Immunoscore Prognostic in Colon Cancer CCO Independent Conference Coverage* of the 2016 ASCO Annual Meeting, June 3-7, 2016 *CCO is an independent medical education company that provides state-of-the-art medical information to healthcare professionals through conference coverage and other educational programs. This activity is supported by educational grants from Amgen, Ariad, Bayer Healthcare Pharmaceuticals, Celgene Corporation, Genentech, Incyte, Merck, and Taiho Pharmaceuticals.

Immunoscore in Colon Cancer Conventional risk prediction methods for cancer provide little information on postoperative outcome[1,2] Histopathologic tissue analysis focuses only on tumor cells Host immune response not incorporated Comprehensive evaluation of molecular features and immune reactions, including systemic and local immunologic biomarkers, critical for improving prognostic methods[1,2] Immunoscore evaluates immune infiltrate in center and invasive margin of tumor and may be useful as prognostic marker[1,2] Worldwide Immunoscore Consortium evaluated standardized Immunoscore assay in routine clinical settings[3] 1. Galon J, et al. J Transl Med. 2012;10:205. 2. Galon J, et al. J Pathol. 2014;232:199-209. 3. Galon J, et al. ASCO 2016. Abstract 3500. Slide credit: clinicaloptions.com

Immunoscore in Colon Cancer: Study Design Inclusion criteria Colon cancer (excluding rectum cancer), stage I/II/III (T1-T4, N0-N2, M0) No neoadjuvant treatments Clinical data and follow-up 3855 pts quantified for Immunoscore study; 2667 analyzed after quality control and exclusion criteria Primary endpoint: TTR for Immunoscore (high/low) Secondary endpoint: TTR for Immunoscore (high/intermediate/low) all stages and stage II Limitations: heterogeneity of pts between centers, in patient care between countries, in IHC staining, and in clinical data and follow-up between centers TTR, time to recurrence. Slide credit: clinicaloptions.com Galon J, et al. ASCO 2016. Abstract 3500.

Immunoscore in Colon Cancer: Study Design Reference center sent SOP to all participating centers to standardize tissue collection and IHC procedures Participating centers obtained tissue samples and performed IHC before sending data to reference center (in Europe) or clinical data center (in US) External statistician analyzed all data using digital quantification of whole slide FFPE for 3 cohorts: Training set Internal validation set External validation set SOP, standard operating procedure. Slide credit: clinicaloptions.com Galon J, et al. ASCO 2016. Abstract 3500.

Immunoscore in Colon Cancer: Pt Population Characteristics Training Set (n = 700) Internal Validation Set (n = 636) External Validation Set (n = 969) Median age, yrs 68.3 (±12.6) 68.3 (±12.2) 68.2 (± 32.7) Male, n (%) 346 (49.4) 339 (53.3) 497 (51.3) T stage, n (%) T1 T2 T3 T4 37 (5.3) 109 (15.6) 452 (64.6) 102 (14.6) 34 (5.3) 97 (15.3) 427 (67.1) 78 (12.3) 32 (3.3) 153 (15.8) 635 (65.5) 149 (15.4) N stage, n (%) N0 N1 N2 508 (73.4) 124 (17.9) 60 (8.7) 482 (76.3) 107 (16.9) 43 (6.8) 608 (64.1) 223 (23.5) 117 (12.3) Median lymph nodes, n 22.1 (±15.2) 21.8 (±16.9) 16.4 (±11.8) Proximal colon cancer, n (%) Distal colon cancer, n (%) Missing, n (%) 349 (49.9) 2 (0.3) 307 (48.3) 327 (51.5) 1 (0.2) 527 (54.9) 431 (44.9) 2 (0.2) Slide credit: clinicaloptions.com Galon J, et al. ASCO 2016. Abstract 3500.

Immunoscore in Colon Cancer: Pt Population Characteristics Training Set Internal Validation Set External Validation Set Time to end of follow-up Median survival, mos 5-yr survival rate, % 143.6 (127.3-162.2) 74.9 (71.6-78.2) 180.7 (147.7-197.6) 77.8 (74.5-81.1) 160.1 (124.5-191.4) 68.8 (65.6-72.0) Recurrence-free survival time 122.3 (107.6-132.8) 68.3 (64.7-71.9) 140.2 (116.6-150.4) 71.3 (67.6-75.0) 95.1 (80.0-106.9) 58.3 (54.9-61.8) All multivariate models adjusted for Immunoscore, age, sex, T stage, N stage, and stratified by city center Slide credit: clinicaloptions.com Galon J, et al. ASCO 2016. Abstract 3500.

Immunoscore in Colon Cancer: Time to Recurrence, Training Set Outcome Training Set High Intermediate Low Primary objective (high/low): 5-yr time to recurrence, % (95% CI) 67.3 (59.4-76.2) -- 85.3 (82.1-88.6) HR (95% CI) 0.41 (0.28-0.61) P value C-index < .0001 0.60 Secondary objective (high/int/low): 81.9 (77.7-86.3) 92.3 (88.2-96.6) Int vs high: 0.51 (0.34-0.77) Low vs high: 0.19 (0.10-0.37) 0.64 Slide credit: clinicaloptions.com Galon J, et al. ASCO 2016. Abstract 3500.

Internal Validation Set Immunoscore in Colon Cancer: Time to Recurrence, Internal Validation Set Outcome Internal Validation Set High Intermediate Low Primary objective (high/low): 5-yr time to recurrence, % (95% CI) 74.3 (67.1-82.3) -- 88.0 (84.8-91.3) HR (95% CI) 0.41 (0.27-0.65) P value C-index < .0001 0.60 Secondary objective (high/int/low): 86.1 (82.1-90.4) 91.4 (86.6-96.5) Int vs high: 0.48 (0.30-0.77) Low vs high: 0.27 (0.14-0.53) .0001 0.63 Slide credit: clinicaloptions.com Galon J, et al. ASCO 2016. Abstract 3500.

External Validation Set Immunoscore in Colon Cancer: Time to Recurrence, External Validation Set Outcome External Validation Set High Intermediate Low Primary objective (high/low): 5-yr time to recurrence, % (95% CI) 58.3 (51.2-66.4) -- 76.2 (72.8-79.9) HR (95% CI) 0.51 (0.38-0.68) P value C-index < .0001 0.56 Secondary objective (high/int/low): 72.0 (67.5-76.9) 83.1 (78.1-88.4) Int vs high: 0.62 (0.46-0.84) Low vs high: 0.33 (0.22-0.49) 0.60 CI, confidence interval; FRS, recurrence free survival. Slide credit: clinicaloptions.com Galon J, et al. ASCO 2016. Abstract 3500.

Immunoscore in Colon Cancer: Secondary Objectives Efficacy Endpoints Outcome High Intermediate Low 5-yr time to recurrence, % (95% CI) 69.0 (65.2-72.9) 80.6 (78.3-83.0) 88.9 (86.3-91.6) HR (95% CI) P value Int vs high: 0.58 (0.48-0.71) Low vs high: 0.29 (0.21-0.38) < .0001 5-yr DFS, % (95% CI) 57.6 (53.8-61.7) 69.2 (66.6-71.9) 75.4 (72.0-79.0) Int vs high: 0.69 (0.60-0.80) Low vs high: 0.52 (0.43-0.62) 5-yr OS, % (95% CI) 66.9 (63.4-70.7) 77.3 (75.0-79.7) 81.5 (78.5-84.6) Int vs high: 0.73 (0.63-0.85) Low vs high: 0.59 (0.49-0.71) Slide credit: clinicaloptions.com Galon J, et al. ASCO 2016. Abstract 3500.

Immunoscore in Colon Cancer: Conclusions Primary endpoint reached: TTR significantly longer in pts with high Immunoscore at all stages Significance remained in multivariate analysis in all cohorts (TS, IVS, EVS) Immunoscore predicts TTR, DFS, and OS Base on findings, authors suggest that Immunoscore may be incorporated as new classification for cancer, TNM- Immune (TNM-I) First standardized immune-based assay for classifying cancer, which authors suggest may help to individualize immunotherapy in the future EVS, external validation set; DFS, disease-free survival; IVS, internal validation set; TS, training set; TTR, time to recurrence. Slide credit: clinicaloptions.com Galon J, et al. ASCO 2016. Abstract 3500.

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