Tumor Biology

, Volume 37, Issue 1, pp 541–552 | Cite as

Contribution of resident and recruited macrophages to the photodynamic intervention of colorectal tumor microenvironment

  • María Florencia Pansa
  • María Julia Lamberti
  • Ingrid Sol Cogno
  • Silvia Graciela Correa
  • Natalia Belén Rumie Vittar
  • Viviana Alicia Rivarola
Original Article

Abstract

The study of cellular interactions in the tumor microenvironment has become one of the main areas of research in the fight against cancer. Tumor-associated macrophages (TAMs) influence tumor progression and therapy response due to its functional plasticity. Regarding cancer treatment, photodynamic therapy (PDT) is a minimally invasive and clinically approved procedure that involves the administration of a photosensitizer (PS), a nontoxic photosensitizing drug which is selectively retained in neoplastic tissue. Here, we investigated the role of resident and nonresident macrophages in the context of a PDT-treated colorectal tumor by developing a combination of 2-D and three-dimensional (3-D) experimental platform, recreating tumor-stroma interactions in vitro. Enhancement of cytotoxicity of PDT was achieved in the presence of nonresident macrophages which had a strong anti-tumor phenotype mediated by the production of nitric oxide, IL-6, and tumor necrosis factor alpha (TNF-α). On the contrary, tumor resident macrophages induced a pro-tumor phenotype promoting tumor cell migration and endothelial stimulation. Due to their plasticity, tumor-resident or tumor-recruited macrophages can differentially influence the response of tumors to PDT, so their multifactorial roles should be considered in the overall design of anti-tumor therapeutic.

Keywords

Photodynamic therapy Tumor microenvironment Macrophages TAMs Colorectal cancer 

Abbreviations

TME

Tumor microenvironment

TAMs

Tumor-associated macrophages

NO

Nitric oxide

iNOS

Inducible nitric oxide synthase

TNF-α

Tumor necrosis factor alpha

IL-1

Interleukin-1

IL-6

Interleukin-6

IL-12

Interleukin-12

IL-23

Interleukin-23

PDT

Photodynamic therapy

Me-ALA

Methyl-aminolevulinic acid

3-D

Three-dimensional

GFP

Green fluorescent protein

FBS

Fetal bovine serum

PS

Photosensitizer

CM

Conditioned media

MTT

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

DMSO

Dimethyl sulfoxide

homTME

Homotypic tumor microenvironment

hetTME/Res

Heterotypic tumor microenvironment with resident macrophages

hetTME/NRes

Heterotypic tumor microenvironment with nonresident macrophages

PI

Propidium iodide

Notes

Acknowledgments

This work was supported by grants from Consejo Nacional de Investigaciones Científicas y Técnicas (CONICET), Agencia Nacional de Promoción Científica y Tecnológica (PICT), Secretaría de Ciencia y Técnica (SECyT), Universidad Nacional de Rio Cuarto, Argentina. VR, SGC, ISC, and NBRV are members of the Scientific Researcher Career at CONICET. MFP and MJL hold fellowship from CONICET.

Conflicts of interest

None

Supplementary material

13277_2015_3768_Fig6_ESM.gif (60 kb)
Figure S1

Disruption of heterotypic spheroid. Spheroids composed by SW480-G and RAW 264.7 cells lost tumor-stroma physical interaction during experimental manipulation. For nuclear staining, spheroids were incubated with Hoechst dye (blue). SW480-G tumor cells (green) constitutively expressed GFP. Bar = 100 μm. HÖ: Hoechst. (GIF 60 kb)

13277_2015_3768_MOESM1_ESM.tif (261 kb)
High Resolution Image (TIFF 260 kb)
13277_2015_3768_Fig7_ESM.gif (107 kb)
Figure S2

Experimental schedule of photodynamic treatments on homTME, hetTME/Res and hetTME/NRes. (A) homTME: SW480-G cells (20.000/spheroid) were seeded on agarose-coated round-bottom 96-multiwell to allow 3D structure generation, for 72 h. Next, spheroids were incubated with 0.3 mM Me-ALA during 24 h (PDT-treated) or DMEM without FBS (untreated). Then, PDT-treated spheroids were irradiated with a dose of 0.7 J/cm2. Immediately after that, fresh complete medium was added to both untreated and PDT-treated spheroids. Viability, spheroid size and tumor migration were analyzed 1 or 10 days later. (B) hetTME/Res: SW480-G cells (20.000/spheroid) were seeded on agarose-coated round-bottom 96-multiwell to allow 3D structure generation, for 72 h. Next, spheroids were incubated on macrophage monolayer with 0.3 mM Me-ALA during 24 h (PDT-treated) or DMEM without FBS (untreated). Then, PDT-treated co-cultures were irradiated with a dose of 0.7 J/cm2. Immediately after that, fresh complete medium was added to both untreated and PDT-treated co-cultures. Viability, spheroid size and tumor migration were analyzed 1 or 10 days later. (C) hetTME/NRes: SW480-G cells (20.000/spheroid) were seeded on agarose-coated round-bottom 96-multiwell to allow 3-D structure generation, for 72 h. Next, spheroids were incubated with 0.3 mM Me-ALA during 24 h (PDT-treated) or DMEM without FBS (untreated). Then, PDT-treated spheroids were irradiated with a dose of 0.7 J/cm2. Immediately after that, fresh complete medium was added to both untreated and PDT-treated co-cultures, and seeded on macrophage monolayers. Viability, spheroid size and tumor migration were analyzed 1 or 10 days later. homTME: homotypic tumor microenvironment, hetTME/Res: heterotypic tumor microenvironment with resident macrophages, hetTME/NRes: heterotypic tumor microenvironment with non-resident macrophages, PDT: Photodynamic therapy, FBS: fetal bovine serum. (GIF 106 kb)

13277_2015_3768_MOESM2_ESM.tif (541 kb)
High Resolution Image (TIFF 540 kb)

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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  1. 1.Departamento de Biología Molecular, Facultad de Ciencias Exactas Físico-Químicas y NaturalesUniversidad Nacional de Río CuartoRío CuartoArgentina
  2. 2.Departamento de Bioquímica Clínica, CIBICI-CONICET, Facultad de Ciencias QuímicasUniversidad Nacional de CórdobaCórdobaArgentina

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