41 results on '"Ponstingl, Hannes"'
Search Results
2. SETBP1 overexpression acts in the place of class-defining mutations to drive FLT3-ITD–mutant AML
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Pacharne, Suruchi, Dovey, Oliver M., Cooper, Jonathan L., Gu, Muxin, Friedrich, Mathias J., Rajan, Sandeep S., Barenboim, Maxim, Collord, Grace, Vijayabaskar, M.S., Ponstingl, Hannes, De Braekeleer, Etienne, Bautista, Ruben, Mazan, Milena, Rad, Roland, Tzelepis, Konstantinos, Wright, Penny, Gozdecka, Malgorzata, and Vassiliou, George S.
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- 2021
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3. Genetic perturbation of PU.1 binding and chromatin looping at neutrophil enhancers associates with autoimmune disease
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Watt, Stephen, Vasquez, Louella, Walter, Klaudia, Mann, Alice L., Kundu, Kousik, Chen, Lu, Sims, Ying, Ecker, Simone, Burden, Frances, Farrow, Samantha, Farr, Ben, Iotchkova, Valentina, Elding, Heather, Mead, Daniel, Tardaguila, Manuel, Ponstingl, Hannes, Richardson, David, Datta, Avik, Flicek, Paul, Clarke, Laura, Downes, Kate, Pastinen, Tomi, Fraser, Peter, Frontini, Mattia, Javierre, Biola-Maria, Spivakov, Mikhail, and Soranzo, Nicole
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- 2021
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4. Correction to: PiggyBac mutagenesis and exome sequencing identify genetic driver landscapes and potential therapeutic targets of EGFR-mutant gliomas
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Noorani, Imran, de la Rosa, Jorge, Choi, Yoon Ha, Strong, Alexander, Ponstingl, Hannes, Vijayabaskar, M. S., Lee, Jusung, Lee, Eunmin, Richard-Londt, Angela, Friedrich, Mathias, Furlanetto, Federica, Fuente, Rocio, Banerjee, Ruby, Yang, Fengtang, Law, Frances, Watts, Colin, Rad, Roland, Vassiliou, George, Kim, Jong Kyoung, Santarius, Thomas, Brandner, Sebastian, and Bradley, Allan
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- 2020
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5. PiggyBac mutagenesis and exome sequencing identify genetic driver landscapes and potential therapeutic targets of EGFR-mutant gliomas
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Noorani, Imran, de la Rosa, Jorge, Choi, Yoon Ha, Strong, Alexander, Ponstingl, Hannes, Vijayabaskar, M. S., Lee, Jusung, Lee, Eunmin, Richard-Londt, Angela, Friedrich, Mathias, Furlanetto, Federica, Fuente, Rocio, Banerjee, Ruby, Yang, Fengtang, Law, Frances, Watts, Colin, Rad, Roland, Vassiliou, George, Kim, Jong Kyoung, Santarius, Thomas, Brandner, Sebastian, and Bradley, Allan
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- 2020
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6. Comprehensive annotation and evolutionary insights into the canine (Canis lupus familiaris) antigen receptor loci
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Martin, Jolyon, Ponstingl, Hannes, Lefranc, Marie-Paule, Archer, Joy, Sargan, David, and Bradley, Allan
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- 2018
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7. Development and validation of a comprehensive genomic diagnostic tool for myeloid malignancies
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McKerrell, Thomas, Moreno, Thaidy, Ponstingl, Hannes, Bolli, Niccolo, Dias, João M.L., Tischler, German, Colonna, Vincenza, Manasse, Bridget, Bench, Anthony, Bloxham, David, Herman, Bram, Fletcher, Danielle, Park, Naomi, Quail, Michael A., Manes, Nicla, Hodkinson, Clare, Baxter, Joanna, Sierra, Jorge, Foukaneli, Theodora, Warren, Alan J., Chi, Jianxiang, Costeas, Paul, Rad, Roland, Huntly, Brian, Grove, Carolyn, Ning, Zemin, Tyler-Smith, Chris, Varela, Ignacio, Scott, Mike, Nomdedeu, Josep, Mustonen, Ville, and Vassiliou, George S.
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- 2016
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8. ATM orchestrates the DNA-damage response to counter toxic non-homologous end-joining at broken replication forks
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Balmus, Gabriel, Pilger, Domenic, Coates, Julia, Demir, Mukerrem, Sczaniecka-Clift, Matylda, Barros, Ana C., Woods, Michael, Fu, Beiyuan, Yang, Fengtang, Chen, Elisabeth, Ostermaier, Matthias, Stankovic, Tatjana, Ponstingl, Hannes, Herzog, Mareike, Yusa, Kosuke, Martinez, Francisco Munoz, Durant, Stephen T., Galanty, Yaron, Beli, Petra, Adams, David J., Bradley, Allan, Metzakopian, Emmanouil, Forment, Josep V., and Jackson, Stephen P.
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- 2019
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9. SRPK1 maintains acute myeloid leukemia through effects on isoform usage of epigenetic regulators including BRD4
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Tzelepis, Konstantinos, De Braekeleer, Etienne, Aspris, Demetrios, Barbieri, Isaia, Vijayabaskar, M. S., Liu, Wen-Hsin, Gozdecka, Malgorzata, Metzakopian, Emmanouil, Toop, Hamish D., Dudek, Monika, Robson, Samuel C., Hermida-Prado, Francisco, Yang, Yu Hsuen, Babaei-Jadidi, Roya, Garyfallos, Dimitrios A., Ponstingl, Hannes, Dias, Joao M. L., Gallipoli, Paolo, Seiler, Michael, Buonamici, Silvia, Vick, Binje, Bannister, Andrew J., Rad, Roland, Prinjha, Rab K., Marioni, John C., Huntly, Brian, Batson, Jennifer, Morris, Jonathan C., Pina, Cristina, Bradley, Allan, Jeremias, Irmela, Bates, David O., Yusa, Kosuke, Kouzarides, Tony, and Vassiliou, George S.
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- 2018
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10. A single-copy Sleeping Beauty transposon mutagenesis screen identifies new PTEN-cooperating tumor suppressor genes
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de la Rosa, Jorge, Weber, Julia, Friedrich, Mathias Josef, Li, Yilong, Rad, Lena, Ponstingl, Hannes, Liang, Qi, de Quirós, Sandra Bernaldo, Noorani, Imran, Metzakopian, Emmanouil, Strong, Alexander, Li, Meng Amy, Astudillo, Aurora, Fernández-García, María Teresa, Fernández-García, María Soledad, Hoffman, Gary J, Fuente, Rocío, Vassiliou, George S, Rad, Roland, López-Otín, Carlos, Bradley, Allan, and Cadiñanos, Juan
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- 2017
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11. Genome-wide transposon screening and quantitative insertion site sequencing for cancer gene discovery in mice
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Friedrich, Mathias J, Rad, Lena, Bronner, Iraad F, Strong, Alexander, Wang, Wei, Weber, Julia, Mayho, Matthew, Ponstingl, Hannes, Engleitner, Thomas, Grove, Carolyn, Pfaus, Anja, Saur, Dieter, Cadiñanos, Juan, Quail, Michael A, Vassiliou, George S, Liu, Pentao, Bradley, Allan, and Rad, Roland
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- 2017
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12. Promoter-bound METTL3 maintains myeloid leukaemia by m6A-dependent translation control
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Barbieri, Isaia, Tzelepis, Konstantinos, Pandolfini, Luca, Shi, Junwei, Millán-Zambrano, Gonzalo, Robson, Samuel C., Aspris, Demetrios, Migliori, Valentina, Bannister, Andrew J., Han, Namshik, De Braekeleer, Etienne, Ponstingl, Hannes, Hendrick, Alan, Vakoc, Christopher R., Vassiliou, George S., and Kouzarides, Tony
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- 2017
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13. A Genetic Progression Model of BrafV600E-Induced Intestinal Tumorigenesis Reveals Targets for Therapeutic Intervention
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Rad, Roland, Cadiñanos, Juan, Rad, Lena, Varela, Ignacio, Strong, Alexander, Kriegl, Lydia, Constantino-Casas, Fernando, Eser, Stefan, Hieber, Maren, Seidler, Barbara, Price, Stacey, Fraga, Mario F., Calvanese, Vincenzo, Hoffman, Gary, Ponstingl, Hannes, Schneider, Günter, Yusa, Kosuke, Grove, Carolyn, Schmid, Roland M., Wang, Wei, Vassiliou, George, Kirchner, Thomas, McDermott, Ultan, Liu, Pentao, Saur, Dieter, and Bradley, Allan
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- 2013
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14. A conditional piggyBac transposition system for genetic screening in mice identifies oncogenic networks in pancreatic cancer
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Rad, Roland, Rad, Lena, Wang, Wei, Strong, Alexander, Ponstingl, Hannes, Bronner, Iraad F, Mayho, Matthew, Steiger, Katja, Weber, Julia, Hieber, Maren, Veltkamp, Christian, Eser, Stefan, Geumann, Ulf, Öllinger, Rupert, Zukowska, Magdalena, Barenboim, Maxim, Maresch, Roman, Cadiñanos, Juan, Friedrich, Mathias, Varela, Ignacio, Constantino-Casas, Fernando, Sarver, Aaron, ten Hoeve, Jelle, Prosser, Haydn, Seidler, Barbara, Bauer, Judith, Heikenwälder, Mathias, Metzakopian, Emmanouil, Krug, Anne, Ehmer, Ursula, Schneider, Günter, Knösel, Thomas, Rümmele, Petra, Aust, Daniela, Grützmann, Robert, Pilarsky, Christian, Ning, Zemin, Wessels, Lodewyk, Schmid, Roland M, Quail, Michael A, Vassiliou, George, Esposito, Irene, Liu, Pentao, Saur, Dieter, and Bradley, Allan
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- 2015
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15. Rapid Measurement of Scalar Three-bond 1HN-1Hα spin coupling constants in 15N-labelled proteins
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Ponstingl, Hannes and Otting, Gottfried
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- 1998
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16. Detection of protein–ligand NOEs with small, weakly binding ligands by combined relaxation and diffusion filtering
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Ponstingl, Hannes and Otting, Gottfried
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- 1997
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17. Resonance assignment and structural analysis of acid denatured E. coli [U-15N]-glutaredoxin 3: Use of 3D 15N-HSQC-(TOCSY-NOESY)-15N-HSQC
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Nordstrand, Kerstin, Ponstingl, Hannes, Holmgren, Arne, and Otting, Gottfried
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- 1996
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18. A CRISPR Dropout Screen Identifies Genetic Vulnerabilities and Therapeutic Targets in Acute Myeloid Leukemia
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Tzelepis, Konstantinos, Koike-Yusa, Hiroko, De Braekeleer, Etienne, Li, Yilong, Metzakopian, Emmanouil, Dovey, Oliver M., Mupo, Annalisa, Grinkevich, Vera, Li, Meng, Mazan, Milena, Gozdecka, Malgorzata, Ohnishi, Shuhei, Cooper, Jonathan, Patel, Miten, McKerrell, Thomas, Chen, Bin, Domingues, Ana Filipa, Gallipoli, Paolo, Teichmann, Sarah, Ponstingl, Hannes, McDermott, Ultan, Saez-Rodriguez, Julio, Huntly, Brian J.P., Iorio, Francesco, Pina, Cristina, Vassiliou, George S., Yusa, Kosuke, Tzelepis, Konstantinos [0000-0002-4865-7648], Mupo, Annalisa [0000-0002-2771-0462], McKerrell, Thomas [0000-0003-4235-0650], Gallipoli, Paolo [0000-0001-7254-2253], Teichmann, Sarah [0000-0002-6294-6366], Huntly, Brian [0000-0003-0312-161X], Correia Antunes Pina, Cristina [0000-0002-2575-6301], Vassiliou, George [0000-0003-4337-8022], and Apollo - University of Cambridge Repository
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Adult ,Resource ,Apoptosis ,genetic vulnerability ,KAT2A ,Genetic vulnerability ,acute myeloid leukemia ,Genetic screen ,AML ,Cell Line, Tumor ,hemic and lymphatic diseases ,Humans ,Clustered Regularly Interspaced Short Palindromic Repeats ,Genetic Testing ,Molecular Targeted Therapy ,lcsh:QH301-705.5 ,neoplasms ,health care economics and organizations ,Cell Proliferation ,Histone Acetyltransferases ,Acute myeloid leukemia ,Reproducibility of Results ,Cell Differentiation ,Leukemia, Myeloid, Acute ,MB-3 ,lcsh:Biology (General) ,genetic screen ,CRISPR ,KAT2A MB-3 - Abstract
Summary Acute myeloid leukemia (AML) is an aggressive cancer with a poor prognosis, for which mainstream treatments have not changed for decades. To identify additional therapeutic targets in AML, we optimize a genome-wide clustered regularly interspaced short palindromic repeats (CRISPR) screening platform and use it to identify genetic vulnerabilities in AML cells. We identify 492 AML-specific cell-essential genes, including several established therapeutic targets such as DOT1L, BCL2, and MEN1, and many other genes including clinically actionable candidates. We validate selected genes using genetic and pharmacological inhibition, and chose KAT2A as a candidate for downstream study. KAT2A inhibition demonstrated anti-AML activity by inducing myeloid differentiation and apoptosis, and suppressed the growth of primary human AMLs of diverse genotypes while sparing normal hemopoietic stem-progenitor cells. Our results propose that KAT2A inhibition should be investigated as a therapeutic strategy in AML and provide a large number of genetic vulnerabilities of this leukemia that can be pursued in downstream studies., Graphical Abstract, Highlights • Optimized CRISPR platform for identification of genome-wide genetic vulnerabilities • Catalog of genetic vulnerabilities in acute myeloid leukemia cell lines • KAT2A inhibition induces myeloid differentiation and apoptosis • KAT2A inhibition arrests the growth of primary AML cells, but not of normal progenitors, Tzelepis et al. optimize a CRISPR-Cas9-based platform for the performance of genome-wide recessive screens and apply it to identify genetic vulnerabilities of human AML cells. They identify several known therapeutic targets including BRD4, DOT1L, and MEN1, and numerous additional candidates. They provide data proposing KAT2A as a potential therapeutic target.
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- 2016
19. Geometrical representation of coherence transfer selection by pulsed field gradients in high-resolution nuclear magnetic resonance.
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Mitschang, Lorenz, Ponstingl, Hannes, Grindrod, David, and Oschkinat, Hartmut
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NUCLEAR magnetic resonance spectroscopy , *COHERENCE (Nuclear physics) , *CYCLOSPORINE - Abstract
A formalism for the calculation of suitable sequences of pulsed field gradients for signal selection in high-resolution NMR spectroscopy is presented. It is based on a geometrical interpretation of coherence transfer pathway selection by pulsed field gradients. The formalism allows the calculation of the suppression rates for undesired pathways and the determination of the most efficient sequence of pulsed field gradients. As an example, sequences for multiplicity filtered 13C/1H correlation experiments are calculated and analyzed. © 1995 American Institute of Physics. [ABSTRACT FROM AUTHOR]
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- 1995
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20. Comprehensive annotation and evolutionary insights into the canine (<italic>Canis lupus familiaris</italic>) antigen receptor loci.
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Martin, Jolyon, Ponstingl, Hannes, Lefranc, Marie-Paule, Archer, Joy, Sargan, David, and Bradley, Allan
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DIAGNOSIS of dog diseases ,DOG diseases ,VETERINARY therapeutics ,ANTIGEN receptors ,IMMUNOGLOBULINS ,T cell receptors - Abstract
Dogs are an excellent model for human disease. For example, the treatment of canine lymphoma has been predictive of the human response to that treatment. However, an incomplete picture of canine (
Canis lupus familiaris ) immunoglobulin (IG) and T cell receptor (TR)—or antigen receptor (AR)—gene loci has restricted their utility. This work advances the annotation of the canine AR loci and looks into breed-specific features of the loci. Bioinformatic analysis of unbiased RNA sequence data was used to complete the annotation of the canine AR genes. This annotation was used to query 107 whole genome sequences from 19 breeds and identified over 5500 alleles across the 550 genes of the seven AR loci: the IG heavy, kappa, and lambda loci; and the TR alpha, beta, gamma, and delta loci. Of note was the discovery that half of the IGK variable (V) genes were located downstream of, and inverted with respect to, the rest of the locus. Analysis of the germline sequences of all the AR V genes identified greater conservation between dog and human than mouse with either. This work brings our understanding of the genetic diversity and expression of AR in dogs to the same completeness as that of mice and men, making it the third species to have all AR loci comprehensively and accurately annotated. The large number of germline sequences serves as a reference for future studies, and has allowed statistically powerful conclusions to be drawn on the pressures that have shaped these loci. [ABSTRACT FROM AUTHOR]- Published
- 2018
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21. Promoter-bound METTL3 maintains myeloid leukaemia by m6A-dependent translation control.
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Barbieri, Isaia, Tzelepis, Konstantinos, Pandolfini, Luca, Shi, Junwei, Millán-Zambrano, Gonzalo, Robson, Samuel C., Aspris, Demetrios, Migliori, Valentina, Bannister, Andrew J., Han, Namshik, De Braekeleer, Etienne, Ponstingl, Hannes, Hendrick, Alan, Vakoc, Christopher R., Vassiliou, George S., and Kouzarides, Tony
- Abstract
N
6 -methyladenosine (m6 A) is an abundant internal RNA modification in both coding and non-coding RNAs that is catalysed by the METTL3-METTL14 methyltransferase complex. However, the specific role of these enzymes in cancer is still largely unknown. Here we define a pathway that is specific for METTL3 and is implicated in the maintenance of a leukaemic state. We identify METTL3 as an essential gene for growth of acute myeloid leukaemia cells in two distinct genetic screens. Downregulation of METTL3 results in cell cycle arrest, differentiation of leukaemic cells and failure to establish leukaemia in immunodeficient mice. We show that METTL3, independently of METTL14, associates with chromatin and localizes to the transcriptional start sites of active genes. The vast majority of these genes have the CAATT-box binding protein CEBPZ present at the transcriptional start site, and this is required for recruitment of METTL3 to chromatin. Promoter-bound METTL3 induces m6 A modification within the coding region of the associated mRNA transcript, and enhances its translation by relieving ribosome stalling. We show that genes regulated by METTL3 in this way are necessary for acute myeloid leukaemia. Together, these data define METTL3 as a regulator of a chromatin-based pathway that is necessary for maintenance of the leukaemic state and identify this enzyme as a potential therapeutic target for acute myeloid leukaemia. [ABSTRACT FROM AUTHOR]- Published
- 2017
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22. A Crispr/Cas9 Drop-out Screen Identifies Genome-Wide Genetic Valnerubilities in Acute Myeloid Leukaemia
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Tzelepis, Konstantinos, Koike-Yusa, Hiroko, De Braekeleer, Etienne, Li, Yilong, Metzakopian, Emmanouil, Dovey, Oliver M., Mupo, Annalisa, Grinkevich, Vera, Mazan, Milena M., Gozdecka, Malgorzata, Cooper, Jonathan L., Patel, Miten, McKerrell, Thomas David Hardman, Chen, Bin, Ponstingl, Hannes, Huntly, Brian J.P, McDermott, Ultan, Saez-Rodriguez, Julio, Iorio, Francesco, Yusa, Kosuke, and Vassiliou, George S.
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- 2015
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23. Leukemia-Associated Somatic Mutations Drive Distinct Patterns of Age-Related Clonal Hemopoiesis.
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McKerrell, Thomas, Park, Naomi, Moreno, Thaidy, Grove, Carolyn S., Ponstingl, Hannes, Stephens, Jonathan, Crawley, Charles, Craig, Jenny, Scott, Mike A., Hodkinson, Clare, Baxter, Joanna, Rad, Roland, Forsyth, Duncan R., Quail, Michael A., Zeggini, Eleftheria, Ouwehand, Willem, Varela, Ignacio, and Vassiliou, George S.
- Abstract
Summary Clonal hemopoiesis driven by leukemia-associated gene mutations can occur without evidence of a blood disorder. To investigate this phenomenon, we interrogated 15 mutation hot spots in blood DNA from 4,219 individuals using ultra-deep sequencing. Using only the hot spots studied, we identified clonal hemopoiesis in 0.8% of individuals under 60, rising to 19.5% of those ≥90 years, thus predicting that clonal hemopoiesis is much more prevalent than previously realized. DNMT3A- R882 mutations were most common and, although their prevalence increased with age, were found in individuals as young as 25 years. By contrast, mutations affecting spliceosome genes SF3B1 and SRSF 2, closely associated with the myelodysplastic syndromes, were identified only in those aged >70 years, with several individuals harboring more than one such mutation. This indicates that spliceosome gene mutations drive clonal expansion under selection pressures particular to the aging hemopoietic system and explains the high incidence of clonal disorders associated with these mutations in advanced old age. [ABSTRACT FROM AUTHOR]
- Published
- 2015
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24. Morphological aspects of oligomeric protein structures
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Ponstingl, Hannes, Thomas Kabir, Gorse, Denise, and Thornton, Janet M.
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PROTEINS , *OLIGOMERS , *MONOMERS , *BIOMOLECULES - Abstract
Abstract: Features of multimeric proteins are reviewed to shed light on the formation of protein assemblies from a structural perspective. The features comprise biochemical and geometric properties. They are compiled on new low-redundancy sets of crystal structures of homomeric proteins with different symmetry and subunit multiplicity, as well as on a set of heteromeric proteins. Crystal structures of likely monomers provide a control group. [Copyright &y& Elsevier]
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- 2005
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25. A Structure-based Anatomy of the E. coli Metabolome
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Nobeli, Irene, Ponstingl, Hannes, Krissinel, Eugene B., and Thornton, Janet M.
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ESCHERICHIA coli , *METABOLITES , *LIGANDS (Biochemistry) , *ANATOMY - Abstract
The Escherichia coli metabolome has been characterised using the two-dimensional structures of 745 metabolites, obtained from the EcoCyc and KEGG databases. Physicochemical properties of the metabolome have been calculated to provide an overview of this set of cognate ligands. A library of fragments commonly found among these molecules has been employed to reveal the main constituents of metabolites, and to assist a broad classification of the metabolome into biochemically relevant classes. Fragment-based fingerprints reveal the metabolome as a continuum in the two-dimensional structural space, where clusters of molecules sharing similar scaffolds can be identified, but are generally overlapping. Nucleotide, carbohydrate and amino acid-like molecules are the most prominent, but at high levels of similarity, a more detailed classification is possible. Classification schemes for the metabolome are a promising tool for understanding the chemical diversity of the metabolome. When used in conjunction with existing classifications of the proteome, they can help to elucidate the binding preferences and promiscuity of proteins and their cognate substrates. [Copyright &y& Elsevier]
- Published
- 2003
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26. Automatic inference of protein quaternary structure from crystals.
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Ponstingl, Hannes, kabir, Thomas, and Thornton, Janet M.
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CRYSTALLOGRAPHY , *CRYSTALS - Abstract
The arrangement of the subunits in an oligomeric protein often cannot be inferred without ambiguity from crystallographic studies. The annotation of the functional assembly of protein structures in the Protein Data Bank (PDB) is incomplete and frequently inconsistent. Instructions for the reconstruction, by symmetry, of the functional assembly from the deposited coordinates are often absent. An automatic procedure is proposed for the inference of assembly structures that are likely to be physiologically relevant. The method scores crystal contacts by their contact size and chemi8cal complementarity. The submit assembly is then inferred from these scored contacts by a clustering procedure involving a single adjustable parameter. When predicting the oligomeric state for a non-redundant set of 55 monomeric and 163 oligomeric protein from dimmers up to hexamers, a classification error rate of 16% was observed. [ABSTRACT FROM AUTHOR]
- Published
- 2003
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27. Discriminating between homodimeric and monomeric proteins in the crystalline state.
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Ponstingl, Hannes, Henrick, Kim, and Thornton, Janet M.
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- 2000
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28. NMR assignments, secondary structure and hydration of oxidized <em>Escherichia coli</em> flavodoxin.
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Ponstingl, Hannes and Otting, Gottfried
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HYDRATION , *ESCHERICHIA coli , *NUCLEAR magnetic resonance spectroscopy , *ISOTOPE separation , *OXIDIZING agents - Abstract
Recombinant flavodoxin from Escherichia coli was uniformly enriched with 15N and 13C isotopes and its oxidized form in aqueous solution investigated by three-dimensional NMR spectroscopy. Nearly complete 1H, 15N and 13C resonance assignments were obtained. The secondary structure was determined from chemical shift, NOE and 3JHNHα coupling constant data. Like homologous long-chain flavodoxins, E. coli flavodoxin contains a five-stranded parallel β-sheet and five helices. The β-strands were found to comprise the residues 3-8, 29-34, 48-56, 80-89, 114-116 and 141-145. The helices comprise residues 12- 25, 40-45, 62-73, 98--108 and 152-166. The FMN-binding site was determined by intermolecular NOEs and low-field shifted amide proton resonances induced by the phosphoester group of the cofactor. The data are in good agreement with a previously predicted model of E. coli flavodoxin [Havel, T. F. (1993) Mol. Sim. 10, 175-210]. The analysis of of water-flavodoxin NOEs revealed the presence of two, possibly three, buried hydration water molecules which are located at sites, where homologous flavodoxins from Anacystis nidulans and Anabena 7/20 contain conserved hydration water molecules. One of these water molecules mediates hydrogen bonds between the protein backbone and the ribityl chain of the FMN cofactor. [ABSTRACT FROM AUTHOR]
- Published
- 1997
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29. MYO5B mutations cause microvillus inclusion disease and disrupt epithelial cell polarity.
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Müller, Thomas, Hess, Michael W., Schiefermeier, Natalia, Pfaller, Kristian, Ebner, Hannes L., Heinz-Erian, Peter, Ponstingl, Hannes, Partsch, Joachim, Röllinghoff, Barbara, Köhler, Henrik, Berger, Thomas, Lenhartz, Henning, Schlenck, Barbara, Houwen, Roderick J., Taylor, Christopher J., Zoller, Heinz, Lechner, Silvia, Goulet, Olivier, Utermann, Gerd, and Ruemmele, Frank M.
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EPITHELIAL cells ,CELL polarity ,GENE mapping ,GENETIC mutation ,GENETIC code ,MYOSIN ,MUSCLE proteins - Abstract
Following homozygosity mapping in a single kindred, we identified nonsense and missense mutations in MYO5B, encoding type Vb myosin motor protein, in individuals with microvillus inclusion disease (MVID). MVID is characterized by lack of microvilli on the surface of enterocytes and occurrence of intracellular vacuolar structures containing microvilli. In addition, mislocalization of transferrin receptor in MVID enterocytes suggests that MYO5B deficiency causes defective trafficking of apical and basolateral proteins in MVID. [ABSTRACT FROM AUTHOR]
- Published
- 2008
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30. A CRISPR Dropout Screen Identifies Genetic Vulnerabilities and Therapeutic Targets in Acute Myeloid Leukemia
- Author
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Tzelepis, Konstantinos, Koike-Yusa, Hiroko, De Braekeleer, Etienne, Li, Yilong, Metzakopian, Emmanouil, Dovey, Oliver M., Mupo, Annalisa, Grinkevich, Vera, Li, Meng, Mazan, Milena, Gozdecka, Malgorzata, Ohnishi, Shuhei, Cooper, Jonathan, Patel, Miten, McKerrell, Thomas, Chen, Bin, Domingues, Ana Filipa, Gallipoli, Paolo, Teichmann, Sarah, Ponstingl, Hannes, McDermott, Ultan, Saez-Rodriguez, Julio, Huntly, Brian J P., Iorio, Francesco, Pina, Cristina, Vassiliou, George S., and Yusa, Kosuke
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3. Good health - Abstract
Cell reports 17(4), 1193-1205 (2016). doi:10.1016/j.celrep.2016.09.079, Published by Cell Press, Maryland Heights, MO
31. A Genetic Progression Model of BrafV600E-Induced Intestinal Tumorigenesis Reveals Targets for Therapeutic Intervention.
- Author
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Rad, Roland, Cadiñanos, Juan, Rad, Lena, Varela, Ignacio, Strong, Alexander, Kriegl, Lydia, Constantino-Casas, Fernando, Eser, Stefan, Hieber, Maren, Seidler, Barbara, Price, Stacey, Fraga, Mario?F., Calvanese, Vincenzo, Hoffman, Gary, Ponstingl, Hannes, Schneider, Günter, Yusa, Kosuke, Grove, Carolyn, Schmid, Roland?M., and Wang, Wei
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CANCER invasiveness , *GENETICS of colon cancer , *RAF genes , *HYPERPLASIA , *ADENOMA , *CARCINOMA , *TARGETED drug delivery , *CELLULAR signal transduction - Abstract
Summary: We show that BRAFV600E initiates an alternative pathway to colorectal cancer (CRC), which progresses through a hyperplasia/adenoma/carcinoma sequence. This pathway underlies significant subsets of CRCs with distinctive pathomorphologic/genetic/epidemiologic/clinical characteristics. Genetic and functional analyses in mice revealed a series of stage-specific molecular alterations driving different phases of tumor evolution and uncovered mechanisms underlying this stage specificity. We further demonstrate dose-dependent effects of oncogenic signaling, with physiologic BrafV600E expression being sufficient for hyperplasia induction, but later stage intensified Mapk-signaling driving both tumor progression and activation of intrinsic tumor suppression. Such phenomena explain, for example, the inability of p53 to restrain tumor initiation as well as its importance in invasiveness control, and the late stage specificity of its somatic mutation. Finally, systematic drug screening revealed sensitivity of this CRC subtype to targeted therapeutics, including Mek or combinatorial PI3K/Braf inhibition. [Copyright &y& Elsevier]
- Published
- 2013
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32. A single-copy Sleeping Beauty transposon mutagenesis screen identifies new PTEN-cooperating tumor suppressor genes
- Author
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Sandra Bernaldo de Quirós, Qi Liang, Hannes Ponstingl, María Soledad Fernández-García, Alexander Strong, George S. Vassiliou, Jorge de la Rosa, Julia Weber, Carlos López-Otín, Rocio Fuente, Emmanouil Metzakopian, Mathias J Friedrich, Juan Cadiñanos, Gary J. Hoffman, Lena Rad, María Teresa Fernández-García, Imran Noorani, Meng Amy Li, Aurora Astudillo, Yang Li, Roland Rad, Allan Bradley, Ponstingl, Hannes [0000-0001-7573-1703], Vassiliou, George S [0000-0003-4337-8022], Cadiñanos, Juan [0000-0001-7561-7759], and Apollo - University of Cambridge Repository
- Subjects
0301 basic medicine ,Cancer genome sequencing ,Male ,Candidate gene ,Gene Dosage ,Mice, Transgenic ,Kaplan-Meier Estimate ,Genome ,Article ,Cell Line ,Insertional mutagenesis ,03 medical and health sciences ,Cell Movement ,Genetics ,medicine ,PTEN ,Animals ,Humans ,Genes, Tumor Suppressor ,Genetic Predisposition to Disease ,Mice, Knockout ,biology ,PTEN Phosphohydrolase ,Prostate ,Cancer ,Prostatic Neoplasms ,Epithelial Cells ,Chromoplexy ,medicine.disease ,Sleeping Beauty transposon system ,3. Good health ,Mutagenesis, Insertional ,030104 developmental biology ,Mutation ,biology.protein ,DNA Transposable Elements ,RNA Interference ,Signal Transduction - Abstract
The overwhelming number of genetic alterations identified through cancer genome sequencing requires complementary approaches to interpret their significance and interactions. Here we developed a novel whole-body insertional mutagenesis screen in mice, which was designed for the discovery of Pten-cooperating tumor suppressors. Toward this aim, we coupled mobilization of a single-copy inactivating Sleeping Beauty transposon to Pten disruption within the same genome. The analysis of 278 transposition-induced prostate, breast and skin tumors detected tissue-specific and shared data sets of known and candidate genes involved in cancer. We validated ZBTB20, CELF2, PARD3, AKAP13 and WAC, which were identified by our screens in multiple cancer types, as new tumor suppressor genes in prostate cancer. We demonstrated their synergy with PTEN in preventing invasion in vitro and confirmed their clinical relevance. Further characterization of Wac in vivo showed obligate haploinsufficiency for this gene (which encodes an autophagy-regulating factor) in a Pten-deficient context. Our study identified complex PTEN-cooperating tumor suppressor networks in different cancer types, with potential clinical implications.
- Published
- 2017
33. SRPK1 maintains acute myeloid leukemia through effects on isoform usage of epigenetic regulators including BRD4
- Author
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Emmanouil Metzakopian, Demetrios Aspris, Paolo Gallipoli, Tony Kouzarides, Francisco Hermida-Prado, Rab K. Prinjha, Isaia Barbieri, Roya Babaei-Jadidi, Samuel Robson, Irmela Jeremias, John C. Marioni, Michael Seiler, João M. L. Dias, Cristina Pina, Kosuke Yusa, Malgorzata Gozdecka, Silvia Buonamici, Yu Hsuen Yang, Etienne De Braekeleer, Brian J. P. Huntly, Allan Bradley, Binje Vick, Andrew J. Bannister, Hannes Ponstingl, Monika Dudek, Hamish D. Toop, M. S. Vijayabaskar, David O. Bates, Wen-Hsin Liu, George S. Vassiliou, Jennifer Batson, Jonathan C. Morris, Dimitrios A. Garyfallos, Konstantinos Tzelepis, Roland Rad, Toop, Hamish D [0000-0003-4637-4764], Ponstingl, Hannes [0000-0001-7573-1703], Dias, Joao ML [0000-0002-8451-3537], Gallipoli, Paolo [0000-0001-7254-2253], Vick, Binje [0000-0003-1956-2778], Prinjha, Rab K [0000-0002-2666-3326], Marioni, John C [0000-0001-9092-0852], Huntly, Brian [0000-0003-0312-161X], Batson, Jennifer [0000-0001-7005-4336], Morris, Jonathan C [0000-0002-5109-9069], Jeremias, Irmela [0000-0003-1773-7677], Vassiliou, George S [0000-0003-4337-8022], and Apollo - University of Cambridge Repository
- Subjects
0301 basic medicine ,Gene isoform ,BRD4 ,Cellular differentiation ,Science ,RNA Splicing ,General Physics and Astronomy ,Cell Cycle Proteins ,HL-60 Cells ,Biology ,SRPK1 ,Protein Serine-Threonine Kinases ,General Biochemistry, Genetics and Molecular Biology ,Article ,Epigenesis, Genetic ,Paediatric cancer ,03 medical and health sciences ,Targeted therapies ,Cancer epigenetics ,hemic and lymphatic diseases ,Humans ,Protein Isoforms ,MYB ,Epigenetics ,lcsh:Science ,neoplasms ,Multidisciplinary ,Biomedical Sciences ,Myeloid leukemia ,Nuclear Proteins ,Cell Differentiation ,General Chemistry ,Cell Cycle Checkpoints ,Chromatin ,3. Good health ,Hematopoiesis ,Leukemia, Myeloid, Acute ,030104 developmental biology ,Cancer research ,lcsh:Q ,K562 Cells ,Transcription Factors - Abstract
We recently identified the splicing kinase gene SRPK1 as a genetic vulnerability of acute myeloid leukemia (AML). Here, we show that genetic or pharmacological inhibition of SRPK1 leads to cell cycle arrest, leukemic cell differentiation and prolonged survival of mice transplanted with MLL-rearranged AML. RNA-seq analysis demonstrates that SRPK1 inhibition leads to altered isoform levels of many genes including several with established roles in leukemogenesis such as MYB, BRD4 and MED24. We focus on BRD4 as its main isoforms have distinct molecular properties and find that SRPK1 inhibition produces a significant switch from the short to the long isoform at the mRNA and protein levels. This was associated with BRD4 eviction from genomic loci involved in leukemogenesis including BCL2 and MYC. We go on to show that this switch mediates at least part of the anti-leukemic effects of SRPK1 inhibition. Our findings reveal that SRPK1 represents a plausible new therapeutic target against AML., SRPK1, a kinase involved in splicing regulation, is a potential therapeutic target for AML patients. Here, the authors show that SRPK1 inhibition changes isoform levels of key epigenetic regulators, including BRD4, and it has anti-tumor effects specifically against MLL-rearranged AML cells.
- Published
- 2018
34. ATM orchestrates the DNA-damage response to counter toxic non-homologous end-joining at broken replication forks
- Author
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Stephen P. Jackson, Matylda Sczaniecka-Clift, Domenic Pilger, Mukerrem Demir, Michael Woods, Petra Beli, Josep V. Forment, Kosuke Yusa, Stephen T. Durant, Julia Coates, Hannes Ponstingl, Elisabeth Chen, Anna Barros, Matthias Ostermaier, Beiyuan Fu, Gabriel Balmus, Allan Bradley, Fengtang Yang, Francisco Munoz Martinez, Tatjana Stankovic, Mareike Herzog, Emmanouil Metzakopian, David J. Adams, Yaron Galanty, Balmus, Gabriel [0000-0003-2872-4468], Pilger, Domenic [0000-0001-7339-0685], Yang, Fengtang [0000-0002-3573-2354], Chen, Elisabeth [0000-0003-2129-7985], Ponstingl, Hannes [0000-0001-7573-1703], Durant, Stephen T [0000-0003-4209-7499], Galanty, Yaron [0000-0001-7167-9004], Beli, Petra [0000-0001-9507-9820], Forment, Josep V [0000-0002-7797-2583], Jackson, Stephen P [0000-0001-9317-7937], and Apollo - University of Cambridge Repository
- Subjects
DNA End-Joining Repair ,endocrine system diseases ,Ataxia Telangiectasia Mutated Proteins ,Piperazines ,law.invention ,chemistry.chemical_compound ,DNA Ligase ATP ,Mice ,0302 clinical medicine ,law ,Mice, Inbred NOD ,DNA Breaks, Double-Stranded ,lcsh:Science ,chemistry.chemical_classification ,Mice, Knockout ,0303 health sciences ,biology ,BRCA1 Protein ,Mouse Embryonic Stem Cells ,3. Good health ,Non-homologous end joining ,030220 oncology & carcinogenesis ,Female ,medicine.drug ,DNA Replication ,DNA damage ,Cell Survival ,Science ,Antineoplastic Agents ,Article ,Olaparib ,03 medical and health sciences ,Cell Line, Tumor ,medicine ,Animals ,Humans ,030304 developmental biology ,DNA ligase ,Topoisomerase ,Neoplasms, Experimental ,chemistry ,Drug Resistance, Neoplasm ,Mutation ,biology.protein ,Cancer research ,Suppressor ,Phthalazines ,lcsh:Q ,Topotecan ,CRISPR-Cas Systems ,Homologous recombination - Abstract
Mutations in the ATM tumor suppressor gene confer hypersensitivity to DNA-damaging chemotherapeutic agents. To explore genetic resistance mechanisms, we performed genome-wide CRISPR-Cas9 screens in cells treated with the DNA topoisomerase I inhibitor topotecan. Thus, we here establish that inactivating terminal components of the non-homologous end-joining (NHEJ) machinery or of the BRCA1-A complex specifically confer topotecan resistance to ATM-deficient cells. We show that hypersensitivity of ATM-mutant cells to topotecan or the poly-(ADP-ribose) polymerase (PARP) inhibitor olaparib reflects delayed engagement of homologous recombination at DNA-replication-fork associated single-ended double-strand breaks (DSBs), allowing some to be subject to toxic NHEJ. Preventing DSB ligation by NHEJ, or enhancing homologous recombination by BRCA1-A complex disruption, suppresses this toxicity, highlighting a crucial role for ATM in preventing toxic LIG4-mediated chromosome fusions. Notably, suppressor mutations in ATM-mutant backgrounds are different to those in BRCA1-mutant scenarios, suggesting new opportunities for patient stratification and additional therapeutic vulnerabilities for clinical exploitation., Mutations in the ATM tumor suppressor gene confer hypersensitivity to DNA-damaging chemotherapeutic agents. Here, the authors provide evidence that these hypersensitivities reflect a crucial role for ATM at damaged replication forks being to prevent toxic DNA end-joining leading to chromosome fusions and cell death.
- Published
- 2018
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35. Novel stem cell technologies are powerful tools to understand the impact of human factors on Plasmodium falciparum malaria.
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Pance A, Ng BL, Mwikali K, Koutsourakis M, Agu C, Rouhani FJ, Montandon R, Law F, Ponstingl H, and Rayner JC
- Subjects
- Humans, Plasmodium falciparum genetics, Erythrocytes parasitology, Stem Cells, Malaria, Falciparum parasitology, Malaria parasitology
- Abstract
Plasmodium falciparum parasites have a complex life cycle, but the most clinically relevant stage of the disease is the invasion of erythrocytes and the proliferation of the parasite in the blood. The influence of human genetic traits on malaria has been known for a long time, however understanding the role of the proteins involved is hampered by the anuclear nature of erythrocytes that makes them inaccessible to genetic tools. Here we overcome this limitation using stem cells to generate erythroid cells with an in-vitro differentiation protocol and assess parasite invasion with an adaptation of flow cytometry to detect parasite hemozoin. We combine this strategy with reprogramming of patient cells to Induced Pluripotent Stem Cells and genome editing to understand the role of key genes and human traits in malaria infection. We show that deletion of basigin ablates invasion while deletion of ATP2B4 has a minor effect and that erythroid cells from reprogrammed patient-derived HbBart α-thalassemia samples poorly support infection. The possibility to obtain patient-secific and genetically modifed erythoid cells offers an unparalleled opportunity to study the role of human genes and polymorphisms in malaria allowing preservation of the genomic background to demonstrate their function and understand their mechanisms., Competing Interests: The authors declare that the research was conducted in the absence of any commercial or financial relationships that could be construed as a potential conflict of interest. The author(s) declared that they were an editorial board member of Frontiers, at the time of submission. This had no impact on the peer review process and the final decision., (Copyright © 2023 Pance, Ng, Mwikali, Koutsourakis, Agu, Rouhani, Montandon, Law, Ponstingl and Rayner.)
- Published
- 2023
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36. The Polygenic and Monogenic Basis of Blood Traits and Diseases.
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Vuckovic D, Bao EL, Akbari P, Lareau CA, Mousas A, Jiang T, Chen MH, Raffield LM, Tardaguila M, Huffman JE, Ritchie SC, Megy K, Ponstingl H, Penkett CJ, Albers PK, Wigdor EM, Sakaue S, Moscati A, Manansala R, Lo KS, Qian H, Akiyama M, Bartz TM, Ben-Shlomo Y, Beswick A, Bork-Jensen J, Bottinger EP, Brody JA, van Rooij FJA, Chitrala KN, Wilson PWF, Choquet H, Danesh J, Di Angelantonio E, Dimou N, Ding J, Elliott P, Esko T, Evans MK, Felix SB, Floyd JS, Broer L, Grarup N, Guo MH, Guo Q, Greinacher A, Haessler J, Hansen T, Howson JMM, Huang W, Jorgenson E, Kacprowski T, Kähönen M, Kamatani Y, Kanai M, Karthikeyan S, Koskeridis F, Lange LA, Lehtimäki T, Linneberg A, Liu Y, Lyytikäinen LP, Manichaikul A, Matsuda K, Mohlke KL, Mononen N, Murakami Y, Nadkarni GN, Nikus K, Pankratz N, Pedersen O, Preuss M, Psaty BM, Raitakari OT, Rich SS, Rodriguez BAT, Rosen JD, Rotter JI, Schubert P, Spracklen CN, Surendran P, Tang H, Tardif JC, Ghanbari M, Völker U, Völzke H, Watkins NA, Weiss S, Cai N, Kundu K, Watt SB, Walter K, Zonderman AB, Cho K, Li Y, Loos RJF, Knight JC, Georges M, Stegle O, Evangelou E, Okada Y, Roberts DJ, Inouye M, Johnson AD, Auer PL, Astle WJ, Reiner AP, Butterworth AS, Ouwehand WH, Lettre G, Sankaran VG, and Soranzo N
- Subjects
- Female, Gene Regulatory Networks genetics, Genome-Wide Association Study methods, Hematopoiesis genetics, Humans, Male, Phenotype, Polymorphism, Single Nucleotide genetics, Genetic Predisposition to Disease genetics, Multifactorial Inheritance genetics
- Abstract
Blood cells play essential roles in human health, underpinning physiological processes such as immunity, oxygen transport, and clotting, which when perturbed cause a significant global health burden. Here we integrate data from UK Biobank and a large-scale international collaborative effort, including data for 563,085 European ancestry participants, and discover 5,106 new genetic variants independently associated with 29 blood cell phenotypes covering a range of variation impacting hematopoiesis. We holistically characterize the genetic architecture of hematopoiesis, assess the relevance of the omnigenic model to blood cell phenotypes, delineate relevant hematopoietic cell states influenced by regulatory genetic variants and gene networks, identify novel splice-altering variants mediating the associations, and assess the polygenic prediction potential for blood traits and clinical disorders at the interface of complex and Mendelian genetics. These results show the power of large-scale blood cell trait GWAS to interrogate clinically meaningful variants across a wide allelic spectrum of human variation., Competing Interests: Declaration of Interests Adam Butterworth has received grants (outside of this work) from AstraZeneca, Biogen, BioMarin, Bioverativ, Merck, Novartis, and Sanofi; James Floyd has consulted for Shionogi; Qi Guo is a full-time employee of BenevolentAI; Joanna Howson is a full-time employee of Novo Nordisk. Parsa Akbari is a full-time employee of Regeneron Pharmaceuticals., (Copyright © 2020 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2020
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37. A Genome-Wide Knockout Screen in Human Macrophages Identified Host Factors Modulating Salmonella Infection.
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Yeung ATY, Choi YH, Lee AHY, Hale C, Ponstingl H, Pickard D, Goulding D, Thomas M, Gill E, Kim JK, Bradley A, Hancock REW, and Dougan G
- Subjects
- Endocytosis, Host-Derived Cellular Factors genetics, Humans, Macrophages microbiology, Models, Theoretical, Salmonella growth & development, Salmonella Infections immunology, THP-1 Cells, Disease Resistance, Gene Knockout Techniques, Genetic Testing, Host-Derived Cellular Factors immunology, Macrophages immunology, Salmonella immunology
- Abstract
A genome-scale CRISPR knockout library screen of THP-1 human macrophages was performed to identify loss-of-function mutations conferring resistance to Salmonella uptake. The screen identified 183 candidate genes, from which 14 representative genes involved in actin dynamics ( ACTR3 , ARPC4 , CAPZB , TOR3A , CYFIP2 , CTTN , and NHLRC2 ), glycosaminoglycan metabolism ( B3GNT1 ), receptor signaling ( PDGFB and CD27 ), lipid raft formation ( CLTCL1 ), calcium transport ( ATP2A2 and ITPR3 ), and cholesterol metabolism ( HMGCR ) were analyzed further. For some of these pathways, known chemical inhibitors could replicate the Salmonella resistance phenotype, indicating their potential as targets for host-directed therapy. The screen indicated a role for the relatively uncharacterized gene NHLRC2 in both Salmonella invasion and macrophage differentiation. Upon differentiation, NHLRC2 mutant macrophages were hyperinflammatory and did not exhibit characteristics typical of macrophages, including atypical morphology and inability to interact and phagocytose bacteria/particles. Immunoprecipitation confirmed an interaction of NHLRC2 with FRYL, EIF2AK2, and KLHL13. IMPORTANCE Salmonella exploits macrophages to gain access to the lymphatic system and bloodstream to lead to local and potentially systemic infections. With an increasing number of antibiotic-resistant isolates identified in humans, Salmonella infections have become major threats to public health. Therefore, there is an urgent need to identify alternative approaches to anti-infective therapy, including host-directed therapies. In this study, we used a simple genome-wide screen to identify 183 candidate host factors in macrophages that can confer resistance to Salmonella infection. These factors may be potential therapeutic targets against Salmonella infections., (Copyright © 2019 Yeung et al.)
- Published
- 2019
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38. Genome-Scale CRISPRa Screen Identifies Novel Factors for Cellular Reprogramming.
- Author
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Yang J, Rajan SS, Friedrich MJ, Lan G, Zou X, Ponstingl H, Garyfallos DA, Liu P, Bradley A, and Metzakopian E
- Subjects
- Animals, Biomarkers, CRISPR-Cas Systems, Cell Line, Gene Dosage, Germ Layers cytology, Germ Layers metabolism, Humans, Induced Pluripotent Stem Cells, Mice, Octamer Transcription Factor-3 genetics, Octamer Transcription Factor-3 metabolism, Pluripotent Stem Cells cytology, Pluripotent Stem Cells metabolism, Transcription Factors genetics, Transcription Factors metabolism, Cellular Reprogramming genetics, Clustered Regularly Interspaced Short Palindromic Repeats, Genome-Wide Association Study
- Abstract
Primed epiblast stem cells (EpiSCs) can be reverted to a pluripotent embryonic stem cell (ESC)-like state by expression of single reprogramming factor. We used CRISPR activation to perform a genome-scale, reprogramming screen in EpiSCs and identified 142 candidate genes. Our screen validated a total of 50 genes, previously not known to contribute to reprogramming, of which we chose Sall1 for further investigation. We show that Sall1 augments reprogramming of mouse EpiSCs and embryonic fibroblasts and that these induced pluripotent stem cells are indeed fully pluripotent including formation of chimeric mice. We also demonstrate that Sall1 synergizes with Nanog in reprogramming and that overexpression in ESCs delays their conversion back to EpiSCs. Lastly, using RNA sequencing, we identify and validate Klf5 and Fam189a2 as new downstream targets of Sall1 and Nanog. In summary, our work demonstrates the power of using CRISPR technology in understanding molecular mechanisms that mediate complex cellular processes such as reprogramming., (Copyright © 2019 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2019
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39. Promoter-bound METTL3 maintains myeloid leukaemia by m 6 A-dependent translation control.
- Author
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Barbieri I, Tzelepis K, Pandolfini L, Shi J, Millán-Zambrano G, Robson SC, Aspris D, Migliori V, Bannister AJ, Han N, De Braekeleer E, Ponstingl H, Hendrick A, Vakoc CR, Vassiliou GS, and Kouzarides T
- Subjects
- Adenosine genetics, Adenosine metabolism, Animals, CRISPR-Cas Systems, Cell Line, Tumor, Cell Proliferation genetics, Chromatin genetics, Chromatin metabolism, Female, Genes, Neoplasm genetics, Humans, Leukemia, Myeloid, Acute pathology, Methyltransferases chemistry, Methyltransferases deficiency, Methyltransferases genetics, Mice, RNA, Messenger biosynthesis, RNA, Messenger genetics, RNA, Messenger metabolism, Ribosomes metabolism, Transcription Initiation Site, Adenosine analogs & derivatives, Gene Expression Regulation, Neoplastic genetics, Leukemia, Myeloid, Acute enzymology, Leukemia, Myeloid, Acute genetics, Methyltransferases metabolism, Promoter Regions, Genetic genetics, Protein Biosynthesis genetics
- Abstract
N
6 -methyladenosine (m6 A) is an abundant internal RNA modification in both coding and non-coding RNAs that is catalysed by the METTL3-METTL14 methyltransferase complex. However, the specific role of these enzymes in cancer is still largely unknown. Here we define a pathway that is specific for METTL3 and is implicated in the maintenance of a leukaemic state. We identify METTL3 as an essential gene for growth of acute myeloid leukaemia cells in two distinct genetic screens. Downregulation of METTL3 results in cell cycle arrest, differentiation of leukaemic cells and failure to establish leukaemia in immunodeficient mice. We show that METTL3, independently of METTL14, associates with chromatin and localizes to the transcriptional start sites of active genes. The vast majority of these genes have the CAATT-box binding protein CEBPZ present at the transcriptional start site, and this is required for recruitment of METTL3 to chromatin. Promoter-bound METTL3 induces m6 A modification within the coding region of the associated mRNA transcript, and enhances its translation by relieving ribosome stalling. We show that genes regulated by METTL3 in this way are necessary for acute myeloid leukaemia. Together, these data define METTL3 as a regulator of a chromatin-based pathway that is necessary for maintenance of the leukaemic state and identify this enzyme as a potential therapeutic target for acute myeloid leukaemia.- Published
- 2017
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40. JAK2 V617F hematopoietic clones are present several years prior to MPN diagnosis and follow different expansion kinetics.
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McKerrell T, Park N, Chi J, Collord G, Moreno T, Ponstingl H, Dias J, Gerasimou P, Melanthiou K, Prokopiou C, Antoniades M, Varela I, Costeas PA, and Vassiliou GS
- Abstract
Competing Interests: Conflict-of-interest disclosure: G.S.V. is a consultant for KYMAB and receives an educational grant from Celgene. The remaining authors declare no competing financial interests.
- Published
- 2017
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41. A CRISPR Dropout Screen Identifies Genetic Vulnerabilities and Therapeutic Targets in Acute Myeloid Leukemia.
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Tzelepis K, Koike-Yusa H, De Braekeleer E, Li Y, Metzakopian E, Dovey OM, Mupo A, Grinkevich V, Li M, Mazan M, Gozdecka M, Ohnishi S, Cooper J, Patel M, McKerrell T, Chen B, Domingues AF, Gallipoli P, Teichmann S, Ponstingl H, McDermott U, Saez-Rodriguez J, Huntly BJP, Iorio F, Pina C, Vassiliou GS, and Yusa K
- Subjects
- Adult, Apoptosis, Cell Differentiation, Cell Line, Tumor, Cell Proliferation, Histone Acetyltransferases antagonists & inhibitors, Histone Acetyltransferases metabolism, Humans, Reproducibility of Results, Clustered Regularly Interspaced Short Palindromic Repeats genetics, Genetic Testing, Leukemia, Myeloid, Acute genetics, Leukemia, Myeloid, Acute therapy, Molecular Targeted Therapy
- Abstract
Acute myeloid leukemia (AML) is an aggressive cancer with a poor prognosis, for which mainstream treatments have not changed for decades. To identify additional therapeutic targets in AML, we optimize a genome-wide clustered regularly interspaced short palindromic repeats (CRISPR) screening platform and use it to identify genetic vulnerabilities in AML cells. We identify 492 AML-specific cell-essential genes, including several established therapeutic targets such as DOT1L, BCL2, and MEN1, and many other genes including clinically actionable candidates. We validate selected genes using genetic and pharmacological inhibition, and chose KAT2A as a candidate for downstream study. KAT2A inhibition demonstrated anti-AML activity by inducing myeloid differentiation and apoptosis, and suppressed the growth of primary human AMLs of diverse genotypes while sparing normal hemopoietic stem-progenitor cells. Our results propose that KAT2A inhibition should be investigated as a therapeutic strategy in AML and provide a large number of genetic vulnerabilities of this leukemia that can be pursued in downstream studies., (Copyright © 2016 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2016
- Full Text
- View/download PDF
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