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A Biobank of Breast Cancer Explants with Preserved Intra-tumor Heterogeneity to Screen Anticancer Compounds

View Article: PubMed Central - PubMed

ABSTRACT

The inter- and intra-tumor heterogeneity of breast cancer needs to be adequately captured in pre-clinical models. We have created a large collection of breast cancer patient-derived tumor xenografts (PDTXs), in which the morphological and molecular characteristics of the originating tumor are preserved through passaging in the mouse. An integrated platform combining in vivo maintenance of these PDTXs along with short-term cultures of PDTX-derived tumor cells (PDTCs) was optimized. Remarkably, the intra-tumor genomic clonal architecture present in the originating breast cancers was mostly preserved upon serial passaging in xenografts and in short-term cultured PDTCs. We assessed drug responses in PDTCs on a high-throughput platform and validated several ex vivo responses in vivo. The biobank represents a powerful resource for pre-clinical breast cancer pharmacogenomic studies (http://caldaslab.cruk.cam.ac.uk/bcape), including identification of biomarkers of response or resistance.

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Related to Figure 4 and STAR Methods(A) Representative microscopy images from 8 PDTX models of short-term cultures (PDTCs) at day 7 after plating.(B) Box plots showing the percentage of human DNA on PDTX (n = 94 samples) and PDTC (n = 15 samples) models (n = 29). Data estimated from our sequencing-based approach.(C) Changes in cell number and viability of PDTCs at each time-point.(D) Representative FACS image from PKH26 assay. Table showing quantification of PKH26 low cells in 2 PDTCs (STG195 and STG201) and a highly proliferative breast cancer cell line (MDAMB231), shown for comparison purposes, at different time-points.(E) Scatterplots comparing AUC values measuring drug response for 19 drugs in 8 models (AB521, AB555, AB582, STG195, STG316, STG321, STG335 and VHIO093) using 3 different viability assays. 10 different doses were tested. Curve fitting and computation of the AUC was done as described in the STAR Methods. Dots highlighted in color correspond to PI3K pathway inhibitors: GDC032 (PI3Kα) in red, GDC0941 (pan PI3K) in blue, AZD6482 (PI3Kβ) in purple, and AZD8055 (mTOR) in green.
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figs4: Related to Figure 4 and STAR Methods(A) Representative microscopy images from 8 PDTX models of short-term cultures (PDTCs) at day 7 after plating.(B) Box plots showing the percentage of human DNA on PDTX (n = 94 samples) and PDTC (n = 15 samples) models (n = 29). Data estimated from our sequencing-based approach.(C) Changes in cell number and viability of PDTCs at each time-point.(D) Representative FACS image from PKH26 assay. Table showing quantification of PKH26 low cells in 2 PDTCs (STG195 and STG201) and a highly proliferative breast cancer cell line (MDAMB231), shown for comparison purposes, at different time-points.(E) Scatterplots comparing AUC values measuring drug response for 19 drugs in 8 models (AB521, AB555, AB582, STG195, STG316, STG321, STG335 and VHIO093) using 3 different viability assays. 10 different doses were tested. Curve fitting and computation of the AUC was done as described in the STAR Methods. Dots highlighted in color correspond to PI3K pathway inhibitors: GDC032 (PI3Kα) in red, GDC0941 (pan PI3K) in blue, AZD6482 (PI3Kβ) in purple, and AZD8055 (mTOR) in green.

Mentions: The PDTXs described constitute a living biobank of breast cancer explants that retain through passaging the inter- and intra-tumor heterogeneity encountered in the clinic. We therefore developed a method to enable the use of this valuable resource for high-content pre-clinical drug screening, similar to the approach widely used with cell lines (Barretina et al., 2012, Garnett et al., 2012). The method involved optimizing short ex vivo culture of cells isolated from the PDTXs (named PDTCs). These short-term PDTC cultures were successfully generated from all models where attempted (n = 27, at least two different passages from each; Figure S4A; see STAR Methods).


A Biobank of Breast Cancer Explants with Preserved Intra-tumor Heterogeneity to Screen Anticancer Compounds
Related to Figure 4 and STAR Methods(A) Representative microscopy images from 8 PDTX models of short-term cultures (PDTCs) at day 7 after plating.(B) Box plots showing the percentage of human DNA on PDTX (n = 94 samples) and PDTC (n = 15 samples) models (n = 29). Data estimated from our sequencing-based approach.(C) Changes in cell number and viability of PDTCs at each time-point.(D) Representative FACS image from PKH26 assay. Table showing quantification of PKH26 low cells in 2 PDTCs (STG195 and STG201) and a highly proliferative breast cancer cell line (MDAMB231), shown for comparison purposes, at different time-points.(E) Scatterplots comparing AUC values measuring drug response for 19 drugs in 8 models (AB521, AB555, AB582, STG195, STG316, STG321, STG335 and VHIO093) using 3 different viability assays. 10 different doses were tested. Curve fitting and computation of the AUC was done as described in the STAR Methods. Dots highlighted in color correspond to PI3K pathway inhibitors: GDC032 (PI3Kα) in red, GDC0941 (pan PI3K) in blue, AZD6482 (PI3Kβ) in purple, and AZD8055 (mTOR) in green.
© Copyright Policy - CC BY
Related In: Results  -  Collection

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getmorefigures.php?uid=PMC5037319&req=5

figs4: Related to Figure 4 and STAR Methods(A) Representative microscopy images from 8 PDTX models of short-term cultures (PDTCs) at day 7 after plating.(B) Box plots showing the percentage of human DNA on PDTX (n = 94 samples) and PDTC (n = 15 samples) models (n = 29). Data estimated from our sequencing-based approach.(C) Changes in cell number and viability of PDTCs at each time-point.(D) Representative FACS image from PKH26 assay. Table showing quantification of PKH26 low cells in 2 PDTCs (STG195 and STG201) and a highly proliferative breast cancer cell line (MDAMB231), shown for comparison purposes, at different time-points.(E) Scatterplots comparing AUC values measuring drug response for 19 drugs in 8 models (AB521, AB555, AB582, STG195, STG316, STG321, STG335 and VHIO093) using 3 different viability assays. 10 different doses were tested. Curve fitting and computation of the AUC was done as described in the STAR Methods. Dots highlighted in color correspond to PI3K pathway inhibitors: GDC032 (PI3Kα) in red, GDC0941 (pan PI3K) in blue, AZD6482 (PI3Kβ) in purple, and AZD8055 (mTOR) in green.
Mentions: The PDTXs described constitute a living biobank of breast cancer explants that retain through passaging the inter- and intra-tumor heterogeneity encountered in the clinic. We therefore developed a method to enable the use of this valuable resource for high-content pre-clinical drug screening, similar to the approach widely used with cell lines (Barretina et al., 2012, Garnett et al., 2012). The method involved optimizing short ex vivo culture of cells isolated from the PDTXs (named PDTCs). These short-term PDTC cultures were successfully generated from all models where attempted (n = 27, at least two different passages from each; Figure S4A; see STAR Methods).

View Article: PubMed Central - PubMed

ABSTRACT

The inter- and intra-tumor heterogeneity of breast cancer needs to be adequately captured in pre-clinical models. We have created a large collection of breast cancer patient-derived tumor xenografts (PDTXs), in which the morphological and molecular characteristics of the originating tumor are preserved through passaging in the mouse. An integrated platform combining in vivo maintenance of these PDTXs along with short-term cultures of PDTX-derived tumor cells (PDTCs) was optimized. Remarkably, the intra-tumor genomic clonal architecture present in the originating breast cancers was mostly preserved upon serial passaging in xenografts and in short-term cultured PDTCs. We assessed drug responses in PDTCs on a high-throughput platform and validated several ex vivo responses in vivo. The biobank represents a powerful resource for pre-clinical breast cancer pharmacogenomic studies (http://caldaslab.cruk.cam.ac.uk/bcape), including identification of biomarkers of response or resistance.

No MeSH data available.


Related in: MedlinePlus