1. MUC1 Is Associated With Diverse Cellular Functions in Normal Epithelial Cells MUC1 (Mucin 1) Plays a Central Role In Cellular Functions of Both Normal Epithelial Cells and Cancer Cells Role of MUC1 in the Growth, Proliferation and Survivals of Cancer Cells MUC1 Is a Scientifically Valid Target for Mounting Immune Responses HOME  PREVNEXT  © 2013 EMD Serono. All rights reserved. Touch here to begin What Role Does MUC1 Play in the Growth, Proliferation and Survival of Cancer Cells? © 2013 EMD Serono. All rights reserved.

2. MUC1 Is Associated With Diverse Cellular Functions in Normal Epithelial Cells MUC1 (Mucin 1) Plays a Central Role In Cellular Functions of Both Normal Epithelial Cells and Cancer Cells Role of MUC1 in the Growth, Proliferation and Survivals of Cancer Cells MUC1 Is a Scientifically Valid Target for Mounting Immune Responses HOME  PREVNEXT  © 2013 EMD Serono. All rights reserved. What Is the Role of MUC1 in Normal Epithelial Cells? © 2013 EMD Serono. All rights reserved. Touch here to begin

3. MUC1 Is Associated With Diverse Cellular Functions in Normal Epithelial Cells MUC1 (Mucin 1) Plays a Central Role In Cellular Functions of Both Normal Epithelial Cells and Cancer Cells Role of MUC1 in the Growth, Proliferation and Survivals of Cancer Cells MUC1 Is a Scientifically Valid Target for Mounting Immune Responses HOME  PREVNEXT  © 2013 EMD Serono. All rights reserved. What Role May MUC1 Play in the Growth, Proliferation and Survival of Cancer Cells? © 2013 EMD Serono. All rights reserved. Touch here to begin

4. MUC1 Is Associated With Diverse Cellular Functions in Normal Epithelial Cells MUC1 (Mucin 1) Plays a Central Role In Cellular Functions of Both Normal Epithelial Cells and Cancer Cells Role of MUC1 in the Growth, Proliferation and Survivals of Cancer Cells MUC1 Is a Scientifically Valid Target for Mounting Immune Responses HOME  PREVNEXT  © 2013 EMD Serono. All rights reserved. Why Is MUC1 Considered a Scientifically Valid Target for Cancer Immunotherapy? © 2013 EMD Serono. All rights reserved. Touch here to begin

5. MUC1 Is Associated With Diverse Cellular Functions in Normal Epithelial Cells MUC1 (Mucin 1) Plays a Central Role In Cellular Functions of Both Normal Epithelial Cells and Cancer Cells Role of MUC1 in the Growth, Proliferation and Survivals of Cancer Cells MUC1 Is a Scientifically Valid Target for Mounting Immune Responses HOME  PREVNEXT  MUC1 Plays a Central Role in Cellular Functions in Epithelial Cells and Cancer Cells Normally glycosylated MUC1 MUC1 is heavily glycosylated and expressed on the apical surface of secretory epithelial cells 1,2 Cancer-associated MUC1 Cancer-associated MUC1 is underglycosylated and overexpressed on cancer cells 1-4 1. Ho SB, et al. 1993;53:641-651. 2. Kufe DW. 2009;9(12):874-885. 3. Hollingsworth MA, et al. 2004;4:45-60. 4. Singh PK, et al. 2006;16:467-476. © 2013 EMD Serono. All rights reserved. MUC1 (Mucin 1) Plays a Central Role In Cellular Functions of Both Normal Epithelial Cells and Cancer Cells HOME NEXTNEXT 

6. MUC1 Is Associated With Diverse Cellular Functions in Normal Epithelial Cells MUC1 (Mucin 1) Plays a Central Role In Cellular Functions of Both Normal Epithelial Cells and Cancer Cells Role of MUC1 in the Growth, Proliferation and Survivals of Cancer Cells MUC1 Is a Scientifically Valid Target for Mounting Immune Responses HOME  PREVNEXT  MUC Is Overexpressed in Many Cancers 1. Zhong XY, et al. 1993;139:281-286. 2. Data on file. EMD Serono, Inc. 3. Rakha EA, et al. 2005;18:1295-1304. 4. Langner C, et al. 2004;17:180-188. 5. Chauhan SC, et al. 2006;19:1386-1394. 6. Croce MV, et al. 2001;7:284-291. 7. Baldus SE, et al. 2002;40:440-449. 8. Qu CF, et al. 2004;91:2086- 2093. 9. DeNardo SJ, et al. 2005;11:7187s-7194s. 10. Utsunomiya T, et al. 1998;4:2605-2614. 11. Saad RS, et al. 2005;32:156-159. 12. Cloosen S, et al. 2006;135:513-516. 13. Kijima H, et al. 2001;21:1285-1289. © 2013 EMD Serono. All rights reserved. MUC1 (Mucin 1) Plays a Central Role In Cellular Functions of Both Normal Epithelial Cells and Cancer Cells  PREVPREVHOME NEXTNEXT 

7. MUC1 Is Associated With Diverse Cellular Functions in Normal Epithelial Cells MUC1 (Mucin 1) Plays a Central Role In Cellular Functions of Both Normal Epithelial Cells and Cancer Cells Role of MUC1 in the Growth, Proliferation and Survivals of Cancer Cells MUC1 Is a Scientifically Valid Target for Mounting Immune Responses HOME  PREVNEXT  What Is MUC1? MUC1 is a large transmembrane glycoprotein found on the apical surface of most secretory epithelial cells 1. Ho SB, et al. 1993;53:641-651. © 2013 EMD Serono. All rights reserved. MUC1 Is Associated With Diverse Cellular Functions in Normal Epithelial Cells  PREVPREVHOME NEXTNEXT 

8. MUC1 Is Associated With Diverse Cellular Functions in Normal Epithelial Cells MUC1 (Mucin 1) Plays a Central Role In Cellular Functions of Both Normal Epithelial Cells and Cancer Cells Role of MUC1 in the Growth, Proliferation and Survivals of Cancer Cells MUC1 Is a Scientifically Valid Target for Mounting Immune Responses HOME  PREVNEXT  What Is the Structure of MUC1? MUC1 is comprised of three domains: 1-3 –Entracellular domain –Transmembrane domain –Cytoplasmic domain (or tail) The extra cellular domain of MUC1 is characterized by many tandem repeat peptides that are extensively glycosylated 1-3 1. Ho SB, et al. 1993;53:641-651. 2. Kufe DW. 2009;9(12):874-885. 3. Hollingsworth MA, et al. 2004;4:45-60. Extracellular domain Transmembrane domain Intracellular domain © 2013 EMD Serono. All rights reserved. MUC1 Is Associated With Diverse Cellular Functions in Normal Epithelial Cells  PREVPREVHOME NEXTNEXT 

9. MUC1 Is Associated With Diverse Cellular Functions in Normal Epithelial Cells MUC1 (Mucin 1) Plays a Central Role In Cellular Functions of Both Normal Epithelial Cells and Cancer Cells Role of MUC1 in the Growth, Proliferation and Survivals of Cancer Cells MUC1 Is a Scientifically Valid Target for Mounting Immune Responses HOME  PREVNEXT  MUC1 Protects Normal Epithelial Cells as Part of the Innate Immune System A central role of MUC1 is to maintain the integrity of the epithelial barrier 1 Extracellular MUC1 forms a protective layer on the apical epithelial cell surface that: 1-3 –Provides lubrication –Acts as physical barrier against pathogens and toxins 1. Hollingsworth MA, et al. Nature Rev Cancer. 2004;4:45-60. 2. Ho SB, et al. Cancer Res. 1993;53:641-651. 3. Kufe DW. Nat Rev Cancer. 2009;9(12):874-885. © 2013 EMD Serono. All rights reserved.  PREVPREV MUC1 Is Associated With Diverse Cellular Functions in Normal Epithelial Cells HOME NEXTNEXT 

10. MUC1 Is Associated With Diverse Cellular Functions in Normal Epithelial Cells MUC1 (Mucin 1) Plays a Central Role In Cellular Functions of Both Normal Epithelial Cells and Cancer Cells Role of MUC1 in the Growth, Proliferation and Survivals of Cancer Cells MUC1 Is a Scientifically Valid Target for Mounting Immune Responses HOME  PREVNEXT  MUC1 Is Involved in Intracellular Signaling Pathways Which Regulate Epithelial Cell Growth, Proliferation and Survival The extracellular domain of MUC1 is a sensor of the external environment through ligand binding and as a consequence of changes in biochemical conditions outside the cell 1,2 Signals are transmitted by conformational changes mediated by the extracellular domain to the intracellular domain 1-3 The intracellular domain in turn activates several intracellular signaling pathways that promote the growth, proliferation and survival of normal epithelial cells 1-3 1. Hollingsworth MA, et al. Nature Rev Cancer. 2004;4:45-60. 2. Ho SB, et al. Cancer Res. 1993;53:641-651. 3. Kufe DW. Nat Rev Cancer. 2009;9(12):874-885. Key extracellular molecules interacting with MUC1: ICAM-1 galectin-3 EGFR Mesothelin © 2013 EMD Serono. All rights reserved. Key intracellular pathways activated by MUC1 PI3K/AKT/mTOR IKKB/NF-kB β-catenin/Wnt RAS/RAF/MEK/ MAPK (ERK 1/2) MUC1 Is Associated With Diverse Cellular Functions in Normal Epithelial Cells  PREVPREVHOME NEXTNEXT 

11. MUC1 Is Associated With Diverse Cellular Functions in Normal Epithelial Cells MUC1 (Mucin 1) Plays a Central Role In Cellular Functions of Both Normal Epithelial Cells and Cancer Cells Role of MUC1 in the Growth, Proliferation and Survivals of Cancer Cells MUC1 Is a Scientifically Valid Target for Mounting Immune Responses HOME  PREVNEXT  Role of MUC1 in the Growth, Proliferation and Survival of Cancer Cells © 2013 EMD Serono. All rights reserved. Cancer-associated MUC1 Cancer-associated MUC1 is underglycosylated and overexpressed on cancer cells 1-4 1. Ho SB, et al. 1993;53:641-651. 2. Kufe DW. 2009;9(12):874-885. 3. Hollingsworth MA, et al. 2004;4:45-60. 4. Singh PK, et al. 2006;16:467-476.  PREVPREVHOME NEXTNEXT  Role of MUC1 in the Growth, Proliferation and Survivals of Cancer Cells

12. MUC1 Is Associated With Diverse Cellular Functions in Normal Epithelial Cells MUC1 (Mucin 1) Plays a Central Role In Cellular Functions of Both Normal Epithelial Cells and Cancer Cells Role of MUC1 in the Growth, Proliferation and Survivals of Cancer Cells MUC1 Is a Scientifically Valid Target for Mounting Immune Responses HOME  PREVNEXT  Cancer-associated MUC1 Promtotes the Growth, Proliferations and Survival of Cancer Cells Malignant transformation leads to: 1,2 –Irreversible loss of epithelial cell polarity –Aberrant expression of MUC1 over the entire cell surface –Over expression and underglycosylation of MUC1 © 2013 EMD Serono. All rights reserved.  PREVPREVHOME NEXTNEXT  Role of MUC1 in the Growth, Proliferation and Survivals of Cancer Cells 1. Hollingsworth MA, et al. 2004;4:45-60. 2. Singh PK, et al. 2006;16:467-476.

13. MUC1 Is Associated With Diverse Cellular Functions in Normal Epithelial Cells MUC1 (Mucin 1) Plays a Central Role In Cellular Functions of Both Normal Epithelial Cells and Cancer Cells Role of MUC1 in the Growth, Proliferation and Survivals of Cancer Cells MUC1 Is a Scientifically Valid Target for Mounting Immune Responses HOME  PREVNEXT  Cancer-associated MUC1 Promotes the Growth, Proliferation and Survival of Cancer Cells Aberrant distribution and overexpression of cancer-associated MUC1 activate both cell surface molecules and intracellular signaling pathways which promote the growth, proliferation and survival of cancer cells 1-4 © 2013 EMD Serono. All rights reserved.  PREVPREVHOME NEXTNEXT  NEXTNEXT  Role of MUC1 in the Growth, Proliferation and Survivals of Cancer Cells 1. Hollingsworth MA, et al. 2004;4:45-60. 2. Singh PK, et al. 2006;16:467-476. 3. Pochampalli MR, et al. 2007;26:1693-1701. 4. Bafna S, et al. 2010;29:2893-2904.

14. MUC1 Is Associated With Diverse Cellular Functions in Normal Epithelial Cells MUC1 (Mucin 1) Plays a Central Role In Cellular Functions of Both Normal Epithelial Cells and Cancer Cells Role of MUC1 in the Growth, Proliferation and Survivals of Cancer Cells MUC1 Is a Scientifically Valid Target for Mounting Immune Responses HOME  PREVNEXT  Cancer-associated MUC1 Is Underglycosylated Compared to Glycosylated on Normal Epithelial Cells Cancer-associated MUC1 is underglycosylated compared with MUC1 expressed on normal epithelial cells 1 –Both the length and number of carbohydrate chains are reduced 1,2 © 2013 EMD Serono. All rights reserved.  PREVPREVHOME NEXTNEXT  NEXTNEXT  Role of MUC1 in the Growth, Proliferation and Survivals of Cancer Cells Abnormal carbohydrate chain Cancer-associated MUC1 Normally glycosylated MUC1 Core peptide Normal carbohydrate chain 1. Ho SB, et al. 1993;53:641-651. 2. Taylor-Papadimitriou J, et al. 1994;12(6):227-233.

15. MUC1 Is Associated With Diverse Cellular Functions in Normal Epithelial Cells MUC1 (Mucin 1) Plays a Central Role In Cellular Functions of Both Normal Epithelial Cells and Cancer Cells Role of MUC1 in the Growth, Proliferation and Survivals of Cancer Cells MUC1 Is a Scientifically Valid Target for Mounting Immune Responses HOME  PREVNEXT  Cancer-associated MUC1 Is a Scientfically Valid Target for Immune Responses Against Cancers © 2013 EMD Serono. All rights reserved. MUC1 Is a Scientifically Valid Target for Mounting Immune Responses  PREVPREVHOMENEXTNEXT  1. Hollingsworth MA, et al. 2004;4:45-60. 2. Singh PK, et al. 2006;16:467-476. 3. Pochampalli MR, et al. 2007;26:1693-1701. 4. Bafna S, et al. 2010;29:2893-2904. MUC1-bound HLA class I molecule Cancer-associated MUC1

16. MUC1 Is Associated With Diverse Cellular Functions in Normal Epithelial Cells MUC1 (Mucin 1) Plays a Central Role In Cellular Functions of Both Normal Epithelial Cells and Cancer Cells Role of MUC1 in the Growth, Proliferation and Survivals of Cancer Cells MUC1 Is a Scientifically Valid Target for Mounting Immune Responses HOME  PREVNEXT  Dense Glycosylation of MUC1 Inhibits Antigen Processing by Normal Epithelial Cells In normal epithelial cells, the dense glycosylation and long carbohydrate chain of MUC1 interfere with antigen processing and results in a compromised presentation of MUC1 peptides bound to human leukocyte antigen (HLA) class I molecules 1,2 © 2013 EMD Serono. All rights reserved. MUC1 Is a Scientifically Valid Target for Mounting Immune Responses  PREVPREVHOMENEXTNEXT  1. Werdelin O, et al. 2002;99(15):9611-9613. 2. Karsten U, et al. 2004;14(8):681-692. Epthielial cell Normally glycosylated MUC1 HLA class I molecule HLA complex transported via Golgi to cell surface Peptides are combined With HLA molecules Inside the ER Processed peptides are transported Into the ER Endoplasmic reticulum (ER) Normally glycosylated MUC1

17. MUC1 Is Associated With Diverse Cellular Functions in Normal Epithelial Cells MUC1 (Mucin 1) Plays a Central Role In Cellular Functions of Both Normal Epithelial Cells and Cancer Cells Role of MUC1 in the Growth, Proliferation and Survivals of Cancer Cells MUC1 Is a Scientifically Valid Target for Mounting Immune Responses HOME  PREVNEXT  Underglycosylation of Cancer-associated MUC1 Facilitates Antigen Processing by CancerCells In cancer cells, the underglycosylated cancer-associated MUC1 is more efficiently processed leading to a much larger repertoire of MUC1-bound HLA class I molecules on cancer cells 3,5 © 2013 EMD Serono. All rights reserved. MUC1 Is a Scientifically Valid Target for Mounting Immune Responses  PREVPREVHOMENEXTNEXT  3. Hiltbold EM, et al. 1999;194:143-149. 4. Purcell AW, et al. 2008;86(7):574-579. 5. Hanish FG, et al. 2003;33:3242-3254. Cancer cell Cancer-associated MUC1 HLA class I molecule HLA complex transported via Golgi to cell surface Peptides are combined With HLA molecules Inside the ER Processed peptides are transported Into the ER Endoplasmic reticulum (ER) Cancer-associated MUC1

18. MUC1 Is Associated With Diverse Cellular Functions in Normal Epithelial Cells MUC1 (Mucin 1) Plays a Central Role In Cellular Functions of Both Normal Epithelial Cells and Cancer Cells Role of MUC1 in the Growth, Proliferation and Survivals of Cancer Cells MUC1 Is a Scientifically Valid Target for Mounting Immune Responses HOME  PREVNEXT  Intracellular Processing of Cancer-Associated Leads to the Presentation of a Novel Epitope Repertoire on HLA Class I Molecules on Cancer Cells Intracellular antigen processing of cancer-associated MUC1 by cancer cells 1–3 : Yields a novel and large T cell epitope repertoire of antignes bound to HLA class I molecules Allows for recognition of cancer cells by MUC1-specific cytotoxic T lymphocytes It is therefore reasonable to speculate that anti-tumor T Cell specific responses against the underglycosylated form of MUC1 bound to HLA class I molecules would be selectively directed at cancer cells. © 2013 EMD Serono. All rights reserved. MUC1 Is a Scientifically Valid Target for Mounting Immune Responses  PREVPREVHOMENEXTNEXT  3. Hiltbold EM, et al. 1999;194:143-149. 4. Purcell AW, et al. 2008;86(7):574-579. 5. Hanish FG, et al. 2003;33:3242-3254.

19. MUC1 Is Associated With Diverse Cellular Functions in Normal Epithelial Cells MUC1 (Mucin 1) Plays a Central Role In Cellular Functions of Both Normal Epithelial Cells and Cancer Cells Role of MUC1 in the Growth, Proliferation and Survivals of Cancer Cells MUC1 Is a Scientifically Valid Target for Mounting Immune Responses HOME  PREVNEXT  MUC1 Is a Scientifically Valid Target for Enhancing Immune responses Against Cancers MUC1 is considered a valid target for cancer immunotherapy 1 NCI objective criteria used to evaluate potentially useful cancer antigens included: 1 –Therapeutic function –Immunogenicity –Specificity –Oncogenicity –Number of patients with antigen positive cancers –Expression level and percent positive cells –Stem cell expression –Number of epitopes –Cellular location of expression © 2013 EMD Serono. All rights reserved. MUC1 Is a Scientifically Valid Target for Mounting Immune Responses  PREVPREVHOMENEXTNEXT  1. Cheever MA, et al. 2009;15(17):5323-5337.