Inflammation and Cancer

Alexander Wu, Ph.D.

Ph.D Program for Translational Medicine, TMU
Sep 26th 2017
Tumor-stroma interactions at the primary site
•Tissues contain many different types of cells to maintain normal physiology

•Most of these cells have positional identity – their locations are defined and
number constrained

•Cancer cells have lost these constraints via mutations in oncogenes and
tumor suppressor genes

•BUT, these tumor cells are still surrounded by these non-malignant

residential cells

•These interactions are IMPORTANT!

Analogy: Tumors are like the gangsters in the neighborhood  recruit good kids from
the neighborhood and turn them into bad kids so they can work for the gangsters.
Recruitment of stromal cells to developing tumors
• primary tumors= cancerous cells + different stromal cell types

• these stromal cells are:

•Endothelial cells – make up the blood and lymphatic
circulatory system
•Pericytes - support small vessels, but it can differentiate into
a fibroblast, smooth muscle cell, or macrophage as well if
required.
•Fibroblasts : connective tissue cells – synthesize ECM and
collagen.
•Various BMDCs (bone marrow derived cells)
•Macrophages
•Neutrophils
•Mast cells
•Myeloid cell-derived suppressor cells (MDSCs)
•Mesenchymal stem cells (MSCs)
 The Primary Tumor microenvironment

A solid tumor consists of heterogenous populations of cells

Joyce JA and Pollard JW. Nature Reviews. (2009), 9:239
 The big Q is:
How are these cells attracted to the tumor
site, and why?

Inflammation
V

Vakkila J and Lotze MT. Nat. Reviews. Immunol. (2004) 4:641-647

Tumor-related inflammations
• Inflammatory diseases increase the risk of developing many types of cancer
(including bladder, cervical, gastric, intestinal, oesophageal, ovarian, prostate and
thyroid cancer).

• Non-steroidal anti-inflammatory drugs reduce the risk of developing certain

cancers (such as colon and breast cancer) and reduce the mortality caused by these
cancers (i.e. COX2 inhibitors, TNF-α antagonists, anti-CXCR4, anti-CCL2/MCP-1).

• Signalling pathways involved in inflammation operate downstream of oncogenic

mutations (such as mutations in the genes encoding RAS, MYC and RET).

• Inflammatory cells, chemokines and cytokines are present in the microenvironment

of all tumors in experimental animal models and humans from the earliest stages of
development.

• The targeting of inflammatory mediators (chemokines and cytokines, such as TNF-α

and IL-1β), key transcription factors involved in inflammation (such as NF-κB and
STAT3) or inflammatory cells decreases the incidence and spread of cancer.
Chronic inflammation and BMDC recruitment
• leukocytes often found in tumor:
•failed attempt of our body to kill cancer cells?
•Detection of cancer antigens? Debatable…
•Due to tumor-related tissue disruption  cell repair
•Inflammation

• Our body sees tumor like a never healing wound (chronic inflammation)
•Continued recruitment of bone marrow derived inflammatory cells to the
tumor mass amplified and de-regulated inflammatory responses

Promote angiogenesis & tissue remodeling

•Induction of angiogenesis: important early stage in the development and growth of
most solid tumors
•Also important for dissemination of cancer cells
Oncogenes & cancer-related inflammation

Mantovani A, Allavena P, Sica A & Balkwill F. Nature (2008); 454(24): 436

The involvement of stromal cells in promoting
tumorigenesis

Focus on macrophages
1. Tumor-associated (or activated) macrophages (TAMs)
• Macrophages are
•Plastic cells
•Very adaptive
•One of the different types of immune cells recruited by tumor cells
•Differentiate into different types of macrophages under the influence of
tumor cells

Mosser DM & Edwards JP. Nature Rev. Immunol (2008), 8: 958-969

1. Classically activated
macrophages

2. Wound healing
macrophages

3. Regulatory
macrophages

Mosser DM & Edwards JP. Nature Rev. Immunol (2008), 8: 958-969

Th1 cytokine

Th2 cytokine
1. Classically activated macrophages :

• arise in response to interferon-γ (IFNγ), which can be produced during an

adaptive immune response by T helper 1 (TH1) cells or CD8+ T cells (not shown)

• or during an innate immune response by natural killer (NK) cells, and tumour-
necrosis factor (TNF), which is produced by antigen-presenting cells (APCs).
2. Wound-healing (alternatively activated) macrophages:

•arise in response to interleukin-4 (IL-4), which can be produced during an adaptive

immune response by TH2 cells or during an innate immune response by granulocytes.

IL-4: stimulation of activated B-cell and T-cell proliferation, and the differentiation of CD4+ T-
cells into Th2 cells.
3. Regulatory macrophages:
•are generated in response to various stimuli, including immune complexes,
prostaglandins, G-protein coupled receptor (GPCR) ligands, glucocorticoids,
apoptotic cells or IL-10.

•Regulatory macrophages produce high levels of IL-10 to suppress immune

responses.

To restore homeostasis
•Macrophages are among the first cells to arrive at the sites of wounding and/or
infection
•Produce cytokines and chemokines  recruit other immune cells
•Produce growth factors, angiogenic factors and proteases  promote tissue
repair
•Kill pathogens via production of ROS and NO
•Present antigens to cytotoxic T cells

•Also important in tissue morphogenesis (developmentally)

•Colony-stimulating factor 1, Csf-1 (-/-) mice: lack macrophages
•Developmental defects: osteopetrosis, dermal hypoplasia, delayed and
aberrant pancreatic morphogenesis and impaired branching
morphogenesis of the mammary gland

Macrophages are important in development: tissue remodeling and trophic functions

Dual function of macrophages in tumor progression. During the early steps of carcinogenesis,
M1 macrophages predominate. These cells exhibit anti-tumor activity as well as elicit specific
anti-tumor adaptive immunity, which, in turn, contributes to their M1 polarization.

In established and advanced neoplasia, when persistent tumor cells have escaped the immune
attack, M2-polarized macrophages predominate and suppress adaptive immunity, which, in
turn, contributes to their M2 polarization
Tumor “uses” macrophages to promote their growth and invasion via the surrounding
stroma
•Tumor (transforming mutations) –lost positional identity and they continue to send out
“help me” signals the result in invasion into vasculature.

Pollard JW. Nature Reviews. Cancer (2004), 4:71-78

 Macrophage-depletion leads to the reduced tumor volume

CTRL (Clod-Lip): clodronate-laden liposome

4 days post
inoculation

CD11b TRITC
Labeling of TAM

Gazzaniga S et al. J. Invest. Dermatology (2007), 27: 2031-2041

How do the TAMs promote tumorigenesis?
1.Angiogenesis
2.Suppression of immunity
3.Invasion & metastasis
I. Angiogenesis:
• The formation of new blood vessels
• requires a wide range of soluble factors.
• examples:
–Basic fibroblast growth factor (bFGF), VEGF, angiopoietins (ANG1 and 2),
IL-1, IL-8, tumor necrosis factor-α (TNF- α), MMP-9, MMP-2 and NO.

TAMs:
• in human breast cancer,
TAMs are found clustered
in ‘hot spots’ in avasular
areas (correlates with high
level of angiogenesis
•Hypoxic condition in
tumor upregulates HIF-
2α in TAMs and increase
VEFG expression.
•TAMs also produce
TNFαinduces MMP-9
expression and release
VEGF from ECM
High number of TAMs located
near COX-2-expressing epithelial
cell nests and increased
microvessel in BCC. Serial sections
showed that aggressive TAM
(micronodular) subtype BCC had marker
increased number of CD68-
positive macrophages aggregated
adjacent to COX-2-expressing
epithelial cancer nests.
Increased number of CD31-
positive microvessels located in
area of dense macrophages
infiltration.

angiogenesis

BCC: basal cell carcinoma

(type of skin cancer)

Tjiu et al., Journal of Investigative Dermatology (2009) 129, 1016–1025

II. TAMs & adaptive immunity

• TAMs – M2 macrophages in established tumors

•M2 cells are characterized

• IL-12low IL-10high phenotype
• poor antigen-presenting ability
• does not produce ROS and NO efficiently – don’t kill tumor cells
efficiently
• non-responsive to LPS stimulation and defective NF-kB
expression – leading to the failure of triggering M1 type
inflammatory responses;
 Phenotypic Switch of Macrophages
•Macrophages are multifunctional cells phenotype switch depending on tumor
microenvironment
How M2 Macrophages help tumor escape immune surveillance
Over-recruitment
of naïve T cells –
preventing T cells
from maturation

Decreased Th1
responses –
decreased tumor
suppression

Suppression of
Th cells – so no
tumor killing
effects – leading
to tumor escape
DC cannot recognize from immune
tumor antigens – no system
tumor killing effects
 Mechanistic explanations of M2-medicated immune escape

Normal

M2 responses

M2 macrophages
III. M2 macrophages and metastases

• TAMs enhance metastases of tumor cells.

•HOW?
• Increase MMP expression – degradation of ECM
• Increase migratory ability in cancer cells
•In breast cancer:
•Paracrine loop between cancer cells and TAMs :
• TAMs produce EGF - increases the invasiveness and migration of
neighbouring breast cancer cells that express the EGF receptor
(EGFR).
• Cancer cells in turn express CSF1, which acts as a potent
chemoattractant and chemokinetic molecule for CSF1R-expressing
TAMs.
•This reciprocal cross-talk can be blocked by either EGFR or CSF1R
antagonists, resulting in a decrease in migration and invasion of
both cancer cells and macrophages.

•Csf1op/op pyMT mice, which have a reduction in macrophage

infiltration, and a decrease in circulating cancer cells.
Tumor-Macrophage interaction in the lung using ex vivo imaging
•Tumor cell
•Macrophage
•Vessel (CD31)
Tumor i.v injection
Lung mets

Extravasated part of
tumor cell

L-Clodronate treatment

Qian et al (2009) Plos One. 4(8): e6562

 The concept of Tumor microenvironment of metastasis

EC: endothelial cells

M : macrophages
TC: Tumor cells

Macrophages around the vessels

Robinson BD et al. Clin. Cancer Res (2009); 15(7): 2433

TMEM density correlates to the metastatic ability of tumor cells

Robinson BD et al. Clin. Cancer Res (2009); 15(7): 2433

 Summary III: TAMs promote tumor metastases

Other
stromal
cells

ECM degradation by MMP production Green: TAMs ;Red: vessels; Blue: collagen
Summary: Why TAMs are the bad guys
Targeting TAMs for cancer treatment

bad

good
Thank you for your attention