60 results on '"Schramek, D."'
Search Results
2. RANK und RANKL - Vom Knochen zum Mammakarzinom
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Sigl V, Schramek D, and Penninger JM
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Stammzellen ,Progesteron ,Knochen ,lcsh:R ,lcsh:Medicine ,Onkologie - Abstract
RANK (Receptor Activator of NF-κB) und sein Ligand RANKL sind Schlüsselmoleküle im Knochenmetabolismus und spielen eine essenzielle Rolle in der Entstehung von pathologischen Knochenveränderungen. Die Deregulation des RANK/RANKL-Systems ist zum Beispiel ein Hauptgrund für das Auftreten von postmenopausaler Osteoporose bei Frauen. Eine weitere wesentliche Funktion von RANK und RANKL liegt in der Entwicklung von milchsekretierenden Drüsen während der Schwangerschaft. Dabei regulieren Sexualhormone, wie zum Beispiel Progesteron, die Expression von RANKL und induzieren dadurch die Proliferation von epithelialen Zellen der Brust. Seit Längerem war schon bekannt, dass RANK und RANKL in der Metastasenbildung von Brustkrebszellen im Knochengewebe beteiligt sind. Wir konnten nun das RANK/RANKLSystem auch als essenziellen Mechanismus in der Entstehung von hormonellem Brustkrebs identifizieren. In diesem Beitrag werden wir daher den neuesten Erkenntnissen besondere Aufmerksamkeit schenken und diese kritisch in Bezug auf Brustkrebsentwicklung betrachten.
- Published
- 2012
3. MKK7 and ARF: New players in the DNA damage response scenery
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Kotsinas, A. Papanagnou, P. Galanos, P. Schramek, D. Townsend, P. Penninger, J.M. Bartek, J. Gorgoulis, V.G.
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body regions - Abstract
Sensing, integrating, and processing of stressogenic signals must be followed by accurate differential response(s) for a cell to survive and avoid malignant transformation. The DNA damage response (DDR) pathway is vital in this process, as it deals with genotoxic/oncogenic insults, having p53 as a nodal effector that performs most of the above tasks. Accumulating data reveal that other pathways are also involved in the same or similar processes, conveying also to p53. Emerging questions are if, how, and when these additional pathways communicate with the DDR axis. Two such stress response pathways, involving the MKK7 stress-activated protein kinase (SAPK) and ARF, have been shown to be interlocked with the ATM/ATR-regulated DDR axis in a highly ordered manner. This creates a new landscape in the DDR orchestrated response to genotoxic/oncogenic insults that is currently discussed. © 2014 Landes Bioscience.
- Published
- 2014
4. A Multi-Hit mouse model to identify cooperating Ras effector pathways in lung cancer
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Musteanu M, Blaas L, Zenz R, Svinka J, Hoffmann T, Grabner B, Schramek D, Kantner HP, Mxfcller M, Kolbe T, Rxfclicke T, Moriggl R, Kenner L, Stoiber D, Penninger JM, Popper H, Casanova E, and Eferl R
- Published
- 2012
5. The stress kinase MKK7 couples oncogenic stress to p53 stability and tumor suppression
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Schramek, D. Kotsinas, A. Meixner, A. Wada, T. Elling, U. Pospisilik, J.A. Neely, G.G. Zwick, R.-H. Sigl, V. Forni, G. Serrano, M. Gorgoulis, V.G. Penninger, J.M.
- Abstract
Most preneoplastic lesions are quiescent and do not progress to form overt tumors. It has been proposed that oncogenic stress activates the DNA damage response and the key tumor suppressor p53, which prohibits tumor growth. However, the molecular pathways by which cells sense a premalignant state in vivo are largely unknown. Here we report that tissue-specific inactivation of the stress signaling kinase MKK7 in KRas G12D-driven lung carcinomas and NeuT-driven mammary tumors markedly accelerates tumor onset and reduces overall survival. Mechanistically, MKK7 acts through the kinases JNK1 and JNK2, and this signaling pathway directly couples oncogenic and genotoxic stress to the stability of p53, which is required for cell cycle arrest and suppression of epithelial cancers. These results show that MKK7 functions as a major tumor suppressor in lung and mammary cancer in mouse and identify MKK7 as a vital molecular sensor to set a cellular anti-cancer barrier. © 2011 Nature America, Inc. All rights reserved.
- Published
- 2011
6. Genome-Wide RNAi Screen Identifies Genes Involved in Intestinal Pathogenic Bacterial Infection
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Cronin SJ, Nehme NT, Limmer S, Liegeois S, Pospisilik JA, Schramek D, Leibbrandt A, Simoes Rde M, Gruber S, Puc U, Ebersberger I, Zoranovic T, Neely GG, von Haeseler A, Ferrandon D, and Penninger JM
- Published
- 2009
7. Gab2 signaling in chronic myeloid leukemia cells confers resistance to multiple Bcr-Abl inhibitors
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Wöhrle, F U, primary, Halbach, S, additional, Aumann, K, additional, Schwemmers, S, additional, Braun, S, additional, Auberger, P, additional, Schramek, D, additional, Penninger, J M, additional, Laßmann, S, additional, Werner, M, additional, Waller, C F, additional, Pahl, H L, additional, Zeiser, R, additional, Daly, R J, additional, and Brummer, T, additional
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- 2012
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8. The docking protein and proto-oncogene product Gab2 is regulated via a novel negative feedback mechanism mediated by 14-3-3 binding
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Brummer, T, primary, Larance, M, additional, Abreu, MT Herrera, additional, Lyons, RJ, additional, Timpson, P, additional, Emmerich, CH, additional, Fleuren, EDG, additional, Lehrbach, GM, additional, Schramek, D, additional, Guilhaus, M, additional, James, DE, additional, and Daly, RJ, additional
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- 2009
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9. Metabolic Regulation of the Epigenome Drives Lethal Infantile Ependymoma
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Samuel Weiss, Leo J.Y. Kim, Xiaochong Wu, Randy Van Ommeren, Yanqing Jiang, Kaitlin Kharas, Evgeny Kanshin, Moloud Ahmadi, Alberto Delaidelli, Geneviève Deblois, David Przelicki, Stephane Angers, Hiromichi Suzuki, Sameer Agnihotri, Bradly G. Wouters, Graham MacLeod, Ricky Tsai, Pasqualino De Antonellis, Michelle Ly, Stacey L. Krumholtz, Paul Guilhamon, James Loukides, Ravi N. Vellanki, Alex Rasnitsyn, Hamza Farooq, Daniel Schramek, Nada Jabado, María Sánchez-Osuna, Laura K. Donovan, Vijay Ramaswamy, Ibrahim El-Hamamy, Joonas Haapasalo, Jeremy N. Rich, Michael D. Taylor, Benjamin A. Garcia, Mike Tyers, Kyle Juraschka, Winnie Ong, Olivier Saulnier, Panagiotis Prinos, John J.Y. Lee, Borja L. Holgado, Olga Sirbu, Craig Daniels, Cheryl H. Arrowsmith, Cory Richman, Poul H. Sorensen, Kulandaimanuvel Antony Michealraj, Sheila K. Singh, Andrea Bajic, Polina Balin, Stephen C. Mack, Betty Luu, Fiona J. Coutinho, Dilakshan Srikanthan, Florence M.G. Cavalli, Sachin Kumar, Evan Y. Wang, Mathieu Lupien, Peter B. Dirks, Maria C. Vladoiu, Lincoln Stein, Livia Garzia, Ahmad Malik, John Wojcik, Avesta Rastan, Michealraj, K. A., Kumar, S. A., Kim, L. J. Y., Cavalli, F. M. G., Przelicki, D., Wojcik, J. B., Delaidelli, A., Bajic, A., Saulnier, O., Macleod, G., Vellanki, R. N., Vladoiu, M. C., Guilhamon, P., Ong, W., Lee, J. J. Y., Jiang, Y., Holgado, B. L., Rasnitsyn, A., Malik, A. A., Tsai, R., Richman, C. M., Juraschka, K., Haapasalo, J., Wang, E. Y., De Antonellis, P., Suzuki, H., Farooq, H., Balin, P., Kharas, K., Van Ommeren, R., Sirbu, O., Rastan, A., Krumholtz, S. L., Ly, M., Ahmadi, M., Deblois, G., Srikanthan, D., Luu, B., Loukides, J., Wu, X., Garzia, L., Ramaswamy, V., Kanshin, E., Sanchez-Osuna, M., El-Hamamy, I., Coutinho, F. J., Prinos, P., Singh, S., Donovan, L. K., Daniels, C., Schramek, D., Tyers, M., Weiss, S., Stein, L. D., Lupien, M., Wouters, B. G., Garcia, B. A., Arrowsmith, C. H., Sorensen, P. H., Angers, S., Jabado, N., Dirks, P. B., Mack, S. C., Agnihotri, S., Rich, J. N., and Taylor, M. D.
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Epigenomics ,Ependymoma ,Male ,ependymoma ,Epigenomic ,Somatic cell ,cancer metabolism ,Infratentorial Neoplasms ,Biology ,General Biochemistry, Genetics and Molecular Biology ,Cell Line ,Histones ,Brain Neoplasm ,03 medical and health sciences ,Epigenome ,0302 clinical medicine ,Histone demethylation ,Histone methylation ,medicine ,Animals ,Humans ,Epigenetics ,030304 developmental biology ,hindbrain development ,Cell Proliferation ,Infratentorial Neoplasm ,0303 health sciences ,Brain Neoplasms ,Animal ,Lysine ,Infant ,DNA Methylation ,medicine.disease ,microenvironment ,Mice, Inbred C57BL ,Histone ,Acetylation ,paediatric cancer ,Mutation ,biology.protein ,Cancer research ,030217 neurology & neurosurgery ,epigenetic ,Human - Abstract
Posterior fossa A (PFA) ependymomas are lethal malignancies of the hindbrain in infants and toddlers. Lacking highly recurrent somatic mutations, PFA ependymomas are proposed to be epigenetically driven tumors for which model systems are lacking. Here we demonstrate that PFA ependymomas are maintained under hypoxia, associated with restricted availability of specific metabolites to diminish histone methylation, and increase histone demethylation and acetylation at histone 3 lysine 27 (H3K27). PFA ependymomas initiate from a cell lineage in the first trimester of human development that resides in restricted oxygen. Unlike other ependymomas, transient exposure of PFA cells to ambient oxygen induces irreversible cellular toxicity. PFA tumors exhibit a low basal level of H3K27me3, and, paradoxically, inhibition of H3K27 methylation specifically disrupts PFA tumor growth. Targeting metabolism and/or the epigenome presents a unique opportunity for rational therapy for infants with PFA ependymoma. Hypoxia reprograms the cellular metabolome and epigenome to promote growth of the most lethal ependymomas.
- Published
- 2020
10. In vivo CRISPR screens reveal SCAF1 and USP15 as drivers of pancreatic cancer.
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Martinez S, Wu S, Geuenich M, Malik A, Weber R, Woo T, Zhang A, Jang GH, Dervovic D, Al-Zahrani KN, Tsai R, Fodil N, Gros P, Gallinger S, Neely GG, Notta F, Sendoel A, Campbell K, Elling U, and Schramek D
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- Animals, Humans, Mice, Cell Line, Tumor, Deoxycytidine analogs & derivatives, Deoxycytidine pharmacology, Deoxycytidine therapeutic use, Gemcitabine, Gene Expression Regulation, Neoplastic, Mutation, Ubiquitin-Specific Proteases genetics, Ubiquitin-Specific Proteases metabolism, CRISPR-Cas Systems, Pancreatic Neoplasms genetics, Pancreatic Neoplasms pathology
- Abstract
Functionally characterizing the genetic alterations that drive pancreatic cancer is a prerequisite for precision medicine. Here, we perform somatic CRISPR/Cas9 mutagenesis screens to assess the transforming potential of 125 recurrently mutated pancreatic cancer genes, which revealed USP15 and SCAF1 as pancreatic tumor suppressors. Mechanistically, we find that USP15 functions in a haploinsufficient manner and that loss of USP15 or SCAF1 leads to reduced inflammatory TNFα, TGF-β and IL6 responses and increased sensitivity to PARP inhibition and Gemcitabine. Furthermore, we find that loss of SCAF1 leads to the formation of a truncated, inactive USP15 isoform at the expense of full-length USP15, functionally coupling SCAF1 and USP15. Notably, USP15 and SCAF1 alterations are observed in 31% of pancreatic cancer patients. Our results highlight the utility of in vivo CRISPR screens to integrate human cancer genomics and mouse modeling for the discovery of cancer driver genes with potential prognostic and therapeutic implications., (© 2024. The Author(s).)
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- 2024
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11. Genome-wide CRISPR screens identify novel regulators of wild-type and mutant p53 stability.
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Lü Y, Cho T, Mukherjee S, Suarez CF, Gonzalez-Foutel NS, Malik A, Martinez S, Dervovic D, Oh RH, Langille E, Al-Zahrani KN, Hoeg L, Lin ZY, Tsai R, Mbamalu G, Rotter V, Ashton-Prolla P, Moffat J, Chemes LB, Gingras AC, Oren M, Durocher D, and Schramek D
- Subjects
- Animals, Humans, Mice, Female, CRISPR-Cas Systems, Breast Neoplasms genetics, Breast Neoplasms metabolism, Cell Line, Tumor, Ubiquitin Thiolesterase genetics, Ubiquitin Thiolesterase metabolism, Gene Expression Regulation, Neoplastic, Clustered Regularly Interspaced Short Palindromic Repeats, Tumor Suppressor Protein p53 genetics, Tumor Suppressor Protein p53 metabolism, Protein Stability, Mutation
- Abstract
Tumor suppressor p53 (TP53) is frequently mutated in cancer, often resulting not only in loss of its tumor-suppressive function but also acquisition of dominant-negative and even oncogenic gain-of-function traits. While wild-type p53 levels are tightly regulated, mutants are typically stabilized in tumors, which is crucial for their oncogenic properties. Here, we systematically profiled the factors that regulate protein stability of wild-type and mutant p53 using marker-based genome-wide CRISPR screens. Most regulators of wild-type p53 also regulate p53 mutants, except for p53 R337H regulators, which are largely private to this mutant. Mechanistically, FBXO42 emerged as a positive regulator for a subset of p53 mutants, working with CCDC6 to control USP28-mediated mutant p53 stabilization. Additionally, C16orf72/HAPSTR1 negatively regulates both wild-type p53 and all tested mutants. C16orf72/HAPSTR1 is commonly amplified in breast cancer, and its overexpression reduces p53 levels in mouse mammary epithelium leading to accelerated breast cancer. This study offers a network perspective on p53 stability regulation, potentially guiding strategies to reinforce wild-type p53 or target mutant p53 in cancer., (© 2024. The Author(s).)
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- 2024
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12. Proteome-scale discovery of protein degradation and stabilization effectors.
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Poirson J, Cho H, Dhillon A, Haider S, Imrit AZ, Lam MHY, Alerasool N, Lacoste J, Mizan L, Wong C, Gingras AC, Schramek D, and Taipale M
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- Humans, Substrate Specificity, Proteolysis Targeting Chimera metabolism, Von Hippel-Lindau Tumor Suppressor Protein metabolism, Deubiquitinating Enzymes analysis, Deubiquitinating Enzymes metabolism, Protein Stability, Proteolysis, Proteome metabolism, Proteomics, Ubiquitin-Protein Ligases analysis, Ubiquitin-Protein Ligases metabolism
- Abstract
Targeted protein degradation and stabilization are promising therapeutic modalities because of their potency, versatility and their potential to expand the druggable target space
1,2 . However, only a few of the hundreds of E3 ligases and deubiquitinases in the human proteome have been harnessed for this purpose, which substantially limits the potential of the approach. Moreover, there may be other protein classes that could be exploited for protein stabilization or degradation3-5 , but there are currently no methods that can identify such effector proteins in a scalable and unbiased manner. Here we established a synthetic proteome-scale platform to functionally identify human proteins that can promote the degradation or stabilization of a target protein in a proximity-dependent manner. Our results reveal that the human proteome contains a large cache of effectors of protein stability. The approach further enabled us to comprehensively compare the activities of human E3 ligases and deubiquitinases, identify and characterize non-canonical protein degraders and stabilizers and establish that effectors have vastly different activities against diverse targets. Notably, the top degraders were more potent against multiple therapeutically relevant targets than the currently used E3 ligases cereblon and VHL. Our study provides a functional catalogue of stability effectors for targeted protein degradation and stabilization and highlights the potential of induced proximity screens for the discovery of new proximity-dependent protein modulators., (© 2024. The Author(s), under exclusive licence to Springer Nature Limited.)- Published
- 2024
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13. Differential DNA damage repair and PARP inhibitor vulnerability of the mammary epithelial lineages.
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Kim H, Aliar K, Tharmapalan P, McCloskey CW, Kuttanamkuzhi A, Grünwald BT, Palomero L, Mahendralingam MJ, Waas M, Mer AS, Elliott MJ, Zhang B, Al-Zahrani KN, Langille ER, Parsons M, Narala S, Hofer S, Waterhouse PD, Hakem R, Haibe-Kains B, Kislinger T, Schramek D, Cescon DW, Pujana MA, Berman HK, and Khokha R
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- 2023
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14. Mutational processes of tobacco smoking and APOBEC activity generate protein-truncating mutations in cancer genomes.
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Adler N, Bahcheli AT, Cheng KCL, Al-Zahrani KN, Slobodyanyuk M, Pellegrina D, Schramek D, and Reimand J
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- Humans, Mutation, Cytidine Deaminase genetics, APOBEC Deaminases genetics, APOBEC Deaminases metabolism, Tobacco Smoking, Neoplasms genetics, Neoplasms pathology
- Abstract
Mutational signatures represent a genomic footprint of endogenous and exogenous mutational processes through tumor evolution. However, their functional impact on the proteome remains incompletely understood. We analyzed the protein-coding impact of single-base substitution (SBS) signatures in 12,341 cancer genomes from 18 cancer types. Stop-gain mutations (SGMs) (i.e., nonsense mutations) were strongly enriched in SBS signatures of tobacco smoking, APOBEC cytidine deaminases, and reactive oxygen species. These mutational processes alter specific trinucleotide contexts and thereby substitute serines and glutamic acids with stop codons. SGMs frequently affect cancer hallmark pathways and tumor suppressors such as TP53 , FAT1 , and APC . Tobacco-driven SGMs in lung cancer correlate with smoking history and highlight a preventable determinant of these harmful mutations. APOBEC-driven SGMs are enriched in YTCA motifs and associate with APOBEC3A expression. Our study exposes SGM expansion as a genetic mechanism by which endogenous and carcinogenic mutational processes directly contribute to protein loss of function, oncogenesis, and tumor heterogeneity.
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- 2023
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15. Author Correction: In vivo CRISPR screens reveal Serpinb9 and Adam2 as regulators of immune therapy response in lung cancer.
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Dervovic D, Malik AA, Chen ELY, Narimatsu M, Adler N, Afiuni-Zadeh S, Krenbek D, Martinez S, Tsai R, Boucher J, Berman JM, Teng K, Ayyaz A, Lü Y, Mbamalu G, Loganathan SK, Lee J, Zhang L, Guidos C, Wrana J, Valipour A, Roux PP, Reimand J, Jackson HW, and Schramek D
- Published
- 2023
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16. In vivo CRISPR screens reveal Serpinb9 and Adam2 as regulators of immune therapy response in lung cancer.
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Dervovic D, Malik AA, Chen ELY, Narimatsu M, Adler N, Afiuni-Zadeh S, Krenbek D, Martinez S, Tsai R, Boucher J, Berman JM, Teng K, Ayyaz A, Lü Y, Mbamalu G, Loganathan SK, Lee J, Zhang L, Guidos C, Wrana J, Valipour A, Roux PP, Reimand J, Jackson HW, and Schramek D
- Subjects
- Animals, Humans, Male, Mice, Antigens, Neoplasm, Immunotherapy, Membrane Proteins genetics, T-Lymphocytes, Cytotoxic, Tumor Microenvironment, Antineoplastic Agents, Fertilins genetics, Lung Neoplasms genetics, Lung Neoplasms therapy, Serpins genetics
- Abstract
How the genetic landscape governs a tumor's response to immunotherapy remains poorly understood. To assess the immune-modulatory capabilities of 573 genes associated with altered cytotoxicity in human cancers, here we perform CRISPR/Cas9 screens directly in mouse lung cancer models. We recover the known immune evasion factors Stat1 and Serpinb9 and identify the cancer testis antigen Adam2 as an immune modulator, whose expression is induced by Kras
G12D and further elevated by immunotherapy. Using loss- and gain-of-function experiments, we show that ADAM2 functions as an oncogene by restraining interferon and TNF cytokine signaling causing reduced presentation of tumor-associated antigens. ADAM2 also restricts expression of the immune checkpoint inhibitors PDL1, LAG3, TIGIT and TIM3 in the tumor microenvironment, which might explain why ex vivo expanded and adoptively transferred cytotoxic T-cells show enhanced cytotoxic efficacy in ADAM2 overexpressing tumors. Together, direct in vivo CRISPR/Cas9 screens can uncover genetic alterations that control responses to immunotherapies., (© 2023. The Author(s).)- Published
- 2023
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17. In Vivo Screening Unveils Pervasive RNA-Binding Protein Dependencies in Leukemic Stem Cells and Identifies ELAVL1 as a Therapeutic Target.
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Vujovic A, de Rooij L, Chahi AK, Chen HT, Yee BA, Loganathan SK, Liu L, Chan DCH, Tajik A, Tsao E, Moreira S, Joshi P, Xu J, Wong N, Balde Z, Jahangiri S, Zandi S, Aigner S, Dick JE, Minden MD, Schramek D, Yeo GW, and Hope KJ
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- Humans, Cell Differentiation, Hematopoietic Stem Cells metabolism, RNA-Binding Proteins genetics, RNA-Binding Proteins metabolism, RNA-Binding Proteins therapeutic use, Mitochondrial Precursor Protein Import Complex Proteins, ELAV-Like Protein 1 genetics, ELAV-Like Protein 1 metabolism, Leukemia, Myeloid, Acute genetics, Leukemia, Myeloid, Acute drug therapy
- Abstract
Acute myeloid leukemia (AML) is fueled by leukemic stem cells (LSC) whose determinants are challenging to discern from hematopoietic stem cells (HSC) or uncover by approaches focused on general cell properties. We have identified a set of RNA-binding proteins (RBP) selectively enriched in human AML LSCs. Using an in vivo two-step CRISPR-Cas9 screen to assay stem cell functionality, we found 32 RBPs essential for LSCs in MLL-AF9;NrasG12D AML. Loss-of-function approaches targeting key hit RBP ELAVL1 compromised LSC-driven in vivo leukemic reconstitution, and selectively depleted primitive malignant versus healthy cells. Integrative multiomics revealed differentiation, splicing, and mitochondrial metabolism as key features defining the leukemic ELAVL1-mRNA interactome with mitochondrial import protein, TOMM34, being a direct ELAVL1-stabilized target whose repression impairs AML propagation. Altogether, using a stem cell-adapted in vivo CRISPR screen, this work demonstrates pervasive reliance on RBPs as regulators of LSCs and highlights their potential as therapeutic targets in AML., Significance: LSC-targeted therapies remain a significant unmet need in AML. We developed a stem-cell-adapted in vivo CRISPR screen to identify key LSC drivers. We uncover widespread RNA-binding protein dependencies in LSCs, including ELAVL1, which we identify as a novel therapeutic vulnerability through its regulation of mitochondrial metabolism. This article is highlighted in the In This Issue feature, p. 171., (©2023 The Authors; Published by the American Association for Cancer Research.)
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- 2023
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18. The NOTCH-RIPK4-IRF6-ELOVL4 Axis Suppresses Squamous Cell Carcinoma.
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Yan Y, Gauthier MA, Malik A, Fotiadou I, Ostrovski M, Dervovic D, Ghadban L, Tsai R, Gish G, Loganathan SK, and Schramek D
- Abstract
Receptor-interacting serine/threonine protein kinase 4 (RIPK4) and its kinase substrate the transcription factor interferon regulatory factor 6 (IRF6) play critical roles in the development and maintenance of the epidermis. In addition, ourselves and others have previously shown that RIPK4 is a NOTCH target gene that suppresses the development of cutaneous and head and neck squamous cell carcinomas (HNSCCs). In this study, we used autochthonous mouse models, where the expression of Pik3ca
H1047R oncogene predisposes the skin and oral cavity to tumor development, and show that not only loss of Ripk4 , but also loss of its kinase substrate Irf6 , triggers rapid SCC development. In vivo rescue experiments using Ripk4 or a kinase-dead Ripk4 mutant showed that the tumor suppressive function of Ripk4 is dependent on its kinase activity. To elucidate critical mediators of this tumor suppressive pathway, we performed transcriptional profiling of Ripk4 -deficient epidermal cells followed by multiplexed in vivo CRISPR screening to identify genes with tumor suppressive capabilities. We show that Elovl4 is a critical Notch-Ripk4-Irf6 downstream target gene, and that Elovl4 loss itself triggers SCC development. Importantly, overexpression of Elovl4 suppressed tumor growth of Ripk4 -deficient keratinocytes. Altogether, our work identifies a potent Notch1-Ripk4-Irf6-Elovl4 tumor suppressor axis.- Published
- 2023
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19. A CRISPR Path to Finding Vulnerabilities and Solving Drug Resistance: Targeting the Diverse Cancer Landscape and Its Ecosystem.
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McLean B, Istadi A, Clack T, Vankan M, Schramek D, Neely GG, and Pajic M
- Abstract
Cancer is the second leading cause of death globally, with therapeutic resistance being a major cause of treatment failure in the clinic. The dynamic signaling that occurs between tumor cells and the diverse cells of the surrounding tumor microenvironment actively promotes disease progression and therapeutic resistance. Improving the understanding of how tumors evolve following therapy and the molecular mechanisms underpinning de novo or acquired resistance is thus critical for the identification of new targets and for the subsequent development of more effective combination regimens. Simultaneously targeting multiple hallmark capabilities of cancer to circumvent adaptive or evasive resistance may lead to significantly improved treatment response in the clinic. Here, the latest applications of functional genomics tools, such as clustered regularly interspaced short palindromic repeats (CRISPR) editing, to characterize the dynamic cancer resistance mechanisms, from improving the understanding of resistance to classical chemotherapeutics, to deciphering unique mechanisms that regulate tumor responses to new targeted agents and immunotherapies, are discussed. Potential avenues of future research in combating therapeutic resistance, the contribution of tumor-stroma signaling in this setting, and how advanced functional genomics tools can help streamline the identification of key molecular determinants of drug response are explored., Competing Interests: The authors declare no conflict of interest., (© 2022 The Authors. Advanced Genetics published by Wiley Periodicals LLC.)
- Published
- 2022
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20. Cross-talk between mutant p53 and p62/SQSTM1 augments cancer cell migration by promoting the degradation of cell adhesion proteins.
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Mukherjee S, Maddalena M, Lü Y, Martinez S, Nataraj NB, Noronha A, Sinha S, Teng K, Cohen-Kaplan V, Ziv T, Arandkar S, Hassin O, Chatterjee R, Pirona AC, Shreberk-Shaked M, Gershoni A, Aylon Y, Elazar Z, Yarden Y, Schramek D, and Oren M
- Subjects
- Cell Adhesion genetics, Cell Line, Tumor, Genes, p53, Humans, Mutation, Sequestosome-1 Protein genetics, Sequestosome-1 Protein metabolism, Cell Movement genetics, Pancreatic Neoplasms genetics, Tumor Suppressor Protein p53 genetics, Tumor Suppressor Protein p53 metabolism
- Abstract
Missense mutations in the p53 tumor suppressor abound in human cancer. Common (“hotspot”) mutations endow mutant p53 (mutp53) proteins with oncogenic gain of function (GOF), including enhanced cell migration and invasiveness, favoring cancer progression. GOF is usually attributed to transcriptional effects of mutp53. To elucidate transcription-independent effects of mutp53, we characterized the protein interactome of the p53R273H mutant in cells derived from pancreatic ductal adenocarcinoma (PDAC), where p53R273H is the most frequent p53 mutant. We now report that p53R273H, but not the p53R175H hotspot mutant, interacts with SQSTM1/p62 and promotes cancer cell migration and invasion in a p62-dependent manner. Mechanistically, the p53R273H-p62 axis drives the proteasomal degradation of several cell junction–associated proteins, including the gap junction protein Connexin 43, facilitating scattered cell migration. Concordantly, down-regulation of Connexin 43 augments PDAC cell migration, while its forced overexpression blunts the promigratory effect of the p53R273H-p62 axis. These findings define a mechanism of mutp53 GOF.
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- 2022
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21. CDK/cyclin dependencies define extreme cancer cell-cycle heterogeneity and collateral vulnerabilities.
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Knudsen ES, Kumarasamy V, Nambiar R, Pearson JD, Vail P, Rosenheck H, Wang J, Eng K, Bremner R, Schramek D, Rubin SM, Welm AL, and Witkiewicz AK
- Subjects
- Cell Cycle genetics, Cell Cycle Proteins genetics, Cell Division, Cyclin-Dependent Kinase 4, Cyclin-Dependent Kinase Inhibitor p27 genetics, Humans, Cyclin-Dependent Kinases metabolism, Neoplasms genetics
- Abstract
Progression through G1/S phase of the cell cycle is coordinated by cyclin-dependent kinase (CDK) activities. Here, we find that the requirement for different CDK activities and cyclins in driving cancer cell cycles is highly heterogeneous. The differential gene requirements associate with tumor origin and genetic alterations. We define multiple mechanisms for G1/S progression in RB-proficient models, which are CDK4/6 independent and elicit resistance to FDA-approved inhibitors. Conversely, RB-deficient models are intrinsically CDK4/6 independent, but exhibit differential requirements for cyclin E. These dependencies for CDK and cyclins associate with gene expression programs that denote intrinsically different cell-cycle states. Mining therapeutic sensitivities shows that there are reciprocal vulnerabilities associated with RB1 or CCND1 expression versus CCNE1 or CDKN2A. Together, these findings illustrate the complex nature of cancer cell cycles and the relevance for precision therapeutic intervention., Competing Interests: Declaration of interests A.L.W. has received royalties from licenses of patient-derived xenograft or organoid models issued by the University of Utah. The university may issue new licenses in the future at its discretion, which may result in additional royalties. E.S.K. and A.K.W. have served on the BioVica scientific advisory board., (Copyright © 2022 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2022
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22. Pan-cancer analysis of non-coding transcripts reveals the prognostic onco-lncRNA HOXA10-AS in gliomas.
- Author
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Isaev K, Jiang L, Wu S, Lee CA, Watters V, Fort V, Tsai R, Coutinho FJ, Hussein SMI, Zhang J, Wu J, Dirks PB, Schramek D, and Reimand J
- Subjects
- Animals, Biomarkers, Tumor genetics, Brain Neoplasms genetics, Brain Neoplasms pathology, Cell Line, Tumor, Cell Movement, Cell Proliferation, Databases, Genetic, Gene Expression Regulation, Neoplastic, Glioma genetics, Glioma pathology, Humans, Isocitrate Dehydrogenase genetics, Isocitrate Dehydrogenase metabolism, Machine Learning, Mice, Inbred NOD, Mice, SCID, Mutation, Neoplasm Invasiveness, Predictive Value of Tests, Prognosis, RNA, Long Noncoding genetics, Reproducibility of Results, Signal Transduction, Mice, Biomarkers, Tumor metabolism, Brain Neoplasms metabolism, Gene Expression Profiling, Glioma metabolism, RNA, Long Noncoding metabolism, Transcriptome
- Abstract
Long non-coding RNAs (lncRNAs) are increasingly recognized as functional units in cancer and powerful biomarkers; however, most remain uncharacterized. Here, we analyze 5,592 prognostic lncRNAs in 9,446 cancers of 30 types using machine learning. We identify 166 lncRNAs whose expression correlates with survival and improves the accuracy of common clinical variables, molecular features, and cancer subtypes. Prognostic lncRNAs are often characterized by switch-like expression patterns. In low-grade gliomas, HOXA10-AS activation is a robust marker of poor prognosis that complements IDH1/2 mutations, as validated in another retrospective cohort, and correlates with developmental pathways in tumor transcriptomes. Loss- and gain-of-function studies in patient-derived glioma cells, organoids, and xenograft models identify HOXA10-AS as a potent onco-lncRNA that regulates cell proliferation, contact inhibition, invasion, Hippo signaling, and mitotic and neuro-developmental pathways. Our study underscores the pan-cancer potential of the non-coding transcriptome for identifying biomarkers and regulators of cancer progression., Competing Interests: Declaration of interests The authors declare no competing interests., (Copyright © 2021 The Author(s). Published by Elsevier Inc. All rights reserved.)
- Published
- 2021
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23. Binary pan-cancer classes with distinct vulnerabilities defined by pro- or anti-cancer YAP/TEAD activity.
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Pearson JD, Huang K, Pacal M, McCurdy SR, Lu S, Aubry A, Yu T, Wadosky KM, Zhang L, Wang T, Gregorieff A, Ahmad M, Dimaras H, Langille E, Cole SPC, Monnier PP, Lok BH, Tsao MS, Akeno N, Schramek D, Wikenheiser-Brokamp KA, Knudsen ES, Witkiewicz AK, Wrana JL, Goodrich DW, and Bremner R
- Subjects
- Animals, Antineoplastic Agents pharmacology, Cell Line, Tumor, Enhancer Elements, Genetic, Gene Expression Regulation, Neoplastic, Humans, Integrins metabolism, Male, Mice, Transgenic, Prostatic Neoplasms genetics, Prostatic Neoplasms pathology, Retinal Neoplasms genetics, Retinal Neoplasms pathology, Retinoblastoma genetics, Retinoblastoma pathology, Retinoblastoma Binding Proteins genetics, TEA Domain Transcription Factors metabolism, Ubiquitin-Protein Ligases genetics, Xenograft Model Antitumor Assays, Lung Neoplasms genetics, Small Cell Lung Carcinoma genetics, TEA Domain Transcription Factors genetics, Transcriptional Coactivator with PDZ-Binding Motif Proteins genetics, YAP-Signaling Proteins genetics
- Abstract
Cancer heterogeneity impacts therapeutic response, driving efforts to discover over-arching rules that supersede variability. Here, we define pan-cancer binary classes based on distinct expression of YAP and YAP-responsive adhesion regulators. Combining informatics with in vivo and in vitro gain- and loss-of-function studies across multiple murine and human tumor types, we show that opposite pro- or anti-cancer YAP activity functionally defines binary YAP
on or YAPoff cancer classes that express or silence YAP, respectively. YAPoff solid cancers are neural/neuroendocrine and frequently RB1-/- , such as retinoblastoma, small cell lung cancer, and neuroendocrine prostate cancer. YAP silencing is intrinsic to the cell of origin, or acquired with lineage switching and drug resistance. The binary cancer groups exhibit distinct YAP-dependent adhesive behavior and pharmaceutical vulnerabilities, underscoring clinical relevance. Mechanistically, distinct YAP/TEAD enhancers in YAPoff or YAPon cancers deploy anti-cancer integrin or pro-cancer proliferative programs, respectively. YAP is thus pivotal across cancer, but in opposite ways, with therapeutic implications., Competing Interests: Declaration of interests B.H.L. reports honoraria and non-financial support from AstraZeneca, and has received research funding from Pfizer and AstraZeneca not related to this work., (Copyright © 2021 Elsevier Inc. All rights reserved.)- Published
- 2021
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24. Prevalence of the Brazilian TP53 Founder c.1010G>A (p.Arg337His) in Lung Adenocarcinoma: Is Genotyping Warranted in All Brazilian Patients?
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Vieira IA, Andreis TF, Fernandes BV, Achatz MI, Macedo GS, Schramek D, and Ashton-Prolla P
- Abstract
In Southern and Southeastern Brazil, there is a germline pathogenic variant with incomplete penetrance located in the oligomerization domain of TP53 , c.1010G>A (p.Arg337His). Due to a founder effect, the variant is present in 0.3% of the general population of the region. Recently, this variant was identified in 4.4 and 8.9% of two apparently unselected, single center case series of Brazilian lung adenocarcinoma (LUAD) patients from the Southeastern and Central regions of the country, respectively. In the present study, our aim was to examine TP53 c.1010G>A allele and genotype frequencies in LUAD samples obtained from patients diagnosed in Southern Brazil. A total of 586 LUAD samples (tumor DNA) recruited from multiple centers in the region were tested, and the mutant allele was identified using TaqMan
® assays in seven cases (7/586, 1.2%) which were submitted to next generation sequencing analyses for confirmation. Somatic EGFR mutations were more frequent in TP53 c.1010G>A carriers than in non-carriers (57.1 vs. 17.6%, respectively). Further studies are needed to confirm if TP53 c.1010G>A is a driver in LUAD carcinogenesis and to verify if there is a combined effect of EGFR and germline TP53 c.1010G>A. Although variant frequency was higher than observed in the general population, it is less than previously reported in LUAD patients from other Brazilian regions. Additional data, producing regional allele frequency information in larger series of patients and including cost-effectiveness analyses, are necessary to determine if TP53 c.1010G>A screening in all Brazilian LUAD patients is justified., 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., (Copyright © 2021 Vieira, Andreis, Fernandes, Achatz, Macedo, Schramek and Ashton-Prolla.)- Published
- 2021
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25. Copper bioavailability is a KRAS-specific vulnerability in colorectal cancer.
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Aubert L, Nandagopal N, Steinhart Z, Lavoie G, Nourreddine S, Berman J, Saba-El-Leil MK, Papadopoli D, Lin S, Hart T, Macleod G, Topisirovic I, Gaboury L, Fahrni CJ, Schramek D, Meloche S, Angers S, and Roux PP
- Subjects
- Animals, Biological Availability, CRISPR-Cas Systems, Cell Line, Tumor, Cell Proliferation, Colorectal Neoplasms genetics, Colorectal Neoplasms pathology, Copper-Transporting ATPases metabolism, Female, Humans, Intestinal Mucosa pathology, Mice, Mice, Knockout, Mice, Nude, Mice, SCID, Mutation, Colorectal Neoplasms metabolism, Copper metabolism, Proto-Oncogene Proteins p21(ras) genetics, Proto-Oncogene Proteins p21(ras) metabolism
- Abstract
Despite its importance in human cancers, including colorectal cancers (CRC), oncogenic KRAS has been extremely challenging to target therapeutically. To identify potential vulnerabilities in KRAS-mutated CRC, we characterize the impact of oncogenic KRAS on the cell surface of intestinal epithelial cells. Here we show that oncogenic KRAS alters the expression of a myriad of cell-surface proteins implicated in diverse biological functions, and identify many potential surface-accessible therapeutic targets. Cell surface-based loss-of-function screens reveal that ATP7A, a copper-exporter upregulated by mutant KRAS, is essential for neoplastic growth. ATP7A is upregulated at the surface of KRAS-mutated CRC, and protects cells from excess copper-ion toxicity. We find that KRAS-mutated cells acquire copper via a non-canonical mechanism involving macropinocytosis, which appears to be required to support their growth. Together, these results indicate that copper bioavailability is a KRAS-selective vulnerability that could be exploited for the treatment of KRAS-mutated neoplasms.
- Published
- 2020
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26. Recognition of Semaphorin Proteins by P. sordellii Lethal Toxin Reveals Principles of Receptor Specificity in Clostridial Toxins.
- Author
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Lee H, Beilhartz GL, Kucharska I, Raman S, Cui H, Lam MHY, Liang H, Rubinstein JL, Schramek D, Julien JP, Melnyk RA, and Taipale M
- Subjects
- Animals, Bacterial Toxins chemistry, Bacterial Toxins toxicity, Binding Sites, CRISPR-Cas Systems genetics, Cell Line, Cryoelectron Microscopy, Edema pathology, Edema prevention & control, Female, Humans, Lung drug effects, Lung pathology, Mice, Mice, Inbred C57BL, Molecular Dynamics Simulation, Mutagenesis, Site-Directed, Protein Binding, Protein Structure, Tertiary, Recombinant Proteins biosynthesis, Recombinant Proteins isolation & purification, Recombinant Proteins therapeutic use, Semaphorins chemistry, Semaphorins genetics, Bacterial Toxins metabolism, Clostridium sordellii metabolism, Semaphorins metabolism
- Abstract
Pathogenic clostridial species secrete potent toxins that induce severe host tissue damage. Paeniclostridium sordellii lethal toxin (TcsL) causes an almost invariably lethal toxic shock syndrome associated with gynecological infections. TcsL is 87% similar to C. difficile TcdB, which enters host cells via Frizzled receptors in colon epithelium. However, P. sordellii infections target vascular endothelium, suggesting that TcsL exploits another receptor. Here, using CRISPR/Cas9 screening, we establish semaphorins SEMA6A and SEMA6B as TcsL receptors. We demonstrate that recombinant SEMA6A can protect mice from TcsL-induced edema. A 3.3 Å cryo-EM structure shows that TcsL binds SEMA6A with the same region that in TcdB binds structurally unrelated Frizzled. Remarkably, 15 mutations in this evolutionarily divergent surface are sufficient to switch binding specificity of TcsL to that of TcdB. Our findings establish semaphorins as physiologically relevant receptors for TcsL and reveal the molecular basis for the difference in tissue targeting and disease pathogenesis between highly related toxins., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)
- Published
- 2020
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27. Metabolic Regulation of the Epigenome Drives Lethal Infantile Ependymoma.
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Michealraj KA, Kumar SA, Kim LJY, Cavalli FMG, Przelicki D, Wojcik JB, Delaidelli A, Bajic A, Saulnier O, MacLeod G, Vellanki RN, Vladoiu MC, Guilhamon P, Ong W, Lee JJY, Jiang Y, Holgado BL, Rasnitsyn A, Malik AA, Tsai R, Richman CM, Juraschka K, Haapasalo J, Wang EY, De Antonellis P, Suzuki H, Farooq H, Balin P, Kharas K, Van Ommeren R, Sirbu O, Rastan A, Krumholtz SL, Ly M, Ahmadi M, Deblois G, Srikanthan D, Luu B, Loukides J, Wu X, Garzia L, Ramaswamy V, Kanshin E, Sánchez-Osuna M, El-Hamamy I, Coutinho FJ, Prinos P, Singh S, Donovan LK, Daniels C, Schramek D, Tyers M, Weiss S, Stein LD, Lupien M, Wouters BG, Garcia BA, Arrowsmith CH, Sorensen PH, Angers S, Jabado N, Dirks PB, Mack SC, Agnihotri S, Rich JN, and Taylor MD
- Subjects
- Animals, Brain Neoplasms genetics, Brain Neoplasms metabolism, Cell Line, Cell Proliferation genetics, DNA Methylation genetics, Epigenomics methods, Histones genetics, Histones metabolism, Humans, Infant, Lysine genetics, Lysine metabolism, Male, Mice, Inbred C57BL, Mutation genetics, Ependymoma genetics, Ependymoma metabolism, Epigenome genetics, Infratentorial Neoplasms genetics, Infratentorial Neoplasms metabolism
- Abstract
Posterior fossa A (PFA) ependymomas are lethal malignancies of the hindbrain in infants and toddlers. Lacking highly recurrent somatic mutations, PFA ependymomas are proposed to be epigenetically driven tumors for which model systems are lacking. Here we demonstrate that PFA ependymomas are maintained under hypoxia, associated with restricted availability of specific metabolites to diminish histone methylation, and increase histone demethylation and acetylation at histone 3 lysine 27 (H3K27). PFA ependymomas initiate from a cell lineage in the first trimester of human development that resides in restricted oxygen. Unlike other ependymomas, transient exposure of PFA cells to ambient oxygen induces irreversible cellular toxicity. PFA tumors exhibit a low basal level of H3K27me3, and, paradoxically, inhibition of H3K27 methylation specifically disrupts PFA tumor growth. Targeting metabolism and/or the epigenome presents a unique opportunity for rational therapy for infants with PFA ependymoma., Competing Interests: Declaration of Interests The authors declare no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)
- Published
- 2020
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28. In vivo CRISPR screens reveal potent driver mutations in head and neck cancers.
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Loganathan SK and Schramek D
- Abstract
We have recently tested the transforming potential of 484 'long-tail' genes, which are recurrently albeit infrequently mutated in head and neck cancers (HNSCC). We identified 15 novel tumor suppressors and our top hits converge on regulating the NOTCH signaling pathway. Therapeutic approaches activating NOTCH signaling could be a promising strategy to treat two-thirds of human HNSCC patients., (© 2020 Taylor & Francis Group, LLC.)
- Published
- 2020
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29. Mutant ACVR1 Arrests Glial Cell Differentiation to Drive Tumorigenesis in Pediatric Gliomas.
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Fortin J, Tian R, Zarrabi I, Hill G, Williams E, Sanchez-Duffhues G, Thorikay M, Ramachandran P, Siddaway R, Wong JF, Wu A, Apuzzo LN, Haight J, You-Ten A, Snow BE, Wakeham A, Goldhamer DJ, Schramek D, Bullock AN, Dijke PT, Hawkins C, and Mak TW
- Subjects
- Activin Receptors, Type I antagonists & inhibitors, Activin Receptors, Type I metabolism, Animals, Basic Helix-Loop-Helix Transcription Factors genetics, Basic Helix-Loop-Helix Transcription Factors metabolism, Bone Morphogenetic Proteins genetics, Bone Morphogenetic Proteins metabolism, Brain Neoplasms drug therapy, Brain Neoplasms genetics, Cell Differentiation genetics, Cell Line, Tumor, Class I Phosphatidylinositol 3-Kinases genetics, Class I Phosphatidylinositol 3-Kinases metabolism, Female, Glioma drug therapy, Glioma genetics, Histones genetics, Histones metabolism, Humans, Lactones pharmacology, Male, Mice, Transgenic, Neoplasms, Experimental genetics, Neoplasms, Experimental pathology, Neuroglia metabolism, Neuroglia pathology, Oligodendroglia pathology, Receptor, Platelet-Derived Growth Factor alpha genetics, Receptor, Platelet-Derived Growth Factor alpha metabolism, SOXC Transcription Factors genetics, SOXC Transcription Factors metabolism, Activin Receptors, Type I chemistry, Activin Receptors, Type I genetics, Brain Neoplasms pathology, Glioma pathology, Mutation
- Abstract
Diffuse intrinsic pontine gliomas (DIPGs) are aggressive pediatric brain tumors for which there is currently no effective treatment. Some of these tumors combine gain-of-function mutations in ACVR1, PIK3CA, and histone H3-encoding genes. The oncogenic mechanisms of action of ACVR1 mutations are currently unknown. Using mouse models, we demonstrate that Acvr1
G328V arrests the differentiation of oligodendroglial lineage cells, and cooperates with Hist1h3bK27M and Pik3caH1047R to generate high-grade diffuse gliomas. Mechanistically, Acvr1G328V upregulates transcription factors which control differentiation and DIPG cell fitness. Furthermore, we characterize E6201 as a dual inhibitor of ACVR1 and MEK1/2, and demonstrate its efficacy toward tumor cells in vivo. Collectively, our results describe an oncogenic mechanism of action for ACVR1 mutations, and suggest therapeutic strategies for DIPGs., Competing Interests: Declarations of Interests The authors declare no competing interests., (Copyright © 2020 Elsevier Inc. All rights reserved.)- Published
- 2020
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30. CRISPR-Switch regulates sgRNA activity by Cre recombination for sequential editing of two loci.
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Chylinski K, Hubmann M, Hanna RE, Yanchus C, Michlits G, Uijttewaal ECH, Doench J, Schramek D, and Elling U
- Subjects
- Animals, CRISPR-Cas Systems, Clustered Regularly Interspaced Short Palindromic Repeats, Genetic Engineering, Homologous Recombination, Integrases, Mice, Mutagenesis, RNA Polymerase III, Gene Editing methods, Mouse Embryonic Stem Cells metabolism, RNA, Guide, CRISPR-Cas Systems genetics
- Abstract
CRISPR-Cas9 is an efficient and versatile tool for genome engineering in many species. However, inducible CRISPR-Cas9 editing systems that regulate Cas9 activity or sgRNA expression often suffer from significant limitations, including reduced editing capacity, off-target effects, or leaky expression. Here, we develop a precisely controlled sgRNA expression cassette that can be combined with widely-used Cre systems, termed CRISPR-Switch (SgRNA With Induction/Termination by Cre Homologous recombination). Switch-ON facilitates controlled, rapid induction of sgRNA activity. In turn, Switch-OFF-mediated termination of editing improves generation of heterozygous genotypes and can limit off-target effects. Furthermore, we design sequential CRISPR-Switch-based editing of two loci in a strictly programmable manner and determined the order of mutagenic events that leads to development of glioblastoma in mice. Thus, CRISPR-Switch substantially increases the versatility of gene editing through precise and rapid switching ON or OFF sgRNA activity, as well as switching OVER to secondary sgRNAs.
- Published
- 2019
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31. Toronto Workshop on Late Recurrence in Estrogen Receptor-Positive Breast Cancer: Part 1: Late Recurrence: Current Understanding, Clinical Considerations.
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Dowling RJO, Kalinsky K, Hayes DF, Bidard FC, Cescon DW, Chandarlapaty S, Deasy JO, Dowsett M, Gray RJ, Henry NL, Meric-Bernstam F, Perlmutter J, Sledge GW, Bratman SV, Carey LA, Chang MC, DeMichele A, Ennis M, Jerzak KJ, Korde LA, Lohmann AE, Mamounas EP, Parulekar WR, Regan MM, Schramek D, Stambolic V, Thorat MA, Whelan TJ, Wolff AC, Woodgett JR, Sparano JA, and Goodwin PJ
- Abstract
Disease recurrence (locoregional, distant) exerts a significant clinical impact on the survival of estrogen receptor-positive breast cancer patients. Many of these recurrences occur late, more than 5 years after original diagnosis, and represent a major obstacle to the effective treatment of this disease. Indeed, methods to identify patients at risk of late recurrence and therapeutic strategies designed to avert or treat these recurrences are lacking. Therefore, an international workshop was convened in Toronto, Canada, in February 2018 to review the current understanding of late recurrence and to identify critical issues that require future study. In this article, the major issues surrounding late recurrence are defined and current approaches that may be applicable to this challenge are discussed. Specifically, diagnostic tests with potential utility in late-recurrence prediction are described as well as a variety of patient-related factors that may influence recurrence risk. Clinical and therapeutic approaches are also reviewed, with a focus on patient surveillance and the implementation of extended endocrine therapy in the context of late-recurrence prevention. Understanding and treating late recurrence in estrogen receptor-positive breast cancer is a major unmet clinical need. A concerted effort of basic and clinical research is required to confront late recurrence and improve disease management and patient survival., (© The Author(s) 2019. Published by Oxford University Press.)
- Published
- 2019
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32. Toronto Workshop on Late Recurrence in Estrogen Receptor-Positive Breast Cancer: Part 2: Approaches to Predict and Identify Late Recurrence, Research Directions.
- Author
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Dowling RJO, Sparano JA, Goodwin PJ, Bidard FC, Cescon DW, Chandarlapaty S, Deasy JO, Dowsett M, Gray RJ, Henry NL, Meric-Bernstam F, Perlmutter J, Sledge GW, Thorat MA, Bratman SV, Carey LA, Chang MC, DeMichele A, Ennis M, Jerzak KJ, Korde LA, Lohmann AE, Mamounas EP, Parulekar WR, Regan MM, Schramek D, Stambolic V, Whelan TJ, Wolff AC, Woodgett JR, Kalinsky K, and Hayes DF
- Abstract
Late disease recurrence (more than 5 years after initial diagnosis) represents a clinical challenge in the treatment and management of estrogen receptor-positive breast cancer (BC). An international workshop was convened in Toronto, Canada, in February 2018 to review the current understanding of late recurrence and to identify critical issues that require future study. The underlying biological causes of late recurrence are complex, with the processes governing cancer cell dormancy, including immunosurveillance, cell proliferation, angiogenesis, and cellular stemness, being integral to disease progression. These critical processes are described herein as well as their role in influencing risk of recurrence. Moreover, observational and interventional clinical trials are proposed, with a focus on methods to identify patients at risk of recurrence and possible strategies to combat this in patients with estrogen receptor-positive BC. Because the problem of late BC recurrence of great importance, recent advances in disease detection and patient monitoring should be incorporated into novel clinical trials to evaluate approaches to enhance patient management. Indeed, future research on these issues is planned and will offer new options for effective late recurrence treatment and prevention strategies., (© The Author(s) 2019. Published by Oxford University Press.)
- Published
- 2019
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33. Apelin inhibition prevents resistance and metastasis associated with anti-angiogenic therapy.
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Uribesalgo I, Hoffmann D, Zhang Y, Kavirayani A, Lazovic J, Berta J, Novatchkova M, Pai TP, Wimmer RA, László V, Schramek D, Karim R, Tortola L, Deswal S, Haas L, Zuber J, Szűcs M, Kuba K, Dome B, Cao Y, Haubner BJ, and Penninger JM
- Subjects
- Animals, Apelin antagonists & inhibitors, Apelin deficiency, Apelin genetics, Apelin Receptors antagonists & inhibitors, Apelin Receptors deficiency, Apelin Receptors genetics, Cell Line, Tumor, Endothelial Cells drug effects, Endothelial Cells metabolism, Endothelial Cells pathology, Female, Gene Expression Regulation, Neoplastic, Humans, Lung Neoplasms genetics, Lung Neoplasms metabolism, Lung Neoplasms pathology, Mammary Neoplasms, Experimental genetics, Mammary Neoplasms, Experimental metabolism, Mammary Neoplasms, Experimental pathology, Mice, Mice, Inbred C57BL, Mice, Knockout, Mouse Embryonic Stem Cells drug effects, Mouse Embryonic Stem Cells metabolism, Mouse Embryonic Stem Cells pathology, Neoplasm Metastasis, Signal Transduction, Tumor Burden drug effects, Tumor Microenvironment, Angiogenesis Inhibitors pharmacology, Apelin metabolism, Apelin Receptors metabolism, Cell Movement drug effects, Drug Resistance, Neoplasm, Lung Neoplasms drug therapy, Mammary Neoplasms, Experimental drug therapy, Neovascularization, Pathologic, Protein Kinase Inhibitors pharmacology, Sunitinib pharmacology
- Abstract
Angiogenesis is a hallmark of cancer, promoting growth and metastasis. Anti-angiogenic treatment has limited efficacy due to therapy-induced blood vessel alterations, often followed by local hypoxia, tumor adaptation, progression, and metastasis. It is therefore paramount to overcome therapy-induced resistance. We show that Apelin inhibition potently remodels the tumor microenvironment, reducing angiogenesis, and effectively blunting tumor growth. Functionally, targeting Apelin improves vessel function and reduces polymorphonuclear myeloid-derived suppressor cell infiltration. Importantly, in mammary and lung cancer, Apelin prevents resistance to anti-angiogenic receptor tyrosine kinase (RTK) inhibitor therapy, reducing growth and angiogenesis in lung and breast cancer models without increased hypoxia in the tumor microenvironment. Apelin blockage also prevents RTK inhibitor-induced metastases, and high Apelin levels correlate with poor prognosis of anti-angiogenic therapy patients. These data identify a druggable anti-angiogenic drug target that reduces tumor blood vessel densities and normalizes the tumor vasculature to decrease metastases., (© 2019 The Authors. Published under the terms of the CC BY 4.0 license.)
- Published
- 2019
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34. AIF-regulated oxidative phosphorylation supports lung cancer development.
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Rao S, Mondragón L, Pranjic B, Hanada T, Stoll G, Köcher T, Zhang P, Jais A, Lercher A, Bergthaler A, Schramek D, Haigh K, Sica V, Leduc M, Modjtahedi N, Pai TP, Onji M, Uribesalgo I, Hanada R, Kozieradzki I, Koglgruber R, Cronin SJ, She Z, Quehenberger F, Popper H, Kenner L, Haigh JJ, Kepp O, Rak M, Cai K, Kroemer G, and Penninger JM
- Subjects
- Animals, Carcinoma, Non-Small-Cell Lung pathology, Cell Line, Tumor, Datasets as Topic, Disease Progression, Glycolysis, Humans, Lung Neoplasms pathology, Mice, Mice, Inbred C57BL, Oxidative Phosphorylation, Apoptosis Inducing Factor physiology, Carcinogenesis metabolism, Carcinoma, Non-Small-Cell Lung metabolism, Lung Neoplasms metabolism
- Abstract
Cancer is a major and still increasing cause of death in humans. Most cancer cells have a fundamentally different metabolic profile from that of normal tissue. This shift away from mitochondrial ATP synthesis via oxidative phosphorylation towards a high rate of glycolysis, termed Warburg effect, has long been recognized as a paradigmatic hallmark of cancer, supporting the increased biosynthetic demands of tumor cells. Here we show that deletion of apoptosis-inducing factor (AIF) in a Kras
G12D -driven mouse lung cancer model resulted in a marked survival advantage, with delayed tumor onset and decreased malignant progression. Mechanistically, Aif deletion leads to oxidative phosphorylation (OXPHOS) deficiency and a switch in cellular metabolism towards glycolysis in non-transformed pneumocytes and at early stages of tumor development. Paradoxically, although Aif-deficient cells exhibited a metabolic Warburg profile, this bioenergetic change resulted in a growth disadvantage of KrasG12D -driven as well as Kras wild-type lung cancer cells. Cell-autonomous re-expression of both wild-type and mutant AIF (displaying an intact mitochondrial, but abrogated apoptotic function) in Aif-knockout KrasG12D mice restored OXPHOS and reduced animal survival to the same level as AIF wild-type mice. In patients with non-small cell lung cancer, high AIF expression was associated with poor prognosis. These data show that AIF-regulated mitochondrial respiration and OXPHOS drive the progression of lung cancer.- Published
- 2019
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35. A Phase 1/1b Study Evaluating Trametinib Plus Docetaxel or Pemetrexed in Patients With Advanced Non-Small Cell Lung Cancer.
- Author
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Gandara DR, Leighl N, Delord JP, Barlesi F, Bennouna J, Zalcman G, Infante JR, Reckamp KL, Kelly K, Shepherd FA, Mazieres J, Janku F, Gardner OS, Mookerjee B, Wu Y, Cox DS, Schramek D, Peddareddigari V, Liu Y, D'Amelio AM Jr, and Blumenschein G Jr
- Subjects
- Adenocarcinoma secondary, Adult, Aged, Aged, 80 and over, Carcinoma, Large Cell secondary, Carcinoma, Non-Small-Cell Lung pathology, Carcinoma, Squamous Cell secondary, Docetaxel, Female, Follow-Up Studies, Humans, Lung Neoplasms pathology, Male, Middle Aged, Neoplasm Staging, Pemetrexed administration & dosage, Prognosis, Pyridones administration & dosage, Pyrimidinones administration & dosage, Survival Rate, Taxoids administration & dosage, Adenocarcinoma drug therapy, Antineoplastic Combined Chemotherapy Protocols therapeutic use, Carcinoma, Large Cell drug therapy, Carcinoma, Non-Small-Cell Lung drug therapy, Carcinoma, Squamous Cell drug therapy, Lung Neoplasms drug therapy
- Abstract
Objectives: This two-part study evaluated trametinib, a MEK1/2 inhibitor, in combination with anticancer agents. Inhibition of MEK, a downstream effector of KRAS, demonstrated preclinical synergy with chemotherapy in KRAS-mutant NSCLC cell lines. Part 1 of this study identified recommended phase 2 doses of trametinib combinations. Part 2, reported herein, evaluated the safety, tolerability, pharmacokinetics, and efficacy of trametinib combinations in patients with NSCLC with and without KRAS mutations., Methods: Phase 1b evaluated trametinib plus docetaxel with growth factor support (trametinib, 2.0 mg once daily, and docetaxel, 75 mg/m
2 every 3 weeks) or pemetrexed (trametinib, 1.5 mg once daily, and pemetrexed, 500 mg/m2 every 3 weeks). Eligibility criteria for the expansion cohorts included metastatic NSCLC with measurable disease, known KRAS mutation status, Eastern Cooperative Oncology Group performance status of 1 or lower, and no more than two prior regimens., Results: The primary end point of overall response rate (ORR) was met for both combinations. A confirmed partial response (PR) was observed in 10 of the 47 patients with NSCLC who received trametinib plus docetaxel (21%). The ORR was 18% (four PRs in 22 patients) in those with KRAS wild-type NSCLC versus 24% (six PRs in 25 patients) in those with KRAS-mutant NSCLC. Of the 42 patients with NSCLC treated with trametinib plus pemetrexed, six (14%) had a PR; the ORR was 17% (four of 23) in patients with KRAS-mutated NSCLC versus 11% (two of 19) in KRAS wild-type NSCLC. Adverse events-most commonly diarrhea, nausea, and fatigue-were manageable., Conclusions: Trametinib-plus-chemotherapy combinations were tolerable. Clinical activity exceeding the ORRs previously reported with docetaxel or pemetrexed alone in KRAS-mutated NSCLC and meeting prespecified criteria was observed., (Copyright © 2016 International Association for the Study of Lung Cancer. Published by Elsevier Inc. All rights reserved.)- Published
- 2017
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36. Translation from unconventional 5' start sites drives tumour initiation.
- Author
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Sendoel A, Dunn JG, Rodriguez EH, Naik S, Gomez NC, Hurwitz B, Levorse J, Dill BD, Schramek D, Molina H, Weissman JS, and Fuchs E
- Subjects
- Animals, Carcinogenesis pathology, Carcinoma, Squamous Cell metabolism, Disease Models, Animal, Disease Progression, Epidermis embryology, Epidermis metabolism, Epidermis pathology, Eukaryotic Initiation Factor-2 metabolism, Female, Humans, Keratinocytes, Male, Mice, Oncogenes genetics, Precancerous Conditions genetics, Precancerous Conditions metabolism, Precancerous Conditions pathology, Prognosis, RNA Interference, RNA, Messenger genetics, RNA, Messenger metabolism, Ribosomes metabolism, SOXB1 Transcription Factors genetics, SOXB1 Transcription Factors metabolism, Skin Neoplasms metabolism, 5' Untranslated Regions genetics, Carcinogenesis genetics, Carcinoma, Squamous Cell genetics, Carcinoma, Squamous Cell pathology, Open Reading Frames genetics, Peptide Chain Initiation, Translational genetics, Skin Neoplasms genetics, Skin Neoplasms pathology
- Abstract
We are just beginning to understand how translational control affects tumour initiation and malignancy. Here we use an epidermis-specific, in vivo ribosome profiling strategy to investigate the translational landscape during the transition from normal homeostasis to malignancy. Using a mouse model of inducible SOX2, which is broadly expressed in oncogenic RAS-associated cancers, we show that despite widespread reductions in translation and protein synthesis, certain oncogenic mRNAs are spared. During tumour initiation, the translational apparatus is redirected towards unconventional upstream initiation sites, enhancing the translational efficiency of oncogenic mRNAs. An in vivo RNA interference screen of translational regulators revealed that depletion of conventional eIF2 complexes has adverse effects on normal but not oncogenic growth. Conversely, the alternative initiation factor eIF2A is essential for cancer progression, during which it mediates initiation at these upstream sites, differentially skewing translation and protein expression. Our findings unveil a role for the translation of 5' untranslated regions in cancer, and expose new targets for therapeutic intervention., Competing Interests: The authors declare no competing financial interests. Readers are welcome to comment on the online version of the paper.
- Published
- 2017
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37. Aberrant regulation of RANKL/OPG in women at high risk of developing breast cancer.
- Author
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Kiechl S, Schramek D, Widschwendter M, Fourkala EO, Zaikin A, Jones A, Jaeger B, Rack B, Janni W, Scholz C, Willeit J, Weger S, Mayr A, Teschendorff A, Rosenthal A, Fraser L, Philpott S, Dubeau L, Keshtgar M, Roylance R, Jacobs IJ, Menon U, Schett G, and Penninger JM
- Subjects
- Biomarkers, Tumor blood, Breast Neoplasms blood, Early Detection of Cancer, Female, Gene Expression Regulation, Neoplastic, Humans, Postmenopause, Prospective Studies, Receptor Activator of Nuclear Factor-kappa B blood, Up-Regulation, Breast Neoplasms metabolism, Neoplastic Cells, Circulating metabolism, Osteoprotegerin blood, Progesterone blood, RANK Ligand blood
- Abstract
Breast cancer is the most common female cancer, affecting approximately one in eight women during their lifetime in North America and Europe. Receptor Activator of NF-kB Ligand (RANKL), its receptor RANK and the natural antagonist osteoprotegerin (OPG) are essential regulators of bone resorption. We have initially shown that RANKL/RANK are essential for hormone-driven mammary epithelial proliferation in pregnancy and RANKL/RANK have been implicated in mammary stem cell biology. Using genetic mouse-models, we and others identified the RANKL/RANK system as a key regulator of sex hormone, BRCA1-mutation, and oncogene-driven breast cancer and we proposed that RANKL/RANK might be involved in the initiation of breast tumors. We now report that in postmenopausal women without known genetic predisposition, high RANKL and progesterone serum levels stratify a subpopulation of women at high risk of developing breast cancer 12-24 months before diagnosis (5.33-fold risk, 95%CI 1.5-25.4; P=0.02). In women with established breast cancer, we demonstrate that RANKL/OPG ratios change dependent on the presence of circulating tumor cells (CTCs). Finally, we show in a prospective human breast cancer cohort that alterations in RANKL/OPG ratios are significantly associated with breast cancer manifestation. These data indicate that the RANKL/RANK/OPG system is deregulated in post-menopausal women at high risk for breast cancer and in women with circulating tumor cells. Thus, serum levels of RANKL/OPG are potentially indicative of predisposition and progression of breast cancer in humans. Advancement of our findings towards clinical application awaits prior validation in independent patient cohorts.
- Published
- 2017
- Full Text
- View/download PDF
38. RANKL/RANK control Brca1 mutation- .
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Sigl V, Owusu-Boaitey K, Joshi PA, Kavirayani A, Wirnsberger G, Novatchkova M, Kozieradzki I, Schramek D, Edokobi N, Hersl J, Sampson A, Odai-Afotey A, Lazaro C, Gonzalez-Suarez E, Pujana MA, Cimba F, Heyn H, Vidal E, Cruickshank J, Berman H, Sarao R, Ticevic M, Uribesalgo I, Tortola L, Rao S, Tan Y, Pfeiler G, Lee EY, Bago-Horvath Z, Kenner L, Popper H, Singer C, Khokha R, Jones LP, and Penninger JM
- Subjects
- Animals, BRCA2 Protein genetics, Breast Neoplasms drug therapy, Breast Neoplasms metabolism, Breast Neoplasms pathology, Cell Proliferation drug effects, Cells, Cultured, DNA Damage drug effects, Epithelial Cells cytology, Epithelial Cells drug effects, Epithelial Cells metabolism, Estrogen Receptor alpha metabolism, Female, Genotype, Humans, Mice, Mice, Inbred C57BL, Mice, Transgenic, RANK Ligand antagonists & inhibitors, RANK Ligand genetics, Receptor Activator of Nuclear Factor-kappa B genetics, Receptors, Progesterone metabolism, Recombinant Fusion Proteins pharmacology, Recombinant Fusion Proteins therapeutic use, Stem Cells cytology, Stem Cells metabolism, Tumor Suppressor Protein p53 genetics, Tumor Suppressor Protein p53 metabolism, BRCA1 Protein genetics, Breast Neoplasms genetics, RANK Ligand metabolism, Receptor Activator of Nuclear Factor-kappa B metabolism
- Abstract
Breast cancer is the most common female cancer, affecting approximately one in eight women during their life-time. Besides environmental triggers and hormones, inherited mutations in the breast cancer 1 (BRCA1) or BRCA2 genes markedly increase the risk for the development of breast cancer. Here, using two different mouse models, we show that genetic inactivation of the key osteoclast differentiation factor RANK in the mammary epithelium markedly delayed onset, reduced incidence, and attenuated progression of Brca1;p53 mutation-driven mammary cancer. Long-term pharmacological inhibition of the RANK ligand RANKL in mice abolished the occurrence of Brca1 mutation-driven pre-neoplastic lesions. Mechanistically, genetic inactivation of Rank or RANKL/RANK blockade impaired proliferation and expansion of both murine Brca1;p53 mutant mammary stem cells and mammary progenitors from human BRCA1 mutation carriers. In addition, genome variations within the RANK locus were significantly associated with risk of developing breast cancer in women with BRCA1 mutations. Thus, RANKL/RANK control progenitor cell expansion and tumorigenesis in inherited breast cancer. These results present a viable strategy for the possible prevention of breast cancer in BRCA1 mutant patients.
- Published
- 2016
- Full Text
- View/download PDF
39. ETS family transcriptional regulators drive chromatin dynamics and malignancy in squamous cell carcinomas.
- Author
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Yang H, Schramek D, Adam RC, Keyes BE, Wang P, Zheng D, and Fuchs E
- Subjects
- Cell Proliferation, Epigenesis, Genetic, Humans, Carcinoma, Squamous Cell pathology, Chromatin metabolism, Gene Regulatory Networks
- Abstract
Tumor-initiating stem cells (SCs) exhibit distinct patterns of transcription factors and gene expression compared to healthy counterparts. Here, we show that dramatic shifts in large open-chromatin domain (super-enhancer) landscapes underlie these differences and reflect tumor microenvironment. By in vivo super-enhancer and transcriptional profiling, we uncover a dynamic cancer-specific epigenetic network selectively enriched for binding motifs of a transcription factor cohort expressed in squamous cell carcinoma SCs (SCC-SCs). Many of their genes, including Ets2 and Elk3, are themselves regulated by SCC-SC super-enhancers suggesting a cooperative feed-forward loop. Malignant progression requires these genes, whose knockdown severely impairs tumor growth and prohibits progression from benign papillomas to SCCs. ETS2-deficiency disrupts the SCC-SC super-enhancer landscape and downstream cancer genes while ETS2-overactivation in epidermal-SCs induces hyperproliferation and SCC super-enhancer-associated genes Fos, Junb and Klf5. Together, our findings unearth an essential regulatory network required for the SCC-SC chromatin landscape and unveil its importance in malignant progression.
- Published
- 2015
- Full Text
- View/download PDF
40. Disruption of STAT3 signalling promotes KRAS-induced lung tumorigenesis.
- Author
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Grabner B, Schramek D, Mueller KM, Moll HP, Svinka J, Hoffmann T, Bauer E, Blaas L, Hruschka N, Zboray K, Stiedl P, Nivarthi H, Bogner E, Gruber W, Mohr T, Zwick RH, Kenner L, Poli V, Aberger F, Stoiber D, Egger G, Esterbauer H, Zuber J, Moriggl R, Eferl R, Győrffy B, Penninger JM, Popper H, and Casanova E
- Subjects
- Adenocarcinoma genetics, Adenocarcinoma metabolism, Animals, Chromatin Immunoprecipitation, Enzyme-Linked Immunosorbent Assay, Gene Knockdown Techniques, Heterografts, Humans, Immunoblotting, In Situ Hybridization, Interleukin-8 metabolism, Lung Neoplasms genetics, Lung Neoplasms metabolism, Mice, NF-kappa B metabolism, Proto-Oncogene Proteins p21(ras) genetics, Real-Time Polymerase Chain Reaction, STAT3 Transcription Factor genetics, Statistics, Nonparametric, Tissue Array Analysis, Adenocarcinoma drug therapy, Carcinogenesis, Gene Expression Regulation, Neoplastic physiology, Lung Neoplasms drug therapy, Proto-Oncogene Proteins p21(ras) metabolism, STAT3 Transcription Factor metabolism, Signal Transduction physiology
- Abstract
STAT3 is considered to play an oncogenic role in several malignancies including lung cancer; consequently, targeting STAT3 is currently proposed as therapeutic intervention. Here we demonstrate that STAT3 plays an unexpected tumour-suppressive role in KRAS mutant lung adenocarcinoma (AC). Indeed, lung tissue-specific inactivation of Stat3 in mice results in increased Kras(G12D)-driven AC initiation and malignant progression leading to markedly reduced survival. Knockdown of STAT3 in xenografted human AC cells increases tumour growth. Clinically, low STAT3 expression levels correlate with poor survival and advanced malignancy in human lung AC patients with smoking history, which are prone to KRAS mutations. Consistently, KRAS mutant lung tumours exhibit reduced STAT3 levels. Mechanistically, we demonstrate that STAT3 controls NF-κB-induced IL-8 expression by sequestering NF-κB within the cytoplasm, thereby inhibiting IL-8-mediated myeloid tumour infiltration and tumour vascularization and hence tumour progression. These results elucidate a novel STAT3-NF-κB-IL-8 axis in KRAS mutant AC with therapeutic and prognostic relevance.
- Published
- 2015
- Full Text
- View/download PDF
41. A dual role for autophagy in a murine model of lung cancer.
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Rao S, Tortola L, Perlot T, Wirnsberger G, Novatchkova M, Nitsch R, Sykacek P, Frank L, Schramek D, Komnenovic V, Sigl V, Aumayr K, Schmauss G, Fellner N, Handschuh S, Glösmann M, Pasierbek P, Schlederer M, Resch GP, Ma Y, Yang H, Popper H, Kenner L, Kroemer G, and Penninger JM
- Subjects
- Animals, Autophagy-Related Protein 5, Disease Progression, Female, Gene Deletion, Gene Expression Profiling, Male, Mice, Mice, Inbred BALB C, Microtubule-Associated Proteins genetics, Mutation genetics, T-Lymphocytes, Regulatory pathology, T-Lymphocytes, Regulatory physiology, Tumor Suppressor Protein p53 genetics, Tumor Suppressor Protein p53 physiology, Autophagy physiology, Disease Models, Animal, Lung Neoplasms pathology, Lung Neoplasms physiopathology, Microtubule-Associated Proteins physiology
- Abstract
Autophagy is a mechanism by which starving cells can control their energy requirements and metabolic states, thus facilitating the survival of cells in stressful environments, in particular in the pathogenesis of cancer. Here we report that tissue-specific inactivation of Atg5, essential for the formation of autophagosomes, markedly impairs the progression of KRas(G12D)-driven lung cancer, resulting in a significant survival advantage of tumour-bearing mice. Autophagy-defective lung cancers exhibit impaired mitochondrial energy homoeostasis, oxidative stress and a constitutively active DNA damage response. Genetic deletion of the tumour suppressor p53 reinstates cancer progression of autophagy-deficient tumours. Although there is improved survival, the onset of Atg5-mutant KRas(G12D)-driven lung tumours is markedly accelerated. Mechanistically, increased oncogenesis maps to regulatory T cells. These results demonstrate that, in KRas(G12D)-driven lung cancer, Atg5-regulated autophagy accelerates tumour progression; however, autophagy also represses early oncogenesis, suggesting a link between deregulated autophagy and regulatory T cell controlled anticancer immunity.
- Published
- 2014
- Full Text
- View/download PDF
42. MKK7 and ARF: new players in the DNA damage response scenery.
- Author
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Kotsinas A, Papanagnou P, Galanos P, Schramek D, Townsend P, Penninger JM, Bartek J, and Gorgoulis VG
- Subjects
- Animals, Ataxia Telangiectasia Mutated Proteins metabolism, Humans, Oncogenes, Phosphorylation, Signal Transduction, Tumor Suppressor Protein p53 metabolism, p38 Mitogen-Activated Protein Kinases metabolism, DNA Damage, DNA Repair, MAP Kinase Kinase 7 metabolism, Reading Frames
- Abstract
Sensing, integrating, and processing of stressogenic signals must be followed by accurate differential response(s) for a cell to survive and avoid malignant transformation. The DNA damage response (DDR) pathway is vital in this process, as it deals with genotoxic/oncogenic insults, having p53 as a nodal effector that performs most of the above tasks. Accumulating data reveal that other pathways are also involved in the same or similar processes, conveying also to p53. Emerging questions are if, how, and when these additional pathways communicate with the DDR axis. Two such stress response pathways, involving the MKK7 stress-activated protein kinase (SAPK) and ARF, have been shown to be interlocked with the ATM/ATR-regulated DDR axis in a highly ordered manner. This creates a new landscape in the DDR orchestrated response to genotoxic/oncogenic insults that is currently discussed.
- Published
- 2014
- Full Text
- View/download PDF
43. Progesterone drives mammary secretory differentiation via RankL-mediated induction of Elf5 in luminal progenitor cells.
- Author
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Lee HJ, Gallego-Ortega D, Ledger A, Schramek D, Joshi P, Szwarc MM, Cho C, Lydon JP, Khokha R, Penninger JM, and Ormandy CJ
- Subjects
- Animals, DNA-Binding Proteins metabolism, Epithelial Cells drug effects, Epithelial Cells metabolism, Epithelial Cells physiology, Female, Gene Expression Regulation, Developmental drug effects, Gene Expression Regulation, Developmental genetics, Mammary Glands, Animal growth & development, Mammary Glands, Animal metabolism, Mice, Mice, Transgenic, RANK Ligand metabolism, RANK Ligand physiology, Receptors, Progesterone genetics, Receptors, Progesterone metabolism, Receptors, Progesterone physiology, Stem Cells physiology, Transcription Factors metabolism, Up-Regulation genetics, Up-Regulation physiology, DNA-Binding Proteins genetics, Mammary Glands, Animal drug effects, Progesterone pharmacology, RANK Ligand pharmacology, Stem Cells metabolism, Transcription Factors genetics
- Abstract
Progesterone-RankL paracrine signaling has been proposed as a driver of stem cell expansion in the mammary gland, and Elf5 is essential for the differentiation of mammary epithelial progenitor cells. We demonstrate that Elf5 expression is induced by progesterone and that Elf5 and progesterone cooperate to promote alveolar development. The progesterone receptor and Elf5 are expressed in a mutually exclusive pattern, and we identify RankL as the paracrine mediator of the effects of progesterone on Elf5 expression in CD61+ progenitor cells and their consequent differentiation. Blockade of RankL action prevented progesterone-induced side branching and the expansion of Elf5(+) mature luminal cells. These findings describe a mechanism by which steroid hormones can produce the expansion of steroid hormone receptor-negative mammary epithelial cells.
- Published
- 2013
- Full Text
- View/download PDF
44. Forward and reverse genetics through derivation of haploid mouse embryonic stem cells.
- Author
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Elling U, Taubenschmid J, Wirnsberger G, O'Malley R, Demers SP, Vanhaelen Q, Shukalyuk AI, Schmauss G, Schramek D, Schnuetgen F, von Melchner H, Ecker JR, Stanford WL, Zuber J, Stark A, and Penninger JM
- Subjects
- Animals, Biomarkers metabolism, Cell Differentiation genetics, Cell Line, Embryo, Mammalian cytology, Embryo, Mammalian physiology, Embryonic Stem Cells cytology, Embryonic Stem Cells drug effects, Genome-Wide Association Study, Mice, Mice, Inbred C57BL, Parthenogenesis genetics, Ricin toxicity, Embryonic Stem Cells physiology, Haploidy, Reverse Genetics methods
- Abstract
All somatic mammalian cells carry two copies of chromosomes (diploidy), whereas organisms with a single copy of their genome, such as yeast, provide a basis for recessive genetics. Here we report the generation of haploid mouse ESC lines from parthenogenetic embryos. These cells carry 20 chromosomes, express stem cell markers, and develop into all germ layers in vitro and in vivo. We also developed a reversible mutagenesis protocol that allows saturated genetic recessive screens and results in homozygous alleles. This system allowed us to generate a knockout cell line for the microRNA processing enzyme Drosha. In a forward genetic screen, we identified Gpr107 as a molecule essential for killing by ricin, a toxin being used as a bioweapon. Our results open the possibility of combining the power of a haploid genome with pluripotency of embryonic stem cells to uncover fundamental biological processes in defined cell types at a genomic scale., (Copyright © 2011 Elsevier Inc. All rights reserved.)
- Published
- 2011
- Full Text
- View/download PDF
45. RANKL/RANK-beyond bones.
- Author
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Hanada R, Hanada T, Sigl V, Schramek D, and Penninger JM
- Subjects
- Animals, Humans, Immune System immunology, Immune System metabolism, Lactation metabolism, Neoplasms metabolism, RANK Ligand immunology, Receptor Activator of Nuclear Factor-kappa B immunology, Signal Transduction physiology, Bone and Bones metabolism, RANK Ligand metabolism, Receptor Activator of Nuclear Factor-kappa B metabolism
- Abstract
Receptor-activator of NF-κB ligand (TNFSF11, also known as RANKL, OPGL, TRANCE, and ODF) and its tumor necrosis factor (TNF)-family receptor RANK are essential regulators of bone remodeling, lymph node formation, establishment of the thymic microenvironment, mammary gland development during pregnancy, and bone metastasis in cancer. We have recently also reported that the RANKL/RANK system controls the incidence and onset of sex hormone, progestin-driven breast cancer. RANKL and RANK are also expressed in the central nervous systems where they play an essential role in body temperature regulation. RANKL activates brain regions involved in thermoregulation and induces fever via the COX2-PGE(2)/EP3R pathway. Moreover, female mice with a RANK gene deleted in neurons and astrocytes exhibit increased basal body temperature, suggesting that the RANKL/RANK system also controls physiological thermoregulation in females under the control of sex hormones. This review will summarize the recently emerging role of the RANKL/RANK signaling axis in mammary gland development, cancer metastasis, hormone-derived breast cancer development, and thermal regulation. Furthermore, we will highlight the striking therapeutic potential of this pathway and provide a molecular rationale for consideration of targeting RANKL/RANK in diseases such as breast cancer.
- Published
- 2011
- Full Text
- View/download PDF
46. A genome-wide Drosophila screen for heat nociception identifies α2δ3 as an evolutionarily conserved pain gene.
- Author
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Neely GG, Hess A, Costigan M, Keene AC, Goulas S, Langeslag M, Griffin RS, Belfer I, Dai F, Smith SB, Diatchenko L, Gupta V, Xia CP, Amann S, Kreitz S, Heindl-Erdmann C, Wolz S, Ly CV, Arora S, Sarangi R, Dan D, Novatchkova M, Rosenzweig M, Gibson DG, Truong D, Schramek D, Zoranovic T, Cronin SJ, Angjeli B, Brune K, Dietzl G, Maixner W, Meixner A, Thomas W, Pospisilik JA, Alenius M, Kress M, Subramaniam S, Garrity PA, Bellen HJ, Woolf CJ, and Penninger JM
- Subjects
- Adult, Animals, Back Pain genetics, Calcium Channels metabolism, Drosophila Proteins metabolism, Gene Knockdown Techniques, Genome-Wide Association Study, Hot Temperature, Humans, Mice, Polymorphism, Single Nucleotide, RNA Interference, Calcium Channels genetics, Drosophila genetics, Drosophila Proteins genetics, Pain genetics
- Abstract
Worldwide, acute, and chronic pain affects 20% of the adult population and represents an enormous financial and emotional burden. Using genome-wide neuronal-specific RNAi knockdown in Drosophila, we report a global screen for an innate behavior and identify hundreds of genes implicated in heat nociception, including the α2δ family calcium channel subunit straightjacket (stj). Mice mutant for the stj ortholog CACNA2D3 (α2δ3) also exhibit impaired behavioral heat pain sensitivity. In addition, in humans, α2δ3 SNP variants associate with reduced sensitivity to acute noxious heat and chronic back pain. Functional imaging in α2δ3 mutant mice revealed impaired transmission of thermal pain-evoked signals from the thalamus to higher-order pain centers. Intriguingly, in α2δ3 mutant mice, thermal pain and tactile stimulation triggered strong cross-activation, or synesthesia, of brain regions involved in vision, olfaction, and hearing., (Copyright © 2010 Elsevier Inc. All rights reserved.)
- Published
- 2010
- Full Text
- View/download PDF
47. Osteoclast differentiation factor RANKL controls development of progestin-driven mammary cancer.
- Author
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Schramek D, Leibbrandt A, Sigl V, Kenner L, Pospisilik JA, Lee HJ, Hanada R, Joshi PA, Aliprantis A, Glimcher L, Pasparakis M, Khokha R, Ormandy CJ, Widschwendter M, Schett G, and Penninger JM
- Subjects
- Animals, Apoptosis radiation effects, Cell Differentiation, Cell Proliferation drug effects, DNA Damage, Epithelial Cells cytology, Epithelial Cells drug effects, Epithelial Cells metabolism, Epithelial Cells radiation effects, Female, Gamma Rays, Integrin alpha6 metabolism, Mammary Neoplasms, Experimental genetics, Mammary Neoplasms, Experimental metabolism, Medroxyprogesterone Acetate administration & dosage, Medroxyprogesterone Acetate adverse effects, Mice, NF-kappa B metabolism, Osteoclasts cytology, Phosphoproteins analysis, Phosphoproteins immunology, Progestins administration & dosage, RANK Ligand deficiency, RANK Ligand genetics, Receptor Activator of Nuclear Factor-kappa B deficiency, Receptor Activator of Nuclear Factor-kappa B genetics, Receptor Activator of Nuclear Factor-kappa B metabolism, Signal Transduction, Stem Cells cytology, Stem Cells drug effects, Stem Cells metabolism, Mammary Neoplasms, Experimental chemically induced, Mammary Neoplasms, Experimental pathology, Progestins adverse effects, RANK Ligand metabolism
- Abstract
Breast cancer is one of the most common cancers in humans and will on average affect up to one in eight women in their lifetime in the United States and Europe. The Women's Health Initiative and the Million Women Study have shown that hormone replacement therapy is associated with an increased risk of incident and fatal breast cancer. In particular, synthetic progesterone derivatives (progestins) such as medroxyprogesterone acetate (MPA), used in millions of women for hormone replacement therapy and contraceptives, markedly increase the risk of developing breast cancer. Here we show that the in vivo administration of MPA triggers massive induction of the key osteoclast differentiation factor RANKL (receptor activator of NF-κB ligand) in mammary-gland epithelial cells. Genetic inactivation of the RANKL receptor RANK in mammary-gland epithelial cells prevents MPA-induced epithelial proliferation, impairs expansion of the CD49f(hi) stem-cell-enriched population, and sensitizes these cells to DNA-damage-induced cell death. Deletion of RANK from the mammary epithelium results in a markedly decreased incidence and delayed onset of MPA-driven mammary cancer. These data show that the RANKL/RANK system controls the incidence and onset of progestin-driven breast cancer.
- Published
- 2010
- Full Text
- View/download PDF
48. Respiratory inductive plethysmography as a method for measuring ventilatory parameters in conscious, non-restrained dogs.
- Author
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Murphy DJ, Renninger JP, and Schramek D
- Subjects
- Acepromazine administration & dosage, Acepromazine pharmacology, Analgesics, Opioid administration & dosage, Analgesics, Opioid pharmacology, Animals, Consciousness, Dogs, Dopamine Antagonists administration & dosage, Dopamine Antagonists pharmacology, Doxapram administration & dosage, Doxapram pharmacology, Male, Morphine administration & dosage, Morphine pharmacology, Posture, Respiration, Respiratory Insufficiency physiopathology, Respiratory Rate drug effects, Respiratory System Agents administration & dosage, Respiratory System Agents pharmacology, Telemetry, Tidal Volume drug effects, Plethysmography, Respiratory Physiological Phenomena
- Abstract
Introduction: Assessing the effects of new chemical entities on respiratory function in animal models is an essential component of preclinical drug safety evaluation. Methods currently available for measuring ventilatory parameters in conscious dogs generally utilize a plethysmograph chamber or a face mask equipped with a pneumotachograph attached to the snout of the animal. These methods require restraint and allow for only short, periodic measurements. Because of these limitations, respiratory inductive plethysmography (RIP) was evaluated as a possible new methodology that will allow for continuous monitoring of respiratory parameters in non-restrained dogs for extended periods of time., Methods: Straps containing inductive coils were placed around the thorax and abdomen to measure thoracic and abdominal excursions. The straps were contained within a protective jacket that was placed on the dogs and the electrical signals from the inductive coils were transmitted by telemetry to a receiver. The data were acquired and analyzed using the Vivometrics(R) LifeShirt(R) PreClinical System. Because postural changes can alter tidal volume measurements using RIP, the jacket also contained an accelerometer that was used to record postural changes during ventilatory measurements., Results: Measurement of ventilatory parameters in dogs following manual placement in different positions (e.g., standing, sitting, lateral recumbent) or during the different postures in non-restrained dogs demonstrated that changes in posture had only a minimal influence (=10% difference) on tidal volume measurements in conscious dogs. Conscious, restrained male beagle dogs given a single intravenous dose of 0.25mg/kg morphine (respiratory depressant) or exposed to a gas mixture containing 8% CO(2) (respiratory stimulant) had values for tidal volume, respiratory rate and minute volume obtained using RIP that were within 7, 3 and 9% of values obtained using a facemask with an attached pneumotachograph. All of the expected changes in tidal volume, respiratory rate and minute volume were also detected in conscious, non-restrained male dogs using RIP following a single intravenous dose of 10mg/kg doxapram (respiratory stimulant), a single intravenous dose of 0.5mg/kg acepromazine (respiratory stimulant) and a single subcutaneous dose of 2mg/kg morphine (respiratory depressant)., Discussion: The results of this study demonstrate that RIP is an acceptable method for measuring ventilatory parameters in conscious non-restrained dogs. RIP expands current methodologies in that it allows for continuous monitoring of ventilatory parameters over extended periods of time. This added capability will allow for respiratory monitoring during both the awake and sleep states, which is significant since control of respiratory drive differs between the awake and sleep states and treatment related effects such as sleep apnea or sleep disordered breathing can have adverse health consequences. Combined with cardiovascular telemetry, this methodology will also allow for the combined monitoring of cardiovascular and respiratory parameters in conscious, non-restrained dogs., (Copyright 2010 Elsevier Inc. All rights reserved.)
- Published
- 2010
- Full Text
- View/download PDF
49. Drosophila genome-wide obesity screen reveals hedgehog as a determinant of brown versus white adipose cell fate.
- Author
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Pospisilik JA, Schramek D, Schnidar H, Cronin SJ, Nehme NT, Zhang X, Knauf C, Cani PD, Aumayr K, Todoric J, Bayer M, Haschemi A, Puviindran V, Tar K, Orthofer M, Neely GG, Dietzl G, Manoukian A, Funovics M, Prager G, Wagner O, Ferrandon D, Aberger F, Hui CC, Esterbauer H, and Penninger JM
- Subjects
- Adipocytes, Brown metabolism, Adipocytes, White metabolism, Adipogenesis, Animals, Cyclic AMP metabolism, Glucocorticoids metabolism, Humans, Mice, Mice, Knockout, Muscle Cells metabolism, Repressor Proteins genetics, Drosophila Proteins metabolism, Hedgehog Proteins metabolism, Obesity genetics
- Abstract
Over 1 billion people are estimated to be overweight, placing them at risk for diabetes, cardiovascular disease, and cancer. We performed a systems-level genetic dissection of adiposity regulation using genome-wide RNAi screening in adult Drosophila. As a follow-up, the resulting approximately 500 candidate obesity genes were functionally classified using muscle-, oenocyte-, fat-body-, and neuronal-specific knockdown in vivo and revealed hedgehog signaling as the top-scoring fat-body-specific pathway. To extrapolate these findings into mammals, we generated fat-specific hedgehog-activation mutant mice. Intriguingly, these mice displayed near total loss of white, but not brown, fat compartments. Mechanistically, activation of hedgehog signaling irreversibly blocked differentiation of white adipocytes through direct, coordinate modulation of early adipogenic factors. These findings identify a role for hedgehog signaling in white/brown adipocyte determination and link in vivo RNAi-based scanning of the Drosophila genome to regulation of adipocyte cell fate in mammals.
- Published
- 2010
- Full Text
- View/download PDF
50. Phosphorylation-dependent binding of 14-3-3 terminates signalling by the Gab2 docking protein.
- Author
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Brummer T, Larance M, Herrera Abreu MT, Lyons RJ, Timpson P, Emmerich CH, Fleuren ED, Lehrbach GM, Schramek D, Guilhaus M, James DE, and Daly RJ
- Subjects
- 14-3-3 Proteins chemistry, Adaptor Proteins, Signal Transducing chemistry, Adaptor Proteins, Signal Transducing genetics, Amino Acid Sequence, Binding Sites genetics, Cell Line, Feedback, Physiological, Humans, Models, Biological, Molecular Sequence Data, Multiprotein Complexes chemistry, Mutagenesis, Site-Directed, Phosphorylation, Protein Binding, Recombinant Proteins chemistry, Recombinant Proteins genetics, Recombinant Proteins metabolism, Sequence Homology, Amino Acid, Signal Transduction, 14-3-3 Proteins metabolism, Adaptor Proteins, Signal Transducing metabolism
- Abstract
Grb2-associated binder (Gab)2 functions downstream of a variety of receptor and cytoplasmic tyrosine kinases as a docking platform for specific signal transducers and performs important functions in both normal physiology and oncogenesis. Gab2 signalling is promoted by its association with specific receptors through the adaptor Grb2. However, the molecular mechanisms that attenuate Gab2 signals have remained unclear. We now demonstrate that growth factor-induced phosphorylation of Gab2 on two residues, S210 and T391, leads to recruitment of 14-3-3 proteins. Together, these events mediate negative-feedback regulation, as Gab2(S210A/T391A) exhibits sustained receptor association and signalling and promotes cell proliferation and transformation. Importantly, introduction of constitutive 14-3-3-binding sites into Gab2 renders it refractory to receptor activation, demonstrating that site-selective binding of 14-3-3 proteins is sufficient to terminate Gab2 signalling. Furthermore, this is associated with reduced binding of Grb2. This leads to a model where signal attenuation occurs because 14-3-3 promotes dissociation of Gab2 from Grb2, and thereby uncouples Gab2 from the receptor complex. This represents a novel regulatory mechanism with implications for diverse tyrosine kinase signalling systems.
- Published
- 2008
- Full Text
- View/download PDF
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