Nat. Rev. Canc, 7, 834

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Nat. Rev. Canc, 7, 834
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Looking at secondary tumor sites
Premetastatic Micrometastases (pre
angiogenic) Metastases
lt-- From where? --gt Dormancy?
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What is it that decides which organ shall suffer
in a case of disseminated cancer? - 1889,
Paget

• Seed and Soil
• organ specificity depends on tumor cell and 2
  site
• Blood Flow Patterns determine sites (1920s)
• 66 consistent with blood flow
• 14 fewer mets than expected
• 20 more mets than expected
• Therefore a combined theory
• Blood flow then compatibility
• -1992, Weiss
• Premetastatic Niche conditioning

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Chambers et al, Nature Reviews Cancer (2002), 2,
563
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What is it that decides which organ shall suffer
in a case of disseminated cancer? - 1889,
Paget

• Seed and Soil
• organ specificity depends on tumor cell and 2
  site
• Blood Flow Patterns determine sites (1920s)
• 66 consistent with blood flow
• 14 fewer mets than expected
• 20 more mets than expected
• Therefore a combined theory
• Blood flow then compatibility
• -1992, Weiss
• Premetastatic Niche conditioning

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Class Action/ Community effects
Heterogeniety of cells in tumor
population Clustered Migration of Tumor
Cells ECM degradation Paracrine Loops (signaling
/adhesion) Temporal Cooperation successive waves
of cancer cells passing by induce progressive
changes
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Head-Neck Tumor
Reacquisition/ Presence of uPAR on cells- growth
of disseminated cells
Aguirre-Ghiso, J., Nat. Rev. Canc, 7, 834
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premetastatic niche - do metastatic tumors
affect host tissues, either locally around the
tumor, or systemically? tumor cells and -
endothelium (vascular, lymphatic) - immune
cells (local, from BM) - fibroblasts/stroma
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Tumor cells and endothelium -lots of
experiments from the 1990s show that cytokine
injection (IL-1, IL-6, TNFa) activate endothelim
and promote subsequent tumor (tail vein met
assays) formation
Orr FW et.al., Am J Pathol 190 310-329
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Tumor cells and endothelium integrins
adhesion at rest
adhesion under flow
Orr FW et.al., Am J Pathol 190 310-329
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Tumor cells and endothelium selectins
Lots of data on sel. upregulation by cytokines
and tumor cells, but this is nice a mouse model
with transgenic E-selectin B16 cells engineered
to express selectin ligands changes the pattern
of tumor metastasis
Biancone L et.al., J Exp Med 183 581-587
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Tumor cells and endothelium direct activation
IL-1
TNFa
lung
liver
both
E-selectin
Injected tumor cells can induce cytokine
production and endothelial changes in the liver
(met site)
Khatib A et.al., Cancer Res 59 1356-61
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Tumor cells and endothelium direct activation
Cytokines actually made by immune cells
(Kupffer?) in the liver By confocal microscopy,
adhesion molecule induction leads to tumor cell
extravasation
Met. tumor cells
GFP tumor
F4/80 (Kupffer?)
TNFa
Non-met. tumor cells
Point though Known that most of these
extravasating cells will not survive
Khatib A et.al., Am J Pathol 167 749-759
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Tumor cells and lymphatic metastasis/niche
Rinderknecht M, Detmar M J Cell Physiol 216
347-354
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Tumor cells and lymphatic metastasis/niche
Keratin 14-driven GFPVEGFA or VEGFC chemical-ind
uced melanoma model
Rinderknecht M, Detmar M J Cell Physiol 216
347-354
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Tumor cells and lymphatic metastasis/niche
Tumor can influence downstream tissue and make it
more hospitable for metastasis. Argue secreted
factors, but do they really exclude circulating
cells?
Hirakawa S et.al., J Exp Med 201 1089-1099
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Tumor cells and bone marrow-derived cells
Whetton AD, Graham GJ Trends Cell Biol 9 233-238
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Systemic tumor effect metastatic niche in the
lung
-benign F2 B16 LLC 3LL highly metastatic cells
inj. s.c. into nude mice, extract lungs on d.14,
microarray - what is upregulated in
pre-metastatic (section staining PCR) lungs?
-top two proteins are secreted chemokine-like
molecules, S100A8 and A9 -expressed by Mac1
myeloidy cells and by lung endothelial cell, not
by tumor cells -more Mac1 cells in
pre-metastatic lungs
Hiratsuka S et.al., Nature Cell Biol 8 (12)
1369-1375
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Systemic tumor effect metastatic niche in the
lung
-what does tumor secrete to induce S100A8/9
expression? apparently, TNFa/TGFb/VEGFA
Hiratsuka S et.al., Nature Cell Biol 8 (12)
1369-1375
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Systemic tumor effect metastatic niche in the
lung
-S100A8/9 promote mac. and tumor cell migration
in vitro -Mac and endothelial cells stimulated
with S100A8 promote tumor cell migration (TNFa,
MIP2, TGFb - p38 for migration)
Hiratsuka S et.al., Nature Cell Biol 8 (12)
1369-1375
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Systemic tumor effect metastatic niche in the
lung
-blocking S100A8/9 reduces myeloid cells and
metastasis (never show late time points, though
- 24hr maximum)
Hiratsuka S et.al., Nature Cell Biol 8 (12)
1369-1375
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Systemic tumor effect metastatic niche in the
lung
TNFa, MIP2, TGFb, S100A8/9
Hiratsuka S et.al., Nature Cell Biol 8 (12)
1369-1375
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Premetastatic Niche Timing of molecular and
cellular changes HPC express VEGFR1 Induction
not dependent on presence of tumor cells
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Intradermal, SubQ tail vein injection tumor
models LLC- metastasize to lung B16-
metastasize to lung, liver, spleen,
kidney using 2 x 106 cells Focus on the
recruitment of HPCs in the premetastatic
niche Histology/Immunohistochemistry FACS
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FACS
FN expression in lungs BMDC/HPCs
Solitary Tumor Cells EPCs Micrometastas
es
Day 3 Day 8 Day 14 Day 18 Day 23
-2 days -1-2 days
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Day 14 BMD clusters
Day 23 µmets
gtirradiation ltLLC implantn
Day 18
lung
?-gal BM
met
BM B16
VEGFR1
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VEGFR1 BM
VEGFR1 CD133
VEGFR1 CD117
Day 14
BMDC are HPCs with stem/progenitor
markers similar VEGFR1 expression in cellular
clusters in pre-metastatic human tissue (lymph
nodes) breast, lung, gastrointestinal
Blockade of VEGFR1 cells (Ab) decrease
clusters, µmet
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VEGFR1 ????
VEGFR1 Id3
Day 14
gt cluster ???? ????
Id lt integrins
Release s Kit-ligand VEGF-A Xpression
enhanced by ??????FN intxn
Anti alpha 4 integrin MMP-9 KO --gt decrease
clusters, µmet Id3 KO decrease R1 HPC
in circ (654 v 3283) rescue
cluster/µmet w/HPCs from wt
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Rescue in Id3 KO
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LLC
FN
PDGFR?
No tumor Day 3 Day 14
B16
FN
qRT-PCR lungs
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Conditioned media experiments Established
clusters after MCM treatment/ Tail vein B16
1 day
4 days
Media alone B16 conditioned media (lone Tumor
cell) BMDCs/Tumor
HPC clusters
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Treat mice with conditioned B16 media before and
after Intradermal LLC- redirect metastases
Spleen Kidney Intestine Oviduct
What redirects?
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Pre-metastatic niche in lymph node
Primary Breast Tumor
Micrometastatic/metastatic niche
Kaplan and Lyden reviews
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Open questions Is the premetastatic niche
real? What tumor derived signals target BMDCs
to tissue-specific sites? Genetic/Chemokine
Expression Profile Is BMDC localization random
or to specific sites within target organs? ie
Selective upregulation of S100A8/A9 or throughout
lungs What exactly is the function of VEGFR1
HPCs in niche? Do they remain
undifferentiated? Do they return to BM? Do they
incorporate into tumor vasculature? What exactly
are the Mac1 cells, are they functioning like
HPCs? How exactly are both HPCs and Mac1 cells
enhancing disseminated tumor cell recruitment?
changes endotheliumMMP activation (is there)
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