Challenges, applications and future directions of precision medicine in prostate cancer – the role of organoids and patient‐derived xenografts

Joshi, Andre; Roberts, Matthew J.; Alinezhad, Saeid; Williams, Elizabeth D.; Vela, Ian

doi:10.1111/bju.15103

Cited by 11 publications

(10 citation statements)

References 49 publications

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“…Pre-clinical patient-derived xenograft (PDX) models are used to understand molecular mechanisms and test potential cancer therapies [ 25 ]. They are generated by implanting clinical cancer biospecimens (most commonly obtained from biopsy or surgical specimens) in immuno-compromised hosts (typically mice), and may sometimes by subsequently serially passaged by transferring a small piece of the resultant tumour to a new host.…”

Section: Introductionmentioning

confidence: 99%

Diversity of Epithelial-Mesenchymal Phenotypes in Circulating Tumour Cells from Prostate Cancer Patient-Derived Xenograft Models

Hassan

Blick

Thompson

et al. 2021

Cancers

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Metastasis is the leading cause of cancer-related deaths worldwide. The epithelial-mesenchymal plasticity (EMP) status of primary tumours has relevance to metastatic potential and therapy resistance. Circulating tumour cells (CTCs) provide a window into the metastatic process, and molecular characterisation of CTCs in comparison to their primary tumours could lead to a better understanding of the mechanisms involved in the metastatic cascade. In this study, paired blood and tumour samples were collected from four prostate cancer patient-derived xenograft (PDX) models (BM18, LuCaP70, LuCaP96, LuCaP105) and assessed using an EMP-focused, 42 gene human-specific, nested quantitative RT-PCR assay. CTC burden varied amongst the various xenograft models with LuCaP96 having the highest number of CTCs per mouse (mean: 704; median: 31) followed by BM18 (mean: 101; median: 21), LuCaP70 (mean: 73; median: 16) and LuCaP105 (mean: 57; median: 6). A significant relationship was observed between tumour size and CTC number (p = 0.0058). Decreased levels of kallikrein-related peptidase 3 (KLK3) mRNA (which encodes prostate-specific antigen; PSA) were observed in CTC samples from all four models compared to their primary tumours. Both epithelial- and mesenchymal-associated genes were commonly expressed at higher levels in CTCs compared to the bulk primary tumour, although some common EMT-associated genes (CDH1, VIM, EGFR, EPCAM) remained unchanged. Immunofluorescence co-staining for pan-cytokeratin (KRT) and vimentin (VIM) indicated variable proportions of CTCs across the full EMP axis, even in the same model. EMP hybrids predominated in the BM18 and LuCaP96 models, but were not detected in the LuCaP105 model, and variable numbers of KRT+ and human VIM+ cells were observed in each model. SERPINE1, which encodes plasminogen activator inhibitor-1 (PAI-1), was enriched at the RNA level in CTCs compared to primary tumours and was the most commonly expressed mesenchymal gene in the CTCs. Co-staining for SERPINE1 and KRT revealed SERPINE1+ cells in 7/11 samples, six of which had SERPINE+KRT+ CTCs. Cell size variation was observed in CTCs. The majority of samples (8/11) contained larger CTCs ranging from 15.3 to 37.8 µm, whilst smaller cells (10.7 ± 4.1 µm, similar in size to peripheral blood mononuclear cells (PBMCs)) were identified in 6 of 11 samples. CTC clusters were also identified in 9/11 samples, containing 2–100 CTCs per cluster. Where CTC heterogeneity was observed in the clusters, epithelial-like cells (KRT+VIM−) were located on the periphery of the cluster, forming a layer around hybrid (KRT+VIM+) or mesenchymal-like (KRT−VIM+) cells. The CTC heterogeneity observed in these models emphasises the complexity in CTC isolation and classification and supports the increasingly recognised importance of the epithelial-mesenchymal hybrid state in cancer progression and metastasis.

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Section: Introductionmentioning

confidence: 99%

Diversity of Epithelial-Mesenchymal Phenotypes in Circulating Tumour Cells from Prostate Cancer Patient-Derived Xenograft Models

Hassan

Blick

Thompson

et al. 2021

Cancers

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“…Recent advances in organotypic culture techniques have enabled ex vivo propagation of tumor-derived PDCOs that incorporate primary cell biology, 3D structural complexity, and tumor heterogeneity into in vitro model systems [2,4,16,23,[33][34][35][36][37]. The conventional approach to organoid analysis includes traditional tissue processing like FFPE and sectioning [4,23,24].…”

Section: Discussionmentioning

confidence: 99%

“…Three-dimensional organotypic models have emerged as an in vitro model system that can incorporate unique features of primary tumor biology, structural complexity, and intra-and interpatient tumor heterogeneity [2,3]. In recent years, considerable improvements have been reported in the development of 3D organoid cultures in a variety of solid tumors, particularly in gastrointestinal carcinomas.…”

Section: Introductionmentioning

confidence: 99%

Live cell molecular analysis of primary prostate cancer organoids identifies persistent androgen receptor signaling

et al. 2021

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Prostate Cancer (PC) is a disease with remarkable tumor heterogeneity that often manifests in significant intra-patient variability with regards to clinical outcomes and treatment response. Commonly available PC cell lines do not accurately reflect the complexity of this disease and there is critical need for development of new models to recapitulate the intricate hierarchy of tumor pathogenesis. In current study, we established ex vivo primary patient-derived cancer organoid (PDCO) cultures from prostatectomy specimens of patients with locally advanced PC. We then performed a comprehensive multi-parameter characterization of the cellular composition utilizing a novel approach for live-cell staining and direct imaging in the integrated microfluidic Stacks device. Using orthogonal flow cytometry analysis, we demonstrate that primary PDCOs maintain distinct subsets of epithelial cells throughout culture and that these cells conserve expression of androgen receptor (AR)-related elements. Furthermore, to confirm the tumor-origin of the PDCOs we have analyzed the expression of PC-associated epigenetic biomarkers including promoter methylation of the GSTP1, RASSF1 and APC and RARb genes by employing a novel microfluidic rare-event screening protocol. These results demonstrate that this ex vivo PDCO model recapitulates the complexity of the epithelial tumor microenvironment of multifocal PC using orthogonal analyses. Furthermore, we propose to leverage the Stacks microfluidic device as a high-throughput, translational platform to interrogate phenotypic and molecular endpoints with the capacity to incorporate a complex tumor microenvironment.

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“…Recent advances in organotypic culture techniques have enabled ex vivo propagation of tumor-derived PDCOs that incorporate primary cell biology, 3D structural complexity, and tumor heterogeneity into in vitro model systems [2,4,23,33,16,[34][35][36][37]. The conventional approach to organoid analysis includes traditional tissue processing like FFPE and sectioning [4,23,24].…”

Section: Discussionmentioning

confidence: 99%

Live Cell Molecular Analysis of Primary Prostate Cancer Organoids Identifies Persistent Androgen Receptor Signaling

Héninger

Kosoff

Rodems

et al. 2021

Preprint

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Prostate Cancer (PC) is a disease with remarkable tumor heterogeneity that often manifests in significant intra-patient variability with regards to clinical outcomes and treatment response. Commonly available PC cell lines do not accurately reflect the complexity of this disease and there is critical need for development of new models to recapitulate the intricate hierarchy of tumor pathogenesis. In current study, we established ex vivo primary patient-derived cancer organoid (PDCO) cultures from prostatectomy specimens of patients with locally advanced PC. We then performed a comprehensive multi-parameter characterization of the cellular composition utilizing a novel approach for live-cell staining and direct imaging in the integrated microfluidic Stacks device. Using orthogonal flow cytometry analysis, we demonstrate that primary PDCOs maintain distinct subsets of epithelial cells throughout culture and that these cells conserve expression of androgen receptor (AR) related elements. Furthermore, to confirm the tumor-origin of the PDCOs we have analyzed the expression of PC-associated epigenetic biomarkers including promoter methylation of the GSTP1, RASSF1 and APC and RARb genes by employing a novel microfluidic rare-event screening protocol.These results demonstrate that this ex vivo PDCO model recapitulates the complexity of the epithelial tumor microenvironment of multifocal PC using orthogonal analyses. Furthermore, we propose to leverage the Stacks microfluidic device as a high-throughput, translational platform to interrogate phenotypic and molecular endpoints with the capacity to incorporate a complex tumor microenvironment.

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Challenges, applications and future directions of precision medicine in prostate cancer – the role of organoids and patient‐derived xenografts

Cited by 11 publications

References 49 publications

Diversity of Epithelial-Mesenchymal Phenotypes in Circulating Tumour Cells from Prostate Cancer Patient-Derived Xenograft Models

Diversity of Epithelial-Mesenchymal Phenotypes in Circulating Tumour Cells from Prostate Cancer Patient-Derived Xenograft Models

Live cell molecular analysis of primary prostate cancer organoids identifies persistent androgen receptor signaling

Live Cell Molecular Analysis of Primary Prostate Cancer Organoids Identifies Persistent Androgen Receptor Signaling

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