A Cancer Stem Cell Model for Studying Brain Metastases From Primary Lung Cancer

Nolte, Sara M.; Venugopal, Chitra; McFarlane, Nicole; Morozova, Olena; Hallett, Robin; O’Farrell, Erin; Manoranjan, Branavan; Murty, Naresh K.; Klurfan, Paula; Kachur, Edward; Provias, John; Farrokhyar, Forough; Hassell, John A.; Marra, Marco A.; Singh, Sheila K.

doi:10.1093/jnci/djt022

Cited by 52 publications

(57 citation statements)

References 68 publications

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“…We also performed a traditional and reliable limiting dilution assay (LDA) to determine the fraction of CSCs in each subpopulation. The LDA results revealed the frequency of sphere formation, providing useful information for interpreting the composition of the mixed population of CSCs [23, 46]. Our current data showed that Lgr5-positive subpopulations possessed a higher percentage of CSCs than did Lgr5-negative subpopulations.…”

Section: Discussionmentioning

confidence: 60%

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Lgr5+CD44+EpCAM+ Strictly Defines Cancer Stem Cells in Human Colorectal Cancer

Leng

Xia

Chen

et al. 2018

Cell Physiol Biochem

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Background/Aims: Although EpCAM+CD44+ cells exhibit more stem-like properties than did EpCAM-CD44- cells, the specificity of EpCAM combined with CD44 in defining CSCs needs further improvement. Lgr5 is used as a biomarker to isolate cancer stem cells (CSCs) in colorectal cancer. However, it remains unclear whether Lgr5, along with EpCAM and CD44, can further identify and define CSCs in colorectal cancer. Methods: Lgr5+CD44+EpCAM+, Lgr5+CD44+EpCAM-, Lgr5+CD44-EpCAM+, Lgr5-CD44+EpCAM+, and Lgr5-CD44-EpCAM-cells were separately isolated using fluorescence-activated cell sorting (FACS). Colony formation, self-renewal, differentiation, and tumorigenic properties of these cells were investigated through in vitro experiments and in vivo tumor xenograft models. The expression of stemness genes and CSC- and epithelial-mesenchymal transition (EMT)-related genes, such as KLF4, Oct4, Sox2, Nanog, CD133, CD44, CD166, ALDH1, Lgr5, E-cadherin, ZO-1, Vimentin, Snail, Slug, and Twist, was examined using real-time PCR. Results: Lgr5-positive subpopulations exhibited higher capacities for colony formation, self-renewal, differentiation, and tumorigenicity as well as higher expression of stemness genes and mesenchymal genes and lower expression of epithelial genes than did Lgr5-negative subpopulations. Conclusion: Our data revealed that tumorigenic cells were highly restricted to Lgr5-positive subpopulations. Most importantly, Lgr5+CD44+EpCAM+ cells exhibited more pronounced CSC-like traits than did any other subpopulation, indicating that Lgr5 combined with CD44 and EpCAM can further improve the stem-like traits of CSCs in colorectal cancer.

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Section: Discussionmentioning

confidence: 60%

“…The LDA and tumor sphere-forming assays were performed as previously described, with some modifications [23]. In brief, limiting dilutions of tumor cells were plated (4 to 0.5 cells per well) in 0.2 ml of serum-free DMEM/F12 medium as described for the sphere-forming assay and were cultured for 7 days.…”

Section: Methodsmentioning

confidence: 99%

Lgr5+CD44+EpCAM+ Strictly Defines Cancer Stem Cells in Human Colorectal Cancer

Leng

Xia

Chen

et al. 2018

Cell Physiol Biochem

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“…Many studies have reported the increased invasiveness of CSCs, implicating the involvement of CSCs in the progression and/or development of metastases (46,47). However, most previous in vitro drug screenings on lung cancer patient-derived cancer cells were performed in serum-containing medium (48), whereas tumor spheres in serum-free condition enrich CSC-like populations (49). Herein, PDXs derived from NSCLC brain metastases gave rise to numerous tumor spheres in a few days after mechanical dissociation ex vivo in a reproducible manner, whereas PDXs from NSCLC primary tumor demonstrated poor viability and sphereforming ability in vitro.…”

Section: Discussionmentioning

confidence: 99%

Patient-Derived Xenografts from Non–Small Cell Lung Cancer Brain Metastases Are Valuable Translational Platforms for the Development of Personalized Targeted Therapy

Lee

et al. 2015

Clinical Cancer Research

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Purpose: The increasing prevalence of distant metastases from non-small cell lung cancer (NSCLC) indicates an urgent need for novel therapeutic modalities. Brain metastasis is particularly common in NSCLC, with severe adverse effects on clinical prognosis. Although the molecular heterogeneity of NSCLC and availability of various targeted agents suggest personalized therapeutic approaches for such brain metastases, further development of appropriate preclinical models is needed to validate the strategies.Experimental Design: We established patient-derived xenografts (PDX) using NSCLC brain metastasis surgical samples and elucidated their possible preclinical and clinical implications for personalized treatment.Results: NSCLC brain metastases (n ¼ 34) showed a significantly higher successful PDX establishment rate than primary specimens (n ¼ 64; 74% vs. 23%). PDXs derived from NSCLC brain metastases recapitulated the pathologic, genetic, and functional properties of corresponding parental tumors. Furthermore, tumor spheres established in vitro from the xenografts under serum-free conditions maintained their in vivo brain metastatic potential. Differential phenotypic and molecular responses to 20 targeted agents could subsequently be screened in vitro using these NSCLC PDXs derived from brain metastases. Although PDX establishment from primary NSCLCs was significantly influenced by histologic subtype, clinical aggressiveness, and genetic alteration status, the brain metastases exhibited consistently adequate in vivo tumor take rate and in vitro tumor sphere formation capacity, regardless of clinical and molecular conditions.Conclusions: Therefore, PDXs from NSCLC brain metastases may better represent the heterogeneous advanced NSCLC population and could be utilized as preclinical models to meet unmet clinical needs such as drug screening for personalized treatments.

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“…It has been demonstrated that these CSCs in lung cancer resist to chemotherapeutic agents and have high tumorigenic potential (12,70). Also, the lung CSCs were shown to have augmented ability to move to other tissues (8,12,71). The most important characteristic of CSCs is their ability to generate a new tumor.…”

Section: Cancer Stem Cells (Cscs)mentioning

confidence: 99%

“…Several CSC makers have been used to identify the CSC population in lung cancer, including cellular expression of CD44, CD133 and CD166 (67)(68)(69)(70)(71). Also aldehyde dehydrogenase (ALDH) activity and expression have been shown to indicate stemness in lung cancer (67)(68)(69)(70)(71).…”

Section: Cancer Stem Cells (Cscs)mentioning

confidence: 99%