Hallmarks of Cancer: Immune evasion & tumor promoting inflammation · 2019. 11. 25. · Chronic inflammation: risk factor for most cancers ! Extended exposure to environmental factors

Prof. Dr. Susanne Sebens

Hallmarks of Cancer: Immune evasion &

tumor promoting inflammation

https://antisensescienceblog.files.wordpress.com/2014/07/hallmarks-10.png

2/79 Lecture Inflammation & Cancer WS 2019/2020

| Susanne Sebens 25.11.2019

1. …common features of acute inflammation and inflammation driven carcinogenesis

2. …and definition of the term „inflammatory stroma/tumor stroma“

3. …the role of the microbiome in tumorigenesis

4. …and definition of the process „Cancer Immunoediting“

4. …strategies of tumor immune evasion

5. …examples how inflammatory cells promote tumor development

6. …therapeutic strategies to overcome immunosuppression in cancer patients

Aims of this lecture

Knowledge of…

*Picture of the 1st slide is taken from: Hanahan & Weinberg, Cell 2011

! Written exam 3 Recapitulation breaks



Chronic inflammation: risk factor for most cancers

! Extended exposure to environmental factors might lead to a smouldering (low-grade)

chronic inflammation which is not detectable by current diagnostic measures.

> low-grade inflammation may be a far more important factor than appreciated.

(Aggarwal et al., Bioch. Pharmacology 2006)

!



(Albini & Sporn Nature Reviews Cancer 2007)

Injury

Infiltration of immune cells

Blood vessel formation

Termination/Healing

Tissue remodelling

Progression/tumor development

Rudolf Virchow (1863): „Chronic irritation which is manifested by chronic inflammation is a key promoter of cancer.“

Harold F. Dvorak (1986): Tumors are wounds that never heal.

Common features of acute inflammation and tumorigenesis !



The two sites of inflammation

Inflammation: important response of the body after infection or injury to kill invading pathogens, remove damaged cells and restore tissue homeostasis

(Aggarwal et al., Bioch. Pharmacology 2006)

!



breast cancer Hodgkin´s lymphoma

(The Biology of Cancer © Garland Science 2007)

Tumors are not only composed of tumor cells but also inflammatory cells

(HistoPathologie Kurs Universität Zürich)

PDAC

tumor cells

(Erkan et al. Nat. Rev. Gastroenterol. Hepatol 2012)

PanIN1a



What is the tumor (inflammatory) stroma?

• variable composition in different tumors, stages, sites …

• interactions via -> direct cell cell contact and -> soluble factors

The tumor stroma (inflammatory stroma, tumor microenvironment) defines

the non-neoplastic tissue within a tumor being composed of a

1. non-cellular compartment

extracellular matrix, growth factors,

chemokines, proteases…

2. cellular compartment

immune cells

endothelial cells

fibroblasts + activated fibroblasts

(myofibroblasts/CAFs)

(Kopfstein und Christofori, Cell Mol Life Sci 2006)

!



Patients with tumors enriched in certain inflammatory cells have a poor prognosis

(Hiroaka et al., Clin. Cancer Res. 2006)

(Kurahara et al. 2009)

(Tsujino et al., Clin. Cancer Res. 2007)

Tregs CAFs

macrophages



The physiological microenvironment prevents the onset of tumors

(Bissell & Hines Nature Medicine 2012)



Normal Fibroblasts exert tumor suppressive functions…

Stroma: control control

Epithel: NMU control

... whereas activated fibroblasts promote tumor growth

Stroma: NMU NMU

Epithel: control NMU

(Maffini et al. J. Cell Science 2004)



(Mitsuhashi et al. Oncotarget 2015)

Fusebacterium status associates with survival of patients with pancreatic cancer

(modified from Belkaid & Naik Nature Immunology 2013)

Host-microbiome homeostasis

Role of the microbiome in tumor development

Elevated abundance of intrapancreatic fungi in pancreatic cancer patients

(Aykat et al. Nature 2019)



DSS/AOM

Bedding from healthy dysbiotic community

Dysregulated microbiome promotes tumorigenesis

DSS/AOM DSS/AOM

(Zackular et al. mBio 2013)

Bedding from healthy dysbiotic community

healthy dysregulated microbiome

!



Immune cells prevent tumor outgrowth

(from: The Biology of Cancer © Garland Science 2014)



Cancer immunoediting: From cancer control by the immune system

to immune evasion by cancer cells

Modified from Veseyl et al, Annu Rev immunol 2011

!



Context matters!




Recapitulation break

Please name four features that acute inflammation and carcinogenesis have in common. Answer:

Which is the correct definition of the term “tumor stroma”?

a) The tumor stroma defines the neoplastic tissue within a tumor.

b) The tumor stroma defines the basal membrane of a tumor.

c) The tumor stroma defines the tumor microenvironment composed of non-neoplastic cells

and a non-cellular compartment.

d) The tumor stroma defines the tumor microenvironment in late stage tumors.

e) The tumor stroma defines the immune cell infiltrate in carcinomas.

Answer:



How do inflammatory cells contribute to initiation and progression of tumors?

How do tumor cells evade the attack by the immune system?

Context matters!




(Inflammatory) stroma cells in the tumor

Main tumor stroma cell populations:

Endothelial cells

Fibroblasts/Myofibroblasts

Immune cells

>First interpretation for the presence of inflammatory (immune) cells in the tumor: Attempt of the body to eliminate the tumor



But…tumors evade the attack by the immune system (Immune escape)

(Vesely et al. Annu. Rev. Immunol. 2011)

!



Endothelial cells

1. Supply of the tumor with oxygen and growth promoting factors

> tumor growth and progression

2. Prerequisite for tumor cell dissemination in secondary organs

> metastasis

3. Other functions?

Lining of blood vessels

Formation of blood vessels (Angiogenesis) when tumor size > 100 µm

Angiogenesis is induced by growth factors, cytokines/chemokines

(e.g. VEGF-A, FGF, IL-8) released by tumor cells or stromal cells



An increased VEGF expression and high number of blood

vessels correlate with poor survival of breast cancer patients

VEGF

(Ghosh et al., Hum. Path. 2008)

MVD

(Tynninen et al., Brit. J. Cancer 2002)

! Some tumors are poorly vascularized!



Endothelial cells from normal pancreas

Endothelial cells from pancreatic carcinoma

% C

D3

1+

ce

lls

Tregs

(Nummer et al., JNCI 2007)

Tumor associated endothelium differentially expresses

adhesion molecules contributing to immune evasion



origin: characteristics:

> Smooth muscle actin (SMA)+

> Elevated production and release of

extracellular matrix proteins, cytokines,

chemokines and growth factors

> altered ECM (matrix remodelling)

Tumor associated fibroblasts

(CAFs/myofibroblasts)

(De Wever et al., Int. J. Cancer 2008)

+ TGF-b1/ FGF-2

derived from tumor/stromal cells

!



Myofibroblasts induce EMT in tumor cells...

EMT= Epithelial-Mesenchymal-Transition > tumor cell dissemination, apoptosis resistance, tumor stemness

Morphology

E-Cadherin

(Larue und Bellacosa, Oncogene 2005)

- fibroblasts + fibroblasts



tum

or

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mm

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0

500

1000

1500

2000

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mono NaCl chemo

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ap

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5

4

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T3M4 co + PMF

mono-tumors co-tumors

T3M4 mono

NaCl as control or chemotherapy

analysis of tumors

(Sebens Müerköster et al. Int. J. Cancer 2008)

tum

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mm

3)

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Reduced response towards chemotherapy

in myofibroblast-enriched tumors

http://images.google.de/imgres?imgurl=http://img.stern.de/_content/59/46/594602/maus_250.jpg&imgrefurl=http://www.stern.de/wissenschaft/natur/:Balzverhalten-Die-Nase-M%E4use-Sex/594602.html&h=188&w=250&sz=14&hl=de&start=13&um=1&usg=__PGvo6GsyuaaQf05klIFztYup-xU=&tbnid=Gs3g0-YIWAdFEM:&tbnh=83&tbnw=111&prev=/images?q%3DLabormaus%26um%3D1%26hl%3Dde

http://images.google.de/imgres?imgurl=http://img.stern.de/_content/59/46/594602/maus_250.jpg&imgrefurl=http://www.stern.de/wissenschaft/natur/:Balzverhalten-Die-Nase-M%E4use-Sex/594602.html&h=188&w=250&sz=14&hl=de&start=13&um=1&usg=__PGvo6GsyuaaQf05klIFztYup-xU=&tbnid=Gs3g0-YIWAdFEM:&tbnh=83&tbnw=111&prev=/images?q%3DLabormaus%26um%3D1%26hl%3Dde



Myofibroblasts contribute to immune evasion

by preventing migration of T cells to the tumor cells

(Rahn et al. Oncotarget 2019)



Myofibroblasts (CAFs) contribute to immunosuppression

(Monteran and Evez. Front. Immunol. 2019)



Impact of myofibroblasts on tumorigenesis

(modified from Calorini and Bianchini 2010)

apoptosis/ therapy resistance

!

EMT

+ immune evasion




Please explain the term “immune evasion” and name three escape strategies

that are used by tumor cells.

Answer:



Immune cells

Tumor infiltrating immune cells:

Macrophages

Myeloid derived suppressor cells (MDSC)

T cells (CD4+, CD8+, gd T cells, Tregs)

Neutrophils

Natural killer (NK) cells

Mast cells

Dendritic cells

most cells have an immunsuppressive or impaired effector phenotype



Simplistic classification of macrophages

(Sica et al. Eur. J. Cancer 2006)

HLA-DRhigh HLA-DRlow

CD163 CD206 TGF-b1

!

IFN-g

CD86



M1- and M2-dichotomy does not exist in vivo: Tumor-associated macrophages exhibit a mixed phenotype

(58,9)

(8,0)

(1,0)

(6,9)

M1

-Ma

c

M2

-Ma

c

HLA-DR CD163

(2,2) (44,5)

TA

M

Fluorescence intensity (Helm et al. Int. J. Cancer. 2014 )



Tumor-associated macrophages heterogeneity depends on ontogeny, activation and localization

(Van Overmeire et al. Frontiers Immunology 2014 )

!



Macrophages increase the blood vessel density in tumors

+ macrophages - macrophages + macrophages

( Lin et al., Cancer Res. 2006)

CSF-1+ CSF-1- tg CSF-1



Tumor cells become invasive in the presence of

pro- and anti-inflammatory macrophages

(Helm et al., Int. J. Cancer 2014)

mono co M1 co M2

* *



Macrophages promote accumulation of myofibroblasts

in tumoral lesions thereby preventing infiltration of CD8+ T cells

(Quaranta et al. 2018)



TAMs are central players

in the tumor microenvironment and tumorigenesis

(Ruffell et al. Trends in Immunology 2012)

!



Chronic inflammation increases cancer risk.

Most cancers arise at sites of chronic inflammation.

Smouldering/subclinical inflammation may be as important in increasing cancer risk (e.g. induced by lifestyle factors) .

Inflammatory cells are abundant in tumors.

In early stages, anti- and protumorigenic immune and inflammatory mechanisms coexist, but if the tumor is not rejected protumorigenic effects dominate and the tumor escapes immune control (cancer immunoediting).

Inflammation is an enabling hallmark of cancer impacting on every step in tumorigenesis – from initiation to progression/metastasis – by promoting the acquisition of other hallmarks of cancer.

Summary



Summary: The tumor-promoting inflammation (tumor stroma) is an enabling hallmark of cancer

Hanahan & Coussens 2012




Which answer regarding macrophages is correct?

a) Macrophages are potent inhibitors of blood vessel formation.

b) M2-Macrophages inhibit tumor outgrowth.

c) Macrophages release high amounts of VEGF by which they promote angiogenesis.

d) By elevated release of VEGF, macrophages inhibit angiogenesis.

e) Macrophages are exclusively found in precursor lesions.

Answer:



Therapeutic interfering with tumor stroma interactions aims at…

1. ... inhibiting tumor cell growth.

2. ... increasing chemo-/radiosensitivity.

3. ... normalizing tumor vasculature to improve drug delivery/inhibiting angiogenesis. 4. ... increasing the immunogenicity of tumors and overcoming immunosuppression.

Is the inflammatory stroma a suitable therapeutic target for treatment of cancer patients?

!



Immune checkpoint inhibitors improve cancer therapy

https://www.google.de/url?sa=i&rct=j&q=&esrc=s&source=images&cd=&cad=rja&uact=8&ved=2ahUKEwib6JXht-DeAhVKy6QKHTQYBCQQjRx6BAgBEAU&url=https://visheshbaat.com/nobel-prize-in-physiology-or-medicine-2018-goes-to/&psig=AOvVaw0Kqr4QtQDyo9IKXYDfsrkJ&ust=1542716162380565



Targeting of PD-L1 (e.g. Darvulumab) or PD-1 (e.g. Pembrolizumab)

(modified from Chen and Han, J Clin Invest. 2015)

PD-L1 =programmed death-ligand 1 PD-1 = programmed cell death receptor-1

Darvulumab approved for: Non-small cell lung carcinoma Pembrolizumab approved for e.g.: Hodgkin-lymphoma Melanoma Non-small cell lung carcinoma

Mechanism: 1. Inhibit immunosuppressive immune cells 2. Increase activation and proliferation of

effector T cells

!



Durvalumab prolongs survival of patients with stage III

unresectable NSCLC after treatment with chemoradiotherapy

(Antonia et al., NEJM 2018)



Pembrolizumub prolongs survival of chemotherapy treated

metastatic NSCLC patients

(Gandhi et al., NEJM 2018)

Note: Checkpoint inhibitors have shown limited efficacy in many solid tumors other immunosuppressive mechanisms? other protumorigenic mechanisms of immune checkpoint regulators?



Chronic inflammation increases cancer risk.

Most cancers arise at sites of chronic inflammation.

Smouldering/subclinical inflammation may be as important in increasing cancer risk (e.g. induced by lifestyle factors) .

Summary

Immune checkpoint inhibition is a promising anti-cancer immunotherapy.

Inflammatory cells are abundant in tumors.

In early stages, anti- and protumorigenic immune and inflammatory mechanisms coexist, but if the tumor is not rejected protumorigenic effects dominate and the tumor escapes immune control (cancer immunoediting).

Inflammation is an enabling hallmark of cancer impacting on every step in tumorigenesis – from initiation to progression/metastasis – by promoting the acquisition of other hallmarks of cancer.

!

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Hallmarks of Cancer: Immune evasion & tumor promoting inflammation · 2019. 11. 25. · Chronic inflammation: risk factor for most cancers ! Extended exposure to environmental factors