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5 results on '"Swain, Amanda"'

1. Genome-wide barcoded transposon screen for cancer drug sensitivity in haploid mouse embryonic stem cells.

Author

Pettitt, Stephen J, Krastev, Dragomir B, Pemberton, Helen N, Fontebasso, Yari, Frankum, Jessica, Rehman, Farah L, Brough, Rachel, Song, Feifei, Bajrami, Ilirjana, Rafiq, Rumana, Wallberg, Fredrik, Kozarewa, Iwanka, Fenwick, Kerry, Armisen-Garrido, Javier, Swain, Amanda, Gulati, Aditi, Campbell, James, Ashworth, Alan, and Lord, Christopher J

Subjects
Animals, Mice, Neoplasms, DNA Transposable Elements, Antineoplastic Agents, Haploidy, Genome-Wide Association Study, Mouse Embryonic Stem Cells
Abstract

We describe a screen for cellular response to drugs that makes use of haploid embryonic stem cells. We generated ten libraries of mutants with piggyBac gene trap transposon integrations, totalling approximately 100,000 mutant clones. Random barcode sequences were inserted into the transposon vector to allow the number of cells bearing each insertion to be measured by amplifying and sequencing the barcodes. These barcodes were associated with their integration sites by inverse PCR. We exposed these libraries to commonly used cancer drugs and profiled changes in barcode abundance by Ion Torrent sequencing in order to identify mutations that conferred sensitivity. Drugs tested included conventional chemotherapeutics as well as targeted inhibitors of topoisomerases, poly(ADP-ribose) polymerase (PARP), Hsp90 and WEE1.

Published
2017

2. Brca2 and Trp53 deficiency cooperate in the progression of mouse prostate tumourigenesis.

Author

Francis, Jeffrey C, McCarthy, Afshan, Thomsen, Martin K, Ashworth, Alan, and Swain, Amanda

Subjects
Animals, Mice, Knockout, Mice, Prostatic Intraepithelial Neoplasia, Prostatic Neoplasms, Cell Transformation, Neoplastic, DNA Damage, Disease Progression, BRCA2 Protein, Apoptosis, Male, Tumor Suppressor Protein p53, Cell Transformation, Neoplastic, Knockout, Genetics, Developmental Biology
Abstract

Epidemiological studies have shown that one of the strongest risk factors for prostate cancer is a family history of the disease, suggesting that inherited factors play a major role in prostate cancer susceptibility. Germline mutations in BRCA2 predispose to breast and ovarian cancer with its predominant tumour suppressor function thought to be the repair of DNA double-strand breaks. BRCA2 has also been implicated in prostate cancer etiology, but it is unclear the impact that mutations in this gene have on prostate tumourigenesis. Here we have undertaken a genetic analysis in the mouse to determine the role of Brca2 in the adult prostate. We show that deletion of Brca2 specifically in prostate epithelia results in focal hyperplasia and low-grade prostate intraepithelial neoplasia (PIN) in animals over 12 months of age. Simultaneous deletion of Brca2 and the tumour suppressor Trp53 in prostate epithelia gave rise to focal hyperplasia and atypical cells at 6 months, leading to high-grade PIN in animals from 12 months. Epithelial cells in these lesions show an increase in DNA damage and have higher levels of proliferation, but also elevated apoptosis. Castration of Brca2;Trp53 mutant animals led to regression of PIN lesions, but atypical cells persisted that continued to proliferate and express nuclear androgen receptor. This study provides evidence that Brca2 can act as a tumour suppressor in the prostate, and the model we describe should prove useful in the development of new therapeutic approaches.

Published
2010

3. A mouse SWATH-mass spectrometry reference spectral library enables deconvolution of species-specific proteomic alterations in human tumour xenografts

Author

Krasny, Lukas, Bland, Philip, Burns, Jessica, Lima, Nadia Carvalho, Harrison, Peter T., Pacini, Laura, Elms, Mark L., Ning, Jian, Martinez, Victor Garcia, Yu, Yi-Ru, Acton, Sophie E., Ho, Ping-Chih, Calvo, Fernando, Swain, Amanda, Howard, Beatrice A., Natrajan, Rachael C., Huang, Paul H., The Institute of Cancer Research (UK), Breast Cancer Research Foundation, Medical Research Council (UK), Ministerio de Economía y Competitividad (España), Ministerio de Ciencia, Innovación y Universidades (España), Agencia Estatal de Investigación (España), and European Research Council

Subjects
Male, Proteomics, Proteome, Mouse, DCIS, lcsh:Medicine, Mice, Nude, Mouse Models, Breast Neoplasms, Cell Communication, Mice, SCID, Mice, Breast cancer, Species Specificity, Tandem Mass Spectrometry, Cell Line, Tumor, lcsh:Pathology, Tumor Microenvironment, Animals, Humans, Resource Article, Databases, Protein, Xenografts, Breast Neoplasms/metabolism, Breast Neoplasms/pathology, Carcinoma, Intraductal, Noninfiltrating/metabolism, Carcinoma, Intraductal, Noninfiltrating/pathology, Chromatography, Liquid, Female, Heterografts, Mice, Inbred C57BL, NIH 3T3 Cells, Neoplasm Proteins/metabolism, Neoplasm Transplantation, Stromal Cells/metabolism, Stromal Cells/pathology, Mass spectrometry, SWATH-MS, lcsh:R, Neoplasm Proteins, Carcinoma, Intraductal, Noninfiltrating, Stromal Cells, lcsh:RB1-214
Abstract

© 2020. Published by The Company of Biologists Ltd., SWATH-mass spectrometry (MS) enables accurate and reproducible proteomic profiling in multiple model organisms including the mouse. Here, we present a comprehensive mouse reference spectral library (MouseRefSWATH) that permits quantification of up to 10,597 proteins (62.2% of the mouse proteome) by SWATH-MS. We exploit MouseRefSWATH to develop an analytical pipeline for species-specific deconvolution of proteomic alterations in human tumour xenografts (XenoSWATH). This method overcomes the challenge of high sequence similarity between mouse and human proteins, facilitating the study of host microenvironment-tumour interactions from ‘bulk tumour’ measurements. We apply the XenoSWATH pipeline to characterize an intraductal xenograft model of breast ductal carcinoma in situ and uncover complex regulation consistent with stromal reprogramming, where the modulation of cell migration pathways is not restricted to tumour cells but also operates in the mouse stroma upon progression to invasive disease. MouseRefSWATH and XenoSWATH open new opportunities for in-depth and reproducible proteomic assessment to address wide-ranging biological questions involving this important model organism., P.H.H. is supported by grants from the Institute of Cancer Research, Breast Cancer Now (2013NovPhD185 and 2014NovPR360) and Cancer Research UK (C36478/ A19281). B.A.H. and R.C.N. are supported by Breast Cancer Now programmatic funding. S.E.A. is supported by grants from Cancer Research UK (CRUK/A19763) and the Medical Research Council (MC_U12266B). F.C. is supported by grants from the Institute of Cancer Research, the Ramon y Cajal Research Programme (RYC2016-20352) and Ministerio de Ciencia, Innovación y Universidades/Agencia Estatal de Investigación/European Regional Development Fund (RTI2018-096778- A-I00). P.-C.H. is supported in part by a Schweizerischer Nationalfonds zur Förderung der Wissenschaftlichen Forschung project grant (31003A_182470) and a European Research Council Starting Grant (802773-MitoGuide).

Published
2020

4. Sex specific retinoic acid signaling is required for the initiation of urogenital sinus bud development

Author

Bryant, Sarah L., Francis, Jeffrey C., Lokody, Isabel B., Wang, Hong, Risbridger, Gail P., Loveland, Kate L., and Swain, Amanda

Subjects
Male, Organogenesis, Prostate, Urogenital System, Tretinoin, Cell Biology, Real-Time Polymerase Chain Reaction, beta-Galactosidase, Immunohistochemistry, Article, Activins, Sexual differentiation, Mice, Sex Factors, Animals, Female, Prostate initiation, Molecular Biology, In Situ Hybridization, Developmental Biology, DNA Primers, Inhibin-beta Subunits, Signal Transduction
Abstract

The mammalian urogenital sinus (UGS) develops in a sex specific manner, giving rise to the prostate in the male and the sinus vagina in the embryonic female. Androgens, produced by the embryonic testis, have been shown to be crucial to this process. In this study we show that retinoic acid signaling is required for the initial stages of bud development from the male UGS. Enzymes involved in retinoic acid synthesis are expressed in the UGS mesenchyme in a sex specific manner and addition of ligand to female tissue is able to induce prostate-like bud formation in the absence of androgens, albeit at reduced potency. Functional studies in mouse organ cultures that faithfully reproduce the initiation of prostate development indicate that one of the roles of retinoic acid signaling in the male is to inhibit the expression of Inhba, which encodes the βA subunit of Activin, in the UGS mesenchyme. Through in vivo genetic analysis and culture studies we show that inhibition of Activin signaling in the female UGS leads to a similar phenotype to that of retinoic acid treatment, namely bud formation in the absence of androgens. Our data also reveals that both androgens and retinoic acid have extra independent roles to that of repressing Activin signaling in the development of the prostate during fetal stages. This study identifies a novel role for retinoic acid as a mesenchymal factor that acts together with androgens to determine the position and initiation of bud development in the male UGS epithelia., Highlights • We show that sex specific retinoic acid is required for male UGS bud initiation. • An increase in retinoic acid can lead to prostate-like formation in females. • We find that activin repression is a downstream target of RA signalling. • RA is a novel mesenchymal signal regulating bud initiation along the UGS.

Published
2014

5. Adrenal development is initiated by Cited2 and Wt1 through modulation of Sf-1 dosage.

Author

Val, Pierre, Martinez-Barbera, Juan-Pedro, and Swain, Amanda

Subjects
ADRENAL cortex, TRANSCRIPTION factors, GENES, GONADS, MICE, GENE expression, GENETIC mutation, MORPHOGENESIS
Abstract

It has been proposed that the mammalian adrenal cortex and gonad are derived from the same primordium present during early urogenital development. Molecular pathways involved in the differentiation of the adrenal cortex from the adrenogonadal primordium (AGP) have yet to be determined. Here we show in mice that the transcription co-factor Cited2 is required for the specification of the adrenal cortex from the AGP. We present genetic and molecular evidence demonstrating that Cited2 interacts with the transcription factor Wt1 to stimulate expression of the nuclear hormone receptor Sf-1 (Nr5a1) in the AGP prior to the separation between gonad and adrenal cortex. We show a direct correlation between the expression levels of Sf-1 in the AGP and the defects in adrenal development found in mice with different Cited2 and Wt1 mutant backgrounds. Analysis of embryos heterozygous for mutations in both Sf-1 and Cited2 confirmed that these genes act in the same pathway during adrenal development. Our studies reveal a regulatory mechanism in which Cited2 acts as a Wt1 co-factor to increase, at a critical time in embryogenesis, the levels of the essential transcription factor Sf-1 in the AGP above the threshold required to determine adrenal development. These results highlight the importance of transcription factor dosage in organogenesis and the role of transcription co-factors such as Cited2 in determining the levels of these factors. [ABSTRACT FROM AUTHOR]

Published
2007
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