The physical biology of circulating tumor cells Owen...
Transcript of The physical biology of circulating tumor cells Owen...
The physical biology of circulating tumor cells
Owen McCarty, OHSU
The physical biology of the vasculature
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RBCs: 4-5 million / l
Platelets: 150,000 400,000 / l
WBCs: 4,500-10,000 / l Coagulation factors:
fibrinogen: 2-3 mg/ml (7600 n )
CTCs: 0-800 / ml
Part 2: Cross-talk between blood cells, CTCs and coagulation in metastasis
Part 1: Characterization of the physical parameters of patient-derived CTCs
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Identification of CTCs using CTC-ID & DIC
Characterization of physical parameters using DIC
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Outputs (per cell): volume mass
density
measured intensity as a f(z)
Goal: Quantify basic physical features of thrombi -the-
Zeiss Axio Imager 2
Method for quantitative imaging
Phillips, McCarty, (2012) Physical Review Letters. 109(11):118105
Phillips, McCarty, (2013) JOVE. In Press Baker, Phillips, McCarty, (2013) Journal of Biomedical Optics. 18(1):16014
Validation: Polystyrene spheres, Diam. 1 20 m
Baker, Phillips, McCarty, (2012) Cell. Mol. Bioeng. 5(4), 488-492.
Physical parameters of CTCs and blood cells
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Phillips, et al. Frontiers in Oncology. (2012) 2:72 Phillips, et al. Frontiers in Oncology. (2012) 2:96
Cell density measurements of SW620 cells
Image segmentation of spatial density maps
Damania, Subramanian, Backman (NWU) Wong, McCarty, Phillips (OHSU). Revising for Journal of Biomedical Optics (Sept, 2013)
Identification of CTCs clusters
Characterization of CTC Clusters
Mike King (Cornell)
Characterization of CTC Clusters - Area
0
200
400
600
800
1000
1 2 3 4 5
Are
a [m
m2 ]
Number of cells per cluster
CTC MCF7 MCF10A
Characterization of CTC Clusters - Radius
0 2 4 6 8
10 12 14 16 18 20
1 2 3 4 5
Effe
ctiv
e R
adiu
s [m
m]
Number of cells per cluster
CTC MCF7 MCF10A
Characterization of CTC Clusters - Mass
0 100 200 300 400 500 600 700 800 900
1000
1 2 3 4 5
Mas
s [p
g]
Number of cells per cluster
CTC MCF7 MCF10A
Model of CTC Transport
Paolo DeCuzzi (Methodist)
Model of CTC Transport
Blood velocity field Thrombin generation
Angie Lee and Paul Newton (USC)
Part 2: Cross-talk between blood cells, CTCs and
coagulation in metastasis
Part 1: Characterization of the physical parameters of patient-derived CTCs
Cancer Metastasis
Nat Rev Cancer. 2011 Feb;11(2):123-34
Cancer Metastasis and the Blood Microenvironment
Cancer cell initiated coagulation
What role does cross-talk between CTCs, blood cells and coagulation play in metastasis?
Approach
Fibrinogen Fibrillar Collagen
Anti-coagulated blood Recalcified blood
Fibrinogen Fibrillar Collagen
Recruitment of colon cancer cells to thrombi under shear
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Recruitment of colon cancer cells to thrombi under shear
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Recruitment of colon cancer cells to thrombi under shear
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Rivaroxaban
CTI
Hirudin
Recruitment of colon cancer cells to thrombi under shear
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Recruitment of colon cancer cells to thrombi under shear
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Baker-Groberg, et al. AJP: Cell Phys Jul 31; in Press
Future directions:
Molecular mechanisms of colon cancer cell thrombi interactions under shear
Does pharmacological inhibition of F Xa inhibit metastasis?
Do neutrophil extracellular traps play a role in metastasis?
AJP: Cell 2013; 305(3): C348-54
Part 2: Cross-talk between blood cells, CTCs and
coagulation in metastasis
Part 1: Characterization of the physical parameters of patient-derived CTCs
Cancer Metastasis
Nat Rev Cancer. 2011 Feb;11(2):123-34
Part 3: Cancer and Thrombosis
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Background: Patients with AML are at elevated risk for thrombosis. Risk to develop thrombosis is closely tied to leukemia subtype. Objective: To evaluate the relative roles of cell count and the surface expression of tissue factor (TF) and phosphatidylserine (PS) in the procoagulant phenotype of AML cells.
Thrombosis & acute myelogenous leukemia (AML)
Cancer and the Blood Microenvironment
Cancer cell initiated coagulation
Effects of Cofactors
X---->Xa Efficiency
IXa 1 IXa, Ca 8 IXa, Ca, PL 5,800 IXa, Ca, PL,VIIIa 1,000,000
II---->IIa Xa 1 Xa, Ca 2 Xa, Ca, PL 8,300 Xa, Ca, PL,Va 2,000,000
factorviia.com Annual Review of Medicine 2008;59: 29-41
Goal: Develop a method to characterize the procoagulant phenotype of leukemic cells
Procoagulant phenotype of acute myelogenous leukemia
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Procoagulant phenotype of acute myelogenous leukemia
Procoagulant phenotype of acute myelogenous leukemia
Tormoen GW, Recht O, Gruber A, Levine RL (MSKCC), McCarty OJ. Phosphatidylserine index as a marker of the procoagulant activity of AML cells. Physical Biology 2013, Oct; 10(5):056010
Future Directions
Clinical Impact: - Indicator for personalized anticoagulant
prophylaxis for patients with cancer
The physical biology of circulating tumor cells
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Mc Carty G roup Joe Aslan K evin Phillips JP Pang Asako Itakura Sandra Baker C ristina Puy Laura H ealy Rachel Rigg Norah Verbout Branden K usanto Juliana Porter L iam Wong Zoë Wong Cassie Loren Garth Tormoen K ristina Haley
Peter K uhn (TSRI) K uhn Lab (TSRI) Paul Newton (USC) Newton Lab (USC) Andras G ruber (O HSU) X iaolin Nan (O HSU) Jonathan Chernoff (F C C C) Ed Manser (Singapore) Robert Insall (Beatson) Laura Machesky (Beatson) Mike K ing (Cornell) Kos Konstantopoulos (JH U) Denis Wirtz (JH U) Ross L evine (MSK C C)
Funding A H A 13E I A126300 NI H R01H L101972 NI H U54C A143906
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Special Issue on the Physical Biology in Cancer
-Owen McCarty, Mike King and Paul Insel,
-Nicole Moore and Larry Nagahara,
-Kimberly Stroka and Konstantinos Konstantopoulos,
-Michael J. Mitchell and Michael R. King, glycocalyx
-Kevin G. Phillips, Peter Kuhn, Owen J.T. McCarty,
-Francois Bordeleau, Turi Alcoser, Cynthia Reinhart-King, metastasis: the role of cytoskeletal
mechanics in cell migratory
JHU PS-OC cross-talk
David Kyle Robinson (2010)
Cassandra Loren (2011)
Whitaker International Scholar Award Sandra Baker (OHSU), Xavier Trepat (Barcelona)
Baker-Groberg, et al. AJP (2013) SSC Meeting (Liverpool, UK, 2012) ISTH Meeting (Amsterdam, NL, 2013) ISTH Young Investigator Award (2013)
Role of Rho GTPases in propulsion and navigation in collective cell migration