Organotypic brain explant culture as a drug evaluation system for malignant brain tumors

Noriaki Minami, Yusuke Maeda, Shunsuke Shibao, Yoshimi Arima, Fumiharu Ohka, Yutaka Kondo, Koji Maruyama, Masatoshi Kusuhara, Takashi Sasayama, Eiji Kohmura, Hideyuki Saya, Oltea Sampetrean

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

Therapeutic options for malignant brain tumors are limited, with new drugs being continuously evaluated. Organotypic brain slice culture has been adopted for neuroscience studies as a system that preserves brain architecture, cellular function, and the vascular network. However, the suitability of brain explants for anticancer drug evaluation has been unclear. We here adopted a mouse model of malignant glioma based on expression of H-RasV12 in Ink4a/Arf−/− neural stem/progenitor cells to establish tumor-bearing brain explants from adult mice. We treated the slices with cisplatin, temozolomide, paclitaxel, or tranilast and investigated the minimal assays required to assess drug effects. Serial fluorescence-based tumor imaging was sufficient for evaluation of cisplatin, a drug with a pronounced cytotoxic action, whereas immunostaining of cleaved caspase 3 (a marker of apoptosis) and of Ki67 (a marker of cell proliferation) was necessary for the assessment of temozolomide action and immunostaining for phosphorylated histone H3 (a marker of mitosis) allowed visualization of paclitaxel-specific effects. Staining for cleaved caspase 3 was also informative in the assessment of drug toxicity for normal brain tissue. Incubation of explants with fluorescently labeled antibodies to CD31 allowed real-time imaging of the microvascular network and complemented time-lapse imaging of tumor cell invasion into surrounding tissue. Our results suggest that a combination of fluorescence imaging and immunohistological staining allows a unified assessment of the effects of various classes of drug on the survival, proliferation, and invasion of glioma cells, and that organotypic brain slice culture is therefore a useful tool for evaluation of antiglioma drugs.

Original languageEnglish
Pages (from-to)2635-2645
Number of pages11
JournalCancer Medicine
Volume6
Issue number11
DOIs
Publication statusPublished - 2017 Nov 1

Fingerprint

Drug Evaluation
Brain Neoplasms
temozolomide
Brain
Glioma
Caspase 3
Pharmaceutical Preparations
Time-Lapse Imaging
Staining and Labeling
Neural Stem Cells
Optical Imaging
Neurosciences
Paclitaxel
Microvessels
Drug-Related Side Effects and Adverse Reactions
Mitosis
Histones
Cisplatin
Blood Vessels
Neoplasms

Keywords

  • Drug screening
  • glioma stem cells
  • invasion
  • malignant brain tumors
  • organotypic slice culture

ASJC Scopus subject areas

  • Oncology
  • Radiology Nuclear Medicine and imaging
  • Cancer Research

Cite this

Organotypic brain explant culture as a drug evaluation system for malignant brain tumors. / Minami, Noriaki; Maeda, Yusuke; Shibao, Shunsuke; Arima, Yoshimi; Ohka, Fumiharu; Kondo, Yutaka; Maruyama, Koji; Kusuhara, Masatoshi; Sasayama, Takashi; Kohmura, Eiji; Saya, Hideyuki; Sampetrean, Oltea.

In: Cancer Medicine, Vol. 6, No. 11, 01.11.2017, p. 2635-2645.

Research output: Contribution to journalArticle

Minami, N, Maeda, Y, Shibao, S, Arima, Y, Ohka, F, Kondo, Y, Maruyama, K, Kusuhara, M, Sasayama, T, Kohmura, E, Saya, H & Sampetrean, O 2017, 'Organotypic brain explant culture as a drug evaluation system for malignant brain tumors', Cancer Medicine, vol. 6, no. 11, pp. 2635-2645. https://doi.org/10.1002/cam4.1174
Minami, Noriaki ; Maeda, Yusuke ; Shibao, Shunsuke ; Arima, Yoshimi ; Ohka, Fumiharu ; Kondo, Yutaka ; Maruyama, Koji ; Kusuhara, Masatoshi ; Sasayama, Takashi ; Kohmura, Eiji ; Saya, Hideyuki ; Sampetrean, Oltea. / Organotypic brain explant culture as a drug evaluation system for malignant brain tumors. In: Cancer Medicine. 2017 ; Vol. 6, No. 11. pp. 2635-2645.
@article{1516dcfd39184eefba83f1a1fb5065cd,
title = "Organotypic brain explant culture as a drug evaluation system for malignant brain tumors",
abstract = "Therapeutic options for malignant brain tumors are limited, with new drugs being continuously evaluated. Organotypic brain slice culture has been adopted for neuroscience studies as a system that preserves brain architecture, cellular function, and the vascular network. However, the suitability of brain explants for anticancer drug evaluation has been unclear. We here adopted a mouse model of malignant glioma based on expression of H-RasV12 in Ink4a/Arf−/− neural stem/progenitor cells to establish tumor-bearing brain explants from adult mice. We treated the slices with cisplatin, temozolomide, paclitaxel, or tranilast and investigated the minimal assays required to assess drug effects. Serial fluorescence-based tumor imaging was sufficient for evaluation of cisplatin, a drug with a pronounced cytotoxic action, whereas immunostaining of cleaved caspase 3 (a marker of apoptosis) and of Ki67 (a marker of cell proliferation) was necessary for the assessment of temozolomide action and immunostaining for phosphorylated histone H3 (a marker of mitosis) allowed visualization of paclitaxel-specific effects. Staining for cleaved caspase 3 was also informative in the assessment of drug toxicity for normal brain tissue. Incubation of explants with fluorescently labeled antibodies to CD31 allowed real-time imaging of the microvascular network and complemented time-lapse imaging of tumor cell invasion into surrounding tissue. Our results suggest that a combination of fluorescence imaging and immunohistological staining allows a unified assessment of the effects of various classes of drug on the survival, proliferation, and invasion of glioma cells, and that organotypic brain slice culture is therefore a useful tool for evaluation of antiglioma drugs.",
keywords = "Drug screening, glioma stem cells, invasion, malignant brain tumors, organotypic slice culture",
author = "Noriaki Minami and Yusuke Maeda and Shunsuke Shibao and Yoshimi Arima and Fumiharu Ohka and Yutaka Kondo and Koji Maruyama and Masatoshi Kusuhara and Takashi Sasayama and Eiji Kohmura and Hideyuki Saya and Oltea Sampetrean",
year = "2017",
month = "11",
day = "1",
doi = "10.1002/cam4.1174",
language = "English",
volume = "6",
pages = "2635--2645",
journal = "Cancer Medicine",
issn = "2045-7634",
publisher = "John Wiley and Sons Ltd",
number = "11",

}

TY - JOUR

T1 - Organotypic brain explant culture as a drug evaluation system for malignant brain tumors

AU - Minami, Noriaki

AU - Maeda, Yusuke

AU - Shibao, Shunsuke

AU - Arima, Yoshimi

AU - Ohka, Fumiharu

AU - Kondo, Yutaka

AU - Maruyama, Koji

AU - Kusuhara, Masatoshi

AU - Sasayama, Takashi

AU - Kohmura, Eiji

AU - Saya, Hideyuki

AU - Sampetrean, Oltea

PY - 2017/11/1

Y1 - 2017/11/1

N2 - Therapeutic options for malignant brain tumors are limited, with new drugs being continuously evaluated. Organotypic brain slice culture has been adopted for neuroscience studies as a system that preserves brain architecture, cellular function, and the vascular network. However, the suitability of brain explants for anticancer drug evaluation has been unclear. We here adopted a mouse model of malignant glioma based on expression of H-RasV12 in Ink4a/Arf−/− neural stem/progenitor cells to establish tumor-bearing brain explants from adult mice. We treated the slices with cisplatin, temozolomide, paclitaxel, or tranilast and investigated the minimal assays required to assess drug effects. Serial fluorescence-based tumor imaging was sufficient for evaluation of cisplatin, a drug with a pronounced cytotoxic action, whereas immunostaining of cleaved caspase 3 (a marker of apoptosis) and of Ki67 (a marker of cell proliferation) was necessary for the assessment of temozolomide action and immunostaining for phosphorylated histone H3 (a marker of mitosis) allowed visualization of paclitaxel-specific effects. Staining for cleaved caspase 3 was also informative in the assessment of drug toxicity for normal brain tissue. Incubation of explants with fluorescently labeled antibodies to CD31 allowed real-time imaging of the microvascular network and complemented time-lapse imaging of tumor cell invasion into surrounding tissue. Our results suggest that a combination of fluorescence imaging and immunohistological staining allows a unified assessment of the effects of various classes of drug on the survival, proliferation, and invasion of glioma cells, and that organotypic brain slice culture is therefore a useful tool for evaluation of antiglioma drugs.

AB - Therapeutic options for malignant brain tumors are limited, with new drugs being continuously evaluated. Organotypic brain slice culture has been adopted for neuroscience studies as a system that preserves brain architecture, cellular function, and the vascular network. However, the suitability of brain explants for anticancer drug evaluation has been unclear. We here adopted a mouse model of malignant glioma based on expression of H-RasV12 in Ink4a/Arf−/− neural stem/progenitor cells to establish tumor-bearing brain explants from adult mice. We treated the slices with cisplatin, temozolomide, paclitaxel, or tranilast and investigated the minimal assays required to assess drug effects. Serial fluorescence-based tumor imaging was sufficient for evaluation of cisplatin, a drug with a pronounced cytotoxic action, whereas immunostaining of cleaved caspase 3 (a marker of apoptosis) and of Ki67 (a marker of cell proliferation) was necessary for the assessment of temozolomide action and immunostaining for phosphorylated histone H3 (a marker of mitosis) allowed visualization of paclitaxel-specific effects. Staining for cleaved caspase 3 was also informative in the assessment of drug toxicity for normal brain tissue. Incubation of explants with fluorescently labeled antibodies to CD31 allowed real-time imaging of the microvascular network and complemented time-lapse imaging of tumor cell invasion into surrounding tissue. Our results suggest that a combination of fluorescence imaging and immunohistological staining allows a unified assessment of the effects of various classes of drug on the survival, proliferation, and invasion of glioma cells, and that organotypic brain slice culture is therefore a useful tool for evaluation of antiglioma drugs.

KW - Drug screening

KW - glioma stem cells

KW - invasion

KW - malignant brain tumors

KW - organotypic slice culture

UR - http://www.scopus.com/inward/record.url?scp=85032965678&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85032965678&partnerID=8YFLogxK

U2 - 10.1002/cam4.1174

DO - 10.1002/cam4.1174

M3 - Article

C2 - 28980419

AN - SCOPUS:85032965678

VL - 6

SP - 2635

EP - 2645

JO - Cancer Medicine

JF - Cancer Medicine

SN - 2045-7634

IS - 11

ER -