CURRENT AND EMERGING BIOMARKERS IN IMMUNO-ONCOLOGY … · • Immune cells infiltrate the tumor and give rise to inflammation. Immune evasion. is the process whereby tumors overcome

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CURRENT ANDEMERGING BIOMARKERS IN IMMUNO-ONCOLOGY RESEARCH

INFLAMMATION IS A HALLMARK OF CANCER

• Immune cells infiltrate the tumor and give rise to inflammation. Immune evasion is the process whereby tumors overcome immune cell control1-4

2

Noninflamed2

Biomarkers for an inflamed phenotype, which may indicate a preexisting immune response, may be

predictive for response to I-O therapy6

Inflamed2,5

NK=natural killer.1. Masucci GV et al. J Immunother Cancer. 2016;4:76. 2. Spranger S, Gajewski TF. Oncoimmunology. 2016;5(3):e1086862. 3. Pardoll DM. Nat Rev Cancer. 2012;12:252-264. 4. Keir ME et al. Annu Rev Immunol. 2008;26:677-704. 5. Shalapour S, Karin M. J Clin Invest. 2015;125(9):3347-3355. 6. Yuan J et al. J Immunother Cancer. 2016;4:3.

THE TUMOR AND THE IMMUNE SYSTEM• The tumor microenvironment is comprised of the cellular milieu in which the tumor exists and

includes MDSCs, lymphocytes, surrounding blood vessels, fibroblasts, signaling molecules and the extracellular matrix1,2

3

Understanding tumor biology and the tumor microenvironment is critical for I-O biomarkers*Effector T cell or NK cell.APC=antigen-presenting cell; MDSC=myeloid-derived suppressor cell; NK=natural killer.1. Whiteside TL. Oncogene. 2008;27(45):5904-5912. 2. Joyce J, Pollard JW. Nat Rev Cancer. 2009;9(4):239-252. 3. Warrington R et al. Allergy Asthma Clin Immunol. 2011;7(suppl 1):S1-S8. 4. Krummel MF et al. Nat Rev Immunol. 2016;16(3):193-201. 5. Gutschner T, Diederichs S. RNA Biology. 2012;9(6):703-719. 6. Li H et al. J Cell Biochem. 2007;101(4):805-815. 7. Chaudhary B, Elkord E. Vaccines. 2016;4(3):28.

Effector cell*

Cytotoxic T cells are the main effector cells of the adaptive immune system. Upon activation, T cells proliferate, migrate, and infiltrate the tumor, promoting tumor cell death3,4

Immune regulatory cells andAPCs play critical roles in maintaining immune homeostasis. They can regulate growth and progression of tumors by inhibiting antitumor immunity2,7

The tumor utilizes mechanisms to sustain proliferative signaling, evade growth suppressors, and resist cell death, thereby enabling replicative immortality, inducing angiogenesis, and activating invasion and metastasis5

The stroma consists of many cell types, including fibroblasts/ myofibroblasts, glial, epithelial, fat, vascular, smooth muscle, and immune cells, along with the extracellular matrix and various signaling molecules. Stromal cells in the microenvironment contribute to tumor development and progression6

A PATH TO PERSONALIZED CANCER THERAPY VIA BIOMARKERS AND DIAGNOSTICS

• The aim of biomarker testing is to individualize cancer treatment: right patient, right therapy, right time

4

Biomarker testing

Patient sample sizes depicted are for illustrative purposes only.Yuan J et al. J Immunother Cancer. 2016;4:3.

Biomarkers and diagnostics help us to understand the disease and, in some instances, predict response to therapy and/or aid in clinical trial selection for specific tumor types

EXPLORING PERSONALIZED IMMUNO-ONCOLOGY (I-O)

5

*Effector T cell or NK cell.dMMR=mismatch repair deficient; IDO1=indoleamine 2,3-dioxygenase-1; LAG-3=lymphocyte-antigen gene 3; MDSC=myeloid-derived suppressive cell; MSI-H=microsatellite instability-high; NK=natural killer; PD-1=programmed death receptor-1; PD-L1=programmed death ligand 1; PD-L2=programmed death ligand 2; TIL=tumor-infiltrating lymphocyte; TMB=tumor mutational burden; Treg=regulatory T cell.1. Ma W et al. J Hematol Oncol. 2016;9(1):47. 2. Gibney GT et al. Lancet Oncol. 2016;17(12):e542-e551. 3. Schumacher TN, Schreiber RD. Science. 2015;348(6230):69-74. 4. Eggermont LJ et al. Trends Biotechnol. 2014;32(9):456-465. 5. Masucci GV, et al. J Immunother Cancer. 2016;4:76.91. 6. Yuan J et al. J Immunother Cancer. 2016;4:3. 7. Chen DS, Mellman I. Nature. 2017;541(7637):321-330. 8. Zitvogel L et al. Sci Transl Med. 2015. doi:10.1126/scitranslmed.3010473.

Host environment Many factors within the host environment may play a role in modulating an immune response7,8

Microbiome | Germline mutations

Immune suppression Cells and proteins within the tumor and its microenvironment are associated with inhibition of the antitumor immune response1,2,6

LAG-3 | Tregs | MDSCs | IDO1

Inflamed tumor Inflamed tumors show evidence of immune-cell infiltration and activation in the tumor microenvironment1,2,5

PD-L1 | PD-L2 | TILs | Inflammation gene signatures

Tumor antigens Tumor antigens are recognized as nonself or foreign by the host immune system and can initiate the adaptive immune response1-4

Neoantigens | TMB | MSI-H/dMMR

*

TUMOR ANTIGENS

NEOANTIGENS AND THE IMMUNE-MEDIATED RESPONSE

• Neoantigens are newly formed antigens that have not been previously recognized by the immune system1,2

– Upon recognition of select neoantigens, several cellular activations may occur in the immune system

• An increased presence of tumor-specific neoantigens may lead to an increased number of tumor-infiltrating lymphocytes, making the tumor more immunogenic1,3

• The biomarkers tumor mutational burden (TMB) and microsatellite instability-high/ mismatch repair deficient (MSI-H/dMMR) are associated with high levels of neoantigens in certain tumors1,4

7

The immune system response to the presence of neoantigens1,3

3. APCs activate T cells

4. T cells locate and destroy tumor cells

2. APCs intercept neoantigens

1. Tumor cells produce neoantigens

APC=antigen-presenting cell.1. Schumacher TN, Schreiber RD. Science. 2015;348(6230):69-74. 2. Lu Y-C, Robbins PF. Semin Immunol. 2016;28(1):22-27. 3. Kim JM, Chen DS. Ann Oncol. 2016;27(8):1492-1504. 4. Le DT et al. N Engl J Med. 2015;372(26):2509-2520.

Tumor antigens

Nucleotiderepeat

MUTATIONS WITHIN MMR GENES MAY LEAD TO MSI-H/dMMR TUMORS

• Mismatch repair is a proofreading mechanism that identifies and removes genetic mutations that naturally arise from the DNA replication process1

– dMMR leads to the accumulation of DNA mutations and results in overall genomic instability2

• Microsatellites are tandem repeat DNA sequences frequently found in non-coding regions3

• Microsatellites are particularly susceptible to mutations; deficiencies in the MMR machinery lead to high levels of microsatellite instability4,5

8

Normal MMR: Errors detected and fixed

Deficient MMR: Errors not detected lead to accumulation

MMR=mismatch repair; MSI-H=microsatellite instability-high.1. Richman S. Int J Oncol. 2015;47(4):1189-1202. 2. Lee V et al. Oncologist. 2016;21(10):1200-1211. 3. Hause RJ et al. Nat Med. 2016;22(11):1342-1350. 4. Bogaert J, Prenen H. Ann Gastroenterol. 2014;27(1):9-14. 5. Kawakami H et al. Curr Treat Options Oncol. 2015;16(7):30.

Repeating sequences are more susceptible to errors in DNA replication

Matched base pairs

Mismatch repair proficient (pMMR)

Mismatched base pair

Mismatched base pairs

Nucleotiderepeat

Mismatched base pair

Mismatch repair deficient (dMMR)

MLH1

PMS2

MSH6

MSH2

MLH1 PMS2

MSH6

MSH2

Tumor antigens

PREVALENCE OF THE MSI-H/dMMR PHENOTYPE

9

• Mismatch repair–deficient tumors were more frequently found in early-stage disease (defined as stage <IV)1-6

MSI-H, indicating dMMR, is found across several tumor types1-3

18%16%14%12%10%8%6%4%2%0%

Late StageEarly Stage

dMMR=mismatch repair deficient; MSI-H=microsatellite instability-high.1. Le DT et al. Science. 2017;357(6349):409-413. 2. Lee V et al. Oncologist. 2016;21(10):1200-1211. 3. Popat S et al. J Clin Oncol. 2005;23(3):609-618. 4. Goldstein J et al. Ann Oncol. 2014;25(5):1032-1038. 5. Koopman M et al. Br J Cancer. 2009;100(2):266-273. 6. Kawakami H et al. J Gastrointest Oncol. 2015;6(6):676-684.

Tumor antigens

Reproduced from Le et al 2017.1

Analysis Type Measurement and Classification

MMR protein deficiency (IHC)1

Immunohistochemistry (IHC)• Antibodies to detect MLH1, MSH2, MSH6, and PMS2 proteins;

visual scoring• dMMR defined as loss of expression of 1 or more of these

proteins by IHC

Microsatellite instability (PCR)2,3

Polymerase Chain Reaction (PCR)• Reference panels including mononucleotide repeats (eg, BAT25,

BAT26) and/or dinucleotide repeats (eg, D2S123, D17S250) can be used to assess MSI

• MSI-H defined as the presence of at least 2 unstable markers among 5 microsatellite markers analyzed (or ≥30% of markers if a large panel is used)

CURRENT CLINICAL MSI/dMMR EVALUATION

• As our understanding of MSI and dMMR as biomarkers increases, newer techniques for evaluation, such as next-generation sequencing (NGS), are emerging4-6

10

dMMR=mismatch repair deficient; MLH1=mutL homolog 1; MSH2=mutS homolog 2; MSH6=mutS homolog 6; MSI=microsatellite instability; MSI-H=MSI-high; PMS2=PMS1 homolog 2.1. Hampel H et al. N Engl J Med. 2005;352(10):1851-1860. 2. Murphy KM et al. J Mol Diagn. 2006;8(3):305-311. 3. Buecher B et al. Dig Liver Dis. 2013;45(6):441-449. 4. Stadler ZK et al. J Clin Oncol. 2016;34(18):2141-2147. 5. Salipante SJ et al. Clin Chem. 2014;60(9):1192-1199. 6. Chalmers ZR et al. Genome Med. 2017;9:34.

Present Absent

Tumor antigens

EVALUATION OF CURRENT AND EMERGING TECHNOLOGY OPTIONS IN dMMR AND MSI TESTING

• At this time, there is no preferred method of measurement of dMMR or MSI based on the concordance analyses

11

Methodology IHC PCR NGS

Biomarker dMMR MSI MSI/dMMR*

Assay performance MSI/dMMR ~90% correlation1 Preliminary data show correlation with PCR2,3

Assessment and tissue requirement

Protein expression; least tissue required4

Small DNA regions;requires more tissue than IHC4

Specified DNA regions;requires more tissue than IHC5

Turnaround time 1–2 days6 5–14 days610–14 days7,8

(median, depending on gene panel)†

*While MMR is usually assessed via IHC, it can also be assessed as part of an NGS panel. †Median turnaround time is based on commercially available tests; this may vary based on gene panel size and test provider.dMMR=MMR deficient; IHC=immunohistochemistry; MMR=mismatch repair; MSI=microsatellite instability; NGS=next-generation sequencing; PCR=polymerase chain reaction.1. Shia J et al. Virchows Arch. 2004;445(5):431-441. 2. Salipante SJ et al. Clin Chem. 2014;60(9):1192-1199. 3. Gan C et al. Genes (Basel). 2015;6(1):46-59. 4. Ryan E et al. Crit Rev Oncol Hematol. 2017;116:38-57. 5. Dagogo-Jack I, Shaw AT. Oncologist. 2016;21(6):662-663. 6. Gibson J et al. Clin Gastroenterol Hepatol. 2014;12(2):171-176. 7. Foundation Medicine. FoundationOne Technical Information and Test Overview. 2017. 8. CMI overview. Caris Life Sciences website http://www.carislifesciences.com/platforms/cmi-overview/. Accessed February 23, 2018.

Tumor antigens

TUMOR MUTATIONAL BURDEN IS A MEASURE OF GENOMIC INSTABILITY

• Tumor mutational burden (TMB) is defined as the number of somatic (acquired) mutations in the tumor genome1,2

• Somatic mutations accumulate over time2

– Environmental factors (eg, tobacco smoking, UV light exposure) can lead to an increase in TMB3,4

– Acquired mutations in DNA repair machinery and mismatch repair factors can increase the frequency of somatic mutations2,4

– Deficient mismatch repair can lead to hypermutability or microsatellite instability, which can lead to an increase in TMB5

12

UV=ultraviolet.1. Chalmers ZR et al. Genome Med. 2017;9(1):34. 2. Stratton MR et al. Nature. 2009;458(7239):719-724. 3. Lawrence MS et al. Nature. 2013;499(7457):214-218. 4. Alexandrov LB et al. Nature. 2013;500(7463):415-421. 5. Lin EI et al. Oncotarget. 2015;6(39):42334-42344.

Tumor antigens

CERTAIN CANCERS HAVE HIGHER MUTATIONAL BURDEN THAN OTHERS

13

AD=adenocarcinoma; ALL=acute lymphoblastic leukemia; AML=acute myeloid leukemia; CLL=chronic lymphocytic leukemia; SCLC=small cell lung cancer; SQ=squamous.Alexandrov LB et al. Nature. 2013;500(7463):415‐421.

Tumor antigens

TMB=tumor mutational burden.1. Ng SB et al. Nature. 2009;461(7261):272-276. 2. Warner JL et al. Genome Med. 2016;8(1):113. 3. Choi M et al. Proc Natl Acad Sci USA. 2009;106(45):19096-19101. 4. Alexandrov LB et al. Nature. 2013;500(7463):415‐421. 5. Chalmers ZR et al. Genome Med. 2017;9(1):34. 6. Cummings CA et al. Clin Transl Sci. 2016;9(6):283-292. 7. Garofalo A et al. Genome Med. 2016;8(1):79. 8. Roszik J et al. BMC Med. 2016;14(1):168.

WGS

WES

Panel

TMB CAN BE ASSESSED USING NEXT-GENERATION SEQUENCING

14

Whole-genome sequencing (WGS)1 Whole-exome sequencing (WES)1 Targeted gene panel2

• Assesses intronic, exonic, and intergenic regions2,3

– Allows for full complement of genomic analyses

• Assesses all coding regions (180,000 exons), making up ~1% of the genome– 85% of mutations contributing

to disease are found in the coding region3

• Assesses a prespecified set of genes

• Uses paired germline samples to subtract germline variation4• May use in silico approaches or

paired germline samples to subtract germline variation2,5

• Typically used with whole-transcriptome analyses (RNA sequencing)6• Optimal number of genes to

determine TMB is under investigation7,8

Tumor antigens

Genome

INTERPLAY OF TUMOR MUTATIONAL BURDEN AND MICROSATELLITE INSTABILITY BY TUMOR TYPE• 83% of MSI-H samples demonstrated high TMB

– In contrast, 16% of TMB-high samples demonstrated MSI-H

15

*Tumor type not provided within reference.CRC=colorectal cancer; MSI-H=microsatellite instability-high; NOS=not otherwise specified; TMB=tumor mutational burden.Chalmers ZR et al. Genome Med. 2017;9(1):34.

Perc

enta

ge o

f spe

cim

ens

60

50

40

30

20

10

0

MSI-H and TMB HighTMB High and MSI StableTMB Low and MSI-H

All specimensN=62,150

n=4328

n=699

Tumor antigens

CORRELATION BETWEEN TUMOR MUTATIONAL BURDEN AND NEOANTIGEN LOAD

• Higher levels of TMB have been shown to correlate with higher neoantigen load1,2

16

Pan-cancer2

TMB=tumor mutational burden.1. Rooney MS et al. Cell. 2015;160(1-2):48-61. 2. Rooney MS et al. Cell. 2015;160(1-2):48-61 [supplemental appendix].

Tumor antigens

INFLAMED TUMOR

BIOLOGY OF PROGRAMMED DEATH LIGAND 1 (PD-L1)

• PD-L1 is expressed on both tumor and immune cells1

• Engagement of PD-L1 with the PD-1 receptor results in the inactivation of T cells, which contributes to tumor evasion2,3

• There are several available immunohistochemistry assays that measure PD-L1 as an I-O biomarker4-9

18

PD-1=programmed death receptor-1; PD-L2=programmed death ligand 2.1. Kerr KM et al. J Thorac Oncol. 2015;10(7):985-989. 2. Pardoll DM. Nat Rev Cancer. 2012;12:252-264. 3. Keir ME et al. Annu Rev Immunol. 2008;26:677-704. 4. PD-L1 IHC 28-8 pharmDx [instructions for use]. Carpinteria, CA: Dako North America, Inc.; 2017. 5. PD-L1 IHC 22C3 pharmDx [instructions for use]. Carpinteria, CA: Dako North America, Inc.; 2017. 6. VENTANA PD-L1 (SP142) Assay [instructions for use]. Tucson, AZ: Ventana Medical Systems, Inc.; 2017. 7. VENTANA PD-L1 (SP263) Assay [instructions for use]. Tucson, AZ: Ventana Medical Systems, Inc.; 2017. 8. PD-L1 IHC 28-8 pharmDx [package insert]. Glostrup, Denmark: Dako Denmark A/S; 2017. 9. PD-L1 IHC 22C3 pharmDx [package insert]. Glostrup, Denmark: Dako Denmark A/S; 2017.

Inflamed tumor

PREVALENCE OF QUANTIFIABLE TUMOR PD-L1 EXPRESSION ACROSS TUMOR TYPES

19

35.4 37.7 43.157.6

76.184.412.0

30.3 19.3

14.2

9.99.9

22.8

23.524.1

17.2

9.74.6

29.8

8.5 13.5 11.04.3

PD-L1 can be expressed at any point along a continuum of 0%–100% on tumor cells

PD-L1 expression varies across tumor types and by cell type, location, histology, and/or line of therapy

Tumor PD-L1 expression†

<1%≥1%–4%≥5%–49%≥50%

64.6

15.6

1.1

Tota

l pop

ulat

ion

scre

ened

(%)*

mNSCLC(n=4930)

mMel(n=3407)

mSCCHN(n=274)

mUC(n=703)

aRCC(n=1918)

mSCLC(n=615)

0

10

20

30

40

50

60

7080

90

100

62.3 56.942.4

23.9

Inflamed tumor

*Samples were obtained from treatment-naïve and previously treated patients screened for eligibility in select clinical trials. †Values indicate percentage of population who express PD-L1 (IHC using the Dako 28-8 PD-L1 pharmDx). aRCC=advanced renal cell carcinoma; IHC=immunohistochemistry; mMel=metastatic melanoma; mNSCLC=metastatic non-small cell lung cancer; mSCCHN=metastatic squamous cell carcinoma of the head and neck; mSCLC=metastatic small cell lung cancer; mUC=metastatic urothelial carcinoma; PD-L1=programmed death ligand 1.Krigsfeld G et al. Poster presentation at AACR 2017. Abstract CT143.

PD-L2 AS AN I-O BIOMARKER

• PD-L2 is expressed on tumor cells, APCs (eg, macrophages and dendritic cells), and a variety of other immune and nonimmune cells1

– PD-L2 expression may vary by tumor type, histology, and cell type2-6

• PD-L2 interaction with PD-1 negatively regulates T-cell proliferation, cytokine production, and cytotoxic activity2

• PD-L2 expression can be measured by IHC1

20

APC=antigen-presenting cell; IHC=immunohistochemistry; PD-1=programmed death receptor-1; PD-L1=programmed death ligand 1; PD-L2=programmed death ligand 2.1. Rozali EN et aI. Clin Dev lmmunol. 2012:656340. 2. Latchman Y et al. Nat Immunol. 2001;2(3):261-268. 3. Shi M et al. Am J Surg Pathol. 2014;38(12):1715-1723. 4. Calles A et al. J Thorac Oncol. 2015;10(12):1726-1735. 5. Liu J et al. Exp Ther Med. 2015;10(5):1947-1952. 6. Ohaegbulam KC et al. Trends Mol Med. 2015;21(1):24-33.

Inflamed tumor

TUMOR-INFILTRATING LYMPHOCYTES (TILs)

• Immune cells, both in circulation and in the tumor microenvironment, are responsible for the destruction of cancer1-4

• TILs are immune cells, such as cytotoxic T cells and NK cells, that enter the tumor and its microenvironment to mediate an antitumor immune response1,5

• Tumors can be characterized by the extent of immune-cell infiltration. The level of TILs correlates with the degree of inflammation1,3

• TIL expression can be measured by IHC, flow cytometry, and droplet digital PCR technology2,6

21

T cell

NK cell

IHC=immunohistochemistry; NK=natural killer; PCR=polymerase chain reaction. 1. Masucci GV et al. J Immunother Cancer. 2016;4:76. 2. Ma W et al. J Hematol Oncol. 2016;9(1):47. 3. Hegde PS et al. Clin Cancer Res. 2016;22(8):1865-1874. 4. Yuan J et al. J Immunother Cancer. 2016;4:3. 5. Wein L et al. Front Oncol. 2017. doi:10.3389/fonc.2017.00156.6. Robins H et al. Sci Transl Med. 2013;5(214):214ra169.

Inflamed tumor

INFLAMMATION GENE SIGNATURES

• Tumor inflammation gene signatures are gene expression profiles reflective of the presence or absence of immune cells in the tumor microenvironment1,2

• Inflammation gene signatures can be measured via targeted mRNA expression profiling, microarrays, RNA-seq, and gene expression panels3,4

• Inflammation gene signatures can contain information on the type, amount, functional orientation, and/or location of immune cells5

• Inflammation gene signatures are under investigation as a predictive biomarker for immunotherapies

22

mRNA=messenger RNA; RNA-seq=RNA sequencing.1. Walker MS, Hughes TA. Int J Mol Med. 2008;21(1):13-17. 2. Linsley PS et al. PLoS One. 2015. doi:10.1371/joumal.pone.0138726. 3. Fumagalli D et al. BMC Genomics. 2014;15:1008. 4. Cesano A. J Immunother Cancer. 2015;3.42. 5. Galon J et al. Immunity. 2013;39(1):11-26.

Inflamed tumor

IMMUNE SUPPRESSION

IMMUNE SUPPRESSION

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1. Huang CT et al. Immunity. 2004;21(4):503-513. 2. Baixeras E et al. J Exp Med. 1992;176(2):327-337. 3. Goding SR et al. J Immunol. 2013;190(9):4899-4909. 4. Blackburn SD et al. Nat Immunol. 2009;10(1):29-37. 5. Camisaschi C et al. J Immunol. 2010;184(11):6545-6551. 6. Pardoll DM. Nat Rev Cancer. 2012;12(4):252-264. 7. Melero I et al. Nat Rev Cancer. 2015;15(8):457-472. 8. Joyce JA, Pollard JW. Nat Rev Cancer. 2009;9(4):239-252. 9. Kumar V et al. Trends Immunol. 2016;37(3):208-220. 10. Mellor AL, Munn DH. Immunol Today.1999;20(10):469-473. 11. Munn DH et al. J Exp Med. 1999;189(9):1363-1372. 12. Munn DH et al. Science. 2002;297(5588):1867-1870. 13. Lee GK et al. Immunology. 2002;107(4):452-460. 14. Mellor AL, Munn DH. Nat Rev Immunol. 2004;4(10):762-774.

Lymphocyte-activation gene 3 (LAG-3) is an immune checkpoint receptor expressed on activated cytotoxic T cells and Tregs. Increased LAG-3 expression can directly promote T-cell exhaustion and indirectly suppress cytotoxic T-cell function. This dual function of LAG-3 can promote tumor immune evasion1-5

Myeloid-derived suppressor cells (MDSCs) are recruited to the tumor microenvironment to suppress effector cell responses through mechanisms including the promotion of T-cell exhaustion and dysfunction8,9

Regulatory T cells (Tregs), or suppressor cells, suppress the immune response by modulating the activation of effector T cells and are important in maintaining self-tolerance and preventing autoimmunity6,7

Indoleamine 2,3-dioxygenase-1 (IDO1)is an intracellular enzyme that initiates the breakdown of tryptophan, which is essential for the survival of cells (eg, cytotoxic T cells), into kynurenine in the tumor microenvironment to suppress immune response10-14

Immune suppression

LAG-3 EXPRESSION IN SOLID TUMOR SAMPLES

• For tumor specimens analyzed across 6 different solid tumor types* a range of low to high LAG-3 expression was observed (0.01%–33%)

• LAG-3 expression may be localized to the perinuclear, membrane, or cytoplasmic regions of lymphocytes, as shown by IHC staining

25

20x

Membrane

Cytoplasmic

Perinuclear

Tumor LAG-3 patterns of expression (IHC): Staining of total nucleated cells in a melanoma tumor sample

*N=245: RCC, 43; gastric, 41; NSCLC, 41; melanoma, 40; SCCHN, 40; urothelial, 40.IHC=immunohistochemistry; LAG-3=lymphocyte-activation gene 3; NSCLC=non-small cell lung cancer; RCC=renal cell carcinoma; SCCHN=squamous cell carcinoma of the head and neck.Edwards R et al. Poster presentation at SITC 2017. Abstract 510P.

Immune suppression

INVESTIGATIONAL I-O BIOMARKERS HAVE THE POTENTIAL TO HELP PERSONALIZE CANCER THERAPY1

• MSI-H/dMMR continues to be studied as a predictive biomarker for I-O in CRC and other tumor types2-5

• Concordance efforts across the various testing methodologies are underway6-14

• TMB, an emerging biomarker, is being studied as a surrogate marker for neoantigen load and therefore may provide insight into response to I-O therapy15-19

• The role of PD-L1 as a prognostic and/or predictive biomarker continues to be researched20,21

• Due to the testing landscape, numerous studies have been initiated to compare the analytical performance of PD-L1 assays22-25

• These concordance data do not impact the FDA approval of the examined assays or their uses• Assessing immune cell infiltrates can provide insights into the TME and may serve as a

prognostic marker across different tumor types1,26-28

• Markers of immune suppression may shed light on the interactions between the tumor and the tumor microenvironment1,18,29

• LAG-3 expression varies in prevalence and location in solid tumor samples30

26

CRC=colorectal cancer; dMMR=mismatch repair deficient; LAG-3=lymphocyte-activation gene 3; MSI-H=microsatellite instability-high; PD-L1=programmed death ligand 1; TIL=tumor-infiltrating lymphocyte; TMB=tumor mutational burden; TME=tumor microenvironment.1. Yuan J et al. J Immunother Cancer. 2016;4:3. 2. Rizvi NA et al. Science. 2015;348(6230):124-128. 3. Lee V et al. Oncologist. 2016;21(10):1200-1211. 4. Sepulveda AR et al. J Clin Oncol. 2017;35(13):1453-1486. 5. Le DT et al. Science. 2017;357(6349):409-413. 6. Cicek MS et al. J Mol Diagn. 2011;13(3):271-281. 7. Lindor NM et al. J Clin Oncol. 2002;20(4):1043-1048. 8. Rigau V et al. Arch Pathol Lab Med. 2003;127(6):694-700. 9. Shia J. J Mol Diagn. 2008;10(4):293-300. 10. McConechy MK et al. Gynecol Oncol. 2015;137(2):306-310. 11. Garcia JJ et al. Mod Pathol. 2006;19(7):950-957. 12. Stelloo E et al. Ann Oncol. 2017;28(1):96-102. 13. Lee JH et al. Genet Test MolBiomarkers. 2014;18(4):229-235. 14. Salipante SJ et al. Clin Chem. 2014;60(9):1192-1199. 15. Schumacher TN, Schreiber RD. Science. 2015;348(6230):69-74. 16. Kim JM, Chen DS. Ann Oncol. 2016;27(8):1492-1504. 17. Liontos M et al. Ann Transl Med. 2016;4(14):264. 18. Sharma P, Allison JP. Science. 2015;348(6230):56-61. 19. Giannakis M et al. Cell Rep. 2016;15:857-865. 20. McDermott DF, Atkins MB. Cancer Med. 2013;2(5):662-673. 21. Patel SP, Kurzrock R. Mol Cancer Ther. 2015;14(4):847-856. 22. Hirsch FR et al. J Thorac Oncol. 2017;12(2):208-222. 23. Batenchuk C et al. Poster presentation at AACR 2017. Abstract 4015. 24. Tsao MS et al. Oral presentation at IASLC WCLC 2017. Abstract PL03.03. 25. Scheel AH et al. Histopathology. 2018;72(3):449-459. 26. Jamal-Hanjani M et al. Clin Cancer Res. 2015;21(6):1258-1266. 27. Stanton SE, Disis ML. J Immunother Cancer. 2016;4:59. 28. Masucci GV, et al. J Immunother Cancer. 2016;4:76. 29. Ma W et al. J Hematol Oncol. 2016;9:47. 30. Edwards R et al. Poster presentation at SITC 2017. Abstract 510P. 31. Hamilton PW et al. Methods. 2014;70(1):59-73. 32. Monville F et al. Poster presentation at AACR 2017. Abstract 590. 33. Gorris MAJ et al. J Immunol. 2018;200:347-354. 34. Villarroel-Espindola et al. Clin Cancer Res. 2017. doi:10.1158/1078-0432.CCR-17-2542.

New technologies are being utilized to identify novel biomarkers more easily and conveniently31-34

Tumor antigens

Inflamed tumor

Immune suppression