scholarly journals Adrenocortical Carcinoma Xenograft in Zebrafish Embryos as a Model To Study the In Vivo Cytotoxicity of Abiraterone Acetate

Endocrinology ◽  
2019 ◽  
Vol 160 (11) ◽  
pp. 2620-2629 ◽  
Author(s):  
Alessandra Gianoncelli ◽  
Michela Guarienti ◽  
Martina Fragni ◽  
Michela Bertuzzi ◽  
Elisa Rossini ◽  
...  
Keyword(s):  
Tumor Growth ◽  
Adrenocortical Carcinoma ◽  
Cell Model ◽  
Control Cell ◽  
Abiraterone Acetate ◽  
New Drugs ◽  
Zebrafish Embryos ◽  
Cell Area ◽  
Zebrafish Model ◽  
Immunodeficient Mice

Abstract Abiraterone acetate (AbiAc) inhibits tumor growth when administered to immunodeficient mice engrafted with the in vitro cell model of human adrenocortical carcinoma (ACC). Here, we developed and validated a zebrafish model engrafted with cortisol-secreting ACC cells to study the effects of AbiAc on tumor growth. The experimental conditions for AbiAc absorption in AB zebrafish embryos including embryo number, AbiAc concentration, and absorption time curve by liquid chromatography–tandem mass spectrometry were set up. The AbiAc effect on steroid production in AB zebrafish embryos was measured as well. ACC cells (the NCI-H295R cell line, the primary cell ACC29, and the negative control cell SW13) were treated with drug-induced liver injury fluorescent dye, and ∼240 cells per 4 nL was injected in the subperidermal space of the yolk sac of AB zebrafish embryos (n = 80 ± 10). The cell area was measured with Noldus DanioScopeTM software. AbiAc absorption in AB zebrafish embryos was stage dependent. Abiraterone (Abi) concentration decreased, whereas its main metabolite, Δ4A, increased. Accordingly, we demonstrated that zebrafish expressed mRNA encoding the enzyme 3β-hydroxysteroid dehydrogenase, which converts Abi in Δ4A. Furthermore, ABiAc reduced cortisol production and increased progesterone in zebrafish embryos. Three days after cell injection, the cortisol-secreting ACC cell area in solvent-treated embryos was significantly higher than that in 1 µM AbiAC‒treated embryos, whereas no AbiAc effect was observed in SW13 cells, which lack the Abi target enzyme CYP17A1.Zebrafish embryos xenografted with ACC tumor cells could be a useful, fast, and reproducible experimental model to preclinically test the activity of new drugs in human ACC.

Immunodeficient mice balb/c nude and modeling of various types of tumor growth for preclinical studies

2017 ◽  
Vol 16 (3) ◽  
pp. 6-13 ◽  
Author(s):  
H. M. Treshalina
Keyword(s):  
Tumor Growth ◽  
Nude Mice ◽  
Review Paper ◽  
Control Cell ◽  
Cellular Immunotherapy ◽  
Federal State ◽  
Malignant Growth ◽  
Forecast Performance ◽  
Immunodeficient Mice ◽  
The Russian Federation

Obtaining preclinical models of malignant growth in a variety of xenografts (heterotransplantation), usually the subcutaneous, is the most important area for the use of nude mice in oncology. In a review paper describes the characteristics of Balb/c nude mice obtained from Buffalo (USA) and introduced to the Federal State Budgetary Scientific Institution “N.N. Blokhin Russian Cancer Research Center» of the Russian Ministry of Health of the Russian Federation and systematized their application. There are various variants «palentology» models of tumor growth, allowing you to personalize the forecast performance of selected impacts for a particular patient. Detailed modeling of metastases of various human cancers: colon cancer, pancreas, esophagus, lung, breast and prostate glands, testicles and ovaries, tumors of the head and neck. The characteristic of orthotopic (“MetaMouse” model) and experimental confirmation of the value of the hypothesis of Paget’s “seed and soil”, postulating a paired part of the process of metastasis of cell and tissue components of the tumor. Describes the use of the subcutaneous xenografts as control cell lines in the study of oncogenic potencies of the various drugs that are recommended for cellular immunotherapy of human. Examples and main thematic literature in recent years.

scholarly journals Klotho Exerts an Emerging Role in Cytokinesis

Genes ◽  
2020 ◽  
Vol 11 (9) ◽  
pp. 1048
Author(s):  
Chiao-Yin Sun ◽  
Chi-Yuan Chou ◽  
Yu-Ying Hsieh ◽  
Kang-Chieh Lo ◽  
Yan-Liang Liou ◽  
...  
Keyword(s):  
Spatial Correlation ◽  
Cell Model ◽  
Critical Role ◽  
Aurora Kinase ◽  
Early Embryo ◽  
Mass Spectrometry Analysis ◽  
Zebrafish Embryos ◽  
Zebrafish Model ◽  
Aurora Kinase B ◽  
Spectrometry Analysis

The Klotho gene functions as an anti-aging gene. A previous klotho-knockout mice study indicated that neither male nor female gametocytes could accomplish the first meiotic division. It suggested that Klotho might regulate cell division. In this study, we determined the roles of Klotho in cytokinesis in cultural human cells (HEK293 and HeLa) and in zebrafish embryos. Immunoprecipitation, mass spectrometry analysis, and a zebrafish model were used in this study. The results showed that Klotho is located in the midbody, which correlated with cytokinesis related kinases, Aurora kinase B and citron kinases, in the late stage of cytokinesis. There was a spatial correlation between the abscission site and the location of Klotho in the cytokinesis bridge. A three-dimensional structural reconstruction study demonstrated there was a spatial correlation among Klotho, Aurora kinase B, and citron kinases in the midbody. In addition, Klotho depletion inactivated Aurora kinases; it was also indicated that Klotho depletion caused aberrant cell cycle and delayed cytokinesis in a cell model. The study with zebrafish embryos suggested that klotho knockdown caused early embryo development abnormality due to dysregulated cytokinesis. In conclusion, Klotho might have a critical role in cytokinesis regulation by interacting with the cytokinesis related kinases.

In vitro reproduction and explanation of an abiraterone acetate withdrawal phenomenon.

2014 ◽  
Vol 32 (4_suppl) ◽  
pp. 154-154 ◽  
Author(s):  
Paul Thelen ◽  
Hubertus Jarry ◽  
Lutz Trojan ◽  
Sabine Brookman-May ◽  
Felix Bremmer
Keyword(s):  
Cell Proliferation ◽  
Tumor Cells ◽  
Cell Model ◽  
Combined Treatment ◽  
Specific Antigen ◽  
Abiraterone Acetate ◽  
Androgen Deprivation ◽  
New Drugs ◽  
Cross Resistance ◽  
Western Blots

154 Background: Androgen deprivation and direct androgen receptor (AR) antagonism are standard treatments for castration resistant prostate cancer (CRPC). The advent of new drugs such as abiraterone acetate (AA) and enzalutamide rendered highly efficient implementations of both anti-androgen strategies. However, reports of therapy resistance or cross resistance soon accompanied reports of successful clinical trials of these new drugs. Moreover, case reports of an AA-withdrawal phenomenon surfaced, an outcome normally associated with AR antagonism e.g. by bicalutamide with high AR-affinity and the potential to switch from antagonism to AR-activation. Methods: CRPC cell model VCaP and VCaP resistant to permanent treatment of 5 µmol/L AA (Janssen-Cilag, Germany) were analyzed under AA and for AA withdrawal. Expression of AR, AR-V7, androgen regulated genes, CYP17A1 and AKR1C3 was quantitated by real time RT-PCR. Further analyses were performed by western blots, prostate-specific antigen (PSA) ELISA kits and cell proliferation BrdU tests. Results: VCaP cells resistant to 5 µmol/L AA after 7 months revealed excessive AR and CYP17A1 expression and a marked decline of tumor cell proliferation in response to AA withdrawal or to a similar extent to added 1 nmol/L testosterone. Proliferation dropped to less than 50% when both procedures were combined. Elimination of tumor cells was accompanied by a massive release of PSA which was most pronounced in the combined treatment. Conclusions: Overexpression of AR turning into a liability as a therapeutic target for a testosterone surge has been reported by several research groups. AA withdrawal from AA resistant cells complies with these necessities by overexpression of AR and CYP17A1 formerly arrested by AA. In patients a PSA decline upon AA withdrawal may be explained by an excessive loss of PSA secreting tumor cells. Therefore, AA qualifies uniquely for effective intermittent androgen deprivation of CRPC.

scholarly journals Edelfosine nanoemulsions inhibit tumor growth of triple negative breast cancer in zebrafish xenograft model

2021 ◽  
Vol 11 (1) ◽  
Author(s):  
Sofia M. Saraiva ◽  
Carlha Gutiérrez-Lovera ◽  
Jeannette Martínez-Val ◽  
Sainza Lores ◽  
Belén L. Bouzo ◽  
...  
Keyword(s):  
Breast Cancer ◽  
Tumor Growth ◽  
Triple Negative Breast Cancer ◽  
Triple Negative ◽  
Xenograft Model ◽  
Skin Barrier ◽  
Zebrafish Embryos ◽  
Biorelevant Media

AbstractTriple negative breast cancer (TNBC) is known for being very aggressive, heterogeneous and highly metastatic. The standard of care treatment is still chemotherapy, with adjacent toxicity and low efficacy, highlighting the need for alternative and more effective therapeutic strategies. Edelfosine, an alkyl-lysophospholipid, has proved to be a promising therapy for several cancer types, upon delivery in lipid nanoparticles. Therefore, the objective of this work was to explore the potential of edelfosine for the treatment of TNBC. Edelfosine nanoemulsions (ET-NEs) composed by edelfosine, Miglyol 812 and phosphatidylcholine as excipients, due to their good safety profile, presented an average size of about 120 nm and a neutral zeta potential, and were stable in biorelevant media. The ability of ET-NEs to interrupt tumor growth in TNBC was demonstrated both in vitro, using a highly aggressive and invasive TNBC cell line, and in vivo, using zebrafish embryos. Importantly, ET-NEs were able to penetrate through the skin barrier of MDA-MB 231 xenografted zebrafish embryos, into the yolk sac, leading to an effective decrease of highly aggressive and invasive tumoral cells’ proliferation. Altogether the results demonstrate the potential of ET-NEs for the development of new therapeutic approaches for TNBC.

scholarly journals Lignans from Bursera fagaroides Affect In Vivo Cell Behavior by Disturbing the Tubulin Cytoskeleton in Zebrafish Embryos

Molecules ◽  
2018 ◽  
Vol 24 (1) ◽  
pp. 8 ◽  
Author(s):  
Mayra Antúnez-Mojica ◽  
Andrés Rojas-Sepúlveda ◽  
Mario Mendieta-Serrano ◽  
Leticia Gonzalez-Maya ◽  
Silvia Marquina ◽  
...  
Keyword(s):  
Cell Migration ◽  
Biological Effects ◽  
In Vitro Models ◽  
In Vivo Model ◽  
Zebrafish Embryos ◽  
Microtubule Cytoskeleton ◽  
Zebrafish Model ◽  
Bioassay Guided Fractionation

By using a zebrafish embryo model to guide the chromatographic fractionation of antimitotic secondary metabolites, seven podophyllotoxin-type lignans were isolated from a hydroalcoholic extract obtained from the steam bark of Bursera fagaroides. The compounds were identified as podophyllotoxin (1), β-peltatin-A-methylether (2), 5′-desmethoxy-β-peltatin-A-methylether (3), desmethoxy-yatein (4), desoxypodophyllotoxin (5), burseranin (6), and acetyl podophyllotoxin (7). The biological effects on mitosis, cell migration, and microtubule cytoskeleton remodeling of lignans 1–7 were further evaluated in zebrafish embryos by whole-mount immunolocalization of the mitotic marker phospho-histone H3 and by a tubulin antibody. We found that lignans 1, 2, 4, and 7 induced mitotic arrest, delayed cell migration, and disrupted the microtubule cytoskeleton in zebrafish embryos. Furthermore, microtubule cytoskeleton destabilization was observed also in PC3 cells, except for 7. Therefore, these results demonstrate that the cytotoxic activity of 1, 2, and 4 is mediated by their microtubule-destabilizing activity. In general, the in vivo and in vitro models here used displayed equivalent mitotic effects, which allows us to conclude that the zebrafish model can be a fast and cheap in vivo model that can be used to identify antimitotic natural products through bioassay-guided fractionation.

scholarly journals High-resolution cardiovascular function confirms functional orthology of myocardial contractility pathways in zebrafish

2010 ◽  
Vol 42 (2) ◽  
pp. 300-309 ◽  
Author(s):  
Jordan T. Shin ◽  
Eugene V. Pomerantsev ◽  
John D. Mably ◽  
Calum A. MacRae
Keyword(s):  
High Speed ◽  
Dynamic Range ◽  
Model Organism ◽  
Cardiovascular Physiology ◽  
Zebrafish Embryos ◽  
Cardiovascular Development ◽  
Zebrafish Model ◽  
Ventricular Performance ◽  
Pharmacological Agents

Phenotype-driven screens in larval zebrafish have transformed our understanding of the molecular basis of cardiovascular development. Screens to define the genetic determinants of physiological phenotypes have been slow to materialize as a result of the limited number of validated in vivo assays with relevant dynamic range. To enable rigorous assessment of cardiovascular physiology in living zebrafish embryos, we developed a suite of software tools for the analysis of high-speed video microscopic images and validated these, using established cardiomyopathy models in zebrafish as well as modulation of the nitric oxide (NO) pathway. Quantitative analysis in wild-type fish exposed to NO or in a zebrafish model of dilated cardiomyopathy demonstrated that these tools detect significant differences in ventricular chamber size, ventricular performance, and aortic flow velocity in zebrafish embryos across a large dynamic range. These methods also were able to establish the effects of the classic pharmacological agents isoproterenol, ouabain, and verapamil on cardiovascular physiology in zebrafish embryos. Sequence conservation between zebrafish and mammals of key amino acids in the pharmacological targets of these agents correlated with the functional orthology of the physiological response. These data provide evidence that the quantitative evaluation of subtle physiological differences in zebrafish can be accomplished at a resolution and with a dynamic range comparable to those achieved in mammals and provides a mechanism for genetic and small-molecule dissection of functional pathways in this model organism.

scholarly journals Induction of protein aggregation in zebrafish embryos as a method for the screening of new drugs or mutations against proteinopathies

MethodsX ◽  
2018 ◽  
Vol 5 ◽  
pp. 322-327
Author(s):  
Pablo Lobos-Ruiz ◽  
Gissela Araya ◽  
Octavio Monasterio ◽  
Luis Pouchucq
Keyword(s):  
Protein Aggregation ◽  
New Drugs ◽  
Zebrafish Embryos

Pcgf1 regulates early neural tube development through histone methylation in zebrafish

2020 ◽  
Author(s):  
Xinyue Li ◽  
Guangyu Ji ◽  
Juan Zhou ◽  
Jingyi Du ◽  
Xian Li ◽  
...  
Keyword(s):  
Cell Fate ◽  
Neural Tube ◽  
Histone Methylation ◽  
Control Cell ◽  
Neural Induction ◽  
Induction Phase ◽  
Zebrafish Embryos ◽  
Rna Seq ◽  
Self Renewal ◽  
Neural Tube Development

Abstract Objective Early neural tube development in the embryo includes neural induction and self-renewal of neural stem cells (NSCs). The abnormal of neural tube development could lead to neural tube defects. The research on the mechanism of neural induction is the key to reveal the pathogenesis of the abnormal of neural tube. Though studies have confirmed a genetic component, the responsible mechanisms for the abnormal of neural tube are still largely unknown. Polycomb repressive complex 1 (PRC1) plays an important role in regulating early embryonic development, and has been sub-classified into six major complexes based on the presence of a Pcgf subunit. Pcgf1, as one of six Pcgf paralogs, is an important requirement in early embryonic brain development. Here, we intended to investigate the role and mechanism of Pcgf1 in early neural tube development of zebrafish embryos. Material and methods Morpholino (MO) antisense oligonucleotides were used to construct a Pcgf1 loss-of function zebrafish model. We analyzed the phenotype of zebrafish embryos and the expression of related genes in the process of neural induction by in situ hybridization, immunolabelling and RNA-sEq. The regulation of histone modifications on gene was detected by western blot and chromatin immunoprecipitation. Results In this study, we found that zebrafish embryos exhibited small head and reduced or even absence of telencephalon after inhibiting the expression of Pcgf1. Moreover, the neural induction process of zebrafish embryos was abnormal, and the subsequent NSCs self-renewal was inhibited under the inhibition of Pcgf1. RNA-seq and gene ontology (GO) analysis identified that the differentially expressed genes were enriched in many functional categories which related to the development phenotype. Finally, our results showed that Pcgf1 regulated the trimethylation of histone H3K27 in the Ngn1 and Otx2 promoter regions, and the levels of H3K4me3 at the promoters of Pou5f3 and Nanog. Conclusion Together, our data for the first time demonstrate that Pcgf1 plays an essential role in early neural induction phase through histone methylation in neural tube development. Our findings reveal a critical context-specific function for Pcgf1 in directing PRC1 to control cell fate.

scholarly journals Involvement of 27-Hydroxycholesterol in Mitotane Action on Adrenocortical Carcinoma

Cells ◽  
2020 ◽  
Vol 9 (4) ◽  
pp. 885
Author(s):  
Antonina Germano ◽  
Daniela Rossin ◽  
Valerio Leoni ◽  
Noemi Iaia ◽  
Laura Saba ◽  
...  
Keyword(s):  
Adrenocortical Carcinoma ◽  
Cholesterol Metabolism ◽  
Cell Model ◽  
Gas Chromatography Mass Spectrometry ◽  
Strong Increase ◽  
Vitro Model ◽  
A Minor ◽  
Cholesterol Precursors

Adrenocortical carcinoma (ACC) is a rare cancer with poor prognosis. Mitotane, the standard treatment for ACC, impairs adrenocortical steroid biosynthesis and cholesterol metabolism. In the H295R cell line, a standard ACC in vitro model, mitotane was previously reported to enhance the production of some oxysterols. To verify the possible mechanistic involvement of oxysterols in the anti-ACC effect of mitotane, a gas chromatography mass spectrometry (GC-MS) profiling of oxysterols and the main cholesterol precursors was carried out in H295R cells. Among the oxysterols detected in mitotane-treated cells, 27OHC was markedly produced, as well as lanosterol and lathosterol cholesterol precursors. In this cell model, mitotane was confirmed to affect mitochondrial transmembrane potential and induce apoptosis. Such cytotoxic effects were perfectly matched by H295R cell treatment with a single identical micromolar amount of 27OHC. The mitotane-dependent strong increase in 27OHC was confirmed in vivo, in the plasma of ACC patients under treatment with the drug. Moreover, lanosterol, lathosterol, desmosterol and, to a minor extent, 24-hydroxycholesterol and 25-hydroxycholesterol plasma levels were significantly increased in those patients. The cytotoxic effect of mitotane on ACC cells may be partly related to the increased intracellular level of 27OHC induced by the drug itself.

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