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

10/12/2013 35 Ishan Patel 35

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