Isolation and characterization of tumorspheres from a recurrent pineoblastoma patient: Feasibility of a patient-derived xenograft

Kwak, Jay Jiyong; Shim, Jae Yong; Kim, Do Kyung; Lee, Ji Hyun; Choi, Jihun; Park, Junseong; Shin, Kyoung Jin; Kim, Se Hoon; Kim, Pilnam; Huh, Yong Min; Kim, Eui Hyun; Chang, Jong Hee; Kim, Sun Ho; Kang, Seok‐Gu

doi:10.3892/ijo.2016.3554

Cited by 16 publications

(14 citation statements)

References 45 publications

(41 reference statements)

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“…Finally, very few human PB cell lines have been successfully established in culture. One line, TS13-19, isolated from sporadic PB and propagated as non-adherent tumorspheres in serum-free media 56 , was also highly sensitive to NOR (Fig. 8g).…”

Section: Resultsmentioning

confidence: 99%

“…Our in silico drug prediction analysis identified multiple tricyclic, antidepressant drugs such as NOR, which ranked at the top, as potential therapeutics for Rb/p53-deleted and Rb/p53mutated PBs as well as for Dicer/p53-and Rb/Dicer1/p53-deficient lesions. NOR inhibited growth of primary Rb/p53-and Rb/ Dicer1/p53-deficient PB cells as well as a human PB cell line 56 . NOR suppressed autophagy flux not by blocking assembly of the autolysosome but by disrupting the lysosome.…”

Section: Discussionmentioning

confidence: 97%

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Modeling germline mutations in pineoblastoma uncovers lysosome disruption-based therapy

et al. 2020

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Pineoblastoma is a rare pediatric cancer induced by germline mutations in the tumor suppressors RB1 or DICER1. Presence of leptomeningeal metastases is indicative of poor prognosis. Here we report that inactivation of Rb plus p53 via a WAP-Cre transgene, commonly used to target the mammary gland during pregnancy, induces metastatic pineoblastoma resembling the human disease with 100% penetrance. A stabilizing mutation rather than deletion of p53 accelerates metastatic dissemination. Deletion of Dicer1 plus p53 via WAP-Cre also predisposes to pineoblastoma, albeit with lower penetrance. In silico analysis predicts tricyclic antidepressants such as nortriptyline as potential therapeutics for both pineoblastoma models. Nortriptyline disrupts the lysosome, leading to accumulation of nonfunctional autophagosome, cathepsin B release and pineoblastoma cell death. Nortriptyline further synergizes with the antineoplastic drug gemcitabine to effectively suppress pineoblastoma in our preclinical models, offering new modality for this lethal childhood malignancy.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 97%

Modeling germline mutations in pineoblastoma uncovers lysosome disruption-based therapy

et al. 2020

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“…Tumorspheres (TS) were directly established from fresh GBM tissues as approved by the institutional review board of Severance Hospital, Yonsei University College of Medicine (4-2012-0212). For isolation of TS from GBM specimens, we followed previously published methods for TS isolation from human brain 17 , 34 . The patient-derived GBM cells (pdGCs; TS) were isolated and cultured in 2D, collagen hydrogel, and pdECM environments, respectively.…”

Section: Methodsmentioning

confidence: 99%

The mode and dynamics of glioblastoma cell invasion into a decellularized tissue-derived extracellular matrix-based three-dimensional tumor model

Koh

Cha

Park

et al. 2018

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Glioblastoma multiforme (GBM) is the most common brain tumor with very aggressive and infiltrative. Extracellular matrix (ECM) plays pivotal roles in the infiltrative characteristics of GBM.To understand the invasive characteristic of GBM, it is necessary to study cell-ECM interaction in the physiologically relevant biomimetic model that recapitulates the GBM-specific ECM microenvironment. Here, we propose biomimetic GBM-specific ECM microenvironment for studying mode and dynamics of glioblastoma cell invasion. Using tissue decellularization process, we constructed a patient tissuederived ECM (pdECM)-based three-dimensional in vitro model. In our model, GBM cells exhibited heterogeneous morphology and altered the invasion routes in a microenvironment-adaptive manner. We further elucidate the effects of inhibition of ECM remodeling-related enzymatic activity (Matrix metalloproteinase (MMP) 2/9, hyaluronan synthase (HAS)) on GBM cell invasion. Interestingly, after blocking both enzyme activity, GBM cells underwent morphological transition and switch the invasion mode. Such adaptability could render cell invasion resistant to anti-cancer target therapy. There results provide insight of how organ-specific matrix differentially regulates cancer cell phenotype, and have significant implications for the design of matrix with appropriate physiologically relevant properties for in vitro tumor model.Invasion and dissemination of cancer cells cause migration of neoplastic cells into surrounding tissues, resulting in mortality in tumor patients 1-3 . In particular, cancer cell invasion is of enormous clinical importance, since it involves both distant metastasis and local spreading, whereby cancer cells degrade and migrate through the tissue. The significance of cancer cell invasion is evident in the glioblastoma multiforme (GBM), which shows infiltrative and rapid growth into the surrounding brain tissue. The invasive and migratory characteristics of GBM cells provide a wealth of information about tumors within a patient 3 .Three-dimensional (3D) in vitro culture systems are increasingly employed to assess cell-ECM interactions and invasion of tumor cells. Indeed, cell behavior and responsiveness are dramatically different in a 3D physiological environment versus two-dimensional (2D) Petri dish conditions 4,5 . Importantly, in 3D cultures, ECM is an essential determinant of the cellular response to invasion and migration processes 6 . Brain ECM has a distinct composition from ECM in other tissues and organs, with a low stiffness and loosely connected cellular network. The ECM component of brain tissues contains high amounts of hyaluronic acid (HA), glycosaminoglycans (GAGs), and proteoglycans, but lacks fibrous materials such as collagen, fibronectin, etc 7,8 .In fact, the interaction between GBM cells and unique extracellular environment of the brain could impact on the invasive characteristics of GBM cells. Accumulated experimental and clinical data demonstrate that invasion

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“…GL261-luc cells were constructed using lentivirus-mediated transduction and puromycin-based selection, and cultured in tumorsphere complete media as previously described. 25,26 U87MG and T98G cells were purchased from Korean Cell Line Bank and grown in media according to American Type Culture Collection recommendations. U87MG cells and T98G cells revealed methylated and unmethylated MGMT promoter region, respectively.…”

Section: Cell Linesmentioning

confidence: 99%

Effect of combined anti-PD-1 and temozolomide therapy in glioblastoma

et al. 2018

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Background: Although programmed death-1 (PD-1) blockade is effective in treating several types of cancer, the efficacy of this agent in glioblastoma (GBM) is largely unknown. Methods: We evaluated therapeutic effects of anti-PD-1, temozolomide (TMZ), and their combination in an orthotopic murine GBM model. The phenotype, number, and composition of lymphocytes were evaluated using flow cytometry. Transcriptional profiles of tumor tissues were analyzed using microarrays. Generation of antitumor immunological memory was investigated upon rechallenge. Results: Combined treatment with anti-PD-1 and TMZ yielded synergistic antitumor efficacy in the presence of donor-originated PD-1 + CD8 + T cells in vitro, necessitating in vivo validation. Whereas TMZ did not rescue GBM-implanted mice, anti-PD-1 completely eradicated GBM in 44.4% of mice, and combination of anti-PD-1 and TMZ in all mice. Anti-PD-1 significantly increased the number of tumorinfiltrating lymhpocytes (TILs), and reduced frequencies of exhausted T cells and regulatory T cells. However, combining TMZ with anti-PD-1 significantly decreased the number of TILs, which was also observed with TMZ treatment alone. A transcriptome analysis of tumor tissues revealed that anti-PD-1 monotherapy perturbed immune-related genes, distinctly with combined therapy. Upon rechallenge, tumor growth was not observed in mice cured by anti-PD-1 monotherapy, whereas tumors regrew in the combination group. Furthermore, an analysis of peripheral blood revealed that antitumor memory T cells were generated in mice cured by anti-PD-1 monotherapy, not in the combination group. Conclusion: PD-1 blockade induces long-term therapeutic response, and combination with TMZ further enhances antitumor efficacy. However, immunological memory is provoked by anti-PD-1 monotherapy, not by combined therapy. ARTICLE HISTORY

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Isolation and characterization of tumorspheres from a recurrent pineoblastoma patient: Feasibility of a patient-derived xenograft

Cited by 16 publications

References 45 publications

Modeling germline mutations in pineoblastoma uncovers lysosome disruption-based therapy

Modeling germline mutations in pineoblastoma uncovers lysosome disruption-based therapy

The mode and dynamics of glioblastoma cell invasion into a decellularized tissue-derived extracellular matrix-based three-dimensional tumor model

Effect of combined anti-PD-1 and temozolomide therapy in glioblastoma

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