biology of metastasis - columbia university · dispersion & seeding detec/on and removal of...
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Biology of metastasis
Swarnali Acharyya Nov 11, 2015
Dispersion&seedingDetec/onandremoval
ofprimarytumor
Treatments for primary breast cancer
89%survivalrateforlocalizedbreastcancers
Dispersion&seedingDetec/onandremoval
ofprimarytumor
Advances in breast cancer treatment
Over 90% survival rate for localized, well treated breast cancers
Be#erdetec)onandimaging
Newsurgicalop)ons
Targetedtherapy
Latency(monthstodecades)
Metastasisisresponsiblefor90%ofcancerdeaths
Overtmetastasis
hAp://seer.cancer.gov/staEacts/html/breast.html
Primary tumor
Metastasis
Cell-autonomous mechanisms: Genetic, epigenetic differences?
Howareprimarytumorsandmetastasisdifferent?
Non cell-autonomous mechanisms: Stromal influences
Metastasisaremoredifficulttotreatandareresistanttomostdrugs
Breast Cancer Genes
Shahetal.,Nature,2012
Mutations are highly concordant between primary tumor and metastasis Ref: Colon cancer, Brannon et al., Genome Biology, 2014
Breastcancersubtypes
Modified from: http://massgenomics.org/2012/09/a-comprehensive-atlas-of-breast-cancer-genomes.html
Luminal A slow growth and best prognosis
Luminal ca : ER positive Express luminal markers (ER alpha, cytokeratins 8/18)
Basal like triple negative cancers: worst prognosis Express basal cell markers (CK 5/6, 14, 17)
Basementmembrane
BasalorMyoepithelialcells
Luminalepithelialcells
HER2 positive breast cancer
Modified from Hongkong Breast Cancer Foundation
TargetHER2ProteinTrastuzumab
Signaling downstream of HER family
http://www.nature.com/nrclinonc/journal/v9/n1/fig_tab/nrclinonc.2011.177_F1.html
Reference: Ellis and Perou, Cancer Discovery, Jan 2013
Trends in breast cancer incidence and mortality
Metastasis: Differences in latencies in cancer types
Lung adenocarcinoma: Rapid metastasis to mutiple organs
Nature
Breast adenocarcinoma: Latency and metastatic speciation
. . . . . .
. . . . .
. . . . . .
WNT/TCF LEF1, HOXB9
EREG, COX2, FASCIN1, MMP1,
ANGPTL4, HBEGF, COX2,
FASCIN1, MMP1, ANGPTL4,
ST6GALNAC5
IL11, ADAMTS1,MMP1, TGFß, CTGF, PTHrP
miR335 miR126
Evolution of metastasis Step-wise or parallel?
Timelineofdevelopmentofmetastasisbasedonmathema)calmodeling
Luebeck,Nature,2010
ClonalEvolu)onofheterogeneousprimarytumorsgivingrisetometastasis
Alternative hypothesis: Parallel evolution of metastasis?
Bone marrow cancer cells genomically different from primary tumor mix - Spread early and evolve
OR - Different subpopulation
which might not give rise to overt metastasis?
Clinically- optimal to decide treatments based on DTCs or primary tumors?
Reference:Gray,CancerCell,2003
Phenotypically normal untransformed cells can persist in the lungs for months
Podsypanina et al., Science, 2008
Delayed oncogene induction still forms tumors
When normal mammary cells have reached the distant site in the absence of transformation at the primary site.
Seed or the soil: which matters in metastasis?
Ref: Fidler, Nat Rev Cancer, 2003
4millioncells/gofprimarytumorshed0.01%successful!
Brief history of metastasis research (Ref: Metastasis Research Society)
Metastatic cells spread through lymphatics or blood?
Blood (hematogenous spread)
Modified from www.iopscience.iop.org
Lymphatic spread
Modified from www.cancer.org
Lymphatic spread of breast cancer
• Might depend on physical restrictions on invasive tumors. • Easier to survive in passive, low shear flow. • Lymphatics more leaky, lacking tight interendothelial junctions (higher chances of intravasation).
Lymphatics more accessible to cancer cells?
Peritumoral lymphatics matter more than intratumoral lymphatics
WongandHynes,CellCycle,2006
Potential pathways of reaching circulation from the primary tumor
WongandHynes,CellCycle,2006
Inprostatecancer,84%withlymphnodespreadhavehematogenousspread.
Drug resistance of metastases: yet another problem to solve
Sensi/ve Resistant
Kuczynskietal.,NatRevClinOncol.,2013
Challengestotargetdrugresistantclones
Latency(monthstodecades)
Overtmetastasis
Animal models to study the complex process of metastasis
Transgenic mouse models of breast cancer
Development of Patient derived xenografts
Kopetz et al., Clinical Cancer research, 2012
Fresh tumor implantation in immunocompromised mice
Tumor grafts in mice resemble the donor patient tumors by histology, genomics, growth
Sitesinpa)ents SitesinmiceMETASTASISSITES
Metastasis-associatedgeneexpression
ApproachI:Interrogatetheend-products
Kang et al Cancer Cell 2003 Minn et al Nature 2005 Tavazoie et al Nature 2008 Bos et al Nature 2009
BrM
LM
BoM
43genes
54genes
102genes
Pro
babi
lity
p < 0.0001
1.0
0.8
0.6
0.4
0.2
0 0 50 100 150
LMS+TβRS+
Lung metastasis-free survival (months)
LMS: 18-gene signature associated with lung relapse
EREG COX2 MMP1 ANGPTL4 FSCN1 NEDD9 ID1 CXCL1 TNC VCAM1 CXCR4 LTBP1 ROBO1 KRTHB1 MAN1A1 KYNU C10orf116 RARRES3
BreastCancerLungMetastasisSignature(LMS)
COX2: vascular permeability Epiregulin: cell motility Fascin1: invadopodia MMP1: collagenase ANGPTL4: endothelial disjunction
Tenascin-C
VCAM-1
Gupta et al Nature 2007 Padua et al Cell 2008 Tavazoie et al Nature 2008
Kim et al Cell 2009 Oskarsson et al JACS 2010
Tumor microenvironment interactions that support tumor progression
Quail and Joyce,, Nat Med, 2013
In transit
Role of platelets in the early hours of metastatic colonization
Reference Labelle et al., PNAS, 2014
At the distant site (lung)
Role of VCAM1 in establishing lung metastasis
Chen and Massague, Clinical Cancer Research, 2012
Fighting for survival
Role of extracellular matrix molecules in metastasis
Tenascin C at the invasive edge of tumors
Ref: Oskarsson et al., Nature Medicine, 2011
Depleting Tenascin-C in cancer cells reduces lung metastasis
Role of Tnc in outgrowth of micrometastasis
TNC interaction with cancer cells at the invasive front enhances NOTCH and WNT signaling
Enhances fitness of metastasis Initiating cells
Extended survival of breast cancer cells for decades in the bone
ModifiedfromhAp://www.myxgeva.com/breast-cancer-bone-metastases.html
The undetectable but present Latent metastasis
Zhangetal.,CancerCell,Jul7,2009
Role of Src signaling in the survival of bone metastasis
• No role in homing to lungs or bone. • If these cells reach, then Src signaling becomes critical for their
outgrowth.
Src enhances survival and outgrowth of latent metastatic cells in the bone marrow
Brain metastasis from breast cancer
Modifiedfromwww.braintumors.in
Plasmin- reactive brain stroma for protection of host
Valienteetal.,Cell,2014
Joyce JA and Pollard JW, Nat Rev Cancer, 2009
Tumor microenvironment components
Macrophage/microglia microenvironment targeting in brain tumors
Pyonteck et al., Nat Med, 2013
T regulatory cells are present in growing tumors and metastases
Poor prognosis in breast cancers
Treg cells ablated from established tumors can reduce metastatic growth
Macrophage
Tum
or fa
ctor
s Tumor
Dendritic Cells
Neutrophil Eosinophil Basophil
Modified from Wynn, Nat Immunology, 14(3), 197-99
Myeloid derived suppressor cells in cancer - Another group of poor prognosis immunosuppressive cells
CXCL1/2 mediates mammary tumor progression
sh Control
shCXCL1/2 0
100
200
300
400
0 20 40
Tum
or v
ol. (
mm
3 )
p<0.
001
Days after injection
Tumor growth
0
20
40
Foci
num
ber/F
OV
shControl shCXCL1/2
p<0.0001
Lung
met
asta
sis shControl shCXCL1/2
Lung metastasis
shControl shCXCL1/2
Cle
aved
cas
pase
3
Apo
ptot
ic c
ells
/FO
V
0
10
20
30
shCon shCXCL1/2
* Apoptosis CXCR2
MMTV-PyMT MDA231-LM2
Granulocytic CD11b+Ly6G+ cells
CXCL1/2 recruit granulocytic CD11b+Ly6G+ cells in the tumor microenvironment
0 5 10 15
Unsorted
CD31+
CD11b+Gr1+
CD11b+Ly6C+
CD11b+Ly6G+
F4/80+
Cxcr2 mRNA levels (Relative units)
MDSC levels correlate with accelerated disease progression and poor survival in patients
- Breast Cancer (Diaz-Montero et al, 2009, Walter et al, 2012)
- Colorectal Cancer (Solito et al, 2011)
- Gastrointestinal Cancer (Gabitass et al., 2011)
- Lymphoma (Montero., 2012)
(Gabrilovich, Ostrand-Rosenberg and Bronte, 2012)
MDSC
Suppress Anti-tumor Immunity
Tumor Angiogenesis (Yang et al, 2004, Shojaei et al, 2007) Blood vessel remodelling and tumor cell extravasation (Yan et al, 2010)
0 20 40 60 80
100 120
0 20 40 60 Tum
or v
olum
e (m
m3 )
S100a9+/+bm S100a9-/- bm
p=0.
001
Days after injection
CXCL1/2 promotes tumor progression via myeloid cell S100A8/9
Met
asta
tic c
ells
/FO
V
S100a9-/- 0 2 4 6 8
10 12 14
p<0.0001
S100a9+/+
S100a9+/+ and S100a9-/- bone marrow reconstituted mice as recipients
Cancer cells to gland #4
Tumor growth Metastasis
Donor bone marrow
CXCL1/2
Breast primary
Metastasis Metasta/cstress
CXCL1/2
Cancer cell
CXCR2
CD11b+Gr1+MyeloidcellsSurvival
Survival
S100A8/9
S100A8/9
CXCL1/2 axis mediates tumor growth and metastasis
Invasionstress
Chemotherapy hyperactivates the CXCL1/2-S100A8/9 cycle
Neoadjuvant Rx samples
S10
0A8/
9 IH
C
scor
ing
Score 0 Score 1
Score 2 Score 3
S10
0A8/
9 sc
ore
Before chemo After chemo
p<0.0001
0
1 2
3 4
X
SB-265610
CXCR2 inhibitor enhances chemotherapy
Lung metastasis
Vehicle i-CXCR2 Two drugs
MD
A23
-LM
2 C
N34
-LM
1
AC chemo
Chemo
Met
asta
tic c
ells
/FO
V
CXCR2i +Chemo
0
4
8
12
16 MDA231-LM2
p=0.028
CN34-LM1
Chemo CXCR2i +Chemo
0
20
40
60
80 p=0.024 Days: 26
CXCR2 inhibitor
34 40 46 53 60
Chemo
0
Implant cancer cells
Primary tumor
Distant metastasis Brain
Bone
Vehicle chemo cxcr2i cxcr2i+chemo0
10000
20000
30000
40000Vehiclechemocxcr2icxcr2i+chemo
Nor
mal
ized
Pho
ton
Flux
Chemo Chemo+CXCR2i
Vehicle CXCR2i 0
10000
20000
30000
40000
Cytotoxic chemo, shortlived targeted therapy…..immunotherapy promise
hAp://www.revealtherapies.com/AllPages/Immunotherapy.html
Dispersion&seedingDetec/onandremoval
ofprimarytumor
Treatments for primary breast cancer
89%survivalrateforlocalizedbreastcancers