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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 in: MedlinePlus

Related to STAR Methods and Figures 2A and 2B(A) Representative images from the histopathological analysis of PDTXs. Images from both an ER+ (STG335) and a triple negative (STG139) model at different passages are shown.(B) Calibration curve for estimation of mouse content based on the proportion of mapped reads from WES to the human genome.(C) Estimated percentage of mouse contamination from WES data. Top panel: data for each model at different passages is represented. Bottom panel: Box plots showing the distribution of percentage of human cells per model in all samples tested, including different mice from the same passage and the same model, and different passages of the same model (n = 94 samples from 29 models; estimates from the same model and passage have been averaged).(D) Representative FACS plot of single cells (left) and a FISH image (right) on an FFPE tissue section from an example PDTX sample. Table shows average percentage of mouse cells in different PDTX models tested by FACS.
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figs2: Related to STAR Methods and Figures 2A and 2B(A) Representative images from the histopathological analysis of PDTXs. Images from both an ER+ (STG335) and a triple negative (STG139) model at different passages are shown.(B) Calibration curve for estimation of mouse content based on the proportion of mapped reads from WES to the human genome.(C) Estimated percentage of mouse contamination from WES data. Top panel: data for each model at different passages is represented. Bottom panel: Box plots showing the distribution of percentage of human cells per model in all samples tested, including different mice from the same passage and the same model, and different passages of the same model (n = 94 samples from 29 models; estimates from the same model and passage have been averaged).(D) Representative FACS plot of single cells (left) and a FISH image (right) on an FFPE tissue section from an example PDTX sample. Table shows average percentage of mouse cells in different PDTX models tested by FACS.

Mentions: Histologically, PDTXs (23 models analyzed) showed similar morphology to the originating tumor; tubule formation and associated stroma were present in the xenograft, as seen in the matched patient cancer sample (Figure S2A). Histological review of multiple PDTX passages (Table S2) revealed that tumor tissue morphology remained stable with serial engraftment. Analysis of immunohistochemistry for epithelial markers (CK5, CK8, CK14, CK18, E-cadherin, and epithelial specific antigen) and for clinical biomarkers (ER, PR, Her2, Ki67, and p53) showed these features were similar in matched pairs of PDTX model and originating breast cancer sample and were consistently retained with passaging (Figure S2A and Table S2 for summary of the data).


A Biobank of Breast Cancer Explants with Preserved Intra-tumor Heterogeneity to Screen Anticancer Compounds
Related to STAR Methods and Figures 2A and 2B(A) Representative images from the histopathological analysis of PDTXs. Images from both an ER+ (STG335) and a triple negative (STG139) model at different passages are shown.(B) Calibration curve for estimation of mouse content based on the proportion of mapped reads from WES to the human genome.(C) Estimated percentage of mouse contamination from WES data. Top panel: data for each model at different passages is represented. Bottom panel: Box plots showing the distribution of percentage of human cells per model in all samples tested, including different mice from the same passage and the same model, and different passages of the same model (n = 94 samples from 29 models; estimates from the same model and passage have been averaged).(D) Representative FACS plot of single cells (left) and a FISH image (right) on an FFPE tissue section from an example PDTX sample. Table shows average percentage of mouse cells in different PDTX models tested by FACS.
© Copyright Policy - CC BY
Related In: Results  -  Collection

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figs2: Related to STAR Methods and Figures 2A and 2B(A) Representative images from the histopathological analysis of PDTXs. Images from both an ER+ (STG335) and a triple negative (STG139) model at different passages are shown.(B) Calibration curve for estimation of mouse content based on the proportion of mapped reads from WES to the human genome.(C) Estimated percentage of mouse contamination from WES data. Top panel: data for each model at different passages is represented. Bottom panel: Box plots showing the distribution of percentage of human cells per model in all samples tested, including different mice from the same passage and the same model, and different passages of the same model (n = 94 samples from 29 models; estimates from the same model and passage have been averaged).(D) Representative FACS plot of single cells (left) and a FISH image (right) on an FFPE tissue section from an example PDTX sample. Table shows average percentage of mouse cells in different PDTX models tested by FACS.
Mentions: Histologically, PDTXs (23 models analyzed) showed similar morphology to the originating tumor; tubule formation and associated stroma were present in the xenograft, as seen in the matched patient cancer sample (Figure S2A). Histological review of multiple PDTX passages (Table S2) revealed that tumor tissue morphology remained stable with serial engraftment. Analysis of immunohistochemistry for epithelial markers (CK5, CK8, CK14, CK18, E-cadherin, and epithelial specific antigen) and for clinical biomarkers (ER, PR, Her2, Ki67, and p53) showed these features were similar in matched pairs of PDTX model and originating breast cancer sample and were consistently retained with passaging (Figure S2A and Table S2 for summary of the data).

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