CRIC013M (mutated KRAS) CRIC002P (wt KRAS) 0 200 400 600 800 1000 1200 1400 10 30 50 70 90 1 Tumor volume (mm 3 ) 0 200 400 600 800 1000 1200 1400 20 30 40 50 60 70 Days post implantation Tumor volume (mm 3 ) Cetuximab (12.5 mg/kg/adm) Days post implantation CRLRB022P in mice 0 500 1000 1500 15 20 25 30 35 Days post implantation Median Tumor volume (mm 3 ) Control Cetuximab (12.5 mg/kg/ adm) 5-FU (50.0 mg/kg/ adm) Oxaliplatin (5.0mg/kg/ adm) Cetuximab IP treatment 5-FU IV treatment Oxaliplatin IV treatment 0 500 1000 1500 15 20 25 30 35 Dayspost implantation MedianTumorvolume (mm 3 ) Control Cetuximab (12.5 mg/kg/adm) 5-FU (50.0 mg/kg/ adm) Oxaliplatin (5.0mg/kg/ adm) Cetuximab IP treatment 5-FU IV treatment Oxaliplatin IV treatment 0 500 1000 1500 15 20 25 30 35 Dayspost implantation MedianTumorvolume (mm 3 ) Control CPT-11 (22mg/kg/ Adm) CPT-11 IV treatment 3 CRLRB022P in rats Control Oxaliplatin (4 mg/kg/ adm) CPT-11 (40 mg/kg/ adm) Cetuximab (10 mg/kg/ adm) 5-FU (30 mg/kg/ adm) Days post implantation 70 60 50 40 30 20 10 Median tumour volume (mm 3 ) 10000 9000 8000 7000 6000 5000 4000 3000 2000 1000 0 Treatment start (D23) Control Oxaliplatin (4 mg/kg/ adm) CPT-11 (40 mg/kg/ adm) Cetuximab (10 mg/kg/ adm) 5-FU (30 mg/kg/ adm) (4 mg/kg/ adm) CPT- adm) (10 mg/kg/ 5- adm ) Days post implantation 70 60 50 40 30 20 10 Median tumour volume (mm 3 ) 10000 9000 8000 7000 6000 5000 4000 3000 2000 1000 0 Days post implantation 70 60 50 40 30 20 10 ) 10000 9000 8000 7000 6000 5000 4000 3000 2000 1000 0 Treatment start (D23) - adm) 5- ) - 5- ) - - ) - - ) - CRLRB022P in mice 0 500 1000 1500 15 20 25 30 35 Days post implantation Median Tumor volume (mm 3 ) Control Cetuximab (12.5 mg/kg/ adm) 5-FU (50.0 mg/kg/ adm) Oxaliplatin (5.0mg/kg/ adm) Cetuximab IP treatment 5-FU IV treatment Oxaliplatin IV treatment 0 500 1000 1500 15 20 25 30 35 Dayspost implantation MedianTumorvolume (mm 3 ) Control Cetuximab (12.5 mg/kg/adm) 5-FU (50.0 mg/kg/ adm) Oxaliplatin (5.0mg/kg/ adm) Cetuximab IP treatment 5-FU IV treatment Oxaliplatin IV treatment CRLRB022P in mice 0 500 1000 1500 15 20 25 30 35 Days post implantation Median Tumor volume (mm 3 ) Control Cetuximab (12.5 mg/kg/ adm) 5-FU (50.0 mg/kg/ adm) Oxaliplatin (5.0mg/kg/ adm) Cetuximab IP treatment 5-FU IV treatment Oxaliplatin IV treatment 0 500 1000 1500 15 20 25 30 35 Dayspost implantation MedianTumorvolume (mm 3 ) Control Cetuximab (12.5 mg/kg/adm) 5-FU (50.0 mg/kg/ adm) Oxaliplatin (5.0mg/kg/ adm) Cetuximab IP treatment 5-FU IV treatment Oxaliplatin IV treatment 0 500 1000 1500 15 20 25 30 35 Dayspost implantation MedianTumorvolume (mm 3 ) Control CPT-11 (22mg/kg/ Adm) CPT-11 IV treatment 3 0 500 1000 1500 15 20 25 30 35 Dayspost implantation MedianTumorvolume (mm 3 ) Control CPT-11 (22mg/kg/ Adm) CPT-11 IV treatment 3 CRLRB022P in rats Control Oxaliplatin (4 mg/kg/ adm) CPT-11 (40 mg/kg/ adm) Cetuximab (10 mg/kg/ adm) 5-FU (30 mg/kg/ adm) Days post implantation 70 60 50 40 30 20 10 Median tumour volume (mm 3 ) 10000 9000 8000 7000 6000 5000 4000 3000 2000 1000 0 Treatment start (D23) Control Oxaliplatin (4 mg/kg/ adm) CPT-11 (40 mg/kg/ adm) Cetuximab (10 mg/kg/ adm) 5-FU (30 mg/kg/ adm) (4 mg/kg/ adm) CPT- adm) (10 mg/kg/ 5- adm ) Days post implantation 70 60 50 40 30 20 10 Median tumour volume (mm 3 ) 10000 9000 8000 7000 6000 5000 4000 3000 2000 1000 0 Days post implantation 70 60 50 40 30 20 10 ) 10000 9000 8000 7000 6000 5000 4000 3000 2000 1000 0 Treatment start (D23) - adm) 5- ) - 5- ) - - ) - - ) - CRLRB022P in rats Control Oxaliplatin (4 mg/kg/ adm) CPT-11 (40 mg/kg/ adm) Cetuximab (10 mg/kg/ adm) 5-FU (30 mg/kg/ adm) Days post implantation 70 60 50 40 30 20 10 Median tumour volume (mm 3 ) 10000 9000 8000 7000 6000 5000 4000 3000 2000 1000 0 Days post implantation 70 60 50 40 30 20 10 Median tumour volume (mm 3 ) 10000 9000 8000 7000 6000 5000 4000 3000 2000 1000 0 Treatment start (D23) Control Oxaliplatin (4 mg/kg/ adm) CPT-11 (40 mg/kg/ adm) Cetuximab (10 mg/kg/ adm) 5-FU (30 mg/kg/ adm) (4 mg/kg/ adm) CPT- adm) (10 mg/kg/ 5- adm ) Days post implantation 70 60 50 40 30 20 10 Median tumour volume (mm 3 ) 10000 9000 8000 7000 6000 5000 4000 3000 2000 1000 0 Days post implantation 70 60 50 40 30 20 10 ) 10000 9000 8000 7000 6000 5000 4000 3000 2000 1000 0 Treatment start (D23) - adm) 5- ) - 5- ) - - ) - - ) - INTRODUCTION MATERIAL AND METHODS CReMEC initiative: Characterization of patient-derived colorectal tumor models and correlation with patient profile Sylvia Julien 1 , Ana Merino-Trigo 2 , Ludovic Lacroix 7,5 , Marc Pocard 3,5 , Diane Goéré 7,5 , Pascale Mariani 4,5 , Sophie Landron 8 , Ludovic Bigot 7,5 , Fariba Nemati 4,5 , Louis-Bastien Weiswald 3,5 , Denis Lantuas 3,5 , Loïc Morgand 9 , Grégoire Prévost 1 , Patrick Gonin 7,5 , Virginie Dangles-Marie 3,4,5 , Alain Pierré 8 , Alain Bruno 8 , Hugues De Thé 5,6 , Hany Soliman 5,6 , Manoel Nunes 2 , Loreley Calvet 2 , Patricia Vrignaud 2 , Olivier Duchamp 9 , Cyril Berthet 9 . 1 Ipsen, 2 sanofi-aventis, 3 Hôpital Lariboisière , 4 Institut Curie, 5 Cancéropole d’Ile de France, 6 Institut Universitaire d'Hématologie, Paris, 7 Institut Gustave Roussy, Villejuif, 8 Institut de recherche Servier, Croissy sur Seine, 9 Oncodesign, Dijon, France. RESULTS CONCLUSION #4169 Well characterized models representing the heterogeneity of human colorectal cancers (CRC) are needed to develop effective therapeutic agents. Establishment of such tools will allow a better prediction of the clinical outcome, taking into account the diversity of each patient tumor phenotype and genotype. For this purpose, we have associated efforts from hospitals, academic groups, biotech and pharmaceutical companies. The goal of this consortium is to create an experimental tumor model resource center to improve or strengthen drug development. From May 2007 to January 2009, 86 surgical specimens [59 primary (P) tumors, 19 metastasis (M), and 8 peritoneal carcinomatosis (C)] were collected from CRC patients (with informed consents and negative HBV, HCV, and HIVs serologies). Tumor samples were subcutaneously xenografted in nude mice and characterized as described below. We report here the results on our panel of models. Patients were informed and gave their consent for providing surgical tumor samples to CReMEC and for HIV1, HIV2, HBV and HCV serological status testing. Tumor samples were collected in 3 medical centers of the Paris-Ile-de-France area: Institut Curie, Institut Gustave Roussy, Hôpital Lariboisière. Fresh tumor material was conditioned into RPMI 1640 with 200 U/mL penicillin, 200 μg/mL streptomycin and 2.5 μg/mL fungizone. Initial xenografting was carried out at the site of sampling, within 12 hours after specimen collection. Procedures were performed according to ethical guidelines for animal care and handling. 20-40 mg fragments were xenografted subcutaneously either in the flank or in the interscapular area, in 3-5 immunodeficient SWISS nude mice Grafted mice were kept for a maximum of 4 months without tumor growth. Clinical data collection Relevant clinical information was collected by the attending physician and included in a standardized data sheet. Identification of the data sheet is anonymous. Histological characterization Samples were fixed for a maximum of 48 hours in alcohol-formalin-acetic acid (AFA) and embedded in paraffin. 5 μm sections were stained with hematoxylin-eosin-saffron. Molecular characterization – sequencing CGH array analysis: Evaluation of genome-wide, gene copy number was evaluated using a 244k CGH array Agilent technology. It was carried out on DNA from the patient sample or at an early passages (P0/P1), and at the passage used for the pharmacological evaluation (P8-9). DNA sequencing: The following genetic markers, relevant in CRC, were selected for sequencing: APC (exons 9 & 16), KRAS (exons 2 & 3), BRAF (exons 11 & 15), TP53 (exons 2 to 11), CTNNB1 (exon 3), PIK3CA (exons 10 and 21), FBXW7 (exons 4 to 11), EGFR (exons 18 to 21) and AKT1 (exon 4). Sanger direct sequencing was performed after PCR amplification of exons of genes harboring hotspot mutations as described in Catalogue Of Somatic Mutations In Cancer (COSMIC: http://www.sanger.ac.uk/genetics/CGP/cosmic/ ). Determination of Microsatellite Instability (MSI) status: The MSI status was determined according to the Consensus Conference (or Revised Bethesda Guidelines) (J. Natl Cancer Inst 2004, 96, 261-268) recommendations using the following five quasimonomorphic markers: NR21, BAT26, BAT25, NR24 and NR22. Pharmacological characterization Tested agents: 5-fluorouracil (5-FU), oxaliplatin (L-OHP), irinotecan (CPT-11) or cetuximab, were tested. In vivo determination of antitumor activity: Tumor fragments were subcutaneously xenografted in SCID mice or nude rats. Tumor-bearing animals were randomized when the mean tumor volume reached 100-200 mm 3 in mice or 500-700 mm 3 in rats. Compounds were formulated in glucose 5% in water. 3 cytotoxic drugs were IV administered, and tested at 70% of their respective highest non toxic dose and using the following regimen in mice: 5-FU at 56 mg/kg/adm Q4Dx2 (30 mg/kg/adm Q7Dx3 in rats) , L-OHP at 5 mg/kg/adm Q4Dx2 (4 mg/kg/adm Q4Dx3 in rats), CPT-11 at 22 mg/kg/adm, Q2Dx3 (40 mg/kg/adm, Q7Dx3in rats) ; 1 targeting therapy was IP administrated: cetuximab at 12.5 mg/kg/adm, (Q3Dx2)x2 (10 mg/kg/adm, IV, Q7Dx3 in rats). Antitumor activity was evaluated by calculating the ∆(T/C) ratio: ∆ ∆ ∆(T/C) (%) = X 100 median T VOL- DY – median T VOL-DX median C VOL- DY – median C VOL-DX Preservation of the tumor phenotype and genotype Clinical characteristics and in vivo tumorgraft take rate Scoring criteria: -= ∆(T/C) > 42 % + = 10 < ∆(T/C) ≦ 42 % ++ = 0 ≦ ∆(T/C) ≦10 % (stable disease) +++ = ∆(T/C) <0 % (tumor regression) 8 4 12 5 CPT-11 7 3 4 15 Cetuximab 0 0 2 27 L-OHP 0 5 17 7 5-FU +++ ++ + - Activity score 22 4 4 <10% 10-50% >50% Mucus Secretion 22 4 4 <10% 10-50% >50% Mucus Secretion Tumor Models Differentiation 2 12 13 2 1 WD WD, MD WD, MD, PD MD, PD PD, UD Differentiation 2 12 13 2 1 WD WD, MD WD, MD, PD MD, PD PD, UD Mutations 1 0 0 0 KRAS/BRAF +++ ++ + - Activity score 4 1 3 11 EGFR mutated pathway 3 2 1 3 EGFR wild-type pathway* 0 0 0 2 BRAF 0 0 2 1 KRAS/PIK3CA 3 1 1 8 KRAS Fresh tumors Surgery * Patient identification * Patient clinical history * Patient informed consent * Negative HIV, HBV, HCV serology Paraffin Tumor bank Snap-frozen Database Tumor growth Molecular Histological Pharmacological Characterization 26 31 36 41 46 51 0 200 400 600 800 1000 1200 1400 CPT-11 treatment CPT-11 22 mg/kg IV Control (untreated) Days after tumor inoculation Median tumor size (mg +/- IQR) Tumor grafting P0 P2 – P6 Establishment of in vivo tumorgraft models RPMI/DMSO freezing Fresh tumors Surgery Fresh tumors Surgery * Patient identification * Patient clinical history * Patient informed consent * Negative HIV, HBV, HCV serology * Patient identification * Patient clinical history * Patient informed consent * Negative HIV, HBV, HCV serology Paraffin Tumor bank Snap-frozen Paraffin Tumor bank Snap-frozen Database Tumor growth Molecular Histological Pharmacological Characterization 26 31 36 41 46 51 0 200 400 600 800 1000 1200 1400 CPT-11 treatment CPT-11 22 mg/kg IV Control (untreated) Days after tumor inoculation Median tumor size (mg +/- IQR) Database Database Tumor growth Molecular Histological Pharmacological Characterization Tumor growth Molecular Histological Pharmacological Characterization 26 31 36 41 46 51 0 200 400 600 800 1000 1200 1400 CPT-11 treatment CPT-11 22 mg/kg IV Control (untreated) Days after tumor inoculation Median tumor size (mg +/- IQR) 26 31 36 41 46 51 0 200 400 600 800 1000 1200 1400 CPT-11 treatment CPT-11 22 mg/kg IV Control (untreated) CPT-11 treatment CPT-11 22 mg/kg IV Control (untreated) Days after tumor inoculation Median tumor size (mg +/- IQR) Tumor grafting P0 P2 – P6 Establishment of in vivo tumorgraft models RPMI/DMSO freezing Tumor grafting P0 P2 – P6 Establishment of in vivo tumorgraft models RPMI/DMSO freezing PR: Partial Response CR: Complete Response TFS: Tumor Free Survival Tumor take rate (%) Parameter Class Number Frequency (%) Site of origin Primary 37 66.1 62.7 Peritoneal carcinosis 5 8.9 62.5 Metastasis # 14 25.0 73.7 Sex Female 35 62.5 66.0 Male 21 37.5 63.6 Age 0- 50y 11 20.4 78.6 [ 50y ; 60y] 12 24.5 50.0 [ 60y ; 70y] 12 20.4 60.0 [ 70y ; 80y] 15 24.5 68.2 >80y 6 10.2 100 Stage 0 - I 2 3.7 22.2 II 8 14.8 57.1 III 13 24.1 92.9 IV 31 57.4 70.4 Patient characteristics in the collection # hepatic, splenic and mesenteric lymph node metastasis Tumor take rate (%) Parameter Class Number Frequency (%) Site of origin Primary 37 66.1 62.7 Peritoneal carcinosis 5 8.9 62.5 Metastasis # 14 25.0 73.7 Sex Female 35 62.5 66.0 Male 21 37.5 63.6 Age 0- 50y 11 20.4 78.6 [ 50y ; 60y] 12 24.5 50.0 [ 60y ; 70y] 12 20.4 60.0 [ 70y ; 80y] 15 24.5 68.2 >80y 6 10.2 100 Stage 0 - I 2 3.7 22.2 II 8 14.8 57.1 III 13 24.1 92.9 IV 31 57.4 70.4 Patient characteristics in the collection # hepatic, splenic and mesenteric lymph node metastasis Significant correlation* between CEA serum level of patient and tumor take of all types Significant distribution* of primary tumors in regards of lymph node status (panel A, N1 = 1 to 6, N2 = 7 to 15 positive regional lymph nodes), stages (panel B) and differentiation status (panel C). No other significant correlation were found among the following parameters: gender, age, resection extent, lymphatic embolies, perinervous invasion, initial treatment, genotype * p values were calculated with logical regression test Gene mutation profile and response to chemotherapy Histopathological analyses completed for 30 colon tumor models were in concordance to those observed in the corresponding patient’s tumor. CGH analysis showed very similar profile between early and advance passages The expression and localization of β β β-catenin, CEA, CAIX, EGFR, CD105 and LYVE-1 is ongoing. The majority of tumor models were non-mucinous adenocarcinomas Splenic metastasis (1/86) Mesenteric lymph node (1/86) All models collected (86) Left Right Primary tumor (59/86) Carcinosis (8/86) Hepatic Metastasis (16/86) Ovarian metastasis (1/86) Left Rect. Left Right Left Right Rect. Splenic metastasis (1/86) Mesenteric lymph node (1/86) All models collected (86) Left Right Primary tumor (59/86) Carcinosis (8/86) Hepatic Metastasis (16/86) Ovarian metastasis (1/86) Left Rect. Left Right Left Right Rect. Right colon Left colon Rectum Models growing on nude mice (56) Primary tumor (37/56) Ca r c i n o s i s ( 5 / 5 6 ) Hepatic Metastasis (12/56) Splenic metastasis (1/56) Mesenteric lymph node (1/56) Left Right Rect.Left Right Left Right Rect. Right colon Left colon Rectum Right colon Left colon Rectum Models growing on nude mice (56) Primary tumor (37/56) Ca r c i n o s i s ( 5 / 5 6 ) Hepatic Metastasis (12/56) Splenic metastasis (1/56) Mesenteric lymph node (1/56) Left Right Rect.Left Right Left Right Rect. Only 1 model showed a poor differentiated (PD) to undifferentiated (UD) characteristic, as well as its correspondent patient Similar CGH profiles between early and advance passages Loss in chromosomes 3p and 4q in advance passage Most models with EGFR mutated pathway are resistant to Cetuximab, nevertheless several Cetuximab sensitive KRAS-mutated models have been identified. Patient: CR-IGR-034P Patient: CR-LRB-010P Patient: CR-LRB-008M P10: CR-LRB-010P P11: CR-LRB-008M P11: CR-IGR-034P Patient Tumor Model Well Differentiated Well to Moderate Differentiated Poor to UnDifferentiated Patient: CR-IGR-034P Patient: CR-LRB-010P Patient: CR-LRB-008M P10: CR-LRB-010P P11: CR-LRB-008M P11: CR-IGR-034P Patient: CR-IGR-034P Patient: CR-LRB-010P Patient: CR-LRB-008M Patient: CR-IGR-034P Patient: CR-IGR-034P Patient: CR-LRB-010P Patient: CR-LRB-010P Patient: CR-LRB-008M Patient: CR-LRB-008M P10: CR-LRB-010P P11: CR-LRB-008M P11: CR-IGR-034P P10: CR-LRB-010P P10: CR-LRB-010P P11: CR-LRB-008M P11: CR-LRB-008M P11: CR-IGR-034P P11: CR-IGR-034P Patient Tumor Model Patient Tumor Model Well Differentiated Well to Moderate Differentiated Poor to UnDifferentiated Sensitivity to Cetuximab versus mutated genes involved in EGFR pathway ∆T/C = -41% (Day 31), 9/10 PR & 4/10 CR CRIGR023M GAIN LOSS CRIGR002C GAIN LOSS * Mutations of PTEN or other genes related to EGFR pathway have not been yet characterized Multiple mutation profiles are observed in our tumor panel (n=60), matching the human tumor genetic heterogeneity Pharmacological studies exhibit diversity in the response to chemotherapy 13/29 tumorgrafts show tumor regression after treatment with at least one standard monotherapy Diversity of colorectal cancers is fully addressed in this collection of patient-derived tumor models. Genotype and phenotype are largely preserved throughout the model establishment process. Difference of gene mutation and drug sensitivity profiles are observed between the models. Plan to complete the full correlation analysis between clinical data, gene mutations, transcriptome profile, ex-vivo and in vivo drug sensitivity. Perspectives to fully exploit this new collection for new drug candidate selection. ∆T/C = < 0 % (Day 36), 10/10 CR + 100% TFS 0 CEA (ng/mL) Positive xenograft take 5 10 15 20 25 30 35 40 45 50 170 175 180 4 1 11 2.3 8 27 p = 0.0292 (*) All types of tumor samples / CEA dosing Negative xenograft take 0 5 10 15 20 25 30 35 40 45 50 170 180 4 1 11 2.3 8 27 p = 0.0292 (*) 0 CEA (ng/mL) Positive xenograft take 5 10 15 20 25 30 35 40 45 50 170 175 180 4 1 11 2.3 8 27 p = 0.0292 (*) All types of tumor samples / CEA dosing Negative xenograft take 0 5 10 15 20 25 30 35 40 45 50 170 180 4 1 11 2.3 8 27 p = 0.0292 (*) A Percentage (%) N0 N0 N1 + N2 N1 + N2 0 20 40 60 80 100 120 (n=11) (n=26) (n=5) (n=17) Primary tumor samples / Lymph node status Positive xenograft take Negative xenograft take p = 0.0038 (**) N0 N0 N1 + N2 N1 + N2 0 20 40 60 80 100 120 (n=11) (n=26) (n=5) (n=17) p = 0.0038 (**) A Percentage (%) N0 N0 N1 + N2 N1 + N2 0 20 40 60 80 100 120 (n=11) (n=26) (n=5) (n=17) Primary tumor samples / Lymph node status Positive xenograft take Negative xenograft take p = 0.0038 (**) N0 N0 N1 + N2 N1 + N2 0 20 40 60 80 100 120 (n=11) (n=26) (n=5) (n=17) p = 0.0038 (**) B Stages 0-I Stages 0-I Stage II Stage II Stage III Stage III Stage IV Stage IV 0 20 40 60 80 100 120 (n=7) (n=6) (n=7) (n=6) (n=13) (n=14) Primary tumor samples / Stages Negative xenograft take Positive xenograft take p = 0.0101 (*) Percentage (%) Stages 0-I Stages 0-I Stage II Stage II Stage III Stage III Stage IV Stage IV 0 20 40 60 80 100 120 (n=7) (n=6) (n=7) (n=6) (n=13) (n=14) p = 0.0101 (*) B Stages 0-I Stages 0-I Stage II Stage II Stage III Stage III Stage IV Stage IV 0 20 40 60 80 100 120 (n=7) (n=6) (n=7) (n=6) (n=13) (n=14) Primary tumor samples / Stages Negative xenograft take Positive xenograft take p = 0.0101 (*) Percentage (%) Stages 0-I Stages 0-I Stage II Stage II Stage III Stage III Stage IV Stage IV 0 20 40 60 80 100 120 (n=7) (n=6) (n=7) (n=6) (n=13) (n=14) p = 0.0101 (*) C Primary tumor samples / Differentiation 0 Well Well Fairly Fairly Weakly 20 40 60 80 100 120 (n=10) (n=11) (n=9) (n=25) Percentage (%) p = 0.0287 (*) Negative xenograft take Positive xenograft take 0 Well Well Fairly Fairly Weakly 20 40 60 80 100 120 (n=10) (n=11) (n=9) (n=25) p = 0.0287 (*) C Primary tumor samples / Differentiation 0 Well Well Fairly Fairly Weakly 20 40 60 80 100 120 (n=10) (n=11) (n=9) (n=25) Percentage (%) p = 0.0287 (*) Negative xenograft take Positive xenograft take 0 Well Well Fairly Fairly Weakly 20 40 60 80 100 120 (n=10) (n=11) (n=9) (n=25) p = 0.0287 (*) Similar tumor response ranges have been observed in mice and rats ∆ ∆ ∆T/C in mice L-OHP: 45% 5-FU: 45% Cetuximab: 78% ∆ ∆ ∆T/C in rat L-OHP: 104% 5-FU: 63% Cetuximab: 19% CPT-11: 1% ∆ ∆ ∆T/C in mice CPT-11 < 0% 70% 80% 80% 50% 70% 90% 60% 80% 80% 50% 90% 90% 70% 60% 90% 60% 40% 100% 40% 70% 40% 80% 50% 80% 50% 80% 80% 80% 80% 50% 90% 40% 80% 30% 90% 80% 90% 90% 85% 60% 85% 50% 60% 70% 40% 40% 80% 50% 30% 50% 70% 90% 70% 80% 80% 50% 50% 50% 50% 80% CRIGR023M CRIC004M CRIC007M CRIGR007P CRIGR025P CRLRB010P CRLRB011M CRLRB013P CRLRB014P CRIC018P CRIC002P CRIC003P CRIC025M CRIGR004P CRIGR008P CRLRB007P CRIGR015P CRLRB019C CRIGR021P CRIC020P CRIC021M CRIGR032P CRIGR029P CRIGR012P CRIGR014P CRIC012P CRIC013M CRLRB008M CRLRB009C CRIGR003P CRIGR009P CRIGR016P CRIGR020P CRIGR001M CRIGR038C CRLRB017P CRIGR034P CRLRB018P CRLRB003P CRLRB004P CRLRB015P CRIC006M CRIC019P CRIC029P CRIC028M CRIGR002P CRIGR002C CRIGR052M CRLRB005P CRIGR047P CRIGR048M CRLRB022P CRIC008P CRIC009M CRIC010P CRIGR011C CRIGR039P CRIGR043P CRIC005P CRIC014P TP53 62% 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 1 1 1 1 1 1 1 1 1 0 0 0 1 1 1 1 0 1 1 1 0 1 1 0 0 0 0 0 0 0 0 APC 53% 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 1 1 1 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 1 0 0 0 0 0 KRAS 45% 1 1 1 1 1 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 0 1 1 1 1 1 1 1 1 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 0 0 PIK3CA 10% 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 FBXW7 8% 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 1 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 BRAF 5% 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 * 0 0 0 0 0 0 0 0 0 1 1 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 CTNNB1 3% 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 EGFR 2% 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 AKT1 0% 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 MSI status MSS MSS MSS MSS MSS MSS MSS MSS MSS MSS MSS MSS MSS MSI-L MSI-L MSS MSS MSS MSS MSS MSS MSS MSS MSS MSI-L MSS MSS MSS MSI-L MSS MSS MSS MSS MSI-L MSI-H MSI-H MSS MSI-L MSI-L MSS MSS MSI-L MSI-L MSI-L MSS MSS MSS MSS MSS MSS MSS MSS MSS MSS Xenograft 1 1 1 1 1 1 1 1 1 1 1 1 1 0 1 1 0 1 0 1 1 1 1 1 1 0 1 1 1 1 1 1 0 0 1 1 1 1 1 1 1 1 1 1 1 1 1 1 0 1 1 1 1 1 1 1 1 1 1 0 CGH Analysis 0 1 1 0 0 1 0 0 0 0 0 1 0 1 1 0 0 0 0 0 0 1 0 1 1 0 0 0 1 0 0 1 1 0 0 0 1 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 0 1 80 0 0 0 1 0 Other ERCC1-A138T FCGR2A-F196Y * BRAF-N594D 5-FU + - - - - + + + ++ + + ++ + + + + + ++ + - + ++ + ++ + + - + - l-OHP - - - - - - - - - - - - + - - - + - - - - - - - - - - - - CPT-11 + + ++ + ++ +++ - +++ - + - - +++ - + ++ + +++ + + + + +++ ++ + +++ +++ + +++ CETUXIMAB + ++ +++ + - - +++ +++ +++ - - - +++ + - +++ - - - - - - ++ ++ +++ + - - - Mutation MSI-L Low (MSI-L) or High (MSI-H) microsatellite instability Wild-type No data available Model from the same patient Xenograft model established CGH analysis % above sample name: % of tumor cells in the analyzed sample Scoring criteria: - = ∆(T/C) > 42 % + = 10 < ∆(T/C) ≦ 42 % ++ = 0 ≦ ∆(T/C) ≦ 10 % (stable desease) +++ = ∆(T/C) <0 % (tumor regression) Characterization in progress % beside gene name: % of mutations among all sequenced samples