Using the Chick Embryo Model to Examine the Effects of Hypoxia Pre-conditioning of Uveal Melanoma Cells on Tumor Growth and Metastasis

Draper, James; Alexander, Jodi; Nair, Rohini; Scullion, Nicole; Narayana, Revu Vl; Aughton, Karen; Herrmann, Anne‐Kristin; Vemuganti, Geeta K.; Kalirai, Helen; Coupland, Sarah E.

doi:10.1080/02713683.2022.2152055

Cited by 4 publications

(3 citation statements)

References 36 publications

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“…Our ndings support the ndings of Palmer and colleagues, who investigated the metastasis of the human epidermis carcinoma cell line HEp3 to the liver and lungs of chicks [48]. Another study from the Liverpool Ocular Oncology Group (LOORG) used a chick embryo tumor model and reported the metastasis of uveal melanoma cells to their metastatic sites, which are the liver and eyes [49,50].…”

Section: Discussionsupporting

confidence: 86%

Evaluation of Cancer Stem Cells in Patient-derived Xenografts of Retinoblastoma in a Chick Embryo-Chorioallontoic Membrane Model

VL,

Mishra,

Jakati

et al. 2024

Preprint

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Background: Cancer stem cells (CSCs) in most tumors, including retinoblastoma (Rb), are associated with tumor initiation, metastasis, and drug resistance. Our previous studies identified Rb CSCs as CD44+/CD133- by flowcytometry using size and phenotype. In this study, we further evaluated two prime characteristics of CSCs, i.e., chemoresistance and tumor-initiating properties using Chick embryo-Chorioallantoic Membrane (CE-CAM) model. Methods: After IRB approval was obtained, fresh Rb tumors (n=15) were collected and sorted using antibodies against CD44, followed by CD133. Drug resistance and gene expression were evaluated using the MTT assay and qPCR. Tumor-initiation and metastatic ability were assessed using the CE-CAM assay by transplanting one million cells into a day-7 chick embryo and were evaluated by gross, confocal microscopy, IVIS spectral imaging, alu-qPCR, and histology. Results: The percentage of CSCs in Rb tumors ranged from 4.8 to 28.4%, exhibiting greater drug resistance and enhanced expression of genes related to CSCs, stemness, drug resistance, EMT and invasion, and metastasis-specific genes than tumor non-CSCs (P<0.0001). The CSCs transplanted cells formed white-colored glistening tumor nodules on the CAM and significantly higher localization of fluorescence signals than non-CSCs (p<0.0001). IVIS imaging revealed positive signals in the brain, which confirmed malignant round cells on histopathology and were quantitatively measured by alu-qPCR. Conclusion: Rb CSCs (CD44+/CD133-) are endowed with inherent drug resistance and tumor initiation potential, as demonstrated by their enhanced capacity to form tumor xenografts in the CE-CAM model with evidence of metastasis. The CE-CAM is a valuable, cost-effective pre-clinical model that evaluates the pathogenesis of tumor progression and targets therapies.

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Section: Discussionsupporting

confidence: 86%

Evaluation of Cancer Stem Cells in Patient-derived Xenografts of Retinoblastoma in a Chick Embryo-Chorioallontoic Membrane Model

VL,

Mishra,

Jakati

et al. 2024

Preprint

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show abstract

“…Our ndings support Palmer and colleagues investigation into the metastasis of the Human Epidermoid Carcinoma cell line (HEp3) to the liver and lung of chicks 48 . Another study from the Liverpool Ocular Oncology Group (LOORG) used a chick embryo tumor model and reported the metastasis of uveal melanoma cells to their metastatic sites, which are the liver and eyes 49,50 .…”

Section: Discussionmentioning

confidence: 99%

Evaluation of Cancer Stem Cells in Patient-derived Xenografts of Retinoblastoma in Chick Embryo-Chorioallontoic Membrane model

VL,

Mishra,

Jakati

et al. 2024

Preprint

View full text Add to dashboard Cite

Background Cancer Stem Cells (CSCs) in most tumors, including Retinoblastoma (Rb), are associated with tumor-initiation, metastasis, and drug resistance. Our previous studies identified Rb CSCs as CD44+/CD133- by flowcytometry using size and phenotype. In this study, we further evaluated two prime characteristics of CSC, i.e., chemoresistance and tumor-initiating properties using Chick embryo-Chorioallantoic Membrane (CE-CAM) model. Methods After obtaining IRB approval, fresh Rb tumors (n = 15) was collected and sorted using antibodies against CD44, followed by CD133. Drug resistance and gene expression were evaluated using MTT assay and qPCR. Tumor-initiation and metastatic ability were assessed using the CE-CAM assay by transplanting one million cells into a day-7 chick embryo and were evaluated by gross, confocal microscopy, IVIS spectral imaging, alu-qPCR, and histology. Results The percentage of CSCs in Rb tumors ranges from 4.8 to 28.4%, exhibiting higher drug resistance and enhanced gene expression of CSC, stemness, drug resistance, EMT & invasion, and metastasis-specific genes than tumor non-CSCs (P < 0.0001). The CSC transplanted cells formed white-colored glistening tumor nodules on the CAM and significantly higher localization of fluorescence signals than non-CSCs (p < 0.0001). IVIS imaging showed positive signals in the brain, which confirmed malignant round cells on histopathology and were quantitatively measured by alu-qPCR. Conclusion The Rb CSCs (CD44+/CD133-) are endowed with inherent drug resistance and tumor-initiating, as demonstrated by enhanced capacity to form tumor xenografts in the CE-CAM model with evidence of metastasis. The CE-CAM is a valuable, cost-effective pre-clinical model that evaluates the pathogenesis of tumor progression and also targets therapies.

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“…Hypoxic environment is thought to be associated with malignant tumor progression [7,8]. In UVM, hypoxia has been shown to promote tumor invasion and progression through activation of Notch and MAPK pathways [9,10]. Moreover, HRGs were associated with the prognosis of UVM [11].…”

Section: Introductionmentioning

confidence: 99%

Hypoxia-Related lncRNA Correlates with Prognosis and Immune Microenvironment in Uveal Melanoma

Chen,

Cheng

et al. 2024

Preprint

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Background Genes related to hypoxia are correlated to the prognosis of a variety of solid malignant tumors. However, How hypoxia related lncRNA function in Uveal Melanoma is still rare. Methods From TCGA database, eighty UVM samples were incorporated. Prognostic HRL was screened by COX univariate analysis and Pearson correlation analysis. HRL characteristics were constructed using Lasso analysis. Gene enrichment analysis revealed the linkage of HRLs to immune features. CCK8 assay was adopted to measure the propagation of MuM2B cells, while tumor invasion and migration were appraised by Transwell and Wound-healing experiments. Inflammatory factors and macrophage polarization were evaluated by qPCR. Results 621 prognostic HRLs were screened and was constructed in 12 HRLs. Risk score showed salient correlation with survival time of UVM patients. In addition, HRL was correlated with diverse key immune checkpoints, revealing possible targets for immunotherapy. Immune-related pathways were highly enriched in high-risk group. Protective HRL, LINC02367, was saliently associated with microenvironment and survival time of UVM patients. In vitro assays demonstrated LINC02367 significantly affected the propogation and migration of MuM2B. LINC02367 modulates macrophage polarization through inflammatory factor levels thereby affecting the immune microenvironment Conclusions In our study, a novel HRL signature was developed to predict the prognosis of UVM patients. HRLs are potential biomarkers and therapeutic targets for UVM.

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Using the Chick Embryo Model to Examine the Effects of Hypoxia Pre-conditioning of Uveal Melanoma Cells on Tumor Growth and Metastasis

Cited by 4 publications

References 36 publications

Evaluation of Cancer Stem Cells in Patient-derived Xenografts of Retinoblastoma in a Chick Embryo-Chorioallontoic Membrane Model

Evaluation of Cancer Stem Cells in Patient-derived Xenografts of Retinoblastoma in a Chick Embryo-Chorioallontoic Membrane Model

Evaluation of Cancer Stem Cells in Patient-derived Xenografts of Retinoblastoma in Chick Embryo-Chorioallontoic Membrane model

Hypoxia-Related lncRNA Correlates with Prognosis and Immune Microenvironment in Uveal Melanoma

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