Kinetics of metastatic breast cancer cell trafficking in bone

Pushkar A. Phadke, Robyn R. Mercer, John F. Harms, Yujiang Jia, Andra R. Frost, Jennifer L. Jewell, Karen M. Bussard, Shakira Nelson, Cynthia Moore, John C. Kappes, Carol V. Gay, Andrea M. Mastro, Danny R. Welch

Research output: Contribution to journalArticle

76 Citations (Scopus)

Abstract

Purpose: In vivo studies have focused on the latter stages of the bone metastatic process (osteolysis), whereas little is known about earlier events, e.g., arrival, localization, and initial colonization. Defining these initial steps may potentially identify the critical points susceptible to therapeutic intervention. Experimental Design: MDA-MB-435 human breast cancer cells engineered with green fluorescent protein were injected into the cardiac left ventricle of athymic mice. Femurs were analyzed by fluorescence microscopy, immunohistochemistry, real-time PCR, flow cytometry, and histomorphometry at times ranging from 1 hour to 6 weeks. Results: Single cells were found in distal metaphyses at 1 hour postinjection and remained as single cells up to 72 hours. Diaphyseal arrest occurred rarely and few cells remained there after 24 hours. At 1 week, numerous foci (2-10 cells) were observed, mostly adjacent to osteoblast-like cells. By 2 weeks, fewer but larger foci (≥50 cells) were seen. Most bones had a single large mass at 4 weeks (originating from a colony or coalescing foci) which extended into the diaphysis by 4 to 6 weeks. Little change (<20%) in osteoblast or osteoclast numbers was observed at 2 weeks, but at 4 to 6 weeks, osteoblasts were dramatically reduced (8% of control), whereas osteoclasts were reduced modestly (to ∼ 60% of control). Conclusions: Early arrest in metaphysis and minimal retention in diaphysis highlight the importance of the local milieu in determining metastatic potential. These results extend the Seed and Soil hypothesis by demonstrating both intertissue and intratissue differences governing metastatic location. Ours is the first in vivo evidence that tumor cells influence not only osteoclasts, as widely believed, but also eliminate functional osteoblasts, thereby restructuring the bone microenvironment to favor osteolysis. The data may also explain why patients receiving bisphosphonates fail to heal bone despite inhibiting resorption, implying that concurrent strategies that restore osteoblast function are needed to effectively treat osteolytic bone metastases.

Original languageEnglish (US)
Pages (from-to)1431-1440
Number of pages10
JournalClinical Cancer Research
Volume12
Issue number5
DOIs
StatePublished - Mar 1 2006

Fingerprint

Breast Neoplasms
Bone and Bones
Osteoblasts
Osteoclasts
Diaphyses
Osteolysis
Heart Ventricles
Diphosphonates
Green Fluorescent Proteins
Fluorescence Microscopy
Nude Mice
Femur
Real-Time Polymerase Chain Reaction
Seeds
Flow Cytometry
Research Design
Soil
Immunohistochemistry
Neoplasm Metastasis
Neoplasms

All Science Journal Classification (ASJC) codes

  • Oncology
  • Cancer Research

Cite this

Phadke, P. A., Mercer, R. R., Harms, J. F., Jia, Y., Frost, A. R., Jewell, J. L., ... Welch, D. R. (2006). Kinetics of metastatic breast cancer cell trafficking in bone. Clinical Cancer Research, 12(5), 1431-1440. https://doi.org/10.1158/1078-0432.CCR-05-1806
Phadke, Pushkar A. ; Mercer, Robyn R. ; Harms, John F. ; Jia, Yujiang ; Frost, Andra R. ; Jewell, Jennifer L. ; Bussard, Karen M. ; Nelson, Shakira ; Moore, Cynthia ; Kappes, John C. ; Gay, Carol V. ; Mastro, Andrea M. ; Welch, Danny R. / Kinetics of metastatic breast cancer cell trafficking in bone. In: Clinical Cancer Research. 2006 ; Vol. 12, No. 5. pp. 1431-1440.
@article{29843489ead34a93a3df2dd7a5986192,
title = "Kinetics of metastatic breast cancer cell trafficking in bone",
abstract = "Purpose: In vivo studies have focused on the latter stages of the bone metastatic process (osteolysis), whereas little is known about earlier events, e.g., arrival, localization, and initial colonization. Defining these initial steps may potentially identify the critical points susceptible to therapeutic intervention. Experimental Design: MDA-MB-435 human breast cancer cells engineered with green fluorescent protein were injected into the cardiac left ventricle of athymic mice. Femurs were analyzed by fluorescence microscopy, immunohistochemistry, real-time PCR, flow cytometry, and histomorphometry at times ranging from 1 hour to 6 weeks. Results: Single cells were found in distal metaphyses at 1 hour postinjection and remained as single cells up to 72 hours. Diaphyseal arrest occurred rarely and few cells remained there after 24 hours. At 1 week, numerous foci (2-10 cells) were observed, mostly adjacent to osteoblast-like cells. By 2 weeks, fewer but larger foci (≥50 cells) were seen. Most bones had a single large mass at 4 weeks (originating from a colony or coalescing foci) which extended into the diaphysis by 4 to 6 weeks. Little change (<20{\%}) in osteoblast or osteoclast numbers was observed at 2 weeks, but at 4 to 6 weeks, osteoblasts were dramatically reduced (8{\%} of control), whereas osteoclasts were reduced modestly (to ∼ 60{\%} of control). Conclusions: Early arrest in metaphysis and minimal retention in diaphysis highlight the importance of the local milieu in determining metastatic potential. These results extend the Seed and Soil hypothesis by demonstrating both intertissue and intratissue differences governing metastatic location. Ours is the first in vivo evidence that tumor cells influence not only osteoclasts, as widely believed, but also eliminate functional osteoblasts, thereby restructuring the bone microenvironment to favor osteolysis. The data may also explain why patients receiving bisphosphonates fail to heal bone despite inhibiting resorption, implying that concurrent strategies that restore osteoblast function are needed to effectively treat osteolytic bone metastases.",
author = "Phadke, {Pushkar A.} and Mercer, {Robyn R.} and Harms, {John F.} and Yujiang Jia and Frost, {Andra R.} and Jewell, {Jennifer L.} and Bussard, {Karen M.} and Shakira Nelson and Cynthia Moore and Kappes, {John C.} and Gay, {Carol V.} and Mastro, {Andrea M.} and Welch, {Danny R.}",
year = "2006",
month = "3",
day = "1",
doi = "10.1158/1078-0432.CCR-05-1806",
language = "English (US)",
volume = "12",
pages = "1431--1440",
journal = "Clinical Cancer Research",
issn = "1078-0432",
publisher = "American Association for Cancer Research Inc.",
number = "5",

}

Phadke, PA, Mercer, RR, Harms, JF, Jia, Y, Frost, AR, Jewell, JL, Bussard, KM, Nelson, S, Moore, C, Kappes, JC, Gay, CV, Mastro, AM & Welch, DR 2006, 'Kinetics of metastatic breast cancer cell trafficking in bone', Clinical Cancer Research, vol. 12, no. 5, pp. 1431-1440. https://doi.org/10.1158/1078-0432.CCR-05-1806

Kinetics of metastatic breast cancer cell trafficking in bone. / Phadke, Pushkar A.; Mercer, Robyn R.; Harms, John F.; Jia, Yujiang; Frost, Andra R.; Jewell, Jennifer L.; Bussard, Karen M.; Nelson, Shakira; Moore, Cynthia; Kappes, John C.; Gay, Carol V.; Mastro, Andrea M.; Welch, Danny R.

In: Clinical Cancer Research, Vol. 12, No. 5, 01.03.2006, p. 1431-1440.

Research output: Contribution to journalArticle

TY - JOUR

T1 - Kinetics of metastatic breast cancer cell trafficking in bone

AU - Phadke, Pushkar A.

AU - Mercer, Robyn R.

AU - Harms, John F.

AU - Jia, Yujiang

AU - Frost, Andra R.

AU - Jewell, Jennifer L.

AU - Bussard, Karen M.

AU - Nelson, Shakira

AU - Moore, Cynthia

AU - Kappes, John C.

AU - Gay, Carol V.

AU - Mastro, Andrea M.

AU - Welch, Danny R.

PY - 2006/3/1

Y1 - 2006/3/1

N2 - Purpose: In vivo studies have focused on the latter stages of the bone metastatic process (osteolysis), whereas little is known about earlier events, e.g., arrival, localization, and initial colonization. Defining these initial steps may potentially identify the critical points susceptible to therapeutic intervention. Experimental Design: MDA-MB-435 human breast cancer cells engineered with green fluorescent protein were injected into the cardiac left ventricle of athymic mice. Femurs were analyzed by fluorescence microscopy, immunohistochemistry, real-time PCR, flow cytometry, and histomorphometry at times ranging from 1 hour to 6 weeks. Results: Single cells were found in distal metaphyses at 1 hour postinjection and remained as single cells up to 72 hours. Diaphyseal arrest occurred rarely and few cells remained there after 24 hours. At 1 week, numerous foci (2-10 cells) were observed, mostly adjacent to osteoblast-like cells. By 2 weeks, fewer but larger foci (≥50 cells) were seen. Most bones had a single large mass at 4 weeks (originating from a colony or coalescing foci) which extended into the diaphysis by 4 to 6 weeks. Little change (<20%) in osteoblast or osteoclast numbers was observed at 2 weeks, but at 4 to 6 weeks, osteoblasts were dramatically reduced (8% of control), whereas osteoclasts were reduced modestly (to ∼ 60% of control). Conclusions: Early arrest in metaphysis and minimal retention in diaphysis highlight the importance of the local milieu in determining metastatic potential. These results extend the Seed and Soil hypothesis by demonstrating both intertissue and intratissue differences governing metastatic location. Ours is the first in vivo evidence that tumor cells influence not only osteoclasts, as widely believed, but also eliminate functional osteoblasts, thereby restructuring the bone microenvironment to favor osteolysis. The data may also explain why patients receiving bisphosphonates fail to heal bone despite inhibiting resorption, implying that concurrent strategies that restore osteoblast function are needed to effectively treat osteolytic bone metastases.

AB - Purpose: In vivo studies have focused on the latter stages of the bone metastatic process (osteolysis), whereas little is known about earlier events, e.g., arrival, localization, and initial colonization. Defining these initial steps may potentially identify the critical points susceptible to therapeutic intervention. Experimental Design: MDA-MB-435 human breast cancer cells engineered with green fluorescent protein were injected into the cardiac left ventricle of athymic mice. Femurs were analyzed by fluorescence microscopy, immunohistochemistry, real-time PCR, flow cytometry, and histomorphometry at times ranging from 1 hour to 6 weeks. Results: Single cells were found in distal metaphyses at 1 hour postinjection and remained as single cells up to 72 hours. Diaphyseal arrest occurred rarely and few cells remained there after 24 hours. At 1 week, numerous foci (2-10 cells) were observed, mostly adjacent to osteoblast-like cells. By 2 weeks, fewer but larger foci (≥50 cells) were seen. Most bones had a single large mass at 4 weeks (originating from a colony or coalescing foci) which extended into the diaphysis by 4 to 6 weeks. Little change (<20%) in osteoblast or osteoclast numbers was observed at 2 weeks, but at 4 to 6 weeks, osteoblasts were dramatically reduced (8% of control), whereas osteoclasts were reduced modestly (to ∼ 60% of control). Conclusions: Early arrest in metaphysis and minimal retention in diaphysis highlight the importance of the local milieu in determining metastatic potential. These results extend the Seed and Soil hypothesis by demonstrating both intertissue and intratissue differences governing metastatic location. Ours is the first in vivo evidence that tumor cells influence not only osteoclasts, as widely believed, but also eliminate functional osteoblasts, thereby restructuring the bone microenvironment to favor osteolysis. The data may also explain why patients receiving bisphosphonates fail to heal bone despite inhibiting resorption, implying that concurrent strategies that restore osteoblast function are needed to effectively treat osteolytic bone metastases.

UR - http://www.scopus.com/inward/record.url?scp=33645080421&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=33645080421&partnerID=8YFLogxK

U2 - 10.1158/1078-0432.CCR-05-1806

DO - 10.1158/1078-0432.CCR-05-1806

M3 - Article

C2 - 16533765

AN - SCOPUS:33645080421

VL - 12

SP - 1431

EP - 1440

JO - Clinical Cancer Research

JF - Clinical Cancer Research

SN - 1078-0432

IS - 5

ER -

Phadke PA, Mercer RR, Harms JF, Jia Y, Frost AR, Jewell JL et al. Kinetics of metastatic breast cancer cell trafficking in bone. Clinical Cancer Research. 2006 Mar 1;12(5):1431-1440. https://doi.org/10.1158/1078-0432.CCR-05-1806