42 results on '"Rust, Alistair G."'
Search Results
2. Optimised ARID1A immunohistochemistry is an accurate predictor of ARID1A mutational status in gynaecological cancers.
- Author
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Khalique, Saira, Naidoo, Kalnisha, Attygalle, Ayoma D., Kriplani, Divya, Daley, Frances, Lowe, Anne, Campbell, James, Jones, Thomas, Hubank, Michael, Fenwick, Kerry, Matthews, Nicholas, Rust, Alistair G., Lord, Christopher J., Banerjee, Susana, and Natrajan, Rachael
- Abstract
Abstract: ARID1A is a tumour suppressor gene that is frequently mutated in clear cell and endometrioid carcinomas of the ovary and endometrium and is an important clinical biomarker for novel treatment approaches for patients with ARID1A defects. However, the accuracy of ARID1A immunohistochemistry (IHC) as a surrogate for mutation status has not fully been established for patient stratification in clinical trials. Here we tested whether ARID1A IHC could reliably predict ARID1A mutations identified by next‐generation sequencing. Three commercially available antibodies – EPR13501 (Abcam), D2A8U (Cell Signaling), and HPA005456 (Sigma) – were optimised for IHC using cell line models and human tissue, and screened across a cohort of 45 gynaecological tumours. IHC was scored independently by three pathologists using an immunoreactive score. ARID1A mutation status was assessed using two independent sequencing platforms and the concordance between ARID1A mutation and protein expression was evaluated using Receiver Operating Characteristic statistics. Overall, 21 ARID1A mutations were identified in 14/43 assessable tumours (33%), the majority of which were predicted to be deleterious. Mutations were identified in 6/17 (35%) ovarian clear cell carcinomas, 5/8 (63%) ovarian endometrioid carcinomas, 2/5 (40%) endometrial carcinomas, and 1/7 (14%) carcinosarcomas. ROC analysis identified greater than 95% concordance between mutation status and IHC using a modified immunoreactive score for all three antibodies allowing a definitive cut‐point for ARID1A mutant status to be calculated. Comprehensive assessment of concordance of ARID1A IHC and mutation status identified EPR13501 as an optimal antibody, with 100% concordance between ARID1A mutation status and protein expression, across different gynaecological histological subtypes. It delivered the best inter‐rater agreement between all pathologists, as well as a clear cost‐benefit advantage. This could allow patients to be accurately stratified based on their ARID1A IHC status into early phase clinical trials. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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3. Capture Hi-C identifies putative target genes at 33 breast cancer risk loci.
- Author
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Baxter, Joseph S., Leavy, Olivia C., Dryden, Nicola H., Maguire, Sarah, Johnson, Nichola, Fedele, Vita, Simigdala, Nikiana, Martin, Lesley-Ann, Andrews, Simon, Wingett, Steven W., Assiotis, Ioannis, Fenwick, Kerry, Chauhan, Ritika, Rust, Alistair G., Orr, Nick, Dudbridge, Frank, Haider, Syed, and Fletcher, Olivia
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SINGLE nucleotide polymorphisms ,BREAST cancer ,CHROMATIN ,CANCER cells ,PROGENITOR cells - Abstract
Genome-wide association studies (GWAS) have identified approximately 100 breast cancer risk loci. Translating these findings into a greater understanding of the mechanisms that influence disease risk requires identification of the genes or non-coding RNAs that mediate these associations. Here, we use Capture Hi-C (CHi-C) to annotate 63 loci; we identify 110 putative target genes at 33 loci. To assess the support for these target genes in other data sources we test for associations between levels of expression and SNP genotype (eQTLs), disease-specific survival (DSS), and compare them with somatically mutated cancer genes. 22 putative target genes are eQTLs, 32 are associated with DSS and 14 are somatically mutated in breast, or other, cancers. Identifying the target genes at GWAS risk loci will lead to a greater understanding of the mechanisms that influence breast cancer risk and prognosis. [ABSTRACT FROM AUTHOR]
- Published
- 2018
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4. Transposon mutagenesis identifies genes that cooperate with mutant Pten in breast cancer progression.
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Rangel, Roberto, Guzman-Rojas, Liliana, Mann, Michael B., Newberg, Justin Y., Takahiro Kodama, Jenkins, Nancy A., Copeland, Neal G., Song-Choon Lee, Ward, Jerrold M., Chin, Kuan-Yew, Hon-Kim Ban, Kenneth, McNoe, Leslie A., Selvanesan, Luxmanan, Black, Michael A., and Rust, Alistair G.
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TRANSPOSONS ,MUTAGENESIS ,GENES ,BREAST cancer research ,PTEN protein - Abstract
Triple-negative breast cancer (TNBC) has the worst prognosis of any breast cancer subtype. To better understand the genetic forces driving TNBC, we performed a transposon mutagenesis screen in a phosphatase and tensin homolog (Pten) mutant mice and identified 12 candidate trunk drivers and a much larger number of progression genes. Validation studies identified eight TNBC tumor suppressor genes, including the GATA-like transcriptional repressor TRPS1. Down-regulation of TRPS1 in TNBC cells promoted epithelial-to-mesenchymal transition (EMT) by deregulating multiple EMT pathway genes, in addition to increasing the expression of SERPINE1 and SERPINB2 and the subsequent migration, invasion, and metastasis of tumor cells. Transposon mutagenesis has thus provided a better understanding of the genetic forces driving TNBC and discovered genes with potential clinical importance in TNBC. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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5. Analyzing tumor heterogeneity and driver genes in single myeloid leukemia cells with SBCapSeq.
- Author
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Mann, Karen M, Newberg, Justin Y, Black, Michael A, Jones, Devin J, Amaya-Manzanares, Felipe, Guzman-Rojas, Liliana, Kodama, Takahiro, Ward, Jerrold M, Rust, Alistair G, van der Weyden, Louise, Yew, Christopher Chin Kuan, Waters, Jill L, Leung, Marco L, Rogers, Keith, Rogers, Susan M, McNoe, Leslie A, Selvanesan, Luxmanan, Navin, Nicholas, Jenkins, Nancy A, and Copeland, Neal G
- Published
- 2016
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6. Sleeping Beauty screen reveals Pparg activation in metastatic prostate cancer.
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Ahmada, Imran, Mui, Ernest, Galbraith, Laura, Patel, Rachana, Ee Hong Tana, Salji, Mark, Rust, Alistair G., Repiscak, Peter, Hedley, Ann, Markert, Elke, Loveridge, Carolyn, van der Weyden, Louise, Edwards, Joanne, Sansom, Owen J., Adams, David J., and Leung, Hing Y.
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PROSTATE cancer prognosis ,PEROXISOME proliferator-activated receptors ,ACETYL-CoA carboxylase ,ATP-citrate lyase ,BIOMARKERS ,PHOSPHATIDYLINOSITOL 3-kinases - Abstract
Prostate cancer (CaP) is the most common adult male cancer in the developed world. The paucity of biomarkers to predict prostate tumor biology makes it important to identify key pathways that confer poor prognosis and guide potential targeted therapy. Using a murine forward mutagenesis screen in a Pten-null background, we identified peroxisome proliferator-activated receptor gamma (Pparg), encoding a ligand-activated transcription factor, as a promoter of metastatic CaP through activation of lipid signaling pathways, including up-regulation of lipid synthesis enzymes [fatty acid synthase (FASN), acetyl-CoA carboxylase (ACC), ATP citrate lyase (ACLY)]. Importantly, inhibition of PPARG suppressed tumor growth in vivo, with down-regulation of the lipid synthesis program. We show that elevated levels of PPARG strongly correlate with elevation of FASN in human CaP and that high levels of PPARG/FASN and PI3K/ pAKT pathway activation confer a poor prognosis. These data suggest that CaP patients could be stratified in terms of PPARG/FASN and PTEN levels to identify patients with aggressive CaP who may respond favorably to PPARG/FASN inhibition. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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7. Sleeping Beauty transposon mutagenesis identifies genes that cooperate with mutant Smad4 in gastric cancer development.
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Haruna Takeda, Rust, Alistair G., Ward, Jerrold M., Chin Kuan Yew, Christopher, Jenkins, Nancy A., and Copeland, Neal G.
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GASTROINTESTINAL cancer ,CANCER research ,TRANSPOSONS ,MUTAGENESIS ,PROTEOLYSIS - Abstract
Mutations in SMAD4 predispose to the development of gastrointestinal cancer, which is the third leading cause of cancer-related deaths. To identify genes driving gastric cancer (GC) development, we performed a Sleeping Beauty (SB) transposon mutagenesis screen in the stomach of Smad4
+/- mutant mice. This screen identified 59 candidate GC trunk drivers and a much larger number of candidate GC progression genes. Strikingly, 22 SB-identified trunk drivers are known or candidate cancer genes, whereas four SB-identified trunk drivers, including PTEN, SMAD4, RNF43, and NF1, are known human GC trunk drivers. Similar to human GC, pathway analyses identified WNT, TGF-β, and PI3K-PTEN signaling, ubiquitin- mediated proteolysis, adherens junctions, and RNA degradation in addition to genes involved in chromatin modification and organization as highly deregulated pathways in GC. Comparative oncogenomic filtering of the complete list of SB-identified genes showed that they are highly enriched for genes mutated in human GC and identified many candidate human GC genes. Finally, by comparing our complete list of SB-identified genes against the list of mutated genes identified in five large-scale human GC sequencing studies, we identified LDL receptor-related protein 1B (LRP1B) as a previously unidentified human candidate GC tumor suppressor gene. In LRP1B, 129 mutations were found in 462 human GC samples sequenced, and LRP1B is one of the top 10 most deleted genes identified in a panel of 3,312 human cancers. SB mutagenesis has, thus, helped to catalog the cooperative molecular mechanisms driving SMAD4-induced GC growth and discover genes with potential clinical importance in human GC. [ABSTRACT FROM AUTHOR]- Published
- 2016
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8. Adenoma development in familial adenomatous polyposis and MUTYH-associated polyposis: somatic landscape and driver genes.
- Author
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Rashid, Mamunur, Fischer, Andrej, Wilson, Cathy H, Tiffen, Jessamy, Rust, Alistair G, Stevens, Philip, Idziaszczyk, Shelley, Maynard, Julie, Williams, Geraint T, Mustonen, Ville, Sampson, Julian R, and Adams, David J
- Abstract
Familial adenomatous polyposis ( FAP) and MUTYH-associated polyposis ( MAP) are inherited disorders associated with multiple colorectal adenomas that lead to a very high risk of colorectal cancer. The somatic mutations that drive adenoma development in these conditions have not been investigated comprehensively. In this study we performed analysis of paired colorectal adenoma and normal tissue DNA from individuals with FAP or MAP, sequencing 14 adenoma whole exomes (eight MAP, six FAP), 55 adenoma targeted exomes (33 MAP, 22 FAP) and germline DNA from each patient, and a further 63 adenomas by capillary sequencing (41 FAP, 22 MAP). With these data we examined the profile of mutated genes, the mutational signatures and the somatic mutation rates, observing significant diversity in the constellations of mutated driver genes in different adenomas, and loss-of-function mutations in WTX (9%; p < 9.99e-06), a gene implicated in regulation of the WNT pathway and p53 acetylation. These data extend our understanding of the early events in colorectal tumourigenesis in the polyposis syndromes. © 2015 The Authors. The Journal of Pathology published by John Wiley & Sons Ltd on behalf of Pathological Society of Great Britain and Ireland. [ABSTRACT FROM AUTHOR]
- Published
- 2016
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9. Somatic drivers of B-ALL in a model of ETV6-RUNX1; Pax5+/- leukemia.
- Author
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van der Weyden, Louise, Giotopoulos, George, Kim Wong, Rust, Alistair G., Robles-Espinoza, Carla Daniela, Hikari Osaki, Huntly, Brian J., and Adams, David J.
- Subjects
LYMPHOBLASTIC leukemia in children ,CANCER-related mortality ,B cells ,CHROMOSOME abnormalities ,MUTAGENESIS ,TRANSPOSONS ,DIAGNOSIS - Abstract
Background: B-cell precursor acute lymphoblastic leukemia (B-ALL) is amongst the leading causes of childhood cancer-related mortality. Its most common chromosomal aberration is the ETV6-RUNX1 fusion gene, with ~25% of ETV6-RUNX1 patients also carrying PAX5 alterations. Methods: We have recreated this mutation background by inter-crossing Etv6-RUNX1 (Etv6
RUNX1-SB ) and Pax5+/- mice and performed an in vivo analysis to find driver genes using Sleeping Beauty transposon-mediated mutagenesis and also exome sequencing. Results: Combination of Etv6-RUNX1 and Pax5+/- alleles generated a transplantable B220 + CD19+ B-ALL with a significant disease incidence. RNA-seq analysis showed a gene expression pattern consistent with arrest at the pre-B stage. Analysis of the transposon common insertion sites identified genes involved in B-cell development (Zfp423) and the JAK/STAT signaling pathway (Jak1, Stat5 and Il2rb), while exome sequencing revealed somatic hotspot mutations in Jak1 and Jak3 at residues analogous to those mutated in human leukemias, and also mutation of Trp53. Conclusions: Powerful synergies exists in our model suggesting STAT pathway activation and mutation of Trp53 are potent drivers of B-ALL in the context of Etv6-RUNX1;Pax5+/- . [ABSTRACT FROM AUTHOR]- Published
- 2015
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10. Transposon mutagenesis identifies genetic drivers of BrafV600E melanoma.
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Mann, Michael B, Copeland, Neal G, Jenkins, Nancy A, Black, Michael A, Jones, Devin J, Newberg, Justin Y, Ward, Jerrold M, Yew, Christopher Chin Kuan, Dupuy, Adam J, Rust, Alistair G, Bosenberg, Marcus W, McMahon, Martin, and Print, Cristin G
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TRANSPOSONS ,MUTAGENESIS ,MELANOGENESIS ,ONCOGENIC DNA viruses ,NETRINS - Abstract
Although nearly half of human melanomas harbor oncogenic BRAF
V600E mutations, the genetic events that cooperate with these mutations to drive melanogenesis are still largely unknown. Here we show that Sleeping Beauty (SB) transposon-mediated mutagenesis drives melanoma progression in BrafV600E mutant mice and identify 1,232 recurrently mutated candidate cancer genes (CCGs) from 70 SB-driven melanomas. CCGs are enriched in Wnt, PI3K, MAPK and netrin signaling pathway components and are more highly connected to one another than predicted by chance, indicating that SB targets cooperative genetic networks in melanoma. Human orthologs of >500 CCGs are enriched for mutations in human melanoma or showed statistically significant clinical associations between RNA abundance and survival of patients with metastatic melanoma. We also functionally validate CEP350 as a new tumor-suppressor gene in human melanoma. SB mutagenesis has thus helped to catalog the cooperative molecular mechanisms driving BRAFV600E melanoma and discover new genes with potential clinical importance in human melanoma. [ABSTRACT FROM AUTHOR]- Published
- 2015
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11. BRAF inhibitor resistance mediated by the AKT pathway in an oncogenic BRAF mouse melanoma model.
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Perna, Daniele, Karreth, Florian A., Rust, Alistair G., Perez-Mancera, Pedro A., Mamunur Rashid, Iorio, Francesco, Alifrangis, Constantine, Arends, Mark J., Bosenberg, Marcus W., Bollag, Gideon, Tuveson, David A., and Adams, David J.
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NEUROENDOCRINE tumors ,ONCOGENES ,MOUSE leukemia ,MELANOMA ,MUTAGENESIS - Abstract
BRAF (v-raf murine sarcoma viral oncogene homolog B) inhibitors elicit a transient anti-tumor response in ~80% of BRAF
V600 -mutant melanoma patients that almost uniformly precedes the emergence of resistance. Here we used a mouse model of melanoma in which melanocyte-specific expression of BrafV618E (analogous to the human BRAFV600 mutation) led to the development of skin hyperpigmentation and nevi, as well as melanoma formation with incomplete penetrance. Sleeping Beauty insertional mutagenesis in this model led to accelerated and fully penetrant melanomagenesis and synchronous tumor formation. Treatment of BrafV618E transposon mice with the BRAF inhibitor PLX4720 resulted in tumor regression followed by relapse. Analysis of transposon insertions identified eight genes including Braf, Mitf, and ERas (ES-cell expressed Ras) as candidate resistance genes. Expression of ERAS in human melanoma cell lines conferred resistance to PLX4720 and induced hyperphosphorylation of AKT (v-akt murine thymoma viral oncogene homolog 1), a phenotype reverted by combinatorial treatment with PLX4720 and the AKT inhibitor MK2206. We show that ERAS expression elicits a prosurvival signal associated with phosphorylation/inactivation of BAD, and that the resistance of hepatocyte growth factor-treated human melanoma cells to PLX4720 can be reverted by treatment with the BAD-like BH3 mimetic ABT-737. Thus, we define a role for the AKT/BAD pathway in resistance to BRAF inhibition and illustrate an in vivo approach for finding drug resistance genes. [ABSTRACT FROM AUTHOR]- Published
- 2015
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12. Transposon mutagenesis identifies genes and evolutionary forces driving gastrointestinal tract tumor progression.
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Takeda, Haruna, Yew, Christopher Chin Kuan, Mann, Michael B, Jenkins, Nancy A, Rust, Alistair G, Adams, David J, Copeland, Neal G, Koso, Hideto, Wei, Zhubo, and Ward, Jerrold M
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TRANSPOSONS ,MUTAGENESIS ,GASTROINTESTINAL diseases ,TUMORS ,GENETICS ,MICROBIAL mutation - Abstract
To provide a more comprehensive understanding of the genes and evolutionary forces driving colorectal cancer (CRC) progression, we performed Sleeping Beauty (SB) transposon mutagenesis screens in mice carrying sensitizing mutations in genes that act at different stages of tumor progression. This approach allowed us to identify a set of genes that appear to be highly relevant for CRC and to provide a better understanding of the evolutionary forces and systems properties of CRC. We also identified six genes driving malignant tumor progression and a new human CRC tumor-suppressor gene, ZNF292, that might also function in other types of cancer. Our comprehensive CRC data set provides a resource with which to develop new therapies for treating CRC. [ABSTRACT FROM AUTHOR]
- Published
- 2015
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13. The mutational landscapes of genetic and chemical models of Kras-driven lung cancer.
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Westcott, Peter M. K., Halliwill, Kyle D., To, Minh D., Rashid, Mamunur, Rust, Alistair G., Keane, Thomas M., Delrosario, Reyno, Jen, Kuang-Yu, Gurley, Kay E., Kemp, Christopher J., Fredlund, Erik, Quigley, David A., Adams, David J., and Balmain, Allan
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TUMORS ,NEOPLASTIC cell transformation ,NON-small-cell lung carcinoma ,CARCINOMA ,CANCER - Abstract
Next-generation sequencing of human tumours has refined our understanding of the mutational processes operative in cancer initiation and progression, yet major questions remain regarding the factors that induce driver mutations and the processes that shape mutation selection during tumorigenesis. Here we performed whole-exome sequencing on adenomas from three mouse models of non-small-cell lung cancer, which were induced either by exposure to carcinogens (methyl-nitrosourea (MNU) and urethane) or by genetic activation of Kras (Kras
LA2 ). Although the MNU-induced tumours carried exactly the same initiating mutation in Kras as seen in the KrasLA2 model (G12D), MNU tumours had an average of 192 non-synonymous, somatic single-nucleotide variants, compared with only six in tumours from the KrasLA2 model. By contrast, the KrasLA2 tumours exhibited a significantly higher level of aneuploidy and copy number alterations compared with the carcinogen-induced tumours, suggesting that carcinogen-induced and genetically engineered models lead to tumour development through different routes. The wild-type allele of Kras has been shown to act as a tumour suppressor in mouse models of non-small-cell lung cancer. We demonstrate that urethane-induced tumours from wild-type mice carry mostly (94%) Kras Q61R mutations, whereas those from Kras heterozygous animals carry mostly (92%) Kras Q61L mutations, indicating a major role for germline Kras status in mutation selection during initiation. The exome-wide mutation spectra in carcinogen-induced tumours overwhelmingly display signatures of the initiating carcinogen, while adenocarcinomas acquire additional C > T mutations at CpG sites. These data provide a basis for understanding results from human tumour genome sequencing, which has identified two broad categories of tumours based on the relative frequency of single-nucleotide variations and copy number alterations, and underline the importance of carcinogen models for understanding the complex mutation spectra seen in human cancers. [ABSTRACT FROM AUTHOR]- Published
- 2015
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14. Mutational Genomics for Cancer Pathway Discovery.
- Author
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de Ridder, Jeroen, Kool, Jaap, Uren, Anthony G., Bot, Jan, de Jong, Johann, Rust, Alistair G., Berns, Anton, van Lohuizen, Maarten, Adams, David J., Wessels, Lodewyk, and Reinders, Marcel
- Published
- 2013
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15. Using Varying Negative Examples to Improve Computational Predictions of Transcription Factor Binding Sites.
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Rezwan, Faisal, Sun, Yi, Davey, Neil, Adams, Rod, Rust, Alistair G., and Robinson, Mark
- Published
- 2012
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16. Improving Transcription Factor Binding Site Predictions by Using Randomised Negative Examples.
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Rezwan, Faisal, Sun, Yi, Davey, Neil, Adams, Rod, Rust, Alistair G., and Robinson, Mark
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- 2012
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17. Evolutionary Neural Topiaryt: Growing and Sculpting Artificial Neurons to Order.
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Rust, Alistair G., Adams, Rod, and Bolouri, Hamid
- Published
- 2000
18. Chromatin Landscapes of Retroviral and Transposon Integration Profiles.
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de Jong, Johann, Akhtar, Waseem, Badhai, Jitendra, Rust, Alistair G., Rad, Roland, Hilkens, John, Berns, Anton, van Lohuizen, Maarten, Wessels, Lodewyk F. A., and de Ridder, Jeroen
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RETROVIRUS genetics ,TRANSPOSONS ,MOLECULAR biology ,CANCER research ,GENE therapy ,CANCER genes ,CANCER genetics - Abstract
The ability of retroviruses and transposons to insert their genetic material into host DNA makes them widely used tools in molecular biology, cancer research and gene therapy. However, these systems have biases that may strongly affect research outcomes. To address this issue, we generated very large datasets consisting of to unselected integrations in the mouse genome for the Sleeping Beauty (SB) and piggyBac (PB) transposons, and the Mouse Mammary Tumor Virus (MMTV). We analyzed (epi)genomic features to generate bias maps at both local and genome-wide scales. MMTV showed a remarkably uniform distribution of integrations across the genome. More distinct preferences were observed for the two transposons, with PB showing remarkable resemblance to bias profiles of the Murine Leukemia Virus. Furthermore, we present a model where target site selection is directed at multiple scales. At a large scale, target site selection is similar across systems, and defined by domain-oriented features, namely expression of proximal genes, proximity to CpG islands and to genic features, chromatin compaction and replication timing. Notable differences between the systems are mainly observed at smaller scales, and are directed by a diverse range of features. To study the effect of these biases on integration sites occupied under selective pressure, we turned to insertional mutagenesis (IM) screens. In IM screens, putative cancer genes are identified by finding frequently targeted genomic regions, or Common Integration Sites (CISs). Within three recently completed IM screens, we identified 7%–33% putative false positive CISs, which are likely not the result of the oncogenic selection process. Moreover, results indicate that PB, compared to SB, is more suited to tag oncogenes. [ABSTRACT FROM AUTHOR]
- Published
- 2014
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19. Insertional Mutagenesis and Deep Profiling Reveals Gene Hierarchies and a Myc/p53-Dependent Bottleneck in Lymphomagenesis.
- Author
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Huser, Camille A., Gilroy, Kathryn L., de Ridder, Jeroen, Kilbey, Anna, Borland, Gillian, Mackay, Nancy, Jenkins, Alma, Bell, Margaret, Herzyk, Pawel, van der Weyden, Louise, Adams, David J., Rust, Alistair G., Cameron, Ewan, and Neil, James C.
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RETROVIRUS diseases ,MUTAGENESIS ,MOUSE leukemia ,TRANSGENIC mice ,CANCER invasiveness - Abstract
Retroviral insertional mutagenesis (RIM) is a powerful tool for cancer genomics that was combined in this study with deep sequencing (RIM/DS) to facilitate a comprehensive analysis of lymphoma progression. Transgenic mice expressing two potent collaborating oncogenes in the germ line (CD2-MYC, -Runx2) develop rapid onset tumours that can be accelerated and rendered polyclonal by neonatal Moloney murine leukaemia virus (MoMLV) infection. RIM/DS analysis of 28 polyclonal lymphomas identified 771 common insertion sites (CISs) defining a ‘progression network’ that encompassed a remarkably large fraction of known MoMLV target genes, with further strong indications of oncogenic selection above the background of MoMLV integration preference. Progression driven by RIM was characterised as a Darwinian process of clonal competition engaging proliferation control networks downstream of cytokine and T-cell receptor signalling. Enhancer mode activation accounted for the most efficiently selected CIS target genes, including Ccr7 as the most prominent of a set of chemokine receptors driving paracrine growth stimulation and lymphoma dissemination. Another large target gene subset including candidate tumour suppressors was disrupted by intragenic insertions. A second RIM/DS screen comparing lymphomas of wild-type and parental transgenics showed that CD2-MYC tumours are virtually dependent on activation of Runx family genes in strong preference to other potent Myc collaborating genes (Gfi1, Notch1). Ikzf1 was identified as a novel collaborating gene for Runx2 and illustrated the interface between integration preference and oncogenic selection. Lymphoma target genes for MoMLV can be classified into (a) a small set of master regulators that confer self-renewal; overcoming p53 and other failsafe pathways and (b) a large group of progression genes that control autonomous proliferation in transformed cells. These findings provide insights into retroviral biology, human cancer genetics and the safety of vector-mediated gene therapy. [ABSTRACT FROM AUTHOR]
- Published
- 2014
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20. Inactivating CUX1 mutations promote tumorigenesis.
- Author
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Wong, Chi C, Martincorena, Inigo, Rust, Alistair G, Rashid, Mamunur, Alifrangis, Constantine, Alexandrov, Ludmil B, Tiffen, Jessamy C, Kober, Christina, Green, Anthony R, Massie, Charles E, Nangalia, Jyoti, Lempidaki, Stella, Döhner, Hartmut, Döhner, Konstanze, Bray, Sarah J, McDermott, Ultan, Papaemmanuil, Elli, Campbell, Peter J, and Adams, David J
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GENE silencing ,GENETIC mutation ,NEOPLASTIC cell transformation ,CANCER genetics ,HOMEOBOX proteins ,GENETIC transcription - Abstract
A major challenge in cancer genetics is to determine which low-frequency somatic mutations are drivers of tumorigenesis. Here we interrogate the genomes of 7,651 diverse human cancers and find inactivating mutations in the homeodomain transcription factor gene CUX1 (cut-like homeobox 1) in ∼1-5% of various tumors. Meta-analysis of CUX1 mutational status in 2,519 cases of myeloid malignancies reveals disruptive mutations associated with poor survival, highlighting the clinical significance of CUX1 loss. In parallel, we validate CUX1 as a bona fide tumor suppressor using mouse transposon-mediated insertional mutagenesis and Drosophila cancer models. We demonstrate that CUX1 deficiency activates phosphoinositide 3-kinase (PI3K) signaling through direct transcriptional downregulation of the PI3K inhibitor PIK3IP1 (phosphoinositide-3-kinase interacting protein 1), leading to increased tumor growth and susceptibility to PI3K-AKT inhibition. Thus, our complementary approaches identify CUX1 as a pan-driver of tumorigenesis and uncover a potential strategy for treating CUX1-mutant tumors. [ABSTRACT FROM AUTHOR]
- Published
- 2014
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21. Transposon mutagenesis identifies genes driving hepatocellular carcinoma in a chronic hepatitis B mouse model.
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Bard-Chapeau, Emilie A, Nguyen, Anh-Tuan, Rust, Alistair G, Sayadi, Ahmed, Lee, Philip, Chua, Belinda Q, New, Lee-Sun, de Jong, Johann, Ward, Jerrold M, Chin, Christopher K Y, Chew, Valerie, Toh, Han Chong, Abastado, Jean-Pierre, Benoukraf, Touati, Soong, Richie, Bard, Frederic A, Dupuy, Adam J, Johnson, Randy L, Radda, George K, and Chan, Eric Chun Yong
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TRANSPOSONS ,HEPATITIS B ,LABORATORY mice ,MUTAGENESIS ,LIVER cancer ,GENETIC regulation - Abstract
The most common risk factor for developing hepatocellular carcinoma (HCC) is chronic infection with hepatitis B virus (HBV). To better understand the evolutionary forces driving HCC, we performed a near-saturating transposon mutagenesis screen in a mouse HBV model of HCC. This screen identified 21 candidate early stage drivers and a very large number (2,860) of candidate later stage drivers that were enriched for genes that are mutated, deregulated or functioning in signaling pathways important for human HCC, with a striking 1,199 genes being linked to cellular metabolic processes. Our study provides a comprehensive overview of the genetic landscape of HCC. [ABSTRACT FROM AUTHOR]
- Published
- 2014
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22. A FOXO3-IRF7 gene regulatory circuit limits inflammatory sequelae of antiviral responses.
- Author
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Litvak, Vladimir, Ratushny, Alexander V., Lampano, Aaron E., Schmitz, Frank, Huang, Albert C., Raman, Ayush, Rust, Alistair G., Bergthaler, Andreas, Aitchison, John D., and Aderem, Alan
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GENE regulatory networks ,INFECTION prevention ,INTERFERONS ,TRANSCRIPTION factors ,INTERFERON regulatory factors - Abstract
Antiviral responses must be tightly regulated to defend rapidly against infection while minimizing inflammatory damage. Type 1 interferons (IFN-I) are crucial mediators of antiviral responses and their transcription is regulated by a variety of transcription factors; principal among these is the family of interferon regulatory factors (IRFs). The IRF gene regulatory networks are complex and contain multiple feedback loops. The tools of systems biology are well suited to elucidate the complex interactions that give rise to precise coordination of the interferon response. Here we have used an unbiased systems approach to predict that a member of the forkhead family of transcription factors, FOXO3, is a negative regulator of a subset of antiviral genes. This prediction was validated using macrophages isolated from Foxo3-null mice. Genome-wide location analysis combined with gene deletion studies identified the Irf7 gene as a critical target of FOXO3. FOXO3 was identified as a negative regulator of Irf7 transcription and we have further demonstrated that FOXO3, IRF7 and IFN-I form a coherent feed-forward regulatory circuit. Our data suggest that the FOXO3-IRF7 regulatory circuit represents a novel mechanism for establishing the requisite set points in the interferon pathway that balances the beneficial effects and deleterious sequelae of the antiviral response. [ABSTRACT FROM AUTHOR]
- Published
- 2012
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23. The deubiquitinase USP9X suppresses pancreatic ductal adenocarcinoma.
- Author
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Pérez-Mancera, Pedro A., Rust, Alistair G., van der Weyden, Louise, Kristiansen, Glen, Li, Allen, Sarver, Aaron L., Silverstein, Kevin A. T., Grützmann, Robert, Aust, Daniela, Rümmele, Petra, Knösel, Thomas, Herd, Colin, Stemple, Derek L., Kettleborough, Ross, Brosnan, Jacqueline A., Li, Ang, Morgan, Richard, Knight, Spencer, Yu, Jun, and Stegeman, Shane
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PANCREATIC duct ,ADENOCARCINOMA ,MUTAGENESIS ,GENES ,DISEASE progression ,LABORATORY mice ,CANCER - Abstract
Pancreatic ductal adenocarcinoma (PDA) remains a lethal malignancy despite much progress concerning its molecular characterization. PDA tumours harbour four signature somatic mutations in addition to numerous lower frequency genetic events of uncertain significance. Here we use Sleeping Beauty (SB) transposon-mediated insertional mutagenesis in a mouse model of pancreatic ductal preneoplasia to identify genes that cooperate with oncogenic Kras
G12D to accelerate tumorigenesis and promote progression. Our screen revealed new candidate genes for PDA and confirmed the importance of many genes and pathways previously implicated in human PDA. The most commonly mutated gene was the X-linked deubiquitinase Usp9x, which was inactivated in over 50% of the tumours. Although previous work had attributed a pro-survival role to USP9X in human neoplasia, we found instead that loss of Usp9x enhances transformation and protects pancreatic cancer cells from anoikis. Clinically, low USP9X protein and messenger RNA expression in PDA correlates with poor survival after surgery, and USP9X levels are inversely associated with metastatic burden in advanced disease. Furthermore, chromatin modulation with trichostatin A or 5-aza-2?-deoxycytidine elevates USP9X expression in human PDA cell lines, indicating a clinical approach for certain patients. The conditional deletion of Usp9x cooperated with KrasG12D to accelerate pancreatic tumorigenesis in mice, validating their genetic interaction. We propose that USP9X is a major tumour suppressor gene with prognostic and therapeutic relevance in PDA. [ABSTRACT FROM AUTHOR]- Published
- 2012
- Full Text
- View/download PDF
24. Nuclear receptor binding protein 1 regulates intestinal progenitor cell homeostasis and tumour formation.
- Author
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Wilson, Catherine H, Crombie, Catriona, van der Weyden, Louise, Poulogiannis, George, Rust, Alistair G, Pardo, Mercedes, Gracia, Tannia, Yu, Lu, Choudhary, Jyoti, Poulin, Gino B, McIntyre, Rebecca E, Winton, Douglas J, March, H Nikki, Arends, Mark J, Fraser, Andrew G, and Adams, David J
- Subjects
NUCLEAR receptors (Biochemistry) ,CARRIER proteins ,GENETIC regulation ,PROGENITOR cells ,HOMEOSTASIS ,GENETIC testing - Abstract
Genetic screens in simple model organisms have identified many of the key components of the conserved signal transduction pathways that are oncogenic when misregulated. Here, we identify H37N21.1 as a gene that regulates vulval induction in let-60(n1046gf), a strain with a gain-of-function mutation in the Caenorhabditis elegans Ras orthologue, and show that somatic deletion of Nrbp1, the mouse orthologue of this gene, results in an intestinal progenitor cell phenotype that leads to profound changes in the proliferation and differentiation of all intestinal cell lineages. We show that Nrbp1 interacts with key components of the ubiquitination machinery and that loss of Nrbp1 in the intestine results in the accumulation of Sall4, a key mediator of stem cell fate, and of Tsc22d2. We also reveal that somatic loss of Nrbp1 results in tumourigenesis, with haematological and intestinal tumours predominating, and that nuclear receptor binding protein 1 (NRBP1) is downregulated in a range of human tumours, where low expression correlates with a poor prognosis. Thus NRBP1 is a conserved regulator of cell fate, that plays an important role in tumour suppression. [ABSTRACT FROM AUTHOR]
- Published
- 2012
- Full Text
- View/download PDF
25. Sleeping Beauty mutagenesis reveals cooperating mutations and pathways in pancreatic adenocarcinoma.
- Author
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Mann, Karen M., Ward, Jerrold M., Yew, Christopher Chin Kuan, Kovochich, Anne, Dawson, David W., Black, Michael A., Brett, Benjamin T., Sheetz, Todd E., Dupuy, Adam J., Chang, David K., Biankin, Andrew V., Waddell, Nicola, Kassahn, Karin S., Grimmond, Sean M., Rust, Alistair G., Adams, David J., Jenkins, Nancy A., and Copeland, Neal G.
- Subjects
PANCREATIC cancer ,MUTAGENESIS ,ADENOCARCINOMA ,CANCER invasiveness ,CANCER genes ,GENE frequency ,CHROMATIN ,LABORATORY mice - Abstract
Pancreatic cancer is one of the most deadly cancers affecting the Western world. Because the disease is highly metastatic and difficult to diagnosis until late stages, the 5-y survival rate is around 5%. The identification of molecular cancer drivers is critical for furthering our understanding of the disease and development of improved diagnostic tools and therapeutics. We have conducted a mutagenic screen using Sleeping Beauty (SB) in mice to identify new candidate cancer genes in pancreatic cancer. By combining SB with an oncogenic Kras allele, we observed highly metastatic pancreatic adenocarcinomas. Using two independent statistical methods to identify loci commonly mutated by SB in these tumors, we identified 681 loci that comprise 543 candidate cancer genes (CCGs); 75 of these CCGs, including Mll3 and Ptk2, have known mutations in human pancreatic cancer. We identified point mutations in human pancreatic patient samples for another 11 CCGs, including Acvr2a and Map2k4. Importantly, 10% of the CCGs are involved in chromatin remodeling, including Arid4b, Kdm6a, and Nsd3, and all SB tumors have at least one mutated gene involved in this process; 20 CCGs, including Ctnnd1, Fbxo11, and Vgll4, are also significantly associated with poor patient survival. SB mutagenesis provides a rich resource of mutations in potential cancer drivers for cross-comparative analyses with ongoing sequencing efforts in human pancreatic adenocarcinoma. [ABSTRACT FROM AUTHOR]
- Published
- 2012
- Full Text
- View/download PDF
26. Increased tumorigenesis associated with loss of the tumor suppressor gene Cadm1.
- Author
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van der Weyden, Louise, Arends, Mark J., Rust, Alistair G., Poulogiannis, George, McIntyre, Rebecca E., and Adams, David J.
- Subjects
TUMOR suppressor genes ,CARCINOGENESIS ,LYMPHOMAS ,LABORATORY mice ,CARCINOGENICITY - Abstract
Background: CADM1 encodes an immunoglobulin superfamily (IGSF) cell adhesion molecule. Inactivation of CADM11, either by promoter hypermethylation or loss of heterozygosity, has been reported in a wide variety of tumor types, thus it has been postulated as a tumor suppressor gene.Findings: We show for the first time that Cadm1 homozygous null mice die significantly faster than wildtype controls due to the spontaneous development of tumors at an earlier age and an increased tumor incidence of predominantly lymphomas, but also some solid tumors. Tumorigenesis was accelerated after irradiation of Cadm1mice, with the reduced latency in tumor formation suggesting there are genes that collaborate with loss of Cadm1in tumorigenesis. To identify these co-operating genetic events, we performed a Sleeping Beauty transposonmediatedinsertional mutagenesis screen in Cadm1 mice, and identified several common insertion sites (CIS) found specifically on a Cadm1-null background (and not wildtype background).Conclusion: We confirm that Cadm1 is indeed a bona fide tumor suppressor gene and provide new insights into genetic partners that co-operate in tumorigenesis when Cadm1-expression is lost. [ABSTRACT FROM AUTHOR]
- Published
- 2012
- Full Text
- View/download PDF
27. Insertional mutagenesis identifies multiple networks of cooperating genes driving intestinal tumorigenesis.
- Author
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March, H Nikki, Rust, Alistair G, Wright, Nicholas A, ten Hoeve, Jelle, de Ridder, Jeroen, Eldridge, Matthew, van der Weyden, Louise, Berns, Anton, Gadiot, Jules, Uren, Anthony, Kemp, Richard, Arends, Mark J, Wessels, Lodewyk F A, Winton, Douglas J, and Adams, David J
- Subjects
GENETICS of colon cancer ,MUTAGENESIS ,MUTAGENICITY testing ,CARCINOGENESIS ,COMPARATIVE genomic hybridization ,GENETICS - Abstract
The evolution of colorectal cancer suggests the involvement of many genes. To identify new drivers of intestinal cancer, we performed insertional mutagenesis using the Sleeping Beauty transposon system in mice carrying germline or somatic Apc mutations. By analyzing common insertion sites (CISs) isolated from 446 tumors, we identified many hundreds of candidate cancer drivers. Comparison to human data sets suggested that 234 CIS-targeted genes are also dysregulated in human colorectal cancers. In addition, we found 183 CIS-containing genes that are candidate Wnt targets and showed that 20 CISs-containing genes are newly discovered modifiers of canonical Wnt signaling. We also identified mutations associated with a subset of tumors containing an expanded number of Paneth cells, a hallmark of deregulated Wnt signaling, and genes associated with more severe dysplasia included those encoding members of the FGF signaling cascade. Some 70 genes had co-occurrence of CIS pairs, clustering into 38 sub-networks that may regulate tumor development. [ABSTRACT FROM AUTHOR]
- Published
- 2011
- Full Text
- View/download PDF
28. Acute sensitivity of the oral mucosa to oncogenic K-ras.
- Author
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van der Weyden, Louise, Alcolea, Maria P, Jones, Philip H, Rust, Alistair G, Arends, Mark J, and Adams, David J
- Published
- 2011
- Full Text
- View/download PDF
29. Integrating genomic binding site predictions using real-valued meta classifiers.
- Author
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Yi Sun, Robinson, Mark, Adams, Rod, Te Boekhorst, Rene, Rust, Alistair G., and Davey, Neil
- Subjects
GENOMICS ,BINDING sites ,TRANSCRIPTION factors ,NUCLEOTIDE sequence ,ALGORITHMS - Abstract
Currently the best algorithms for predicting transcription factor binding sites in DNA sequences are severely limited in accuracy. There is good reason to believe that predictions from different classes of algorithms could be used in conjunction to improve the quality of predictions. In this paper, we apply single layer networks, rules sets, support vector machines and the Adaboost algorithm to predictions from 12 key real valued algorithms. Furthermore, we use a ‘window’ of consecutive results as the input vector in order to contextualise the neighbouring results. We improve the classification result with the aid of under- and over-sampling techniques. We find that support vector machines and the Adaboost algorithm outperform the original individual algorithms and the other classifiers employed in this work. In particular they give a better tradeoff between recall and precision. [ABSTRACT FROM AUTHOR]
- Published
- 2009
- Full Text
- View/download PDF
30. Function of C/EBPδ in a regulatory circuit that discriminates between transient and persistent TLR4-induced signals.
- Author
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Litvak, Vladimir, Ramsey, Stephen A, Rust, Alistair G, Zak, Daniel E, Kennedy, Kathleen A, Lampano, Aaron E, Nykter, Matti, Shmulevich, Ilya, and Aderem, Alan
- Abstract
The innate immune system is like a double-edged sword: it is absolutely required for host defense against infection, but when uncontrolled, it can trigger a plethora of inflammatory diseases. Here we use systems-biology approaches to predict and confirm the existence of a gene-regulatory network involving dynamic interaction among the transcription factors NF-κB, C/EBPδ and ATF3 that controls inflammatory responses. We mathematically modeled transcriptional regulation of the genes encoding interleukin 6 and C/EBPδ and experimentally confirmed the prediction that the combination of an initiator (NF-κB), an amplifier (C/EBPδ) and an attenuator (ATF3) forms a regulatory circuit that discriminates between transient and persistent Toll-like receptor 4–induced signals. Our results suggest a mechanism that enables the innate immune system to detect the duration of infection and to respond appropriately. [ABSTRACT FROM AUTHOR]
- Published
- 2009
- Full Text
- View/download PDF
31. Probabilistic Inference of Transcription Factor Binding from Multiple Data Sources.
- Author
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Lähdesmäki, Harri, Rust, Alistair G., and Shmulevich, Ilya
- Subjects
INFERENCE (Logic) ,TRANSCRIPTION factors ,MOLECULAR biology ,CELLS ,STATISTICAL hypothesis testing ,GENES ,DNA ,BIOLOGY ,DATA - Abstract
An important problem in molecular biology is to build a complete understanding of transcriptional regulatory processes in the cell. We have developed a flexible, probabilistic framework to predict TF binding from multiple data sources that differs from the standard hypothesis testing (scanning) methods in several ways. Our probabilistic modeling framework estimates the probability of binding and, thus, naturally reflects our degree of belief in binding. Probabilistic modeling also allows for easy and systematic integration of our binding predictions into other probabilistic modeling methods, such as expression-based gene network inference. The method answers the question of whether the whole analyzed promoter has a binding site, but can also be extended to estimate the binding probability at each nucleotide position. Further, we introduce an extension to model combinatorial regulation by several TFs. Most importantly, the proposed methods can make principled probabilistic inference from multiple evidence sources, such as, multiple statistical models (motifs) of the TFs, evolutionary conservation, regulatory potential, CpG islands, nucleosome positioning, DNase hypersensitive sites, ChIP-chip binding segments and other (prior) sequence-based biological knowledge. We developed both a likelihood and a Bayesian method, where the latter is implemented with a Markov chain Monte Carlo algorithm. Results on a carefully constructed test set from the mouse genome demonstrate that principled data fusion can significantly improve the performance of TF binding prediction methods. We also applied the probabilistic modeling framework to all promoters in the mouse genome and the results indicate a sparse connectivity between transcriptional regulators and their target promoters. To facilitate analysis of other sequences and additional data, we have developed an on-line web tool, ProbTF, which implements our probabilistic TF binding prediction method using multiple data sources. Test data set, a web tool, source codes and supplementary data are available at: http://www.probtf.org. [ABSTRACT FROM AUTHOR]
- Published
- 2008
- Full Text
- View/download PDF
32. Uncovering a Macrophage Transcriptional Program by Integrating Evidence from Motif Scanning and Expression Dynamics.
- Author
-
Ramsey, Stephen A., Klemm, Sandy L., Zak, Daniel E., Kennedy, Kathleen A., Thorsson, Vesteinn, Bin Li, Gilchrist, Mark, Gold, Elizabeth S., Johnson, Carrie D., Litvak, Vladimir, Navarro, Garnet, Roach, Jared C., Rosenberger, Carrie M., Rust, Alistair G., Yudkovsky, Natalya, Aderem, Alan, and Shmulevich, Ilya
- Subjects
MACROPHAGES ,GENE expression ,TRANSCRIPTION factors ,KILLER cells ,IMMUNOCOMPETENT cells ,CONNECTIVE tissue cells ,NUCLEOTIDE sequence - Abstract
Macrophages are versatile immune cells that can detect a variety of pathogen-associated molecular patterns through their Toll-like receptors (TLRs). In response to microbial challenge, the TLR-stimulated macrophage undergoes an activation program controlled by a dynamically inducible transcriptional regulatory network. Mapping a complex mammalian transcriptional network poses significant challenges and requires the integration of multiple experimental data types. In this work, we inferred a transcriptional network underlying TLR-stimulated murine macrophage activation. Microarray-based expression profiling and transcription factor binding site motif scanning were used to infer a network of associations between transcription factor genes and clusters of co-expressed target genes. The time-lagged correlation was used to analyze temporal expression data in order to identify potential causal influences in the network. A novel statistical test was developed to assess the significance of the time-lagged correlation. Several associations in the resulting inferred network were validated using targeted ChIP-on-chip experiments. The network incorporates known regulators and gives insight into the transcriptional control of macrophage activation. Our analysis identified a novel regulator (TGIF1) that may have a role in macrophage activation. [ABSTRACT FROM AUTHOR]
- Published
- 2008
- Full Text
- View/download PDF
33. The Innate Immune Database (IIDB).
- Author
-
Korb, Martin, Rust, Alistair G., Thorsson, Vesteinn, Battail, Christophe, Bin Li, Daehee Hwang, Kennedy, Kathleen A., Roach, Jared C., Rosenberger, Carrie M., Gilchrist, Mark, Zak, Daniel, Johnson, Carrie, Marzolf, Bruz, Aderem, Alan, Shmulevich, Ilya, and Bolouri, Hamid
- Subjects
BONE marrow ,TRANSCRIPTION factors ,IMMUNOLOGY ,MEDICAL sciences ,GENES - Abstract
Background: As part of a National Institute of Allergy and Infectious Diseases funded collaborative project, we have performed over 150 microarray experiments measuring the response of C57/BL6 mouse bone marrow macrophages to toll-like receptor stimuli. These microarray expression profiles are available freely from our project web site http:// www.innateImmunity-systemsbiology.org. Here, we report the development of a database of computationally predicted transcription factor binding sites and related genomic features for a set of over 2000 murine immune genes of interest. Our database, which includes microarray coexpression clusters and a host of web-based query, analysis and visualization facilities, is available freely via the internet. It provides a broad resource to the research community, and a stepping stone towards the delineation of the network of transcriptional regulatory interactions underlying the integrated response of macrophages to pathogens. Description: We constructed a database indexed on genes and annotations of the immediate surrounding genomic regions. To facilitate both gene-specific and systems biology oriented research, our database provides the means to analyze individual genes or an entire genomic locus. Although our focus to-date has been on mammalian toll-like receptor signaling pathways, our database structure is not limited to this subject, and is intended to be broadly applicable to immunology. By focusing on selected immune-active genes, we were able to perform computationally intensive expression and sequence analyses that would currently be prohibitive if applied to the entire genome. Using six complementary computational algorithms and methodologies, we identified transcription factor binding sites based on the Position Weight Matrices available in TRANSFAC. For one example transcription factor (ATF3) for which experimental data is available, over 50% of our predicted binding sites coincide with genome-wide chromatin immnuopreciptation (ChIP-chip) results. Our database can be interrogated via a web interface. Genomic annotations and binding site predictions can be automatically viewed with a customized version of the Argo genome browser. Conclusion: We present the Innate Immune Database (IIDB) as a community resource for immunologists interested in gene regulatory systems underlying innate responses to pathogens. The database website can be freely accessed at http://db.systemsbiology.net/IIDB. [ABSTRACT FROM AUTHOR]
- Published
- 2008
- Full Text
- View/download PDF
34. Systems biology approaches identify ATF3 as a negative regulator of Toll-like receptor 4.
- Author
-
Gilchrist, Mark, Thorsson, Vesteinn, Bin Li, Rust, Alistair G., Korb, Martin, Kennedy, Kathleen, Tsonwin Hai, Bolouri, Hamid, and Aderem, Alan
- Subjects
IMMUNE system ,PROTEINS ,TRANSCRIPTION factors ,CYTOKINES ,DNA microarrays ,MACROPHAGES - Abstract
The innate immune system is absolutely required for host defence, but, uncontrolled, it leads to inflammatory disease. This control is mediated, in part, by cytokines that are secreted by macrophages. Immune regulation is extraordinarily complex, and can be best investigated with systems approaches (that is, using computational tools to predict regulatory networks arising from global, high-throughput data sets). Here we use cluster analysis of a comprehensive set of transcriptomic data derived from Toll-like receptor (TLR)-activated macrophages to identify a prominent group of genes that appear to be regulated by activating transcription factor 3 (ATF3), a member of the CREB/ATF family of transcription factors. Network analysis predicted that ATF3 is part of a transcriptional complex that also contains members of the nuclear factor (NF)-κB family of transcription factors. Promoter analysis of the putative ATF3-regulated gene cluster demonstrated an over-representation of closely apposed ATF3 and NF-κB binding sites, which was verified by chromatin immunoprecipitation and hybridization to a DNA microarray. This cluster included important cytokines such as interleukin (IL)-6 and IL-12b. ATF3 and Rel (a component of NF-κB) were shown to bind to the regulatory regions of these genes upon macrophage activation. A kinetic model of Il6 and Il12b messenger RNA expression as a function of ATF3 and NF-κB promoter binding predicted that ATF3 is a negative regulator of Il6 and Il12b transcription, and this hypothesis was validated using Atf3-null mice. ATF3 seems to inhibit Il6 and Il12b transcription by altering chromatin structure, thereby restricting access to transcription factors. Because ATF3 is itself induced by lipopolysaccharide, it seems to regulate TLR-stimulated inflammatory responses as part of a negative-feedback loop. [ABSTRACT FROM AUTHOR]
- Published
- 2006
- Full Text
- View/download PDF
35. TRANSCRIPTION BINDING SITE PREDICTION USING MARKOV MODELS.
- Author
-
Abnizova, Irina, Rust, Alistair G., Robinson, Mark, Te Boekhorst, Rene, and Gilks, Walter R.
- Subjects
BINDING sites ,MARKOV processes ,GENE expression ,GENETIC regulation ,NUCLEOTIDE sequence ,TRANSCRIPTION factors ,GENETIC algorithms - Abstract
One of the main goals of analysing DNA sequences is to understand the temporal and positional information that specifies gene expression. An important step in this process is the recognition of gene expression regulatory elements. Experimental procedures for this are slow and costly. In this paper we present a computational non-supervised algorithm that facilitates the process by statistically identifying the most likely regions within a putative regulatory sequence. A probabilistic technique is presented, based on the approximation of regulatory DNA with a Markov chain, for the location of putative transcription factor binding sites in a single stretch of DNA. Hereto we developed a procedure to approximate the order of Markov model for a given DNA sequence that circumvents some of the prohibitive assumptions underlying Markov modeling. Application of the algorithm to data from 55 genes in five species shows the high sensitivity of this Markov search algorithm. Our algorithm does not require any prior knowledge in the form of description or cross-genomic comparison; it is context sensitive and takes DNA heterogeneity into account. [ABSTRACT FROM AUTHOR]
- Published
- 2006
- Full Text
- View/download PDF
36. A data integration methodology for systems biology: Experimental verification.
- Author
-
Daehee Hwang, Smith, Jennifer J., Leslie, Deena M., Weston, Andrea D., Rust, Alistair G., Ramsey, Stephen, de Atauri, Pedro, Siegel, Andrew F., Bolouri, Hamid, Aitchison, John D., and Hood, Leroy
- Subjects
GENES ,GLYCOSIDES ,MESSENGER RNA ,DNA ,NUCLEIC acids ,METHODOLOGY - Abstract
The integration of data from multiple global assays is essential to understanding dynamic spatiotemporal interactions within cells. In a companion paper, we reported a data integration methodology, designated Pointillist, that can handle multiple data types from technologies with different noise characteristics. Here we demonstrate its application to the integration of 18 data sets relating to galactose utilization in yeast. These data include global changes in mRNA and protein abundance, genome-wide protein-DNA interaction data, database information, and computational predictions of protein-DNA and protein-protein interactions. We divided the integration task to determine three network components: key system elements (genes and proteins), protein-protein interactions, and protein-DNA interactions. Results indicate that the reconstructed network efficiently focuses on and recapitulates the known biology of galactose utilization. It also provided new insights, some of which were verified experimentally. The methodology described here, addresses a critical need across all domains of molecular and cell biology, to effectively integrate large and disparate data sets. [ABSTRACT FROM AUTHOR]
- Published
- 2005
- Full Text
- View/download PDF
37. A data integration methodology for systems biology.
- Author
-
Daehee Hwang, Rust, Alistair G., Ramsey, Stephen, Smith, Jennifer J., Leslie, Deena M., Weston, Andrea D., de Atauri, Pedro, Aitchison, John D., Hood, Leroy, Siegel, Andrew F., and Bolouri, Hamid
- Subjects
TECHNOLOGY ,BIOTECHNOLOGY ,METHODOLOGY ,FIRE assay ,COMPUTER software ,OPEN source software - Abstract
Different experimental technologies measure different aspects of a system and to differing depth and breadth. High-throughput assays have inherently high false-positive and false-negative rates. Moreover, each technology includes systematic biases of a different nature. These differences make network reconstruction from multiple data sets difficult and error-prone. Additionally, because of the rapid rate of progress in biotechnology, there is usually no curated exemplar data set from which one might estimate data integration parameters. To address these concerns, we have developed data integration methods that can handle multiple data sets differing in statistical power, type, size, and network coverage without requiring a curated training data set. Our methodology is general in purpose and may be applied to integrate data from any existing and future technologies. Here we outline our methods and then demonstrate their performance by applying them to simulated data sets. The results show that these methods select true-positive data elements much more accurately than classical approaches. In an accompanying companion paper, we demonstrate the applicability of our approach to biological data. We have integrated our methodology into a free open source software package named POINTILLIST. [ABSTRACT FROM AUTHOR]
- Published
- 2005
- Full Text
- View/download PDF
38. Cake: a bioinformatics pipeline for the integrated analysis of somatic variants in cancer genomes.
- Author
-
Rashid, Mamunur, Robles-Espinoza, Carla Daniela, Rust, Alistair G., and Adams, David J.
- Subjects
CANCER genetics ,BIOINFORMATICS ,SINGLE nucleotide polymorphisms ,SENSITIVITY analysis ,COMPUTER workstation clusters ,PERFORMANCE evaluation ,COMPUTER algorithms - Abstract
Summary: We have developed Cake, a bioinformatics software pipeline that integrates four publicly available somatic variant-calling algorithms to identify single nucleotide variants with higher sensitivity and accuracy than any one algorithm alone. Cake can be run on a high-performance computer cluster or used as a stand-alone application.Availabilty: Cake is open-source and is available from http://cakesomatic.sourceforge.net/Contact: da1@sanger.ac.ukSupplementary Information: Supplementary data are available at Bioinformatics online. [ABSTRACT FROM AUTHOR]
- Published
- 2013
- Full Text
- View/download PDF
39. Transposon mutagenesis identifies genes that transform neural stem cells into glioma-initiating cells.
- Author
-
Koso, Hideto, Takeda, Haruna, Kuan Yewa, Christopher Chin, Ward, Jerrold M., Nariai, Naoki, Ueno, Kazuko, Nagasaki, Masao, Watanabe, Sumiko, Rust, Alistair G., Adams, David J., Copeland, Neal G., and Jenkins, Nancy A.
- Subjects
TRANSPOSONS ,MUTAGENESIS ,NEURAL stem cells ,GLIOMAS ,CANCER cells ,TUMORS - Abstract
In this article, the authors discuss the role of transposon mutagenesis in identifying genes that transform neural stem cells (NSCs) into glioma-initiating cells (GC). They mention a study which used transposans to identify genes and signaling pathways to transform NSCs into cancer initiating cells (CIC) for GC. The study also examined the combinations of CIS genes in tumors derived from a single immortalized line.
- Published
- 2012
- Full Text
- View/download PDF
40. Somatic drivers of B-ALL in a model of ETV6-RUNX1; Pax5(+/-) leukemia.
- Author
-
van der Weyden, Louise, Giotopoulos, George, Wong, Kim, Rust, Alistair G, Robles-Espinoza, Carla Daniela, Osaki, Hikari, Huntly, Brian J, and Adams, David J
- Abstract
Background: B-cell precursor acute lymphoblastic leukemia (B-ALL) is amongst the leading causes of childhood cancer-related mortality. Its most common chromosomal aberration is the ETV6-RUNX1 fusion gene, with ~25% of ETV6-RUNX1 patients also carrying PAX5 alterations.Methods: We have recreated this mutation background by inter-crossing Etv6-RUNX1 (Etv6 (RUNX1-SB)) and Pax5(+/-) mice and performed an in vivo analysis to find driver genes using Sleeping Beauty transposon-mediated mutagenesis and also exome sequencing.Results: Combination of Etv6-RUNX1 and Pax5(+/-) alleles generated a transplantable B220 + CD19+ B-ALL with a significant disease incidence. RNA-seq analysis showed a gene expression pattern consistent with arrest at the pre-B stage. Analysis of the transposon common insertion sites identified genes involved in B-cell development (Zfp423) and the JAK/STAT signaling pathway (Jak1, Stat5 and Il2rb), while exome sequencing revealed somatic hotspot mutations in Jak1 and Jak3 at residues analogous to those mutated in human leukemias, and also mutation of Trp53.Conclusions: Powerful synergies exists in our model suggesting STAT pathway activation and mutation of Trp53 are potent drivers of B-ALL in the context of Etv6-RUNX1;Pax5(+/-). [ABSTRACT FROM AUTHOR]- Published
- 2015
- Full Text
- View/download PDF
41. Corrigendum: The deubiquitinase USP9X suppresses pancreatic ductal adenocarcinoma.
- Author
-
Pérez-Mancera, Pedro A., Rust, Alistair G., van der Weyden, Louise, Kristiansen, Glen, Li, Allen, Sarver, Aaron L., Silverstein, Kevin A. T., Grützmann, Robert, Aust, Daniela, Rümmele, Petra, Knösel, Thomas, Herd, Colin, Stemple, Derek L., Kettleborough, Ross, Brosnan, Jacqueline A., Li, Ang, Morgan, Richard, Knight, Spencer, Yu, Jun, and Stegeman, Shane
- Subjects
ADENOCARCINOMA - Abstract
A correction to the article "The deubiquitinase USP9X suppresses pancreatic ductal adenocarcinoma" that was published in the 2012 issue is presented.
- Published
- 2013
- Full Text
- View/download PDF
42. Systems biology approaches identify ATF3 as a negative regulator of Toll-like receptor 4.
- Author
-
Gilchrist, Mark, Thorsson, Vesteinn, Li, Bin, Rust, Alistair G., Korb, Martin, Roach, Jared C., Kennedy, Kathleen, Hai, Tsonwin, Bolouri, Hamid, and Aderem, Alan
- Subjects
BIOLOGY - Abstract
A correction to the article "Systems Biology Approaches Identify ATF3 as a Negative Regulator of Toll-like Receptor 4," by Mark Gilchrist and colleagues that was published in a 2006 issue is presented.
- Published
- 2008
- Full Text
- View/download PDF
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