Targeted immunotherapy: unleashing the immune system against cancer

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Targeted immunotherapy: unleashing the immune system against cancer Suzanne L. Topalian, MD Johns Hopkins University School of Medicine Sidney Kimmel Comprehensive Cancer Center 2013 MMS Annual Education Program: Navigating the Currents of Change

Disclosure Information Suzanne L. Topalian, MD I have the following financial relationships to disclose: Consultant for: Bristol-Myers Squibb (uncompensated), Jounce Therapeutics, Sanofi, and Amplimmune (spouse) Research grant support from Bristol-Myers Squibb Royalties through institution: Amplimmune (spouse), Bristol-Myers Squibb (spouse)

Cancer is a genetic disease rationale for personalized medicine limitation: rapid resistance Cancer is an immunological disorder immunotherapy is a common denominator that can activate immune responses against mutant proteins the adaptable immune system can keep pace with tumor evolution

The adaptive immune system: an ideal anti-cancer agent Diversity T cells - 10 18 Antibodies - 10 22 Tumor cell Specificity Can distinguish minute chemical alterations T cell Memory After effective antigen priming, immunity can last for decades

Immune tolerance: Multiple barriers to tumor elimination Regulatory immune cells T-cell inhibitory receptors T reg cells CTLA-4, PD-1, LAG-3, etc. MDSCs Immunosuppressive cytokines IL-6, IL-10, TGF-ß, VEGF Solution: adoptive transfer of T cells optimized/engineered ex vivo Solution: systemic blockade of suppressive cytokines or inhibitory receptors/ligands ( immune checkpoints )

Advances in Immunotherapy 2011 Anti-CTLA-4 (ipilimumab) was approved for treating metastatic melanoma based on improved overall survival in a randomized study. However, the grade 3-4 drug-related toxicity rate approximated the clinical benefit rate Adapted from Hodi et al, NEJM 2010 Control Ipilimumab

Antigen presenting or tumor cell The PD-1/PD-L1 pathway: New strategies for immune checkpoint blockade T cell PD-L1 is expressed by many human tumors and may play a pivotal role in local cancer immunosuppression Pardoll 2012

APC Role of PD-1 in suppressing antitumor immunity B7.1 CD28 T cell Activation (cytokines, lysis, prolif., migration) MHC-Ag Tumor TCR Signal 1 PD-1 PD-L1 Inhibition (anergy, exhaustion, death) (-) (-) Tumor (-)

APC Role of PD-1 in suppressing antitumor immunity B7.1 CD28 T cell Activation (cytokines, lysis, prolif., migration) MHC-Ag Tumor TCR Signal 1 PD-1 PD-L1 Inhibition (anergy, exhaustion, death) (-) (-) Tumor (-) Anti- PD-1

CTLA-4 vs. PD-1: Distinct immune checkpoints B7.1/2 CD28 APC signal 1 CTLA-4 to surface APC + signal 1 Costim. ligand Naïve/resting T cell Costim. receptor CTLA-4 Experienced T cell APC signal 1 Traffic to periphery Tissue signal 1 T cell priming Topalian et al., Curr Opin Immunol 2012 PD-L1 PD-1

Nivolumab (anti-pd-1) (BMS-936558, MDX-1106, ONO-4538) Fully human IgG4 anti-human PD-1 blocking mab High affinity for PD-1 (K D ~ 3 nm), blocks binding to both known ligands: PD-L1 (B7-H1, broadly inducible) and PD-L2 (B7-DC, selective for APCs) Favorable safety profile and preliminary evidence of clinical activity in patients with treatment-refractory solid tumors in a first-in-human study (Brahmer et al., J Clin Oncol 2010)

Two early phase clinical trials of anti-pd-1 (nivolumab) in patients with advanced solid tumors (Brahmer et al., JCO 2010; Topalian et al., NEJM 2012) 1 st Treatment Cycle Follow Up or Additional Treatment Cycle(s) Day 1 Dose 0.3-10 mg/kg Day 57 Scans Day 85 Scans 2 years or until CR/PR or PD 1 st Treatment Cycle Follow Up or Additional Treatment Cycle(s) Days 1, 15, 29 43 Dose 0.1-10 mg/kg Day 57 Scans 2 years or until CR or PD Eligible patients: treatment-refractory metastatic lung cancer, melanoma, kidney, colon, or prostate cancer

CRC Durable anti-pd-1 responses OFF THERAPY: CR Stop Rx immune memory (Lipson et al., CCR 2013) Latest eval.: CR 0 1 yr 2 yr 3 yr 4 yr RCC Stop Rx Best resp.(pr) CR Latest eval.: CR 0 1 yr 2 yr 3 yr 4 yr 5 yr MEL Stop Best Rx resp.(pr) New LN met Resume Rx PR Latest eval. PR 0 1 yr 2 yr 3 yr 4 yr 5 yr

Clinical activity of anti-pd-1 (nivolumab) Tumor Type Dose (mg/kg) No. pts ORR (CR/PR) No. pts (%) SD 24 wk No. pts (%) NSCLC 1-10 122 20 (16) 11 (9) MEL 0.1-10 106 33 (31) 6 (6) RCC 1 or 10 34 10 (29) 9 (27) 294 patients started therapy between 2008-2012 and had 6 mo. follow-up. No ORs in CRC (n=19) or CRPC (n=17) 28/54 responses lasted 1 yr in patients with 1 yr follow-up Toxicity: grade 3-4 drug-related AEs15%, mortality 1%. Topalian et al., ESMO 2012

Change in Target Lesions from Baseline (%) Anti-PD-1 response kinetics and durable tumor control off therapy 100 90 80 70 60 50 40 30 20 10 0-10 -20-30 -40-50 -60-70 -80-90 -100 Maximum duration of therapy 0 10 20 30 40 50 60 70 80 90 100 110 120 130 140 Weeks Since Treatment Initiation Melanoma 1 mg/kg First occurrence of new lesion On study Off study

Partial response of locally advanced primary melanoma to anti-pd-1 Pre Tumor Lymphs 2 mo. 35-year-old patient had disease progression after surgery and IL-2. Response to anti-pd-1 ongoing at 23 months.

Partial regression of metastatic kidney cancer in response to anti-pd-1 Pretreatment 6 months 57-year-old patient developed progressive disease after receiving sunitinib, temsirolimus, sorafenib, and pazopanib Currently in cycle 12 anti-pd-1 therapy (~23 months) with ongoing PR

Response of a non-immunogenic tumor to anti-pd-1: Squamous cell lung cancer Pre-Rx 2 months 12 months History: 61-year-old patient with stage IV NSCLC refractory to multiple surgeries, RT, 2 multidrug chemotherapy regimens, and epigenetic therapy (5-AZA and entinostat). Recently completed 2 yrs anti-pd-1 therapy.

Drug-related AEs of special interest in 3% of 304 patients receiving anti-pd-1 AEOSI ( irae ) All grades (n, %) Grade 3-4 (n, %) Any 138 (45) 18 (6) Rash 41 (14) 0 Diarrhea 36 (12) 3 (1) Pruritis 31 (10) 1 (0.3) ALT 13 (4) 2 (1) AST 11 (4) 2 (1) TSH 11 (4) 1 (0.3) Pneumonitis* 10 (3) 3 (1) Vitiligo 9 (3) 0 Infusion reaction 8 (3) 0 *There were 3 (1%) deaths in patients with pneumonitis (2 NSCLC, 1 CRC). AEOSIs in 1% of pts included colitis, hepatitis, hypophysitis, nephritis and thyroiditis. Analysis as of July 2012. Topalian et al, ESMO 2012

Partial response of metastatic mucosal melanoma to anti-pd-1, associated with nephritis Pre-Rx 8 months Prior therapies included multiple surgeries, RT and temozolomide. Nephritis developed after 8 months of anti- PD-1 therapy, associated with administration of radiographic contrast dye.

Nephritis associated with anti-pd-1 therapy: serologic and cellular immune components H&E, 400 X CD3 tubule glomerulus plasma cell intraepithelial T cell

B7.1? antiapoptotic signal in tumor cells PD-L1 PD-L2 Anti-PD-1 PD-1 (-) signal Anti-PD-L1 B7.1? antiapoptotic signal in tumor cells PD-L1 PD-L2 PD-1 (-) signal

If PD-1 pathway blockade acts locally in the tumor immune microenvironment. Then it follows that the tumor site may hold the key to optimizing anti-pd-1/pd-l1 therapy: Biomarkers to select patients most likely to respond to therapy Early indicators of treatment outcomes Identification of resistance pathways and development of synergistic combinatorial therapies

Proportion of patients Preliminary molecular marker studies: Correlation of PD-L1 expression in pretreatment tumor biopsies with clinical response to anti-pd-1 1 0.8 0.6 0.4 0.2 0 13/31 18/31 PD-L1(+) 0/18 18/18 PD-L1(-) CR/PR PD-L1(+) NR PD-L1(-) p=0.002 49 patients include 20 with melanoma,13 NSCLC, 7 colon, 6 kidney, and 3 prostate cancer. * Melanoma Kidney Lung * Normal renal glomerulus

No. patients Correlation of PD-L1 expression with tumor type in 49 patients treated with anti-pd-1 18 16 14 12 10 8 6 4 2 0 0 MEL NSCLC RCC CRC CRPC Responders/total: 8/20 3/13 2/6 0/7 0/3 PD-L1(+) PD-L1(-) Patients were PD-L1+ if 5% of tumor cells in any tumor biopsy expressed cell surface PD-L1, using mab 5H1 (L. Chen) and manual staining technique.

2 Mechanisms for PD-L1 up-regulation in tumors Innate Resistance TUMOR Oncogenic Pathway MHC-pep TCR PD-1 PD-L1 T Cell Constitutive tumor signaling induces PD-L1 on tumor cells Adaptive Resistance PD-L1 expression reflects immune reaction TUMOR T Cell TUMOR T Cell Stats IFN-g

Focal PD-L1 expression in melanoma: geographic co-localization with TILs creates a shield against immune attack Tumor Lymphs

% of cases % survival PD-L1 expression in melanoma correlates with the presence of TILs and increased overall survival in patients with metastatic disease 100 80 60 TILs No TILs p<0.0001 p=0.032 PD-L1(+) PD-L1(-) 40 20 0 PD-L1(+), n=57 PD-L1(-), n=93 Duration follow-up (mo) TILs are necessary but not sufficient for PD-L1 expression (Taube et al., Science Transl Med 2012)

# patients PD-L1 tumor PD-L1 expression on tumor cells and TAMs in oropharyngeal SCCHN: association with CD8+ TILs, IFN-g and HPV+ tumors CD68 p=0.003 p=0.002 CD3 15 10 5 0 HPV+ (n=20) HPV- (n=7) PD-L1+ PD-L1- Lyford-Pike, Pai, et al., Cancer Res 2013

Survival probability # Tumors PD-L1 tumor PD-L1 expression on tumor cells and TAMs in Merkel cell Ca: association with CD8+ TILs, MCV, and overall survival stroma CD68 20 10 0 MCV+ (n=34) 0 MCV- (n=8) PD-L1+ PD-L1- p=0.01 CD8 Duration of follow-up (years) Lipson, Taube, et al., Cancer Immunol Res 2013

PD-1 pathway in virus-associated cancers: at the crossroads of cancer immunology and microbial immunology Principles of PD-1/PD-L1 mechanism-of-action first demonstrated in preclinical models of infectious disease PD-1 pathway restrains host immune responses against chronic infection Virus-associated cancers account for a large proportion of cancer deaths worldwide PD-1 blockade now in the clinic for virus-associated hepatocellular cancer, in planning for HIV

Potential PD-1 pathway interactions in chronic HIV infection Increased PD-L1 expression on APCs in HIV infection APC PD-1 PD-L1/2?? CD4 T cell HIV+ PD-1 and other coinhibitory molecules are expressed on HIV specific CD8 T cells (-) (-) (-) CD8 T cell CD4 T cell HIV+?? PD-1+ CD4 T cells might be an important latent viral reservoir

Dissecting mechanisms and resistance pathways: Immune infiltrates at the boundary of PD-L1+ melanoma cells PD-L1 H&E H&E IFN-g mrna is overexpressed in PD-L1(+) vs. PD-L1(-) melanomas (Taube et al., Science Transl Med 2012) Gene expression profiling reveals inflammatory signature in PD-L1(+) tumors

-log 10 p value Gene expression profile of TILs from PD-L1(+) vs. (-) melanomas reveals functional groups of differentially expressed genes and potential bypass pathways IL-22R -4.4x CCL5 2.6x PRF1 3.1x CD8a 2.4x IL-10 3.3x TLR3 4.6x IL-21 4.1x PD-1 7.3x CXCL1 8.3x IFN-g 11.3x PD-L1 4.4x LAG-3 10.7x Th1 CD8 T Checkpoint log 2 fold change multiplex qrt-pcr, CD45 normalization

Association of lymphocyte activation gene-3 (LAG-3)+ lymphocytes with PD-L1+ melanoma cells H&E PD-L1 LAG-3 IHC tumor lymphs Young, Taube et al., AACR 2013 abstr. #465 ISH Meeker, Taube, Advanced Cell Diagnostics

Tumor Volume (mm3) Synergistic anti-tumor effects of dual checkpoint blockade: anti-pd-1 and anti-lag-3 in a murine tumor model Control Ab (0/10 TF) Anti- LAG-3 (0/10 TF) Anti-PD-1 (4/10 TF) 2000 Anti-LAG-3 1500 +Anti-PD-1 (8/10 TF) 1000 500 0 0 10 20 30 40 50 Days Post-Treatment Initiation (Shown: MC38 colon cancer) Drake, Vignali, Korman, Pardoll et al., Cancer Res 2012

Therapeutic implications for PD-1 pathway blockade in adaptive resistance model Strong endogenous anti-tumor immune response PD-L1 up-regulation in tumor Anti-PD-1 monotherapy RESPONSE Weak endogenous anti-tumor immune response No PD-L1 up-regulation in tumor 1 Inducer of anti-tumor immunity (vaccine, TKIs, immunogenic chemo, RT) Endogenous anti-tumor immune response 2 PD-L1 expression in tumor Anti-PD-1 RESPONSE

PD-1 pathway blocking agents in clinical testing www.clinicaltrials.gov Target Molecule Company PD-1 PD-L1 (B7-H1) Amplimmune/GSK AMP-224 (PD-L2/Fc fusion protein) Bristol-Myers Squibb Nivolumab/BMS-936558/ MDX-1106/ Ono-4538 (fully human IgG4 mab) CureTech Pidilizumab/CT-011 (humanized IgG1 mab) N/A BMS-936559/MDX- 1105 (fully human IgG4 mab) N/A Genentech N/A MPDL3280A (Fc-modified IgG1 mab) MedImmune N/A Medi4736 (mab) Merck Lambrolizumab/MK-3475 (humanized IgG4 mab) N/A

CTLA-4 Cloning of CTLA-4 gene Demonstration that B7.1/2 are CTLA-4 ligands Demonstration of potent immune regulatory role of CTLA-4 in KO mice Treatment of murine tumors with anti-ctla-4 Anti-CTLA-4 into patients First demonstration of clinical activity of anti-ctla-4 in melanoma Anti-CTLA-4 confers survival benefit in melanoma FDA approval for anti-ctla-4 in melanoma 1985 1990 1995 2000 2005 2010 2015 PD-1 Cloning of PD-1 gene Demonstration of organ-specific autoimmunity in PD-1 KO mice PD-L1/PD-L2 Identified as PD-1 ligands PD-L1 expression in murine tumors confers immune resistance & high PD-L1 expression in human tumors First demonstration of clinical activity of anti-pd-1 Anti-PD-1 into patients Predicted FDA approval of anti-pd-1 Anti-PD-1 & anti-pd-l1 Induce durable tumor responses in melanoma, kidney & lung cancer Pardoll, Nature Immunol 2012

How can the path to clinical approval be accelerated? Regulatory challenges for targeted immunotherapies: Defining appropriate endpoints for clinical efficacy Understanding unique side-effects and developing effective monitoring and management guidelines Identifying biomarkers predicting response, which may be dynamic and tissue-specific

ACKNOWLEDGEMENTS Sponsored by BMS, NIH, Barney Foundation, and Melanoma Research Alliance Thanks to collaborating clinical trial centers. Sponsored by BMS, NIH, Barney Fdn., and Melanoma Research Alliance