Paul Ehlrich

Paul Ehlrich (1854-1915)

William Bradley Cooley

Cooley’s toxin

William Bradley Cooley (1862-1936)

The long path to cancer immunotherapy: tumor immunology

Innate

Adaptive

R. VirchowInflammation

′800

Coley’s Toxin

1920s

MacrophageCytotoxicityAlexander1970

NK cellsHerbermanWigzellmid’70

1910Erhlichdream

1950sBurnetImmunesurveillance

1950sPrehnBaldwinTumorsare immunogenic

′60-70Cr releaseAssay,CTLCerottini

′70StutmanSurveillancechallenged

MAGE-1and tu agidentified

Tumor-associatedMacrophages (TAM) promote cancer Mantovani 78-92

NK - missing selfMorettasKärre

′80

Back to VirchowBalkwill and Mantovani

2001

New paradigm formalized

2011

Targeting TAMTrabectedin, anti-CSF1R

2012 – to date

′90SurveillanceComes back:3 EsSchreiber

′90s - to dateCheckpointsAllison

2011HanahanWeinberg

2011ApprovalAnti-CTLA4

Innate

Adaptive

INFLAMMATION, MACROPHAGES AND CANCER

Balkwill and Mantovani, Lancet 2001

RUDOLF LUDWIG KARLVIRCHOW (1821-1902)

(Mantovani et al. Lancet 2008; Nature 2008)

30

20

10

00 24 48 72

hours

IN VITRO

Control

mFS6 + TAM

Control

mFS6 + TAM

[3 H] T

dRup

take

(cpm

x 10

-3)

In Vitro and in vivo

(Mantovani A. 1978) (Bottazzi et al Science 1983)

Ilya Mechnikov (1845-1916)The Nobel Prize in Physiology and Medicine

1908(TAM: Robert Evans, Transplantation, 1972;

Evans and Alexander, Nature, 1970)

(Mantovani, Sica, Allavena, Balkwill Nature 2008; Mantovani Nature 2009)

Mantovani, Sica, Allavena, Balkwill Nature 2008; Mantovani Nature 2009; Hanahan and Weiberg Cell, 2000; Cell 2011

Farbstudie Quadrate, Wassily Kandinsky, 1913

M1

M2

Mantovani and Allavena , J. Exp . Med 2015

Ontogeny and regulation of cells of the monocyte-macrophage lineage

MACROPHAGE POLARIZATION AS SEENIN THE MILANO METRO MAP

Inspired by Luke O’Neill

TAM in Tumor Progression

Mantovani and Allavena, J Exp Med 2015; Nature Rev Clin Oncol, 2016 under revision

M2M2-like

(Bottazzi et al J Immunol 1990)

TAM, INFLAMMATION AND CANCER: TRANSLATION

1. Pathology: prognostic vs predictive

Marchesi F et al., Gut 2015

TAM IN PANCREATIC CARCINOMA HAVE AN M2-LIKE PHENOTYPE

Marchesi F et al., Gut 2015

THE YIN YANG OF TAM AND CHEMOTHERAPY IN PANCREATIC CARCINOMA

1. Pathology: prognostic vs predictive

2. Prevention

TAM, INFLAMMATION AND CANCER: TRANSLATION

1. Pathology: prognostic vs predictive

2. Prevention

3. Reeducation

TAM, INFLAMMATION AND CANCER: TRANSLATION

TRANSLATING IMMUNOLOGICAL COMPLEXITY INTO CLINICAL BENEFIT

Antibodies: the fulfilment of a dream

Mantovani A., Caprioli V., Gritti P., Spreafico F., Transplantation, 1977

Cittera E., et al J Immunol, 2007

TAM as a therapeutic target: Activation of Function

TLR

CD40

FcR

SIRP-αCD47

TAM

M1-like:Direct

Cytoxicity

Tumor cell

AdaptiveImmunity

ADPCADCC

Anti-CD47 mAb

Therapeutic antibodies

mAb(e.g CD20)

Mantovani et al, Nature Rev Clin Oncol, 2016 under revision

1. Pathology: prognostic vs predictive

2. Prevention

3. Reeducation

4. Targetingcorruptedpolicemen

TAM, INFLAMMATION AND CANCER: TRANSLATION

The dual role of TAM in response to chemotherapy and radiotherapy

Response to immunogenic cell death

Orchestration of a tissue repair response after damage

Synergy in tumor cell killing

Reorientation of effector functions (M1-like)

Abscopal effect

Skewing and suppressing T cells

A nurturing niche for CSC

Mantovani et al, Nature Rev Clin Oncol, 2016 under revision

Clinical patterns of response to Trabectedin

Pre-treatment Cycle 2

Pre-treatment Cycle 3

Classical patternAtypical pattern of response

Adapted from F. Grosso et al, Lancet Oncology2007

Pre-treatment

Cycle 1 Cycle 5

Cycle 8 Cycle 11

TRABECTEDIN

TARGETING CANCER-RELATED INFLAMMATION: TRABECTEDIN AS A FORM OF IMMUNOTHERAPY

Germano et al Cancer Res 2005; 2010; Cancer Cell, 2013; D’Incalci Mol Cancer Ther 2010

*

IC 50 on tumor cells*

EMA and FDA approved

Ries C, … Sica A, … Ruttinger D et al, Cancer Cell, 2014

Fig. 6 RG7155 Decreases CSF-1R CD163 TAM in Patients with VariousTypes of Solid Tumors and Alters Intratumoral T Cell CompositionEmactuzumab

Targeting Tumor-AssociatedMacrophages with Anti-CSF-

1R Antibody Reveals a Strategy for Cancer Therapy

Ries C, … Sica A, … Ruttinger D et al, Cancer Cell, 2014

Fig.5 RG7155 Treatment Reduces Tumor Burden in Patients with Diffuse – Type Giant Cell TumorsEmactuzumab

Targeting Tumor-AssociatedMacrophages with Anti-CSF-1R Antibody Reveals a Strategy for

Cancer Therapy

TAM as a therapeutic target: Inhibition of function

VEGFR1CCL2CCL5

CCR2CCR5

C5aR

TRAIL

Trabectedin

COX1,2HOIDOarginase

TAM

VISTAB7H4

PD-L1,L2

Aspirin

mAbmAb

Polarization

Recruitment

Recruitment, survival

Immunosuppression

mAbKinase

inhibitors

checkpoint

Mantovani et al, Nature Rev Clin Oncol, 2016 under revision

RESISTANCE TO SELECTED MICROBES (eg A. fumigatus, P. aeruginosa) –REGULATION OF INFLAMMATION

ASN220

Chr 3q25mRNA 1866 bp

Prot 381 aa

PTX signature(HxCxS/TWxS)

Ex 1 Ex 2 Ex 3

SP NTD PTX

OmpAMen B

components

C1qFactor H

FcγR P-selectinFicolinsMBL

Microbe recognition

C activationand regulation of

inflammation

Opsonization Regulation ofleukocyte

recruitment and inflammation

Repertoiresynergism

Garlanda et al Nature 2002; Deban al Nature Immunol 2010; Lu et al Nature 2009; Bottazzi et al Annu Rev Immunol 2010

PTX3 TRANSLATION

• Diagnostic/prognostic (ELISA, genetics): earlier marker and better related to prognosis compared to CRP

• Therapy (A. fumigatus; P. aeruginosa)

PTX3 TRANSLATION - GENETICS

• IN HUMANS GENETIC POLYMORPHISMS ASSOCIATED WITH SUSCEPTIBILITY TO INFECTION (TB+, P. AERUGINOSA*, UROPATHOGENIC E. COLI#, A.FUMIGATUS$)

* Chiarini, Genes Immun 2010+ Olesen, Genes Immun. 2007# Jaillon et al Immunity 2014

$ Cunha et al New Engl J Med2014

• PTX3 polymorphisms were associated with susceptibility to A. fumigatusinfection in patients undergoing hematopoietic stem cell transplantation

• Haplotype AC was associated with increased protein expression

N Engl J Med. 2014 Jan 30;370(5):421-32.

Results confirmed and extended in 1101 pts in the Swiss Organ Transplantation cohort, 2015(Wójtowicz A, et al, Clin Infect 2015) and

a lung transplantation cohort (Cunha et al 2015)

Methylation of the PTX3 gene in human cancer

(Bonavita et al, Cell 2015) Esophageal Ca: Wang et al World J Gastro; Colon: Tsuji et al BBRC, 2016

PTX3 as an extrinsic oncosuppressor in mouse and human cancer

Bonavita et al Cell 2015

Genetic or

(NF-κB; STAT3; HIFs)Transcription factors

(NF-κB; STAT3; HIFs)

EXTRINSIC PATHWAY

Inflammatory conditions, Infections

INTRINSIC PATHWAY

Oncogenetic events

Chemokines, Cytokines, Prostaglandins (COX-2)

Inflammatory cells (eg Macrophages)

Infections(Virus, HBV, HCV,HPV;Bacteria, H. pylori,Protozoa, schistosoma)Protozoa S. Mansony)

Autoimmune Autoinflammatory(IBD)Irritants (COPD)

Proliferation, survival, Epithelial-Mesenchymal Transition; neo-angiogenesis; invasion, metastasis; inhibition of adaptive immunity; CSC;

response to hormones and chemotherapeutic agents

ras / raf

Oncosuppressor(TGFβ; VHL/HIF;PTEN; α catenin )RelB

Nuclear WnT/β catenin; Myc

TyrosineKinases(RET/PTC; EGF-R)

CANCER-RELATED INFLAMMATION

Mantovani A, Allavena P, Sica A, and Balkwill F, Nature2008

Two pathways link inflammation and cancer

Humoral innate immunity (PTX3; Complement)

(Bonavita et al Cell 2015)

Mantovani A, Treccani, Il libro dell’anno 2015

CANCER IMMUNO-EDITING

From Dunn and Schreiber Nature Immunology 11, 2002

The Basis of Oncoimmunology

A. Karolina Palucka, Lisa M. Coussens Cell 164, March 10, 2016

The Cancer-Immunity Cycle

Chen DS and Mellman I, Immunity 2015

BLOCKADE OF IMMUNE CHECKPOINTS TO ENHANCE T CELL RESPONSES (CTLA-4; PD-1; PD-L1)

Sharma P and Allison JP, Science 2015

Checkpoint blockade activates antitumour immunity

Wolchok JD and Chan TA, Nature 2014

RELEASING THE BRAKES ON CANCER IMMUNOTHERAPY

Ribas A., NEJM 2015

The Cancer-Immunity Cycle

Chen DS and Mellman I, Immunity 2015

The Promise of Cancer ImmunotherapyO

vera

ll Sur

viva

l 100

• Genetic Instability: an Achilles’ heel?

• Memory

• Repertoire – adaptability

• Synergy

Time

Immunotherapy

Chemo Target

46

APC T cell TUMOR

Not only CTLA4 or PD1 !

IL-1R8 AS A KEY REGULATOR (CHECKPOINT) OF NK CELL DIFFERENTIATION AND FUNCTION

IL-18R

IL-18

Metastasis

Viral infection

NK cells

Solid tumor

IL-1R8

FasL

IFN-γ

GrB

activation

Macrophages

Dendritic cells

IL-18R

IL-18

Metastasis

Viral infection

NK cells

IL-1R8

FasL

Solid tumor

IFN-γ

GrB

activation

Macrophages

Dendritic cells

(Molgora, Bonavita et al unpublished)

X

Liver carcinogenesis

Lung

IL-18

IL-18

NK cells NK cells

Liver carcinogenesis

Lung Metastasis

Mantovani A, Treccani, Il libro dell’anno 2015

Translating immunological complexity into clinical benefit

Cells

OVERVIEW OF CART-cell THERAPY IN THE CLINIC

Maus M.V., June C.H., Clin Cancer Res, 2016

Rosenberg SA and Kochenderfer JN, Nat. Rev. Clin. Oncol. 2013

THE DESIGN OF SUCCESSIVE GENERATIONS OF CARs

Kershaw M.H., Westwood J.A., Darcy P.K., Nat Rev Cancer, 2013

June CH, Riddell SR, Schumacher TN, Sci Transl Med. 2015

Adoptive cell therapy is currently represented by three general approaches

«I HAVE A DREAM», M. L. KING

A. Karolina Palucka, Lisa M. Coussens Cell 164, March 10, 2016

Cancer cells are “Self”

Tumor antigensTumor-specific: expressed only in tumor cells

Tumor-associated: overexpressed in tumor cells

Immune response

Mechanism of evasion from immune surveillance: Lack or loss of immunogenic

peptidesImmunoediting

The Basis of Oncoimmunology

A. Karolina Palucka, Lisa M. Coussens Cell 164, March 10, 2016

The Cancer-Immunity Cycle

Chen DS and Mellman I, Immunity 2015

IMMUNE RESPONSE

REGULATION: the immunological synapses

B7

Costimulatory and coinhibitory receptors in the immune synapse

(Checkpoint Inhibitors)

Mechanisms of evasion from immune surveillance

•Tumor antigens are poorly immunogenic•Lack or loss of immunogenic peptides

•Loss of MHC molecules

•Lack of co-stimulatory molecules•Production of immune-suppressive factors

(IL-10, TGFβ, VEGF ……)

The escape phase of cancer immunoediting

REGULATION of ADAPTIVE IMMUNITY

B7

PD-1 Receptor/PD-1 Ligands

BLOCKADE OF IMMUNE CHECKPOINTS TO ENHANCE T CELL RESPONSES (CTLA-4; PD-1; PD-L1)

Sharma P and Allison JP, Science 2015

RELEASING THE BRAKES ON CANCER IMMUNOTHERAPY

Ribas A., NEJM 2015

Checkpoint blockade activates antitumour immunity

Wolchok JD and Chan TA, Nature 2014

The Promise of Cancer ImmunotherapyO

vera

ll Sur

viva

l 100

• Genetic Instability: an Achilles’ heel?

• Memory

• Repertoire – adaptability

• Synergy

Time

Immunotherapy

Chemo Target