Complex in vitro and in vivo prostate cancer models for the PREDECT consortium: Abstract

Suzana Vidic, Norbert Esser, Ronald de Hoogt, Ingrid Verberne, Ira Kogan-Sakin, Yan Yan Stein, Varda Rotter, Michael Barbier, Yolanda Chong, Sabine D. Breucker, Karine Smans, Malin Akerfelt, Matthias Nees, Peter King, Ian Hickson, Wytske van Weerden, Ralph Graeser

Research output: Contribution to journalReview ArticleScientific

Abstract

The failure of many drugs in the clinic - drugs proven to be active in pre-clinical assays - has raised question marks on the predictive power of these models, often based on cell line data.

The divergence of these cell lines from the original tumors, and their rapid growth as simple monocultures on 2D plastic were highlighted as potentially causing these issues.
Implantation of the cell lines into mice adds complexity to these models, but the crosstalk with mouse stroma also leads to confounding results (eg, HGF, IL-6).

PREDECT, a European consortium funded by the Innovative Medicines Initiative (IMI), has the goal to build and characterise more predictive Oncology model platforms for three pathologies (lung, breast and prostate). Signalling pathways and heterogeneity of these model platforms are compared to each other as well as to primary human tissue using a central TMA-based platform.

For prostate cancer, a 3D tumor growth assay system was established recently using RFP/FLuc-labelled LNCaP, PC346C, or VCaP human prostate cancer cells grown in a matrix of Matrigel, collagen, or a mix of the two, in the presence or absence of GFP/RLuc labelled human stromal cells (WPMY, or an immortalised cancer-associated fibroblast cell line, CAF). Cell densities, ratios, and matrix concentrations/volumes were optimised for cells cultured for 3 weeks.
In an approach to improve tumor-stroma crosstalk in vivo, LNCaP cells pre-grown in 3D cultures with or without human stromal cells were implanted orthotopically into SCID mice.

Growth of the tumor and stromal cells was monitored by in vivo bioluminescence, and the effect of the co-implantation on the primary tumor and metastasis was analysed via ex-vivo luciferase assays and IHC.

Whereas LNCaP and PC346C cells readily formed spheroids in 3D culture, VCaPs remained as single cells. Growth of VCaPs was facilitated by pre-forming spheroids in vitro, followed by matrix embedding. Addition of stromal cells stimulated growth of not only the tumor, but also the stromal cells in some conditions, as monitored using RFP and GFP live cell imaging.
3D, as well as stromal cell-mediated treatment resistance was observed. Using the fluorescent dyes Hoechst, EdU, and NucView, the proliferative status of individual cells within spheroids could be analysed in situ. Finally, paraffin-embedding of the 3D cultures allowed for TMA analysis and comparison to other models and patient samples.
Results will also be shown on the orthotopic growth of LNCaP tumors and the effect of stromal cell co-implantation.

Introducing complexity to cell culture may help to generate more realistic, and thus also more predictive models - in a first step using even standard cell lines.
Original languageEnglish
Article number2023
JournalCancer Research
Volume74
Issue number19 Supplement
DOIs
Publication statusPublished - 2014
MoE publication typeB1 Article in a scientific magazine
Event105th Annual Meeting of the American Association for Cancer Research - San Diego, CA, United States
Duration: 5 Apr 20149 Apr 2014

Fingerprint

Stromal Cells
Prostatic Neoplasms
Growth
Cell Line
Neoplasms
Paraffin Embedding
Somatostatin-Secreting Cells
SCID Mice
In Vitro Techniques
Tumor Cell Line
Luciferases
Fluorescent Dyes
Pharmaceutical Preparations
Plastics
Prostate
Cultured Cells
Interleukin-6
Breast
Collagen
Cell Culture Techniques

Cite this

Vidic, S., Esser, N., de Hoogt, R., Verberne, I., Kogan-Sakin, I., Yan Stein, Y., ... Graeser, R. (2014). Complex in vitro and in vivo prostate cancer models for the PREDECT consortium: Abstract. Cancer Research, 74(19 Supplement), [2023]. https://doi.org/10.1158/1538-7445.AM2014-2023
Vidic, Suzana ; Esser, Norbert ; de Hoogt, Ronald ; Verberne, Ingrid ; Kogan-Sakin, Ira ; Yan Stein, Yan ; Rotter, Varda ; Barbier, Michael ; Chong, Yolanda ; Breucker, Sabine D. ; Smans, Karine ; Akerfelt, Malin ; Nees, Matthias ; King, Peter ; Hickson, Ian ; van Weerden, Wytske ; Graeser, Ralph. / Complex in vitro and in vivo prostate cancer models for the PREDECT consortium : Abstract. In: Cancer Research. 2014 ; Vol. 74, No. 19 Supplement.
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title = "Complex in vitro and in vivo prostate cancer models for the PREDECT consortium: Abstract",
abstract = "The failure of many drugs in the clinic - drugs proven to be active in pre-clinical assays - has raised question marks on the predictive power of these models, often based on cell line data. The divergence of these cell lines from the original tumors, and their rapid growth as simple monocultures on 2D plastic were highlighted as potentially causing these issues.Implantation of the cell lines into mice adds complexity to these models, but the crosstalk with mouse stroma also leads to confounding results (eg, HGF, IL-6).PREDECT, a European consortium funded by the Innovative Medicines Initiative (IMI), has the goal to build and characterise more predictive Oncology model platforms for three pathologies (lung, breast and prostate). Signalling pathways and heterogeneity of these model platforms are compared to each other as well as to primary human tissue using a central TMA-based platform.For prostate cancer, a 3D tumor growth assay system was established recently using RFP/FLuc-labelled LNCaP, PC346C, or VCaP human prostate cancer cells grown in a matrix of Matrigel, collagen, or a mix of the two, in the presence or absence of GFP/RLuc labelled human stromal cells (WPMY, or an immortalised cancer-associated fibroblast cell line, CAF). Cell densities, ratios, and matrix concentrations/volumes were optimised for cells cultured for 3 weeks.In an approach to improve tumor-stroma crosstalk in vivo, LNCaP cells pre-grown in 3D cultures with or without human stromal cells were implanted orthotopically into SCID mice.Growth of the tumor and stromal cells was monitored by in vivo bioluminescence, and the effect of the co-implantation on the primary tumor and metastasis was analysed via ex-vivo luciferase assays and IHC.Whereas LNCaP and PC346C cells readily formed spheroids in 3D culture, VCaPs remained as single cells. Growth of VCaPs was facilitated by pre-forming spheroids in vitro, followed by matrix embedding. Addition of stromal cells stimulated growth of not only the tumor, but also the stromal cells in some conditions, as monitored using RFP and GFP live cell imaging.3D, as well as stromal cell-mediated treatment resistance was observed. Using the fluorescent dyes Hoechst, EdU, and NucView, the proliferative status of individual cells within spheroids could be analysed in situ. Finally, paraffin-embedding of the 3D cultures allowed for TMA analysis and comparison to other models and patient samples.Results will also be shown on the orthotopic growth of LNCaP tumors and the effect of stromal cell co-implantation.Introducing complexity to cell culture may help to generate more realistic, and thus also more predictive models - in a first step using even standard cell lines.",
author = "Suzana Vidic and Norbert Esser and {de Hoogt}, Ronald and Ingrid Verberne and Ira Kogan-Sakin and {Yan Stein}, Yan and Varda Rotter and Michael Barbier and Yolanda Chong and Breucker, {Sabine D.} and Karine Smans and Malin Akerfelt and Matthias Nees and Peter King and Ian Hickson and {van Weerden}, Wytske and Ralph Graeser",
year = "2014",
doi = "10.1158/1538-7445.AM2014-2023",
language = "English",
volume = "74",
journal = "Cancer Research",
issn = "0008-5472",
number = "19 Supplement",

}

Vidic, S, Esser, N, de Hoogt, R, Verberne, I, Kogan-Sakin, I, Yan Stein, Y, Rotter, V, Barbier, M, Chong, Y, Breucker, SD, Smans, K, Akerfelt, M, Nees, M, King, P, Hickson, I, van Weerden, W & Graeser, R 2014, 'Complex in vitro and in vivo prostate cancer models for the PREDECT consortium: Abstract', Cancer Research, vol. 74, no. 19 Supplement, 2023. https://doi.org/10.1158/1538-7445.AM2014-2023

Complex in vitro and in vivo prostate cancer models for the PREDECT consortium : Abstract. / Vidic, Suzana; Esser, Norbert; de Hoogt, Ronald; Verberne, Ingrid; Kogan-Sakin, Ira; Yan Stein, Yan; Rotter, Varda; Barbier, Michael; Chong, Yolanda; Breucker, Sabine D.; Smans, Karine; Akerfelt, Malin; Nees, Matthias; King, Peter; Hickson, Ian; van Weerden, Wytske; Graeser, Ralph.

In: Cancer Research, Vol. 74, No. 19 Supplement, 2023, 2014.

Research output: Contribution to journalReview ArticleScientific

TY - JOUR

T1 - Complex in vitro and in vivo prostate cancer models for the PREDECT consortium

T2 - Abstract

AU - Vidic, Suzana

AU - Esser, Norbert

AU - de Hoogt, Ronald

AU - Verberne, Ingrid

AU - Kogan-Sakin, Ira

AU - Yan Stein, Yan

AU - Rotter, Varda

AU - Barbier, Michael

AU - Chong, Yolanda

AU - Breucker, Sabine D.

AU - Smans, Karine

AU - Akerfelt, Malin

AU - Nees, Matthias

AU - King, Peter

AU - Hickson, Ian

AU - van Weerden, Wytske

AU - Graeser, Ralph

PY - 2014

Y1 - 2014

N2 - The failure of many drugs in the clinic - drugs proven to be active in pre-clinical assays - has raised question marks on the predictive power of these models, often based on cell line data. The divergence of these cell lines from the original tumors, and their rapid growth as simple monocultures on 2D plastic were highlighted as potentially causing these issues.Implantation of the cell lines into mice adds complexity to these models, but the crosstalk with mouse stroma also leads to confounding results (eg, HGF, IL-6).PREDECT, a European consortium funded by the Innovative Medicines Initiative (IMI), has the goal to build and characterise more predictive Oncology model platforms for three pathologies (lung, breast and prostate). Signalling pathways and heterogeneity of these model platforms are compared to each other as well as to primary human tissue using a central TMA-based platform.For prostate cancer, a 3D tumor growth assay system was established recently using RFP/FLuc-labelled LNCaP, PC346C, or VCaP human prostate cancer cells grown in a matrix of Matrigel, collagen, or a mix of the two, in the presence or absence of GFP/RLuc labelled human stromal cells (WPMY, or an immortalised cancer-associated fibroblast cell line, CAF). Cell densities, ratios, and matrix concentrations/volumes were optimised for cells cultured for 3 weeks.In an approach to improve tumor-stroma crosstalk in vivo, LNCaP cells pre-grown in 3D cultures with or without human stromal cells were implanted orthotopically into SCID mice.Growth of the tumor and stromal cells was monitored by in vivo bioluminescence, and the effect of the co-implantation on the primary tumor and metastasis was analysed via ex-vivo luciferase assays and IHC.Whereas LNCaP and PC346C cells readily formed spheroids in 3D culture, VCaPs remained as single cells. Growth of VCaPs was facilitated by pre-forming spheroids in vitro, followed by matrix embedding. Addition of stromal cells stimulated growth of not only the tumor, but also the stromal cells in some conditions, as monitored using RFP and GFP live cell imaging.3D, as well as stromal cell-mediated treatment resistance was observed. Using the fluorescent dyes Hoechst, EdU, and NucView, the proliferative status of individual cells within spheroids could be analysed in situ. Finally, paraffin-embedding of the 3D cultures allowed for TMA analysis and comparison to other models and patient samples.Results will also be shown on the orthotopic growth of LNCaP tumors and the effect of stromal cell co-implantation.Introducing complexity to cell culture may help to generate more realistic, and thus also more predictive models - in a first step using even standard cell lines.

AB - The failure of many drugs in the clinic - drugs proven to be active in pre-clinical assays - has raised question marks on the predictive power of these models, often based on cell line data. The divergence of these cell lines from the original tumors, and their rapid growth as simple monocultures on 2D plastic were highlighted as potentially causing these issues.Implantation of the cell lines into mice adds complexity to these models, but the crosstalk with mouse stroma also leads to confounding results (eg, HGF, IL-6).PREDECT, a European consortium funded by the Innovative Medicines Initiative (IMI), has the goal to build and characterise more predictive Oncology model platforms for three pathologies (lung, breast and prostate). Signalling pathways and heterogeneity of these model platforms are compared to each other as well as to primary human tissue using a central TMA-based platform.For prostate cancer, a 3D tumor growth assay system was established recently using RFP/FLuc-labelled LNCaP, PC346C, or VCaP human prostate cancer cells grown in a matrix of Matrigel, collagen, or a mix of the two, in the presence or absence of GFP/RLuc labelled human stromal cells (WPMY, or an immortalised cancer-associated fibroblast cell line, CAF). Cell densities, ratios, and matrix concentrations/volumes were optimised for cells cultured for 3 weeks.In an approach to improve tumor-stroma crosstalk in vivo, LNCaP cells pre-grown in 3D cultures with or without human stromal cells were implanted orthotopically into SCID mice.Growth of the tumor and stromal cells was monitored by in vivo bioluminescence, and the effect of the co-implantation on the primary tumor and metastasis was analysed via ex-vivo luciferase assays and IHC.Whereas LNCaP and PC346C cells readily formed spheroids in 3D culture, VCaPs remained as single cells. Growth of VCaPs was facilitated by pre-forming spheroids in vitro, followed by matrix embedding. Addition of stromal cells stimulated growth of not only the tumor, but also the stromal cells in some conditions, as monitored using RFP and GFP live cell imaging.3D, as well as stromal cell-mediated treatment resistance was observed. Using the fluorescent dyes Hoechst, EdU, and NucView, the proliferative status of individual cells within spheroids could be analysed in situ. Finally, paraffin-embedding of the 3D cultures allowed for TMA analysis and comparison to other models and patient samples.Results will also be shown on the orthotopic growth of LNCaP tumors and the effect of stromal cell co-implantation.Introducing complexity to cell culture may help to generate more realistic, and thus also more predictive models - in a first step using even standard cell lines.

U2 - 10.1158/1538-7445.AM2014-2023

DO - 10.1158/1538-7445.AM2014-2023

M3 - Review Article

VL - 74

JO - Cancer Research

JF - Cancer Research

SN - 0008-5472

IS - 19 Supplement

M1 - 2023

ER -

Vidic S, Esser N, de Hoogt R, Verberne I, Kogan-Sakin I, Yan Stein Y et al. Complex in vitro and in vivo prostate cancer models for the PREDECT consortium: Abstract. Cancer Research. 2014;74(19 Supplement). 2023. https://doi.org/10.1158/1538-7445.AM2014-2023