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2  2014 Genentech USA, Inc. MBoC Program Common genetic and molecular alterations in lung cancer Point mutation Single base pair mutation in the EGFR gene Chromosome 7 Evasion of immune destruction Translocations in chromosome 2p Amplification of EGFR Signaling pathway alterations A point mutation in the epidermal growth factor receptor (EGFR) gene (L858R) can result in abnormal kinase activation of the EGFR protein 1 Translocations within chromosome 2p result in the fusion of echinoderm microtubule-associated protein-like 4 (EML4) to anaplastic lymphoma kinase (ALK); this mutation has potent oncogenic activity in lung cancer 2 Amplification of a region on chromosome 7—which encodes the EGFR gene—can result in overexpression of this gene 3 Mutation, amplification, deletion, methylation, or post-translational modification along the PI3K/AKT/mTOR signaling pathway can dysregulate cell growth and proliferation 4 Interaction between pathways, such as cross-talk through numerous points of convergence between the PI3K/AKT/mTOR and RAS/RAF/MEK signaling cascades, enable cell growth, division, survival, and the development of therapeutic resistance in lung cancer 4 By the time tumors are detected in the clinic, tumor cells have already evolved mechanisms of dysregulating the immune response and escaping destruction 5 References: 1.Yatabe Y, Mitsudomi T. Epidermal growth factor receptor mutations in lung cancers. Pathol Int. 2007;57: Shaw AT, Yeap BY, Mino-Kenudson M, et al. Clinical features and outcome of patients with non–small-cell lung cancer who harbor EML4-ALK. J Clin Oncol. 2009;27: Hirsch FR, Varella-Garcia M, Bunn PA Jr, et al. Epidermal growth factor receptor in non–small-cell lung carcinomas: correlation between gene copy number and protein expression and impact on prognosis. J Clin Oncol. 2003;21: Heavey S, O’Byrne KJ, Gately K. Strategies for co-targeting the PI3K/AKT/mTOR pathway in NSCLC. Cancer Treat Rev. 2014;40: Kirkwood JM, Butterfield LH, Tarhini AA, Zarour H, Kalinski P, Ferrone S. Immunotherapy of cancer in CA Cancer J Clin. 2012;62: Notes

3  2014 Genentech USA, Inc. MBoC Program EML4-ALK fusion mediates aberrant ALK activation ALK=anaplastic lymphoma kinase; EML4=echinoderm microtubule-associated protein-like 4; Grb2=growth factor receptor-bound protein-2; MAPK=mitogen-activated protein kinase; MEK=mitogen-activated protein kinase kinase; PI3K=phosphatidylinositol 3-kinase; Shc=Src homology 2 domain containing transforming protein 1; Sos=son of sevenless. Camidge DR, Doebele RC. Nat Rev Clin Oncol. 2012;9: ProliferationSurvival MEK Raf Sos Grb2 Shc RAS PI3K MAPK AKT ↓ Apoptosis Inversion The ALK gene is located on chromosome 2 and codes a transmembrane receptor TK in the insulin receptor superfamily Expression of native ALK in adult human tissues seems to be restricted to the small intestine, testes, and nervous system, and some evidence suggests it plays a role in neurological development Under normal conditions, activation of ALK occurs via ligand-induced dimerization, which leads to autophosphorylation Activation of downstream signaling pathways—including Janus kinase (JAK)/signal transducer and activator of transcription (STAT), phosphatidylinositol 3-kinase (PI3K)/AKT, and mitogen-activated protein kinase (MAPK)—leads to cell proliferation and differentiation When fused with EML4, it can lead to constitutive activation, even without the presence of ligands Reference: Camidge DR, Doebele RC. Treating ALK-positive lung cancer—early successes and future challenges. Nat Rev Clin Oncol. 2012;9: Notes

4  2014 Genentech USA, Inc. MBoC Program AKT PDK1 PI3K Cyclin D1p27BADGSK3ß NFκB mTOR MAPK Targeting the EGFR pathway with SMIs: Preventing phosphorylation events Cell-cycle control ↓ Apoptosis Ras Sos Grb2Shc MEK Raf ↑ Survival EGFRHER3 ProliferationAngiogenesis EGFR Sos Grb2Shc EGFR BAD=Bcl-2–associated death promoter; EGFR=epidermal growth factor receptor; Grb2=growth factor receptor-bound protein 2; GSK3β=glycogen synthase kinase 3 beta; HER3=human epidermal growth factor receptor-3; MAPK=mitogen-activated protein kinase; MEK=mitogen-activated protein kinase kinase; mTOR=mammalian target of rapamycin; NFκB=nuclear factor kappa–light-chain enhancer of activated B cells; PDK1=phosphoinositide-dependent kinase-1; PI3K=phosphatidylinositol 3-kinase; Raf=rapidly accelerating fibrosarcoma; Ras=rat sarcoma; SMI=small molecule inhibitor; Shc=Src homology 2 domain containing transforming protein 1; Sos=son of sevenless. Olayioye MA, et al. EMBO J. 2000;19: Members of the HER family are established therapeutic targets Additional therapeutic strategies are being developed that target HER family receptors and intracellular signaling proteins Small-molecule tyrosine kinase inhibitors (TKIs) that prevent signal transduction via the TK domain of the receptor are one mechanism for targeting receptors Other small-molecule inhibitors target intracellular signaling intermediates Reference: Rowinsky EK. The ErbB family: targets for therapeutic development against cancer and therapeutic strategies using monoclonal antibodies and tyrosine kinase inhibitors. Annu Rev Med. 2004;55: Notes