Fluorescence-based experimental model to evaluate the concomitant effect of drugs on the tumour microenvironment and cancer cells

Karthik Ramasamy; Hazera Khatun; Lee Macpherson; Mathew P Caley; Justin Sturge; Ghulam J Mufti; Stephen A Schey; Yolanda Calle

doi:10.1111/j.1365-2141.2012.09103.x

Fluorescence-based experimental model to evaluate the concomitant effect of drugs on the tumour microenvironment and cancer cells

Karthik Ramasamy, Hazera Khatun, Lee Macpherson, Mathew P Caley, Justin Sturge, Ghulam J Mufti, Stephen A Schey, Yolanda Calle

Research output: Contribution to journal › Article › peer-review

Abstract

The response of the tumour microenvironment to anti-cancer drugs can influence treatment efficacy. Current drug-screening methodologies fail to distinguish and quantify simultaneously the concomitant effect of drugs on the tumour stroma and cancer cells. To overcome this limitation we have developed a fluorescence-based experimental model that employs mCherry-labelled stromal cells (e.g. bone marrow fibroblastic stromal cells) co-cultured in direct contact with enhanced green fluorescent protein-labelled tumour cell lines for accurate assessment of proliferation and viability in both cell compartments and adhesion of tumour cells. Additionally, we used fluorescence-based image analysis to determine morphological changes that correlate with cell function (e.g. morphology of the actin cytoskeleton and nuclearity of osteoclasts to predict their bone resorption activity). Using this platform we have revealed that dexamethasone induces HS5 fibroblast proliferation and contact with multiple myeloma cells via a process involving Src/c-Abl kinases. Osteoclasts also inhibited dexamethasone-induced apoptosis in myeloma cells while retaining their normal morphology and functionality in bone resorption. Myeloma resistance to dexamethasone mediated by HS5 cells and osteoclasts was reversed by treatment with the Src/c-Abl inhibitor dasatinib but not with bortezomib. This new experimental platform provides a more precise screening of new therapeutics for improved efficacy of tumour cell killing within the bone marrow microenvironment.

Original language	English
Pages (from-to)	564-79
Number of pages	16
Journal	British Journal of Haematology
Volume	157
Issue number	5
DOIs	https://doi.org/10.1111/j.1365-2141.2012.09103.x
Publication status	Published - Jun 2012

Keywords

Actins
Antineoplastic Agents
Apoptosis
Bone Marrow
Bone Resorption
Cell Line
Cell Proliferation
Coculture Techniques
Dasatinib
Dexamethasone
Drug Screening Assays, Antitumor
High-Throughput Screening Assays
Humans
Multiple Myeloma
Oncogene Proteins v-abl
Osteoclasts
Pyrimidines
Stromal Cells
Thiazoles
Tumor Microenvironment
src-Family Kinases

Access to Document

10.1111/j.1365-2141.2012.09103.x

Cite this

@article{615cac4ec674446da1b07c580d0b2c3c,

title = "Fluorescence-based experimental model to evaluate the concomitant effect of drugs on the tumour microenvironment and cancer cells",

abstract = "The response of the tumour microenvironment to anti-cancer drugs can influence treatment efficacy. Current drug-screening methodologies fail to distinguish and quantify simultaneously the concomitant effect of drugs on the tumour stroma and cancer cells. To overcome this limitation we have developed a fluorescence-based experimental model that employs mCherry-labelled stromal cells (e.g. bone marrow fibroblastic stromal cells) co-cultured in direct contact with enhanced green fluorescent protein-labelled tumour cell lines for accurate assessment of proliferation and viability in both cell compartments and adhesion of tumour cells. Additionally, we used fluorescence-based image analysis to determine morphological changes that correlate with cell function (e.g. morphology of the actin cytoskeleton and nuclearity of osteoclasts to predict their bone resorption activity). Using this platform we have revealed that dexamethasone induces HS5 fibroblast proliferation and contact with multiple myeloma cells via a process involving Src/c-Abl kinases. Osteoclasts also inhibited dexamethasone-induced apoptosis in myeloma cells while retaining their normal morphology and functionality in bone resorption. Myeloma resistance to dexamethasone mediated by HS5 cells and osteoclasts was reversed by treatment with the Src/c-Abl inhibitor dasatinib but not with bortezomib. This new experimental platform provides a more precise screening of new therapeutics for improved efficacy of tumour cell killing within the bone marrow microenvironment.",

keywords = "Actins, Antineoplastic Agents, Apoptosis, Bone Marrow, Bone Resorption, Cell Line, Cell Proliferation, Coculture Techniques, Dasatinib, Dexamethasone, Drug Screening Assays, Antitumor, High-Throughput Screening Assays, Humans, Multiple Myeloma, Oncogene Proteins v-abl, Osteoclasts, Pyrimidines, Stromal Cells, Thiazoles, Tumor Microenvironment, src-Family Kinases",

author = "Karthik Ramasamy and Hazera Khatun and Lee Macpherson and Caley, {Mathew P} and Justin Sturge and Mufti, {Ghulam J} and Schey, {Stephen A} and Yolanda Calle",

note = "{\textcopyright} 2012 Blackwell Publishing Ltd.",

year = "2012",

month = jun,

doi = "10.1111/j.1365-2141.2012.09103.x",

language = "English",

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T1 - Fluorescence-based experimental model to evaluate the concomitant effect of drugs on the tumour microenvironment and cancer cells

AU - Ramasamy, Karthik

AU - Khatun, Hazera

AU - Macpherson, Lee

AU - Caley, Mathew P

AU - Sturge, Justin

AU - Mufti, Ghulam J

AU - Schey, Stephen A

AU - Calle, Yolanda

PY - 2012/6

Y1 - 2012/6

N2 - The response of the tumour microenvironment to anti-cancer drugs can influence treatment efficacy. Current drug-screening methodologies fail to distinguish and quantify simultaneously the concomitant effect of drugs on the tumour stroma and cancer cells. To overcome this limitation we have developed a fluorescence-based experimental model that employs mCherry-labelled stromal cells (e.g. bone marrow fibroblastic stromal cells) co-cultured in direct contact with enhanced green fluorescent protein-labelled tumour cell lines for accurate assessment of proliferation and viability in both cell compartments and adhesion of tumour cells. Additionally, we used fluorescence-based image analysis to determine morphological changes that correlate with cell function (e.g. morphology of the actin cytoskeleton and nuclearity of osteoclasts to predict their bone resorption activity). Using this platform we have revealed that dexamethasone induces HS5 fibroblast proliferation and contact with multiple myeloma cells via a process involving Src/c-Abl kinases. Osteoclasts also inhibited dexamethasone-induced apoptosis in myeloma cells while retaining their normal morphology and functionality in bone resorption. Myeloma resistance to dexamethasone mediated by HS5 cells and osteoclasts was reversed by treatment with the Src/c-Abl inhibitor dasatinib but not with bortezomib. This new experimental platform provides a more precise screening of new therapeutics for improved efficacy of tumour cell killing within the bone marrow microenvironment.

AB - The response of the tumour microenvironment to anti-cancer drugs can influence treatment efficacy. Current drug-screening methodologies fail to distinguish and quantify simultaneously the concomitant effect of drugs on the tumour stroma and cancer cells. To overcome this limitation we have developed a fluorescence-based experimental model that employs mCherry-labelled stromal cells (e.g. bone marrow fibroblastic stromal cells) co-cultured in direct contact with enhanced green fluorescent protein-labelled tumour cell lines for accurate assessment of proliferation and viability in both cell compartments and adhesion of tumour cells. Additionally, we used fluorescence-based image analysis to determine morphological changes that correlate with cell function (e.g. morphology of the actin cytoskeleton and nuclearity of osteoclasts to predict their bone resorption activity). Using this platform we have revealed that dexamethasone induces HS5 fibroblast proliferation and contact with multiple myeloma cells via a process involving Src/c-Abl kinases. Osteoclasts also inhibited dexamethasone-induced apoptosis in myeloma cells while retaining their normal morphology and functionality in bone resorption. Myeloma resistance to dexamethasone mediated by HS5 cells and osteoclasts was reversed by treatment with the Src/c-Abl inhibitor dasatinib but not with bortezomib. This new experimental platform provides a more precise screening of new therapeutics for improved efficacy of tumour cell killing within the bone marrow microenvironment.

KW - Actins

KW - Antineoplastic Agents

KW - Apoptosis

KW - Bone Marrow

KW - Bone Resorption

KW - Cell Line

KW - Cell Proliferation

KW - Coculture Techniques

KW - Dasatinib

KW - Dexamethasone

KW - Drug Screening Assays, Antitumor

KW - High-Throughput Screening Assays

KW - Humans

KW - Multiple Myeloma

KW - Oncogene Proteins v-abl

KW - Osteoclasts

KW - Pyrimidines

KW - Stromal Cells

KW - Thiazoles

KW - Tumor Microenvironment

KW - src-Family Kinases

U2 - 10.1111/j.1365-2141.2012.09103.x

DO - 10.1111/j.1365-2141.2012.09103.x

M3 - Article

C2 - 22428569

SN - 1365-2141

VL - 157

SP - 564

EP - 579

JO - British Journal of Haematology

JF - British Journal of Haematology

IS - 5

ER -