2 86.7 HIGH F 73 right colon na 0 12 S2 ntmsi77 2 142.1 HIGH M 37 left colon na 0 36 S2 ntmsi78 2 14.1 LOW F 53 right colon na 0 44 S2 ntmsi80 2 16.5 LOW F 77 right colon na 0 59 S2 ntmsi82 2 3576.2 HIGH F 86 right colon na 0 69 S2 ntmsi83 2 8.1 LOW F 61 right colon Y FOLFOX 1 16 S2 ntmsi84 2 256.3 HIGH F 82 right colon na 1 13 S2 ntmsi85 2 21.7 LOW M 57 right colon na 0 45 S2 ntmsi86 2 3.9 LOW M 83 right colon na 0 1 S2 ntmsi88 2 17.1 LOW F 77 right colon na 0 19 S2 ntmsi89 2 14.7 LOW F 103 right colon na 1 30 S2 ntmsi90 2 0.6 LOW F 55 right colon na 0 60 S2 ntmsi92 2 162.3 HIGH F 85 right colon na 0 3 S2 ntmsi93 2 9.4 LOW M 39 right colon na 0 72 S2 ntmsi94 2 65.9 HIGH F 89 right colon na 0 48 S2 ntmsi39 3 4.9 LOW F 78 right colon Y XELOX Avastin 1 20 S2 ntmsi40 3 46.6 LOW F 76 right colon Y LV5FU 0 73 S2 ntmsi42 3 651.0 HIGH M 34 right colon Y FOLFOX 0 32 S2 ntmsi43 3 88.5 HIGH F 26 left colon Y FOLFOX 0 16 S2 ntmsi47 3 55.8 HIGH M 55 right colon Y FOLFOX 0 54 S2 ntmsi49 3 57.1 HIGH M 44 right colon Y FOLFOX 0 91 S2 ntmsi50 3 229.0 HIGH F 51 right colon Y XELOX Avastin 0 45 S2 ntmsi51 3 3.5 LOW F 55 right colon Y FOLFOX 0 36 S2 ntmsi52 3 0.4 LOW M 43 right colon Y FOLFOX 1 8 S2 ntmsi59 3 123.7 HIGH M 52 right colon Y FOLFOX 0 60 S2 ntmsi61 3 25.5 LOW F 74 right colon Y FOLFOX 1 6 S2 ntmsi79 3 21.9 LOW M 83 right colon na 1 19 S2 ntmsi81 3 7.6 LOW M 81 right colon Y XELOX Avastin 0 48 S2 ntmsi91 3 371.0 HIGH F 84 right colon Y FOLFOX 0 39 S2 ntmsi95 3 54.0 HIGH M 56 right colon Y FOLFOX 0 26 na : not applicable; M: Male; F: Female; : o; Y: Yes; DFS 0: no relapse; DFS 1: relapse. ature Medicine doi:10.1038/nm.2457

Table S4. Associations between HSP110Δ9 expression and clinicopathological variables in MSI All Patients (=84) Series 1 Patients (=30) Series 2 Patients (=54) TM Stage 2 Patients (=59) TM Stage 3 Patients (=25) Clinical Annotation Modality n=32 n=54 n=8 n=22 n=24 n=30 n=21 n=38 n=11 n=14 0 70 31 (97%) 39 (75%) 25 8 (100%) 17 (77%) 45 23 (96%) 22 (73%) 53 20 (95%) 33 (87%) 17 11 (100%) 6 (43%) DFS event 84 0,013 30 0,29 54 0,033 59 0,41 25 2,9E-03 1 14 1 (3%) 13 (25%) 5 0 (0%) 5 (23%) 9 1 (4%) 8 (27%) 6 1 (5%) 5 (13%) 8 0 (0%) 8 (57%) 0 71 32 (100%) 39 (75%) 21 8 (100%) 13 (59%) 50 24 (100%) 26 (87%) 51 21 (100%) 30 (79%) 1 (100%) 9 (64%) OS event 84 1,30E-03 30 0,067 54 0,12 59 0,041 25 0,046 1 13 0 (0%) 13 (25%) 9 0 (0%) 9 (41%) 4 0 (0%) 4 (13%) 8 0 (0%) 8 (21%) 5 0 (0%) 5 (36%) 2 59 21 (66%) 38 (73%) 20 5 (62%) 15 (68%) 39 16 (67%) 23 (77%) - - - - - - TM Stage 84 0,47 54 0,54 - - - - 3 25 11 (34%) 14 (27%) 10 3 (38%) 7 (32%) 15 8 (33%) 7 (23%) - - - - - - left colon 10 4 (13%) 6 (12%) 6 0 (0%) 6 (29%) 4 4 (17%) 0 (0%) 7 3 (15%) 4 (11%) 3 1 (9%) 2 (15%) Tumor Location 81 1 28 0,29 53 0,036 57 0,69 24 1 right colon 71 27 (87%) 44 (88%) 22 7 (100%) 15 (71%) 49 20 (83%) 29 (100%) 50 17 (85%) 33 (89%) 21 10 (91%) 11 (85%) o 51 19 (66%) 32 (65%) 22 6 (75%) 16 (76%) 29 13 (62%) 16 (57%) 38 14 (74%) 24 (67%) 13 5 (50%) 8 (62%) HPCC 78 1 29 1 49 0,78 55 0,76 23 0,69 Yes 27 10 (34%) 17 (35%) 7 2 (25%) 5 (24%) 20 8 (38%) 12 (43%) 17 5 (26%) 12 (33%) 10 5 (50%) 5 (38%) F 39 15 (47%) 24 (46%) 10 4 (50%) 6 (27%) 29 11 (46%) 18 (60%) 28 10 (48%) 18 (47%) 11 5 (45%) 6 (43%) Gender 84 1 30 0,38 54 0,41 59 1 25 1 M 45 17 (53%) 28 (54%) 20 4 (50%) 16 (73%) 25 13 (54%) 12 (40%) 31 11 (52%) 20 (53%) 14 6 (55%) 8 (57%) <50 16 5 (16%) 11 (21%) 6 0 (0%) 6 (27%) 10 5 (21%) 5 (17%) 10 2 (10%) 8 (21%) 6 3 (27%) 3 (21%) Age [50;70[ 84 29 0,41 14 (44%) 15 (29%) 0 0,097 5 (62%) 5 (23%) 54 19 0,82 9 (38%) 10 (33%) 59 19 0,55 8 (38%) 11 (29%) 25 10 0,27 6 (55%) 4 (29%) >=70 39 13 (41%) 26 (50%) 14 3 (38%) 11 (50%) 25 10 (42%) 15 (50%) 1 (52%) 19 (50%) 9 2 (18%) 7 (50%) Age at diagnosis median 84-0,94 67 70 30-0,09 68 66 54-0,43 60 71 59-0,36 73 70 25-0,21 55 67 : Fisher exact test s, t-test s for Age at diagnosis; -: ot Applicable; DFS 0: no relapse; DFS 1: relapse; OS 0: alive; OS 1: deceased. Series 1 Patients (=30) Series 1 TM Stage 2 Patients (=20) Series 1 TM Stage 3 Patients (=10) Molecular Annotation Modality n=8 n=22 n=5 n=15 n=3 n=7 CI CIMP Low 23 4 (67%) 19 (86%) 16 3 (100%) 13 (87%) 7 1 (33%) 6 (86%) 28 0,29 18 1 10 0,18 High 5 2 (33%) 3 (14%) 2 0 (0%) 2 (13%) 3 2 (67%) 1 (14%) egative 10 2 (25%) 8 (38%) 5 1 (20%) 4 (27%) 5 1 (33%) 4 (67%) 29 0,67 9 0,52 Positive 19 6 (75%) 13 (62%) 15 4 (80%) 11 (73%) 4 2 (67%) 2 (33%) BRAF Gene mutation Status KRAS Gene mutation Status Mutated 9 3 (38%) 6 (29%) 20 7 1 (20%) 6 (40%) 2 2 (67%) 0 (0%) 29 0,67 0,61 9 0,083 Wild-Type 20 5 (62%) 15 (71%) 3 4 (80%) 9 (60%) 7 1 (33%) 6 (100%) Mutated 7 1 (12%) 6 (29%) 20 4 0 (0%) 4 (27%) 3 1 (33%) 2 (33%) 29 0,63 0,53 9 1 Wild-Type 22 7 (88%) 15 (71%) 6 5 (100%) 11 (73%) 6 2 (67%) 4 (67%) Mutated 14 4 (50%) 10 (45%) 10 3 (60%) 7 (47%) 4 1 (33%) 3 (43%) BAX* Gene mutation Status 10 1 Wild-Type 16 4 (50%) 12 (55%) 10 2 (40%) 8 (53%) 6 2 (67%) 4 (57%) Mutated 12 3 (38%) 9 (41%) 7 2 (40%) 5 (33%) 5 1 (33%) 4 (57%) GRB14* Gene mutation Status 10 1 Wild-Type 18 5 (62%) 13 (59%) 13 3 (60%) 10 (67%) 5 2 (67%) 3 (43%) Mutated 15 4 (50%) 11 (50%) 9 2 (40%) 7 (47%) 6 2 (67%) 4 (57%) IGF2R* Gene mutation Status 10 1 Wild-Type 15 4 (50%) 11 (50%) 11 3 (60%) 8 (53%) 4 1 (33%) 3 (43%) Mutated 17 7 (88%) 10 (45%) 12 4 (80%) 8 (53%) 5 3 (100%) 2 (29%) RAD50* Gene mutation Status 30 0,092 20 0,6 10 0,17 Wild-Type 13 1 (12%) 12 (55%) 8 1 (20%) 7 (47%) 5 0 (0%) 5 (71%) Mutated 13 3 (38%) 10 (45%) 9 2 (40%) 7 (47%) 4 1 (33%) 3 (43%) TCF4* Gene mutation Status 10 1 Wild-Type 17 5 (62%) 12 (55%) 11 3 (60%) 8 (53%) 6 2 (67%) 4 (57%) Methylated 21 6 (75%) 15 (71%) 16 4 (80%) 12 (80%) 5 2 (67%) 3 (50%) CACA1G Methylation Status 29 1 9 1 Unmethylated 8 2 (25%) 6 (29%) 4 1 (20%) 3 (20%) 4 1 (33%) 3 (50%) Methylated 14 5 (62%) 9 (43%) 11 3 (60%) 8 (53%) 3 2 (67%) 1 (17%) IGF2 Methylation Status 29 0,43 9 0,23 Unmethylated 15 3 (38%) 12 (57%) 9 2 (40%) 7 (47%) 6 1 (33%) 5 (83%) Methylated 16 4 (50%) 12 (55%) 12 3 (60%) 9 (60%) 4 1 (33%) 3 (43%) MGMT Methylation Status 10 1 Unmethylated 14 4 (50%) 10 (45%) 8 2 (40%) 6 (40%) 6 2 (67%) 4 (57%) Methylated 17 6 (75%) 11 (50%) 12 3 (60%) 9 (60%) 5 3 (100%) 2 (29%) MLH1 Methylation Status 30 0,41 10 0,17 Unmethylated 13 2 (25%) 11 (50%) 8 2 (40%) 6 (40%) 5 0 (0%) 5 (71%) Methylated 21 6 (75%) 15 (71%) 6 4 (80%) 12 (80%) 5 2 (67%) 3 (50%) EUROG1 Methylation Status 29 1 1 9 1 Unmethylated 8 2 (25%) 6 (29%) 20 4 1 (20%) 3 (20%) 4 1 (33%) 3 (50%) Methylated 22 7 (88%) 15 (71%) 17 5 (100%) 12 (80%) 5 2 (67%) 3 (50%) p14 ARF Methylation Status 29 0,63 20 0,54 9 1 Unmethylated 7 1 (12%) 6 (29%) 3 0 (0%) 3 (20%) 4 1 (33%) 3 (50%) Methylated 9 3 (38%) 6 (29%) 8 3 (60%) 5 (33%) 1 0 (0%) 1 (17%) p16 Methylation Status 29 0,67 20 0,35 9 1 Unmethylated 20 5 (62%) 15 (71%) 12 2 (40%) 10 (67%) 8 3 (100%) 5 (83%) Methylated 5 3 (38%) 2 (10%) 4 2 (40%) 2 (13%) 1 1 (33%) 0 (0%) RASSF1 Methylation Status 29 0,11 20 0,25 9 0.33 Unmethylated 24 5 (62%) 19 (90%) 16 3 (60%) 13 (87%) 8 2 (67%) 6 (100%) Methylated 11 4 (50%) 7 (33%) 8 2 (40%) 6 (40%) 3 2 (67%) 1 (17%) RUX3 Methylation Status 29 0,43 9 0,23 Unmethylated 18 4 (50%) 14 (67%) 12 3 (60%) 9 (60%) 6 1 (33%) 5 (83%) Methylated 18 6 (75%) 12 (57%) 12 3 (60%) 9 (60%) 6 3 (100%) 3 (50%) SOCS1 Methylation Status 29 0,67 9 0,46 Unmethylated 11 2 (25%) 9 (43%) 8 2 (40%) 6 (40%) 3 0 (0%) 3 (50%) Fisher exact test s, t-test s for Age; -: on Applicable. * Mutations at coding microsatellite sequences MSI (target genes). ature Medicine doi:10.1038/nm.2457