83 results on '"Heissmeyer, V."'
Search Results
2. A human immune dysregulation syndrome characterized by severe hyperinflammation with a homozygous nonsense Roquin-1 mutation
- Author
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Tavernier, S. J., Athanasopoulos, V., Verloo, P., Behrens, G., Staal, J., Bogaert, D. J., Naesens, L., De Bruyne, M., Van Gassen, S., Parthoens, E., Ellyard, J., Cappello, J., Morris, L. X., Van Gorp, H., Van Isterdael, G., Saeys, Y., Lamkanfi, M., Schelstraete, P., Dehoorne, J., Bordon, V., Van Coster, R., Lambrecht, B. N., Menten, B., Beyaert, R., Vinuesa, C. G., Heissmeyer, V., Dullaers, M., and Haerynck, F.
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- 2019
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3. Author Correction: A human immune dysregulation syndrome characterized by severe hyperinflammation with a homozygous nonsense Roquin-1 mutation
- Author
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Tavernier, S. J., Athanasopoulos, V., Verloo, P., Behrens, G., Staal, J., Bogaert, D. J., Naesens, L., De Bruyne, M., Van Gassen, S., Parthoens, E., Ellyard, J., Cappello, J., Morris, L. X., Van Gorp, H., Van Isterdael, G., Saeys, Y., Lamkanfi, M., Schelstraete, P., Dehoorne, J., Bordon, V., Van Coster, R., Lambrecht, B. N., Menten, B., Beyaert, R., Vinuesa, C. G., Heissmeyer, V., Dullaers, M., and Haerynck, F.
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- 2019
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4. Transcription factor NF-κB is constitutively activated in acute lymphoblastic leukemia cells
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Kordes, U, Krappmann, D, Heissmeyer, V, Ludwig, WD, and Scheidereit, C
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- 2000
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5. Author Correction: A human immune dysregulation syndrome characterized by severe hyperinflammation with a homozygous nonsense Roquin-1 mutation (Nature Communications, (2019), 10, 1, (4779), 10.1038/s41467-019-12704-6)
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Tavernier, S.J. (Simon), Athanasopoulos, V. (V.), Verloo, P. (Patrick), Behrens, G. (G.), Staal, J. (J.), Bogaert, D.J.A. (Delfien), Naesens, L. (L.), De Bruyne, M. (M.), Van Gassen, S. (S.), Parthoens, E. (Eef), Ellyard, J. (J.), Cappello, J. (J.), Morris, L.X. (L. X.), Van Gorp, H. (H.), Van Isterdael, G. (Gert), Saeys, Y. (Yvan), Lamkanfi, M. (Mohamed), Schelstraete, P. (P.), Dehoorne, J. (J.), Bordon, V., Coster, R.N.A. (R. N A) van, Lambrecht, B.N.M. (Bart), Menten, B., Beyaert, R. (Rudi), Vinuesa, C.G. (C. G.), Heissmeyer, V. (V.), Dullaers, M., Haerynck, F. (F.), Tavernier, S.J. (Simon), Athanasopoulos, V. (V.), Verloo, P. (Patrick), Behrens, G. (G.), Staal, J. (J.), Bogaert, D.J.A. (Delfien), Naesens, L. (L.), De Bruyne, M. (M.), Van Gassen, S. (S.), Parthoens, E. (Eef), Ellyard, J. (J.), Cappello, J. (J.), Morris, L.X. (L. X.), Van Gorp, H. (H.), Van Isterdael, G. (Gert), Saeys, Y. (Yvan), Lamkanfi, M. (Mohamed), Schelstraete, P. (P.), Dehoorne, J. (J.), Bordon, V., Coster, R.N.A. (R. N A) van, Lambrecht, B.N.M. (Bart), Menten, B., Beyaert, R. (Rudi), Vinuesa, C.G. (C. G.), Heissmeyer, V. (V.), Dullaers, M., and Haerynck, F. (F.)
- Abstract
The original version of the Supplementary Information associated with this Article included an incorrect Supplementary Information file, in which only the first page of the file was included. The HTML has been updated to include a corrected and complete version of the Supplementary Information file.
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- 2019
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6. A human immune dysregulation syndrome characterized by severe hyperinflammation with a homozygous nonsense Roquin-1 mutation
- Author
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Tavernier, S.J. (Simon), Athanasopoulos, V. (V.), Verloo, P. (Patrick), Behrens, G. (G.), Staal, J. (J.), Bogaert, D.J.A. (Delfien), Naesens, L. (L.), De Bruyne, M. (M.), Van Gassen, S. (S.), Parthoens, E. (Eef), Ellyard, J. (J.), Cappello, J. (J.), Morris, L.X. (L. X.), Van Gorp, H. (H.), Van Isterdael, G. (Gert), Saeys, Y. (Yvan), Lamkanfi, M. (Mohamed), Schelstraete, P. (P.), Dehoorne, J. (J.), Bordon, V., Coster, R.N.A. (R. N A) van, Lambrecht, B.N.M. (Bart), Menten, B., Beyaert, R. (Rudi), Vinuesa, C.G. (C. G.), Heissmeyer, V. (V.), Dullaers, M., Haerynck, F. (F.), Tavernier, S.J. (Simon), Athanasopoulos, V. (V.), Verloo, P. (Patrick), Behrens, G. (G.), Staal, J. (J.), Bogaert, D.J.A. (Delfien), Naesens, L. (L.), De Bruyne, M. (M.), Van Gassen, S. (S.), Parthoens, E. (Eef), Ellyard, J. (J.), Cappello, J. (J.), Morris, L.X. (L. X.), Van Gorp, H. (H.), Van Isterdael, G. (Gert), Saeys, Y. (Yvan), Lamkanfi, M. (Mohamed), Schelstraete, P. (P.), Dehoorne, J. (J.), Bordon, V., Coster, R.N.A. (R. N A) van, Lambrecht, B.N.M. (Bart), Menten, B., Beyaert, R. (Rudi), Vinuesa, C.G. (C. G.), Heissmeyer, V. (V.), Dullaers, M., and Haerynck, F. (F.)
- Abstract
Hyperinflammatory syndromes are life-threatening disorders caused by overzealous immune cell activation and cytokine release, often resulting from defects in negative feedback mechanisms. In the quintessential hyperinflammatory syndrome familial hemophagocytic lymphohistiocytosis (HLH), inborn errors of cytotoxicity result in effector cell accumulation, immune dysregulation and, if untreated, tissue damage and death. Here, we describe a human case with a homozygous nonsense R688* RC3H1 mutation suffering from hyperinflammation, presenting as relapsing HLH. RC3H1 encodes Roquin-1, a posttranscriptional repressor of immune-regulatory proteins such as ICOS, OX40 and TNF. Comparing the R688* variant with the murine M199R variant reveals a phenotypic resemblance, both in immune cell activation, hypercytokinemia and disease development. Mechanistically, R688* Roquin-1 fails to localize to P-bodies and interact with the CCR4-NOT deadenylation complex, impeding mRNA decay and dysregulating cytokine production. The results from this unique case suggest that impaired Roquin-1 function provokes hyperinflammation by a failure to quench immune activation.
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- 2019
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7. A human immune dysregulation syndrome characterized by severe hyperinflammation with a homozygous nonsense Roquin-1 mutation
- Author
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Tavernier, SJ, Athanasopoulos, V, Verloo, P, Behrens, G, Staal, J, Bogaert, DJ, Naesens, L, de Bruyne, M, Van Gassen, S, Parthoens, E, Ellyard, J, Cappello, J, Morris, LX, Van Gorp, H, Van Isterdael, G, Saeys, Y, Lamkanfi, M, Schelstraete, P, Dehoorne, J, Bordon, V, Van Coster, R, Lambrecht, Bart, Menten, B, Beyaert, R, Vinuesa, CG, Heissmeyer, V, Dullaers, M, Haerynck, F, Tavernier, SJ, Athanasopoulos, V, Verloo, P, Behrens, G, Staal, J, Bogaert, DJ, Naesens, L, de Bruyne, M, Van Gassen, S, Parthoens, E, Ellyard, J, Cappello, J, Morris, LX, Van Gorp, H, Van Isterdael, G, Saeys, Y, Lamkanfi, M, Schelstraete, P, Dehoorne, J, Bordon, V, Van Coster, R, Lambrecht, Bart, Menten, B, Beyaert, R, Vinuesa, CG, Heissmeyer, V, Dullaers, M, and Haerynck, F
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- 2019
8. Posttranscriptional Gene Regulation of T Follicular Helper Cells by RNA-Binding Proteins and microRNAs
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Baumjohann, D. and Heissmeyer, V.
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CD4-Positive T-Lymphocytes ,Immunology ,Models, Immunological ,RNA-Binding Proteins ,T follicular regulatory ,Cell Differentiation ,Review ,T-Lymphocytes, Helper-Inducer ,T Follicular Helper ,T Follicular Regulatory ,Roquin ,Regnase-1 ,Micrornas ,Mir-17-92 ,Mir-155 ,Mir-146a ,microRNAs ,miR-155 ,miR-146a ,regnase-1 ,Gene Expression Regulation ,T follicular helper ,Immunology and Allergy ,Animals ,Humans ,miR-17–92 ,Signal Transduction - Abstract
T follicular helper (Tfh) cells are critically involved in the establishment of potent antibody responses against infectious pathogens, such as viruses and bacteria, but their dysregulation may also result in aberrant antibody responses that frequently coincide with autoimmune diseases or allergies. The fate and identity of Tfh cells is tightly controlled by gene regulation on the transcriptional and posttranscriptional level. Here, we provide deeper insights into the posttranscriptional mechanisms that regulate Tfh cell differentiation, function, and plasticity through the actions of RNA-binding proteins (RBPs) and small endogenously expressed regulatory RNAs called microRNAs (miRNAs). The Roquin family of RBPs has been shown to dampen spontaneous activation and differentiation of naive CD4(+) T cells into Tfh cells, since CD4(+) T cells with Roquin mutations accumulate as Tfh cells and provide inappropriate B cell help in the production of autoantibodies. Moreover, Regnase-1, an endoribonuclease that regulates a set of targets, which strongly overlaps with that of Roquin, is crucial for the prevention of autoantibody production. Interestingly, both Roquin and Regnase-1 proteins are cleaved and inactivated after TCR stimulation by the paracaspase MALT1. miRNAs are expressed in naive CD4(+) T cells and help preventing spontaneous differentiation into effector cells. While most miRNAs are downregulated upon T cell activation, several miRNAs have been shown to regulate the fate of these cells by either promoting (e.g., miR-17-92 and miR-155) or inhibiting (e.g., miR-146a) Tfh cell differentiation. Together, these different aspects highlight a complex and dynamic regulatory network of posttranscriptional gene regulation in Tfh cells that may also be active in other T helper cell populations, including Th1, Th2, Th17, and Treg.
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- 2018
9. Roquin targets mRNAs in a 3 '-UTR-specific manner by different modes of regulation
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Essig, K., Kronbeck, N., Guimaraes, J.C., Lohs, C., Schlundt, A., Hoffmann, A.L., Behrens, G., Brenner, S., Kowalska, J., Lopez-Rodriguez, C., Jemielity, J., Holtmann, H., Reiche, K., Hackermüller, J., Sattler, M., Zavolan, M., and Heissmeyer, V.
- Abstract
The RNA-binding proteins Roquin-1 and Roquin-2 redundantly control gene expression and cell-fate decisions. Here, we show that Roquin not only interacts with stem-loop structures, but also with a linear sequence element present in about half of its targets. Comprehensive analysis of a minimal response element of the Nfkbid 3'-UTR shows that six stem-loop structures cooperate to exert robust and profound post-transcriptional regulation. Only binding of multiple Roquin proteins to several stem-loops exerts full repression, which redundantly involved deadenylation and decapping, but also translational inhibition. Globally, most Roquin targets are regulated by mRNA decay, whereas a small subset, including the Nfat5 mRNA, with more binding sites in their 3'-UTRs, are also subject to translational inhibition. These findings provide insights into how the robustness and magnitude of Roqu-inmediated regulation is encoded in complex cis-elements.
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- 2018
10. Binding of NUFIP2 to Roquin promotes recognition and regulation of ICOS mRNA
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Rehage, N. (Nina), Davydova, E. (Elena), Conrad, C. (Christine), Behrens, G. (Gesine), Maiser, A. (Andreas), Stehklein, J.E. (Jenny E.), Brenner, S. (Sven), Klein, J. (Juliane), Jeridi, A. (Aicha), Hoffmann, A. (Anne), Lee, E. (Eunhae), Dianzani, U. (Umberto), Willemsen, R. (Rob), Feederle, R. (Regina), Reiche, K. (Kristin), Hackermüller, J. (Jörg), Leonhardt, H. (Heinrich), Sharma, S. (Sonia), Niessing, D. (Dierk), Heissmeyer, V. (Vigo), Rehage, N. (Nina), Davydova, E. (Elena), Conrad, C. (Christine), Behrens, G. (Gesine), Maiser, A. (Andreas), Stehklein, J.E. (Jenny E.), Brenner, S. (Sven), Klein, J. (Juliane), Jeridi, A. (Aicha), Hoffmann, A. (Anne), Lee, E. (Eunhae), Dianzani, U. (Umberto), Willemsen, R. (Rob), Feederle, R. (Regina), Reiche, K. (Kristin), Hackermüller, J. (Jörg), Leonhardt, H. (Heinrich), Sharma, S. (Sonia), Niessing, D. (Dierk), and Heissmeyer, V. (Vigo)
- Abstract
The ubiquitously expressed RNA-binding proteins Roquin-1 and Roquin-2 are essential for appropriate immune cell function and postnatal survival of mice. Roquin proteins repress target mRNAs by recognizing secondary structures in their 3′-UTRs and by inducing mRNA decay. However, it is unknown if other cellular proteins contribute to target control. To identify cofactors of Roquin, we used RNA interference to screen ~1500 genes involved in RNA-binding or mRNA degradation, and identified NUFIP2 as a cofactor of Roquin-induced mRNA decay. NUFIP2 binds directly and with high affinity to Roquin, which stabilizes NUFIP2 in cells. Post-transcriptional repression of human ICOS by endogenous Roquin proteins requires two neighboring non-canonical stem-loops in the ICOS 3′-UTR. This unconventional cis-element as well as another tandem loop known to confer Roquin-mediated regulation of the Ox40 3′-UTR, are bound cooperatively by Roquin and NUFIP2. NUFIP2 therefore emerges as a cofactor that contributes to mRNA target recognition by Roquin.
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- 2018
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11. A translational silencing function of MCPIP1/Regnase-1 specified by the target site context
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Behrens, G., Winzen, R., Rehage, N., Dörrie, A., Barsch, M., Hoffmann, Anne, Hackermüller, Jörg, Tiedje, C., Heissmeyer, V., Holtmann, H., Behrens, G., Winzen, R., Rehage, N., Dörrie, A., Barsch, M., Hoffmann, Anne, Hackermüller, Jörg, Tiedje, C., Heissmeyer, V., and Holtmann, H.
- Abstract
The expression of proteins during inflammatory and immune reactions is coordinated by post-transcriptional mechanisms. A particularly strong suppression of protein expression is exerted by a conserved translational silencing element (TSE) identified in the 3′ UTR of NFKBIZ mRNA, which is among the targets of the RNA-binding proteins Roquin-1/2 and MCPIP1/Regnase-1. We present evidence that in the context of the TSE MCPIP1, so far known for its endonuclease activity toward mRNAs specified by distinct stem–loop (SL) structures, also suppresses translation. Overexpression of MCPIP1 silenced translation in a TSE-dependent manner and reduced ribosome occupancy of the mRNA. Correspondingly, MCPIP1 depletion alleviated silencing and increased polysomal association of the mRNA. Translationally silenced NFKBIZ or reporter mRNAs were mostly capped, polyadenylated and ribosome associated. Furthermore, MCPIP1 silenced also cap-independent, CrPV-IRES-dependent translation. This suggests that MCPIP1 suppresses a post-initiation step. The TSE is predicted to form five SL structures. SL4 and 5 resemble target structures reported for MCPIP1 and together were sufficient for MCPIP1 binding and mRNA destabilization. Translational silencing, however, required SL1–3 in addition. Thus the NFKBIZ TSE functions as an RNA element in which sequences adjacent to the site of interaction with MCPIP1 and dispensable for accelerated mRNA degradation extend the functional repertoire of MCPIP1 to translational silencing.
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- 2018
12. Binding of NUFIP2 to Roquin promotes recognition and regulation of ICOS mRNA
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Rehage, N., Davydova, E., Conrad, C., Behrens, G., Maiser, A., Stehklein, J.E., Brenner, S., Klein, J., Jeridi, A., Hoffmann, Anne, Lee, E., Dianzani, U., Willemsen, R., Feederle, R., Reiche, K., Hackermüller, Jörg, Leonhardt, H., Sharma, S., Niessing, D., Heissmeyer, V., Rehage, N., Davydova, E., Conrad, C., Behrens, G., Maiser, A., Stehklein, J.E., Brenner, S., Klein, J., Jeridi, A., Hoffmann, Anne, Lee, E., Dianzani, U., Willemsen, R., Feederle, R., Reiche, K., Hackermüller, Jörg, Leonhardt, H., Sharma, S., Niessing, D., and Heissmeyer, V.
- Abstract
The ubiquitously expressed RNA-binding proteins Roquin-1 and Roquin-2 are essential for appropriate immune cell function and postnatal survival of mice. Roquin proteins repress target mRNAs by recognizing secondary structures in their 3′-UTRs and by inducing mRNA decay. However, it is unknown if other cellular proteins contribute to target control. To identify cofactors of Roquin, we used RNA interference to screen ~1500 genes involved in RNA-binding or mRNA degradation, and identified NUFIP2 as a cofactor of Roquin-induced mRNA decay. NUFIP2 binds directly and with high affinity to Roquin, which stabilizes NUFIP2 in cells. Post-transcriptional repression of human ICOS by endogenous Roquin proteins requires two neighboring non-canonical stem-loops in the ICOS 3′-UTR. This unconventional cis-element as well as another tandem loop known to confer Roquin-mediated regulation of the Ox40 3′-UTR, are bound cooperatively by Roquin and NUFIP2. NUFIP2 therefore emerges as a cofactor that contributes to mRNA target recognition by Roquin.
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- 2018
13. Binding of NUFIP2 to Roquin promotes recognition and regulation of ICOS mRNA
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Rehage, N, Davydova, E, Conrad, C, Behrens, G, Maiser, A, Stehklein, JE, Brenner, S, de Klein, Annelies, Jeridi, A, Hoffmann, A, Lee, E, Dianzani, U, Willemsen, Rob, Feederle, R, Reiche, K, Hackermuller, J, Leonhardt, H, Sharma, S, Niessing, D, Heissmeyer, V, Rehage, N, Davydova, E, Conrad, C, Behrens, G, Maiser, A, Stehklein, JE, Brenner, S, de Klein, Annelies, Jeridi, A, Hoffmann, A, Lee, E, Dianzani, U, Willemsen, Rob, Feederle, R, Reiche, K, Hackermuller, J, Leonhardt, H, Sharma, S, Niessing, D, and Heissmeyer, V
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- 2018
14. Roquin recognizes a non-canonical hexaloop structure in the 3'-UTR of Ox40
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Janowski, R., Heinz, G.A., Schlundt, A., Wommelsdorf, N., Brenner, S., Gruber, A.R., Blank, M., Buch, T., Buhmann, R., Zavolan, M., Niessing, D., Heissmeyer, V., and Sattler, M.
- Abstract
The RNA-binding protein Roquin is required to prevent autoimmunity. Roquin controls T-helper cell activation and differentiation by limiting the induced expression of costimulatory receptors such as tumor necrosis factor receptor superfamily 4 (Tnfrs4 or Ox40). A constitutive decay element (CDE) with a characteristic triloop hairpin was previously shown to be recognized by Roquin. Here we use SELEX assays to identify a novel U-rich hexaloop motif, representing an alternative decay element (ADE). Crystal structures and NMR data show that the Roquin-1 ROQ domain recognizes hexaloops in the SELEX-derived ADE and in an ADE-like variant present in the Ox40 3'-UTR with identical binding modes. In cells, ADE-like and CDE-like motifs cooperate in the repression of Ox40 by Roquin. Our data reveal an unexpected recognition of hexaloop cis elements for the posttranscriptional regulation of target messenger RNAs by Roquin.
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- 2016
15. Roquin binds microRNA-146a and Argonaute2 to regulate microRNA homeostasis
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Srivastava, M, Duan, G, Kershaw, NJ, Athanasopoulos, V, Yeo, JHC, Ose, T, Hu, D, Brown, SHJ, Jergic, S, Patel, HR, Pratama, A, Richards, S, Verma, A, Jones, EY, Heissmeyer, V, Preiss, T, Dixon, NE, Chong, MMW, Babon, JJ, Vinuesa, CG, Srivastava, M, Duan, G, Kershaw, NJ, Athanasopoulos, V, Yeo, JHC, Ose, T, Hu, D, Brown, SHJ, Jergic, S, Patel, HR, Pratama, A, Richards, S, Verma, A, Jones, EY, Heissmeyer, V, Preiss, T, Dixon, NE, Chong, MMW, Babon, JJ, and Vinuesa, CG
- Abstract
Roquin is an RNA-binding protein that prevents autoimmunity and inflammation via repression of bound target mRNAs such as inducible costimulator (Icos). When Roquin is absent or mutated (Roquin(san)), Icos is overexpressed in T cells. Here we show that Roquin enhances Dicer-mediated processing of pre-miR-146a. Roquin also directly binds Argonaute2, a central component of the RNA-induced silencing complex, and miR-146a, a microRNA that targets Icos mRNA. In the absence of functional Roquin, miR-146a accumulates in T cells. Its accumulation is not due to increased transcription or processing, rather due to enhanced stability of mature miR-146a. This is associated with decreased 3' end uridylation of the miRNA. Crystallographic studies reveal that Roquin contains a unique HEPN domain and identify the structural basis of the 'san' mutation and Roquin's ability to bind multiple RNAs. Roquin emerges as a protein that can bind Ago2, miRNAs and target mRNAs, to control homeostasis of both RNA species.
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- 2015
16. Wedelolactone suppresses LPS-induced caspase-11 expression by directly inhibiting the IKK Complex
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Kobori, M, primary, Yang, Z, additional, Gong, D, additional, Heissmeyer, V, additional, Zhu, H, additional, Jung, Y-K, additional, Gakidis, M Angelica M, additional, Rao, A, additional, Sekine, T, additional, Ikegami, F, additional, Yuan, C, additional, and Yuan, J, additional
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- 2003
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17. NF-kappa B p105 is a target of Ikappa B kinases and controls signal induction of Bcl-3-p50 complexes
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Heissmeyer, V., primary
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- 1999
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18. Wedelolactone suppresses LPS-induced caspase-11 expression by directly inhibiting the IKK Complex.
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Kobori, M., Yang, Z., Gong, D., Heissmeyer, V., Zhu, H., Jung, Y.-K., Gakidis, M. Angelica M., Rao, A., Sekine, T., Ikegami, F., Yuan, C., and Yuan, J.
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ENDOTOXINS ,MICROBIAL polysaccharides ,CYTOKINES ,APOPTOSIS ,PHOSPHORYLATION ,PROSTAGLANDINS - Abstract
Caspase-11 is a key regulator of proinflammatory cytokine IL-1ß maturation and pathological apoptosis. Caspase-11 is not expressed in most tissues under normal condition, but highly inducible upon pathological stimulation such as in the presence of lipopolysaccharide (LPS). Here, we describe the identification and characterization of wedelolactone, a natural compound that inhibits LPS-induced caspase-11 expression in cultured cells by inhibiting NF-?B-mediated transcription. We demonstrate that wedelolactone is an inhibitor of IKK, a kinase critical for activation of NF-?B by mediating phosphorylation and degradation of I?Ba.Cell Death and Differentiation (2004) 11, 123-130. doi:10.1038/sj.cdd.4401325 Published online 3 October 2003 [ABSTRACT FROM AUTHOR]
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- 2004
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19. Correction to "The thymocyte-specific RNA-binding protein Arpp21 provides TCR repertoire diversity by binding to the 3'-UTR and promoting Rag1 mRNA expression".
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Xu M, Ito-Kureha T, Kang HS, Chernev A, Raj T, Hoefig KP, Hohn C, Giesert F, Wang Y, Pan W, Ziętara N, Straub T, Feederle R, Daniel C, Adler B, König J, Feske S, Tsokos GC, Wurst W, Urlaub H, Sattler M, Kisielow J, Wulczyn FG, Łyszkiewicz M, and Heissmeyer V
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- 2024
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20. Aloperine Suppresses Cancer Progression by Interacting with VPS4A to Inhibit Autophagosome-lysosome Fusion in NSCLC.
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Guo W, Zhou H, Wang J, Lu J, Dong Y, Kang Z, Qiu X, Ouyang X, Chen Q, Li J, Cheng X, Du K, Li M, Lin Z, Jin M, Zhang L, Sarapultsev A, Shi K, Li F, Zhang G, Wu K, Rong Y, Heissmeyer V, Liu Y, Li Y, Huang K, Luo S, and Hu D
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- Animals, Mice, Humans, Cell Line, Tumor, Disease Models, Animal, Vesicular Transport Proteins metabolism, Vesicular Transport Proteins genetics, Disease Progression, Cell Proliferation drug effects, Autophagy drug effects, Apoptosis drug effects, Carcinoma, Non-Small-Cell Lung metabolism, Carcinoma, Non-Small-Cell Lung drug therapy, Carcinoma, Non-Small-Cell Lung genetics, Carcinoma, Non-Small-Cell Lung pathology, Autophagosomes metabolism, Autophagosomes drug effects, Lung Neoplasms metabolism, Lung Neoplasms drug therapy, Lung Neoplasms genetics, Lung Neoplasms pathology, Lysosomes metabolism, Lysosomes drug effects, Quinolizidines pharmacology
- Abstract
Aloperine (ALO), a quinolizidine-type alkaloid isolated from a natural Chinese herb, has shown promising antitumor effects. Nevertheless, its common mechanism of action and specific target remain elusive. Here, it is demonstrated that ALO inhibits the proliferation and migration of non-small cell lung cancer cell lines in vitro and the tumor development in several mouse tumor models in vivo. Mechanistically, ALO inhibits the fusion of autophagosomes with lysosomes and the autophagic flux, leading to the accumulation of sequestosome-1 (SQSTM1) and production of reactive oxygen species (ROS), thereby inducing tumor cell apoptosis and preventing tumor growth. Knockdown of SQSTM1 in cells inhibits ROS production and reverses ALO-induced cell apoptosis. Furthermore, VPS4A is identified as a direct target of ALO, and the amino acids F153 and D263 of VPS4A are confirmed as the binding sites for ALO. Knockout of VPS4A in H1299 cells demonstrates a similar biological effect as ALO treatment. Additionally, ALO enhances the efficacy of the anti-PD-L1/TGF-β bispecific antibody in inhibiting LLC-derived subcutaneous tumor models. Thus, ALO is first identified as a novel late-stage autophagy inhibitor that triggers tumor cell death by targeting VPS4A., (© 2024 The Author(s). Advanced Science published by Wiley‐VCH GmbH.)
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- 2024
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21. Crosstalk between Regnase-1 and -3 shapes mast cell survival and cytokine expression.
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Bataclan M, Leoni C, Moro SG, Pecoraro M, Wong EH, Heissmeyer V, and Monticelli S
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- Animals, Mice, Humans, Ribonuclease, Pancreatic metabolism, Ribonuclease, Pancreatic genetics, Ribonucleases metabolism, Ribonucleases genetics, Gene Expression Regulation, RNA-Binding Proteins metabolism, RNA-Binding Proteins genetics, Mice, Inbred C57BL, Cell Proliferation, Inflammation metabolism, Transcription Factors, Mast Cells metabolism, Cytokines metabolism, Cell Survival genetics
- Abstract
Post-transcriptional regulation of immune-related transcripts by RNA-binding proteins (RBPs) impacts immune cell responses, including mast cell functionality. Despite their importance in immune regulation, the functional role of most RBPs remains to be understood. By manipulating the expression of specific RBPs in murine mast cells, coupled with mass spectrometry and transcriptomic analyses, we found that the Regnase family of proteins acts as a potent regulator of mast cell physiology. Specifically, Regnase-1 is required to maintain basic cell proliferation and survival, whereas both Regnase-1 and -3 cooperatively regulate the expression of inflammatory transcripts upon activation, with Tnf being a primary target in both human and mouse cells. Furthermore, Regnase-3 directly interacts with Regnase-1 in mast cells and is necessary to restrain Regnase-1 expression through the destabilization of its transcript. Overall, our study identifies protein interactors of endogenously expressed Regnase factors, characterizes the regulatory interplay between Regnase family members in mast cells, and establishes their role in the control of mast cell homeostasis and inflammatory responses., (© 2024 Bataclan et al.)
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- 2024
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22. The thymocyte-specific RNA-binding protein Arpp21 provides TCR repertoire diversity by binding to the 3'-UTR and promoting Rag1 mRNA expression.
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Xu M, Ito-Kureha T, Kang HS, Chernev A, Raj T, Hoefig KP, Hohn C, Giesert F, Wang Y, Pan W, Ziętara N, Straub T, Feederle R, Daniel C, Adler B, König J, Feske S, Tsokos GC, Wurst W, Urlaub H, Sattler M, Kisielow J, Wulczyn FG, Łyszkiewicz M, and Heissmeyer V
- Subjects
- Animals, Mice, Cell Differentiation genetics, RNA, Messenger genetics, RNA, Messenger metabolism, RNA-Binding Proteins genetics, RNA-Binding Proteins metabolism, Thymus Gland metabolism, Receptors, Antigen, T-Cell genetics, Receptors, Antigen, T-Cell metabolism, Thymocytes metabolism
- Abstract
The regulation of thymocyte development by RNA-binding proteins (RBPs) is largely unexplored. We identify 642 RBPs in the thymus and focus on Arpp21, which shows selective and dynamic expression in early thymocytes. Arpp21 is downregulated in response to T cell receptor (TCR) and Ca
2+ signals. Downregulation requires Stim1/Stim2 and CaMK4 expression and involves Arpp21 protein phosphorylation, polyubiquitination and proteasomal degradation. Arpp21 directly binds RNA through its R3H domain, with a preference for uridine-rich motifs, promoting the expression of target mRNAs. Analysis of the Arpp21-bound transcriptome reveals strong interactions with the Rag1 3'-UTR. Arpp21-deficient thymocytes show reduced Rag1 expression, delayed TCR rearrangement and a less diverse TCR repertoire. This phenotype is recapitulated in Rag1 3'-UTR mutant mice harboring a deletion of the Arpp21 response region. These findings show how thymocyte-specific Arpp21 promotes Rag1 expression to enable TCR repertoire diversity until signals from the TCR terminate Arpp21 and Rag1 activities., (© 2024. The Author(s).)- Published
- 2024
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23. Unrestrained cleavage of Roquin-1 by MALT1 induces spontaneous T cell activation and the development of autoimmunity.
- Author
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Schmidt H, Raj T, O'Neill TJ, Muschaweckh A, Giesert F, Negraschus A, Hoefig KP, Behrens G, Esser L, Baumann C, Feederle R, Plaza-Sirvent C, Geerlof A, Gewies A, Isay SE, Ruland J, Schmitz I, Wurst W, Korn T, Krappmann D, and Heissmeyer V
- Subjects
- Mice, Animals, Mucosa-Associated Lymphoid Tissue Lymphoma Translocation 1 Protein genetics, Inflammation metabolism, Cell Differentiation, Receptors, Antigen, T-Cell genetics, Ubiquitin-Protein Ligases, Autoimmunity, Encephalomyelitis, Autoimmune, Experimental genetics
- Abstract
Constitutive activation of the MALT1 paracaspase in conventional T cells of Malt1
TBM/TBM (TRAF6 Binding Mutant = TBM) mice causes fatal inflammation and autoimmunity, but the involved targets and underlying molecular mechanisms are unknown. We genetically rendered a single MALT1 substrate, the RNA-binding protein (RBP) Roquin-1, insensitive to MALT1 cleavage. These Rc3h1Mins/Mins mice showed normal immune homeostasis. Combining Rc3h1Mins/Mins alleles with those encoding for constitutively active MALT1 (TBM) prevented spontaneous T cell activation and restored viability of Malt1TBM/TBM mice. Mechanistically, we show how antigen/MHC recognition is translated by MALT1 into Roquin cleavage and derepression of Roquin targets. Increasing T cell receptor (TCR) signals inactivated Roquin more effectively, and only high TCR strength enabled derepression of high-affinity targets to promote Th17 differentiation. Induction of experimental autoimmune encephalomyelitis (EAE) revealed increased cleavage of Roquin-1 in disease-associated Th17 compared to Th1 cells in the CNS. T cells from Rc3h1Mins/Mins mice did not efficiently induce the high-affinity Roquin-1 target IκBNS in response to TCR stimulation, showed reduced Th17 differentiation, and Rc3h1Mins/Mins mice were protected from EAE. These data demonstrate how TCR signaling and MALT1 activation utilize graded cleavage of Roquin to differentially regulate target mRNAs that control T cell activation and differentiation as well as the development of autoimmunity., Competing Interests: Competing interests statement:The authors declare no competing interest.- Published
- 2023
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24. Null and missense mutations of ERI1 cause a recessive phenotypic dichotomy in humans.
- Author
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Guo L, Salian S, Xue JY, Rath N, Rousseau J, Kim H, Ehresmann S, Moosa S, Nakagawa N, Kuroda H, Clayton-Smith J, Wang J, Wang Z, Banka S, Jackson A, Zhang YM, Wei ZJ, Hüning I, Brunet T, Ohashi H, Thomas MF, Bupp C, Miyake N, Matsumoto N, Mendoza-Londono R, Costain G, Hahn G, Di Donato N, Yigit G, Yamada T, Nishimura G, Ansel KM, Wollnik B, Hrabě de Angelis M, Mégarbané A, Rosenfeld JA, Heissmeyer V, Ikegawa S, and Campeau PM
- Subjects
- Humans, Mutation, Missense genetics, RNA, Ribosomal, 5.8S, RNA, RNA, Messenger genetics, Exoribonucleases genetics, Histones genetics
- Abstract
ERI1 is a 3'-to-5' exoribonuclease involved in RNA metabolic pathways including 5.8S rRNA processing and turnover of histone mRNAs. Its biological and medical significance remain unclear. Here, we uncover a phenotypic dichotomy associated with bi-allelic ERI1 variants by reporting eight affected individuals from seven unrelated families. A severe spondyloepimetaphyseal dysplasia (SEMD) was identified in five affected individuals with missense variants but not in those with bi-allelic null variants, who showed mild intellectual disability and digital anomalies. The ERI1 missense variants cause a loss of the exoribonuclease activity, leading to defective trimming of the 5.8S rRNA 3' end and a decreased degradation of replication-dependent histone mRNAs. Affected-individual-derived induced pluripotent stem cells (iPSCs) showed impaired in vitro chondrogenesis with downregulation of genes regulating skeletal patterning. Our study establishes an entity previously unreported in OMIM and provides a model showing a more severe effect of missense alleles than null alleles within recessive genotypes, suggesting a key role of ERI1-mediated RNA metabolism in human skeletal patterning and chondrogenesis., Competing Interests: Declaration of interests The Department of Molecular and Human Genetics at Baylor College of Medicine receives revenue from clinical genetic testing completed at Baylor Genetics Laboratories., (Copyright © 2023 American Society of Human Genetics. Published by Elsevier Inc. All rights reserved.)
- Published
- 2023
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25. The mRNA methyltransferase Mettl3 modulates cytokine mRNA stability and limits functional responses in mast cells.
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Leoni C, Bataclan M, Ito-Kureha T, Heissmeyer V, and Monticelli S
- Subjects
- RNA, Messenger genetics, Quality of Life, Interleukin-13 genetics, RNA Stability genetics, Methyltransferases genetics, Cytokines genetics, Mast Cells
- Abstract
Mast cells are central players in allergy and asthma, and their dysregulated responses lead to reduced quality of life and life-threatening conditions such as anaphylaxis. The RNA modification N
6 -methyladenosine (m6 A) has a prominent impact on immune cell functions, but its role in mast cells remains unexplored. Here, by optimizing tools to genetically manipulate primary mast cells, we reveal that the m6 A mRNA methyltransferase complex modulates mast cell proliferation and survival. Depletion of the catalytic component Mettl3 exacerbates effector functions in response to IgE and antigen complexes, both in vitro and in vivo. Mechanistically, deletion of Mettl3 or Mettl14, another component of the methyltransferase complex, lead to the enhanced expression of inflammatory cytokines. By focusing on one of the most affected mRNAs, namely the one encoding the cytokine IL-13, we find that it is methylated in activated mast cells, and that Mettl3 affects its transcript stability in an enzymatic activity-dependent manner, requiring consensus m6 A sites in the Il13 3'-untranslated region. Overall, we reveal that the m6 A machinery is essential in mast cells to sustain growth and to restrain inflammatory responses., (© 2023. The Author(s).)- Published
- 2023
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26. Critical functions of N 6 -adenosine methylation of mRNAs in T cells.
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Ito-Kureha T and Heissmeyer V
- Subjects
- Methylation, RNA, Messenger genetics, RNA, Messenger metabolism, Adenosine genetics, T-Lymphocytes metabolism
- Abstract
The existence of N
6 -adenosine methylation (m6 A) of mRNA has been known for a long time, but only recently its regulatory potential was uncovered. Current research deciphers the molecular determinants leading to the deposition of this modification and consequences for modified mRNAs. It also evaluates the importance of such modifications for specific cell types and programs. In this review, we summarize the current knowledge on m6 A modification of mRNAs in conventional and regulatory T cells and T-cell-driven immune responses and pathology. We discuss the impact of m6 A modification on T cell activation including cytokine and antigen receptor signaling or sensing of double-stranded RNAs (dsRNA)., Competing Interests: Declaration of competing interest The authors declare that they have no known competing financial interests or personal relationships that could have appeared to influence the work reported in this paper., (Copyright © 2022 The Authors. Published by Elsevier B.V. All rights reserved.)- Published
- 2023
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27. The arginine methyltransferase PRMT7 promotes extravasation of monocytes resulting in tissue injury in COPD.
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Günes Günsel G, Conlon TM, Jeridi A, Kim R, Ertüz Z, Lang NJ, Ansari M, Novikova M, Jiang D, Strunz M, Gaianova M, Hollauer C, Gabriel C, Angelidis I, Doll S, Pestoni JC, Edelmann SL, Kohlhepp MS, Guillot A, Bassler K, Van Eeckhoutte HP, Kayalar Ö, Konyalilar N, Kanashova T, Rodius S, Ballester-López C, Genes Robles CM, Smirnova N, Rehberg M, Agarwal C, Krikki I, Piavaux B, Verleden SE, Vanaudenaerde B, Königshoff M, Dittmar G, Bracke KR, Schultze JL, Watz H, Eickelberg O, Stoeger T, Burgstaller G, Tacke F, Heissmeyer V, Rinkevich Y, Bayram H, Schiller HB, Conrad M, Schneider R, and Yildirim AÖ
- Subjects
- Animals, Arginine metabolism, Histones metabolism, Humans, Intracellular Signaling Peptides and Proteins, Mice, Monocytes metabolism, Protein-Arginine N-Methyltransferases metabolism, Pulmonary Disease, Chronic Obstructive genetics
- Abstract
Extravasation of monocytes into tissue and to the site of injury is a fundamental immunological process, which requires rapid responses via post translational modifications (PTM) of proteins. Protein arginine methyltransferase 7 (PRMT7) is an epigenetic factor that has the capacity to mono-methylate histones on arginine residues. Here we show that in chronic obstructive pulmonary disease (COPD) patients, PRMT7 expression is elevated in the lung tissue and localized to the macrophages. In mouse models of COPD, lung fibrosis and skin injury, reduced expression of PRMT7 associates with decreased recruitment of monocytes to the site of injury and hence less severe symptoms. Mechanistically, activation of NF-κB/RelA in monocytes induces PRMT7 transcription and consequential mono-methylation of histones at the regulatory elements of RAP1A, which leads to increased transcription of this gene that is responsible for adhesion and migration of monocytes. Persistent monocyte-derived macrophage accumulation leads to ALOX5 over-expression and accumulation of its metabolite LTB4, which triggers expression of ACSL4 a ferroptosis promoting gene in lung epithelial cells. Conclusively, inhibition of arginine mono-methylation might offer targeted intervention in monocyte-driven inflammatory conditions that lead to extensive tissue damage if left untreated., (© 2022. The Author(s).)
- Published
- 2022
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28. Cooperation of RNA-Binding Proteins - a Focus on Roquin Function in T Cells.
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Behrens G and Heissmeyer V
- Subjects
- Endoribonucleases metabolism, Gene Expression Regulation, Lymphocyte Activation, RNA, Messenger genetics, RNA-Binding Proteins genetics, RNA-Binding Proteins metabolism, T-Lymphocytes
- Abstract
Post-transcriptional gene regulation by RNA-binding proteins (RBPs) is important in the prevention of inflammatory and autoimmune diseases. With respect to T cell activation and differentiation, the RBPs Roquin-1/2 and Regnase-1 play pivotal roles by inducing degradation and/or translational silencing of target mRNAs. These targets encode important proinflammatory mediators and thus Roquin and Regnase-1 functions dampen cellular programs that can lead to inflammation and autoimmune disease. Recent findings demonstrate direct physical interaction of both RBPs. Here, we propose that cooperativity of trans -acting factors may be more generally used to reinforce the regulatory impact on selected targets and promote specific cell fate decisions. We develop this concept for Roquin and Regnase-1 function in resting and activated T cells and discuss the involvement in autoimmunity as well as how the therapeutic potential can be used in anti-tumor therapies., 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 © 2022 Behrens and Heissmeyer.)
- Published
- 2022
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29. Defining the RBPome of primary T helper cells to elucidate higher-order Roquin-mediated mRNA regulation.
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Hoefig KP, Reim A, Gallus C, Wong EH, Behrens G, Conrad C, Xu M, Kifinger L, Ito-Kureha T, Defourny KAY, Geerlof A, Mautner J, Hauck SM, Baumjohann D, Feederle R, Mann M, Wierer M, Glasmacher E, and Heissmeyer V
- Subjects
- Animals, DNA-Binding Proteins, Gene Expression Regulation, HEK293 Cells, Humans, Inducible T-Cell Co-Stimulator Protein genetics, Mice, Proto-Oncogene Proteins c-vav, STAT1 Transcription Factor, STAT4 Transcription Factor, Signal Transduction, Trans-Activators metabolism, Ubiquitin-Protein Ligases genetics, RNA, Messenger metabolism, RNA-Binding Proteins genetics, RNA-Binding Proteins metabolism, T-Lymphocytes, Helper-Inducer metabolism
- Abstract
Post-transcriptional gene regulation in T cells is dynamic and complex as targeted transcripts respond to various factors. This is evident for the Icos mRNA encoding an essential costimulatory receptor that is regulated by several RNA-binding proteins (RBP), including Roquin-1 and Roquin-2. Here, we identify a core RBPome of 798 mouse and 801 human T cell proteins by utilizing global RNA interactome capture (RNA-IC) and orthogonal organic phase separation (OOPS). The RBPome includes Stat1, Stat4 and Vav1 proteins suggesting unexpected functions for these transcription factors and signal transducers. Based on proximity to Roquin-1, we select ~50 RBPs for testing coregulation of Roquin-1/2 targets by induced expression in wild-type or Roquin-1/2-deficient T cells. Besides Roquin-independent contributions from Rbms1 and Cpeb4 we also show Roquin-1/2-dependent and target-specific coregulation of Icos by Celf1 and Igf2bp3. Connecting the cellular RBPome in a post-transcriptional context, we find contributions from multiple RBPs to the prototypic regulation of mRNA targets by individual trans-acting factors., (© 2021. The Author(s).)
- Published
- 2021
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30. Elevated Exhaustion Levels of NK and CD8 + T Cells as Indicators for Progression and Prognosis of COVID-19 Disease.
- Author
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Li M, Guo W, Dong Y, Wang X, Dai D, Liu X, Wu Y, Li M, Zhang W, Zhou H, Zhang Z, Lin L, Kang Z, Yu T, Tian C, Qin R, Gui Y, Jiang F, Fan H, Heissmeyer V, Sarapultsev A, Wang L, Luo S, and Hu D
- Subjects
- Adult, Aged, CD8-Positive T-Lymphocytes immunology, COVID-19 epidemiology, COVID-19 immunology, COVID-19 virology, China epidemiology, Female, Flow Cytometry, Humans, Killer Cells, Natural immunology, Leukocyte Count, Male, Middle Aged, Pandemics, Prognosis, SARS-CoV-2 genetics, T-Lymphocyte Subsets cytology, T-Lymphocyte Subsets immunology, CD8-Positive T-Lymphocytes cytology, COVID-19 blood, Killer Cells, Natural cytology, SARS-CoV-2 physiology
- Abstract
Background: Severe Acute Respiratory Syndrome Coronavirus 2 (SARS-CoV-2) induced Coronavirus Disease 2019 (COVID-19) has posed a global threat to public health. The immune system is crucial in defending and eliminating the virus and infected cells. However, immune dysregulation may result in the rapid progression of COVID-19. Here, we evaluated the subsets, phenotypic and functional characteristics of natural killer (NK) and T cells in patients with COVID-19 and their associations with disease severity., Methods: Demographic and clinical data of COVID-19 patients enrolled in Wuhan Union Hospital from February 25 to February 27, 2020, were collected and analyzed. The phenotypic and functional characteristics of NK cells and T cells subsets in circulating blood and serum levels of cytokines were analyzed via flow cytometry. Then the LASSO logistic regression model was employed to predict risk factors for the severity of COVID-19., Results: The counts and percentages of NK cells, CD4
+ T cells, CD8+ T cells and NKT cells were significantly reduced in patients with severe symptoms. The cytotoxic CD3- CD56dim CD16+ cell population significantly decreased, while the CD3- CD56dim CD16- part significantly increased in severe COVID-19 patients. More importantly, elevated expression of regulatory molecules, such as CD244 and programmed death-1 (PD-1), on NK cells and T cells, as well as decreased serum cytotoxic effector molecules including perforin and granzyme A, were detected in patients with COVID-19. The serum IL-6, IL-10, and TNF-α were significantly increased in severe patients. Moreover, the CD3- CD56dim CD16- cells were screened out as an influential factor in severe cases by LASSO logistic regression., Conclusions: The functional exhaustion and other subset alteration of NK and T cells may contribute to the progression and improve the prognosis of COVID-19. Surveillance of lymphocyte subsets may in the future enable early screening for signs of critical illness and understanding the pathogenesis of this disease., (Copyright © 2020 Li, Guo, Dong, Wang, Dai, Liu, Wu, Li, Zhang, Zhou, Zhang, Lin, Kang, Yu, Tian, Qin, Gui, Jiang, Fan, Heissmeyer, Sarapultsev, Wang, Luo and Hu.)- Published
- 2020
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31. Validation strategies for antibodies targeting modified ribonucleotides.
- Author
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Weichmann F, Hett R, Schepers A, Ito-Kureha T, Flatley A, Slama K, Hastert FD, Angstman NB, Cardoso MC, König J, Hüttelmaier S, Dieterich C, Canzar S, Helm M, Heissmeyer V, Feederle R, and Meister G
- Subjects
- Nucleotides genetics, RNA genetics, RNA, Messenger genetics, Antibodies genetics, Ribonucleotides genetics
- Abstract
Chemical modifications are found on almost all RNAs and affect their coding and noncoding functions. The identification of m
6 A on mRNA and its important role in gene regulation stimulated the field to investigate whether additional modifications are present on mRNAs. Indeed, modifications including m1 A, m5 C, m7 G, 2'-OMe, and Ψ were detected. However, since their abundances are low and tools used for their corroboration are often not well characterized, their physiological relevance remains largely elusive. Antibodies targeting modified nucleotides are often used but have limitations such as low affinity or specificity. Moreover, they are not always well characterized and due to the low abundance of the modification, particularly on mRNAs, generated data sets might resemble noise rather than specific modification patterns. Therefore, it is critical that the affinity and specificity is rigorously tested using complementary approaches. Here, we provide an experimental toolbox that allows for testing antibody performance prior to their use., (© 2020 Weichmann et al.; Published by Cold Spring Harbor Laboratory Press for the RNA Society.)- Published
- 2020
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32. Immune homeostasis and regulation of the interferon pathway require myeloid-derived Regnase-3.
- Author
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von Gamm M, Schaub A, Jones AN, Wolf C, Behrens G, Lichti J, Essig K, Macht A, Pircher J, Ehrlich A, Davari K, Chauhan D, Busch B, Wurst W, Feederle R, Feuchtinger A, Tschöp MH, Friedel CC, Hauck SM, Sattler M, Geerlof A, Hornung V, Heissmeyer V, Schulz C, Heikenwalder M, and Glasmacher E
- Subjects
- 3' Untranslated Regions, Animals, Autoimmunity, B-Lymphocytes metabolism, Flow Cytometry, Gene Expression Regulation, Macrophages metabolism, Mice, Mice, Inbred C57BL, Mice, Knockout, Myeloid Cells enzymology, Real-Time Polymerase Chain Reaction, Ribonucleases genetics, Signal Transduction, T-Lymphocytes metabolism, Homeostasis immunology, Immunity, Innate, Interferons metabolism, Myeloid Cells metabolism, Ribonucleases metabolism
- Abstract
The RNase Regnase-1 is a master RNA regulator in macrophages and T cells that degrades cellular and viral RNA upon NF-κB signaling. The roles of its family members, however, remain largely unknown. Here, we analyzed Regnase-3 -deficient mice, which develop hypertrophic lymph nodes. We used various mice with immune cell-specific deletions of Regnase-3 to demonstrate that Regnase-3 acts specifically within myeloid cells. Regnase-3 deficiency systemically increased IFN signaling, which increased the proportion of immature B and innate immune cells, and suppressed follicle and germinal center formation. Expression analysis revealed that Regnase-3 and Regnase-1 share protein degradation pathways. Unlike Regnase-1, Regnase-3 expression is high specifically in macrophages and is transcriptionally controlled by IFN signaling. Although direct targets in macrophages remain unknown, Regnase-3 can bind, degrade, and regulate mRNAs, such as Zc3h12a ( Regnase-1 ), in vitro. These data indicate that Regnase-3, like Regnase-1, is an RNase essential for immune homeostasis but has diverged as key regulator in the IFN pathway in macrophages., (© 2019 von Gamm et al.)
- Published
- 2019
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33. Production and Application of Stable Isotope-Labeled Internal Standards for RNA Modification Analysis.
- Author
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Borland K, Diesend J, Ito-Kureha T, Heissmeyer V, Hammann C, Buck AH, Michalakis S, and Kellner S
- Subjects
- Animals, Caenorhabditis elegans, Carbon Isotopes chemistry, Dictyostelium, Escherichia coli, Female, HEK293 Cells, Humans, Mass Spectrometry standards, Mice, Mice, Inbred C57BL, Reference Standards, Saccharomyces cerevisiae, Mass Spectrometry methods, RNA chemistry, RNA Processing, Post-Transcriptional
- Abstract
Post-transcriptional RNA modifications have been found to be present in a wide variety of organisms and in different types of RNA. Nucleoside modifications are interesting due to their already known roles in translation fidelity, enzyme recognition, disease progression, and RNA stability. In addition, the abundance of modified nucleosides fluctuates based on growth phase, external stress, or possibly other factors not yet explored. With modifications ever changing, a method to determine absolute quantities for multiple nucleoside modifications is required. Here, we report metabolic isotope labeling to produce isotopically labeled internal standards in bacteria and yeast. These can be used for the quantification of 26 different modified nucleosides. We explain in detail how these internal standards are produced and show their mass spectrometric characterization. We apply our internal standards and quantify the modification content of transfer RNA (tRNA) from bacteria and various eukaryotes. We can show that the origin of the internal standard has no impact on the quantification result. Furthermore, we use our internal standard for the quantification of modified nucleosides in mouse tissue messenger RNA (mRNA), where we find different modification profiles in liver and brain tissue.
- Published
- 2019
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34. Roquin targets mRNAs in a 3'-UTR-specific manner by different modes of regulation.
- Author
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Essig K, Kronbeck N, Guimaraes JC, Lohs C, Schlundt A, Hoffmann A, Behrens G, Brenner S, Kowalska J, Lopez-Rodriguez C, Jemielity J, Holtmann H, Reiche K, Hackermüller J, Sattler M, Zavolan M, and Heissmeyer V
- Subjects
- Amino Acid Motifs, Animals, Base Sequence, Binding Sites, Cross-Linking Reagents chemistry, HeLa Cells, Humans, Mice, Nucleic Acid Conformation, Protein Binding, Protein Biosynthesis, RNA Stability genetics, RNA, Messenger chemistry, RNA, Messenger genetics, RNA, Messenger metabolism, Response Elements genetics, Ribonucleosides metabolism, Transcriptome genetics, 3' Untranslated Regions genetics, Gene Expression Regulation, Ubiquitin-Protein Ligases metabolism
- Abstract
The RNA-binding proteins Roquin-1 and Roquin-2 redundantly control gene expression and cell-fate decisions. Here, we show that Roquin not only interacts with stem-loop structures, but also with a linear sequence element present in about half of its targets. Comprehensive analysis of a minimal response element of the Nfkbid 3'-UTR shows that six stem-loop structures cooperate to exert robust and profound post-transcriptional regulation. Only binding of multiple Roquin proteins to several stem-loops exerts full repression, which redundantly involved deadenylation and decapping, but also translational inhibition. Globally, most Roquin targets are regulated by mRNA decay, whereas a small subset, including the Nfat5 mRNA, with more binding sites in their 3'-UTRs, are also subject to translational inhibition. These findings provide insights into how the robustness and magnitude of Roquin-mediated regulation is encoded in complex cis-elements.
- Published
- 2018
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- View/download PDF
35. Posttranscriptional regulation of T helper cell fate decisions.
- Author
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Hoefig KP and Heissmeyer V
- Subjects
- Animals, Mice, MicroRNAs physiology, Models, Immunological, Signal Transduction, T-Lymphocytes, Helper-Inducer physiology, Cell Differentiation genetics, RNA Processing, Post-Transcriptional, T-Lymphocytes, Helper-Inducer cytology
- Abstract
T helper cell subsets orchestrate context- and pathogen-specific responses of the immune system. They mostly do so by secreting specific cytokines that attract or induce activation and differentiation of other immune or nonimmune cells. The differentiation of T helper 1 (Th1), Th2, T follicular helper, Th17, and induced regulatory T cell subsets from naive T cells depends on the activation of intracellular signal transduction cascades. These cascades originate from T cell receptor and costimulatory receptor engagement and also receive critical input from cytokine receptors that sample the cytokine milieu within secondary lymphoid organs. Signal transduction then leads to the expression of subset-specifying transcription factors that, in concert with other transcription factors, up-regulate downstream signature genes. Although regulation of transcription is important, recent research has shown that posttranscriptional and posttranslational regulation can critically shape or even determine the outcome of Th cell differentiation. In this review, we describe how specific microRNAs, long noncoding RNAs, RNA-binding proteins, and ubiquitin-modifying enzymes regulate their targets to skew cell fate decisions., (© 2018 Hoefig and Heissmeyer.)
- Published
- 2018
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36. A translational silencing function of MCPIP1/Regnase-1 specified by the target site context.
- Author
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Behrens G, Winzen R, Rehage N, Dörrie A, Barsch M, Hoffmann A, Hackermüller J, Tiedje C, Heissmeyer V, and Holtmann H
- Subjects
- Adaptor Proteins, Signal Transducing, Binding Sites, HeLa Cells, Humans, Peptide Chain Elongation, Translational, Protein Domains, RNA, Messenger metabolism, Receptor, EphB3, Ribonucleases chemistry, Ribosomes metabolism, Transcription Factors chemistry, Gene Silencing, I-kappa B Proteins genetics, Nuclear Proteins genetics, Protein Biosynthesis, Regulatory Sequences, Ribonucleic Acid, Ribonucleases metabolism, Transcription Factors metabolism
- Abstract
The expression of proteins during inflammatory and immune reactions is coordinated by post-transcriptional mechanisms. A particularly strong suppression of protein expression is exerted by a conserved translational silencing element (TSE) identified in the 3' UTR of NFKBIZ mRNA, which is among the targets of the RNA-binding proteins Roquin-1/2 and MCPIP1/Regnase-1. We present evidence that in the context of the TSE MCPIP1, so far known for its endonuclease activity toward mRNAs specified by distinct stem-loop (SL) structures, also suppresses translation. Overexpression of MCPIP1 silenced translation in a TSE-dependent manner and reduced ribosome occupancy of the mRNA. Correspondingly, MCPIP1 depletion alleviated silencing and increased polysomal association of the mRNA. Translationally silenced NFKBIZ or reporter mRNAs were mostly capped, polyadenylated and ribosome associated. Furthermore, MCPIP1 silenced also cap-independent, CrPV-IRES-dependent translation. This suggests that MCPIP1 suppresses a post-initiation step. The TSE is predicted to form five SL structures. SL4 and 5 resemble target structures reported for MCPIP1 and together were sufficient for MCPIP1 binding and mRNA destabilization. Translational silencing, however, required SL1-3 in addition. Thus the NFKBIZ TSE functions as an RNA element in which sequences adjacent to the site of interaction with MCPIP1 and dispensable for accelerated mRNA degradation extend the functional repertoire of MCPIP1 to translational silencing.
- Published
- 2018
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37. Binding of NUFIP2 to Roquin promotes recognition and regulation of ICOS mRNA.
- Author
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Rehage N, Davydova E, Conrad C, Behrens G, Maiser A, Stehklein JE, Brenner S, Klein J, Jeridi A, Hoffmann A, Lee E, Dianzani U, Willemsen R, Feederle R, Reiche K, Hackermüller J, Leonhardt H, Sharma S, Niessing D, and Heissmeyer V
- Subjects
- Amino Acid Sequence, Animals, Binding Sites, CD4-Positive T-Lymphocytes cytology, Gene Expression Regulation, HEK293 Cells, HeLa Cells, Humans, Inducible T-Cell Co-Stimulator Protein antagonists & inhibitors, Inducible T-Cell Co-Stimulator Protein immunology, Inverted Repeat Sequences, Mice, Mice, Inbred C57BL, Nuclear Proteins antagonists & inhibitors, Nuclear Proteins immunology, Nucleic Acid Conformation, Primary Cell Culture, Protein Binding, RNA Stability, RNA, Small Interfering genetics, RNA, Small Interfering metabolism, RNA-Binding Proteins antagonists & inhibitors, RNA-Binding Proteins immunology, Receptors, OX40 antagonists & inhibitors, Receptors, OX40 immunology, Recombinant Proteins genetics, Recombinant Proteins immunology, Repressor Proteins immunology, Sequence Alignment, Sequence Homology, Amino Acid, Ubiquitin-Protein Ligases immunology, CD4-Positive T-Lymphocytes immunology, Inducible T-Cell Co-Stimulator Protein genetics, Nuclear Proteins genetics, RNA-Binding Proteins genetics, Receptors, OX40 genetics, Repressor Proteins genetics, Ubiquitin-Protein Ligases genetics
- Abstract
The ubiquitously expressed RNA-binding proteins Roquin-1 and Roquin-2 are essential for appropriate immune cell function and postnatal survival of mice. Roquin proteins repress target mRNAs by recognizing secondary structures in their 3'-UTRs and by inducing mRNA decay. However, it is unknown if other cellular proteins contribute to target control. To identify cofactors of Roquin, we used RNA interference to screen ~1500 genes involved in RNA-binding or mRNA degradation, and identified NUFIP2 as a cofactor of Roquin-induced mRNA decay. NUFIP2 binds directly and with high affinity to Roquin, which stabilizes NUFIP2 in cells. Post-transcriptional repression of human ICOS by endogenous Roquin proteins requires two neighboring non-canonical stem-loops in the ICOS 3'-UTR. This unconventional cis-element as well as another tandem loop known to confer Roquin-mediated regulation of the Ox40 3'-UTR, are bound cooperatively by Roquin and NUFIP2. NUFIP2 therefore emerges as a cofactor that contributes to mRNA target recognition by Roquin.
- Published
- 2018
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38. Roquin Suppresses the PI3K-mTOR Signaling Pathway to Inhibit T Helper Cell Differentiation and Conversion of Treg to Tfr Cells.
- Author
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Essig K, Hu D, Guimaraes JC, Alterauge D, Edelmann S, Raj T, Kranich J, Behrens G, Heiseke A, Floess S, Klein J, Maiser A, Marschall S, Hrabĕ de Angelis M, Leonhardt H, Calkhoven CF, Noessner E, Brocker T, Huehn J, Krug AB, Zavolan M, Baumjohann D, and Heissmeyer V
- Subjects
- Animals, B-Lymphocytes immunology, B-Lymphocytes pathology, Cell Differentiation, Colitis genetics, Colitis pathology, Disease Models, Animal, Female, Forkhead Box Protein O1 genetics, Forkhead Box Protein O1 immunology, Gene Expression Regulation, Germinal Center immunology, Germinal Center pathology, Interleukin-2 Receptor alpha Subunit genetics, Interleukin-2 Receptor alpha Subunit immunology, Lymphocyte Activation, Mice, Mice, Inbred C57BL, Mice, Transgenic, MicroRNAs genetics, MicroRNAs immunology, PTEN Phosphohydrolase genetics, PTEN Phosphohydrolase immunology, Phosphatidylinositol 3-Kinases genetics, Primary Cell Culture, Repressor Proteins deficiency, Repressor Proteins genetics, Signal Transduction, Spleen immunology, Spleen pathology, T-Lymphocytes, Regulatory immunology, T-Lymphocytes, Regulatory pathology, TOR Serine-Threonine Kinases genetics, Th17 Cells immunology, Th17 Cells pathology, Ubiquitin-Protein Ligases deficiency, Ubiquitin-Protein Ligases genetics, Colitis immunology, Phosphatidylinositol 3-Kinases immunology, Repressor Proteins immunology, TOR Serine-Threonine Kinases immunology, Ubiquitin-Protein Ligases immunology
- Abstract
Roquin proteins preclude spontaneous T cell activation and aberrant differentiation of T follicular helper (Tfh) or T helper 17 (Th17) cells. Here we showed that deletion of Roquin-encoding alleles specifically in regulatory T (Treg) cells also caused the activation of conventional T cells. Roquin-deficient Treg cells downregulated CD25, acquired a follicular Treg (Tfr) cell phenotype, and suppressed germinal center reactions but could not protect from colitis. Roquin inhibited the PI3K-mTOR signaling pathway by upregulation of Pten through interfering with miR-17∼92 binding to an overlapping cis-element in the Pten 3' UTR, and downregulated the Foxo1-specific E3 ubiquitin ligase Itch. Loss of Roquin enhanced Akt-mTOR signaling and protein synthesis, whereas inhibition of PI3K or mTOR in Roquin-deficient T cells corrected enhanced Tfh and Th17 or reduced iTreg cell differentiation. Thereby, Roquin-mediated control of PI3K-mTOR signaling prevents autoimmunity by restraining activation and differentiation of conventional T cells and specialization of Treg cells., (Copyright © 2017 Elsevier Inc. All rights reserved.)
- Published
- 2017
- Full Text
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39. Epstein-Barr viral miRNAs inhibit antiviral CD4+ T cell responses targeting IL-12 and peptide processing.
- Author
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Tagawa T, Albanese M, Bouvet M, Moosmann A, Mautner J, Heissmeyer V, Zielinski C, Lutter D, Hoser J, Hastreiter M, Hayes M, Sugden B, and Hammerschmidt W
- Subjects
- Antigen Presentation, B-Lymphocytes immunology, B-Lymphocytes virology, Cell Death, Cell Differentiation, Cell Membrane metabolism, Cytokines metabolism, HEK293 Cells, Humans, Immunity, Inflammation Mediators metabolism, Lysosomes metabolism, MicroRNAs metabolism, Receptors, Cell Surface metabolism, Species Specificity, Th1 Cells cytology, Th1 Cells immunology, CD4-Positive T-Lymphocytes immunology, Herpesvirus 4, Human genetics, Interleukin-12 metabolism, MicroRNAs genetics, Peptides metabolism
- Abstract
Epstein-Barr virus (EBV) is a tumor virus that establishes lifelong infection in most of humanity, despite eliciting strong and stable virus-specific immune responses. EBV encodes at least 44 miRNAs, most of them with unknown function. Here, we show that multiple EBV miRNAs modulate immune recognition of recently infected primary B cells, EBV's natural target cells. EBV miRNAs collectively and specifically suppress release of proinflammatory cytokines such as IL-12, repress differentiation of naive CD4(+) T cells to Th1 cells, interfere with peptide processing and presentation on HLA class II, and thus reduce activation of cytotoxic EBV-specific CD4(+) effector T cells and killing of infected B cells. Our findings identify a previously unknown viral strategy of immune evasion. By rapidly expressing multiple miRNAs, which are themselves nonimmunogenic, EBV counteracts recognition by CD4(+) T cells and establishes a program of reduced immunogenicity in recently infected B cells, allowing the virus to express viral proteins required for establishment of life-long infection., (© 2016 Tagawa et al.)
- Published
- 2016
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40. OX40L blockade protects against inflammation-driven fibrosis.
- Author
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Elhai M, Avouac J, Hoffmann-Vold AM, Ruzehaji N, Amiar O, Ruiz B, Brahiti H, Ponsoye M, Fréchet M, Burgevin A, Pezet S, Sadoine J, Guilbert T, Nicco C, Akiba H, Heissmeyer V, Subramaniam A, Resnick R, Molberg Ø, Kahan A, Chiocchia G, and Allanore Y
- Subjects
- Animals, Antibodies, Monoclonal pharmacology, Antibodies, Monoclonal therapeutic use, Biomarkers blood, Bleomycin, Case-Control Studies, Cells, Cultured, Drug Evaluation, Preclinical, Fibrosis, Fos-Related Antigen-2 genetics, Humans, Hypertension, Pulmonary prevention & control, Mice, Inbred C57BL, Mice, Transgenic, Middle Aged, Molecular Targeted Therapy, Pulmonary Fibrosis genetics, Scleroderma, Systemic drug therapy, Skin metabolism, Transcription Factor AP-1 metabolism, OX40 Ligand antagonists & inhibitors, OX40 Ligand blood, Pulmonary Fibrosis prevention & control, Scleroderma, Systemic blood, Skin pathology
- Abstract
Treatment for fibrosis represents a critical unmet need, because fibrosis is the leading cause of death in industrialized countries, and there is no effective therapy to counteract the fibrotic process. The development of fibrosis relates to the interplay between vessel injury, immune cell activation, and fibroblast stimulation, which can occur in various tissues. Immunotherapies have provided a breakthrough in the treatment of immune diseases. The glycoprotein OX40-OX40 ligand (OX40L) axis offers the advantage of a targeted approach to costimulatory signals with limited impact on the whole immune response. Using systemic sclerosis (SSc) as a prototypic disease, we report compelling evidence that blockade of OX40L is a promising strategy for the treatment of inflammation-driven fibrosis. OX40L is overexpressed in the fibrotic skin and serum of patients with SSc, particularly in patients with diffuse cutaneous forms. Soluble OX40L was identified as a promising serum biomarker to predict the worsening of lung and skin fibrosis, highlighting the role of this pathway in fibrosis. In vivo, OX40L blockade prevents inflammation-driven skin, lung, and vessel fibrosis and induces the regression of established dermal fibrosis in different complementary mouse models. OX40L exerts potent profibrotic effects by promoting the infiltration of inflammatory cells into lesional tissues and therefore the release of proinflammatory mediators, thereafter leading to fibroblast activation.
- Published
- 2016
- Full Text
- View/download PDF
41. Alternative splicing of MALT1 controls signalling and activation of CD4(+) T cells.
- Author
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Meininger I, Griesbach RA, Hu D, Gehring T, Seeholzer T, Bertossi A, Kranich J, Oeckinghaus A, Eitelhuber AC, Greczmiel U, Gewies A, Schmidt-Supprian M, Ruland J, Brocker T, Heissmeyer V, Heyd F, and Krappmann D
- Subjects
- Animals, Caspases metabolism, Down-Regulation, Enzyme Activation, Exons genetics, HEK293 Cells, Heterogeneous-Nuclear Ribonucleoprotein U metabolism, Humans, Interleukin-2 biosynthesis, JNK Mitogen-Activated Protein Kinases metabolism, Jurkat Cells, Mice, Inbred C57BL, Mucosa-Associated Lymphoid Tissue Lymphoma Translocation 1 Protein, NF-kappa B metabolism, Neoplasm Proteins metabolism, Receptors, Antigen, T-Cell metabolism, TNF Receptor-Associated Factor 6 metabolism, Th17 Cells immunology, Up-Regulation, Alternative Splicing genetics, CD4-Positive T-Lymphocytes immunology, Caspases genetics, Lymphocyte Activation immunology, Neoplasm Proteins genetics, Signal Transduction
- Abstract
MALT1 channels proximal T-cell receptor (TCR) signalling to downstream signalling pathways. With MALT1A and MALT1B two conserved splice variants exist and we demonstrate here that MALT1 alternative splicing supports optimal T-cell activation. Inclusion of exon7 in MALT1A facilitates the recruitment of TRAF6, which augments MALT1 scaffolding function, but not protease activity. Naive CD4(+) T cells express almost exclusively MALT1B and MALT1A expression is induced by TCR stimulation. We identify hnRNP U as a suppressor of exon7 inclusion. Whereas selective depletion of MALT1A impairs T-cell signalling and activation, downregulation of hnRNP U enhances MALT1A expression and T-cell activation. Thus, TCR-induced alternative splicing augments MALT1 scaffolding to enhance downstream signalling and to promote optimal T-cell activation.
- Published
- 2016
- Full Text
- View/download PDF
42. Roquin recognizes a non-canonical hexaloop structure in the 3'-UTR of Ox40.
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Janowski R, Heinz GA, Schlundt A, Wommelsdorf N, Brenner S, Gruber AR, Blank M, Buch T, Buhmann R, Zavolan M, Niessing D, Heissmeyer V, and Sattler M
- Subjects
- Animals, Base Sequence, Crystallography, X-Ray, DNA Mutational Analysis, Electrophoretic Mobility Shift Assay, Ligands, Mice, Models, Molecular, Molecular Sequence Data, Nucleotide Motifs genetics, Protein Binding, Protein Structure, Tertiary, Proton Magnetic Resonance Spectroscopy, RNA chemistry, RNA metabolism, SELEX Aptamer Technique, Ubiquitin-Protein Ligases chemistry, 3' Untranslated Regions genetics, Nucleic Acid Conformation, Receptors, OX40 genetics, Ubiquitin-Protein Ligases metabolism
- Abstract
The RNA-binding protein Roquin is required to prevent autoimmunity. Roquin controls T-helper cell activation and differentiation by limiting the induced expression of costimulatory receptors such as tumor necrosis factor receptor superfamily 4 (Tnfrs4 or Ox40). A constitutive decay element (CDE) with a characteristic triloop hairpin was previously shown to be recognized by Roquin. Here we use SELEX assays to identify a novel U-rich hexaloop motif, representing an alternative decay element (ADE). Crystal structures and NMR data show that the Roquin-1 ROQ domain recognizes hexaloops in the SELEX-derived ADE and in an ADE-like variant present in the Ox40 3'-UTR with identical binding modes. In cells, ADE-like and CDE-like motifs cooperate in the repression of Ox40 by Roquin. Our data reveal an unexpected recognition of hexaloop cis elements for the posttranscriptional regulation of target messenger RNAs by Roquin.
- Published
- 2016
- Full Text
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43. In Vivo Killing Capacity of Cytotoxic T Cells Is Limited and Involves Dynamic Interactions and T Cell Cooperativity.
- Author
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Halle S, Keyser KA, Stahl FR, Busche A, Marquardt A, Zheng X, Galla M, Heissmeyer V, Heller K, Boelter J, Wagner K, Bischoff Y, Martens R, Braun A, Werth K, Uvarovskii A, Kempf H, Meyer-Hermann M, Arens R, Kremer M, Sutter G, Messerle M, and Förster R
- Subjects
- Animals, Calcium Signaling, Cell Communication, Cells, Cultured, Cytotoxicity, Immunologic, Humans, Immune Evasion, Mice, Mice, Inbred C57BL, Mice, Knockout, Microscopy, Fluorescence, Multiphoton, Perforin genetics, T-Lymphocyte Subsets virology, T-Lymphocytes, Cytotoxic virology, Herpesviridae Infections immunology, Muromegalovirus immunology, Perforin metabolism, T-Lymphocyte Subsets immunology, T-Lymphocytes, Cytotoxic immunology, Vaccinia immunology, Vaccinia virus immunology
- Abstract
According to in vitro assays, T cells are thought to kill rapidly and efficiently, but the efficacy and dynamics of cytotoxic T lymphocyte (CTL)-mediated killing of virus-infected cells in vivo remains elusive. We used two-photon microscopy to quantify CTL-mediated killing in mice infected with herpesviruses or poxviruses. On average, one CTL killed 2-16 virus-infected cells per day as determined by real-time imaging and by mathematical modeling. In contrast, upon virus-induced MHC class I downmodulation, CTLs failed to destroy their targets. During killing, CTLs remained migratory and formed motile kinapses rather than static synapses with targets. Viruses encoding the calcium sensor GCaMP6s revealed strong heterogeneity in individual CTL functional capacity. Furthermore, the probability of death of infected cells increased for those contacted by more than two CTLs, indicative of CTL cooperation. Thus, direct visualization of CTLs during killing of virus-infected cells reveals crucial parameters of CD8(+) T cell immunity., (Copyright © 2016 Elsevier Inc. All rights reserved.)
- Published
- 2016
- Full Text
- View/download PDF
44. Induced miR-99a expression represses Mtor cooperatively with miR-150 to promote regulatory T-cell differentiation.
- Author
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Warth SC, Hoefig KP, Hiekel A, Schallenberg S, Jovanovic K, Klein L, Kretschmer K, Ansel KM, and Heissmeyer V
- Subjects
- 3' Untranslated Regions, Animals, Base Sequence, CD4-Positive T-Lymphocytes cytology, CD4-Positive T-Lymphocytes physiology, Cell Differentiation genetics, Cells, Cultured, DEAD-box RNA Helicases genetics, DEAD-box RNA Helicases metabolism, Gene Expression Regulation drug effects, Gene Regulatory Networks, Green Fluorescent Proteins genetics, Green Fluorescent Proteins metabolism, Mice, Inbred C57BL, Mice, Transgenic, Molecular Sequence Data, Ribonuclease III genetics, Ribonuclease III metabolism, T-Lymphocytes, Regulatory drug effects, TOR Serine-Threonine Kinases metabolism, Tretinoin pharmacology, MicroRNAs genetics, T-Lymphocytes, Regulatory cytology, T-Lymphocytes, Regulatory physiology, TOR Serine-Threonine Kinases genetics
- Abstract
Peripheral induction of regulatory T (Treg) cells provides essential protection from inappropriate immune responses. CD4(+) T cells that lack endogenous miRNAs are impaired to differentiate into Treg cells, but the relevant miRNAs are unknown. We performed an overexpression screen with T-cell-expressed miRNAs in naive mouse CD4(+) T cells undergoing Treg differentiation. Among 130 candidates, the screen identified 29 miRNAs with a negative and 10 miRNAs with a positive effect. Testing reciprocal Th17 differentiation revealed specific functions for miR-100, miR-99a and miR-10b, since all of these promoted the Treg and inhibited the Th17 program without impacting on viability, proliferation and activation. miR-99a cooperated with miR-150 to repress the expression of the Th17-promoting factor mTOR. The comparably low expression of miR-99a was strongly increased by the Treg cell inducer "retinoic acid", and the abundantly expressed miR-150 could only repress Mtor in the presence of miR-99a. Our data suggest that induction of Treg cell differentiation is regulated by a miRNA network, which involves cooperation of constitutively expressed as well as inducible miRNAs., (© 2015 The Authors.)
- Published
- 2015
- Full Text
- View/download PDF
45. Roquin binds microRNA-146a and Argonaute2 to regulate microRNA homeostasis.
- Author
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Srivastava M, Duan G, Kershaw NJ, Athanasopoulos V, Yeo JH, Ose T, Hu D, Brown SH, Jergic S, Patel HR, Pratama A, Richards S, Verma A, Jones EY, Heissmeyer V, Preiss T, Dixon NE, Chong MM, Babon JJ, and Vinuesa CG
- Subjects
- Animals, Crystallography, X-Ray, HEK293 Cells, Half-Life, Humans, Mice, Inbred C57BL, MicroRNAs genetics, Protein Binding genetics, Protein Structure, Tertiary, RNA Processing, Post-Transcriptional, RNA Stability, Ribonuclease III metabolism, T-Lymphocytes metabolism, Ubiquitin-Protein Ligases chemistry, Argonaute Proteins metabolism, Homeostasis, MicroRNAs metabolism, Ubiquitin-Protein Ligases metabolism
- Abstract
Roquin is an RNA-binding protein that prevents autoimmunity and inflammation via repression of bound target mRNAs such as inducible costimulator (Icos). When Roquin is absent or mutated (Roquin(san)), Icos is overexpressed in T cells. Here we show that Roquin enhances Dicer-mediated processing of pre-miR-146a. Roquin also directly binds Argonaute2, a central component of the RNA-induced silencing complex, and miR-146a, a microRNA that targets Icos mRNA. In the absence of functional Roquin, miR-146a accumulates in T cells. Its accumulation is not due to increased transcription or processing, rather due to enhanced stability of mature miR-146a. This is associated with decreased 3' end uridylation of the miRNA. Crystallographic studies reveal that Roquin contains a unique HEPN domain and identify the structural basis of the 'san' mutation and Roquin's ability to bind multiple RNAs. Roquin emerges as a protein that can bind Ago2, miRNAs and target mRNAs, to control homeostasis of both RNA species.
- Published
- 2015
- Full Text
- View/download PDF
46. The transcription factor NFAT promotes exhaustion of activated CD8⁺ T cells.
- Author
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Martinez GJ, Pereira RM, Äijö T, Kim EY, Marangoni F, Pipkin ME, Togher S, Heissmeyer V, Zhang YC, Crotty S, Lamperti ED, Ansel KM, Mempel TR, Lähdesmäki H, Hogan PG, and Rao A
- Subjects
- Animals, Cells, Cultured, Clonal Anergy drug effects, Gene Expression Regulation genetics, Listeria monocytogenes immunology, Listeriosis immunology, Listeriosis microbiology, Lymphocyte Activation immunology, Mice, Mice, Transgenic, NFATC Transcription Factors genetics, Neoplasms immunology, Promoter Regions, Genetic genetics, Receptors, Antigen, T-Cell immunology, Recombinant Proteins genetics, CD8-Positive T-Lymphocytes immunology, Clonal Anergy genetics, NFATC Transcription Factors physiology, Recombinant Proteins pharmacology, Transcription Factor AP-1 metabolism
- Abstract
During persistent antigen stimulation, CD8(+) T cells show a gradual decrease in effector function, referred to as exhaustion, which impairs responses in the setting of tumors and infections. Here we demonstrate that the transcription factor NFAT controls the program of T cell exhaustion. When expressed in cells, an engineered form of NFAT1 unable to interact with AP-1 transcription factors diminished T cell receptor (TCR) signaling, increased the expression of inhibitory cell surface receptors, and interfered with the ability of CD8(+) T cells to protect against Listeria infection and attenuate tumor growth in vivo. We defined the genomic regions occupied by endogenous and engineered NFAT1 in primary CD8(+) T cells and showed that genes directly induced by the engineered NFAT1 overlapped with genes expressed in exhausted CD8(+) T cells in vivo. Our data show that NFAT promotes T cell anergy and exhaustion by binding at sites that do not require cooperation with AP-1., (Copyright © 2015 Elsevier Inc. All rights reserved.)
- Published
- 2015
- Full Text
- View/download PDF
47. Uncoupling Malt1 threshold function from paracaspase activity results in destructive autoimmune inflammation.
- Author
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Gewies A, Gorka O, Bergmann H, Pechloff K, Petermann F, Jeltsch KM, Rudelius M, Kriegsmann M, Weichert W, Horsch M, Beckers J, Wurst W, Heikenwalder M, Korn T, Heissmeyer V, and Ruland J
- Subjects
- Animals, B-Lymphocytes immunology, Caspases deficiency, Cell Differentiation immunology, Gene Expression Regulation, Homeostasis immunology, Humans, Immunity, Mucosal immunology, Interferon-gamma biosynthesis, Lymphocyte Activation immunology, Mice, Mutant Strains, Mucosa-Associated Lymphoid Tissue Lymphoma Translocation 1 Protein, Neoplasm Proteins deficiency, Nerve Degeneration immunology, Nerve Degeneration pathology, RNA, Messenger genetics, RNA, Messenger metabolism, Receptors, Antigen, T-Cell metabolism, Signal Transduction immunology, T-Lymphocytes, Regulatory immunology, Autoimmunity, Caspases metabolism, Inflammation immunology, Inflammation pathology, Neoplasm Proteins metabolism
- Abstract
The paracaspase Malt1 is a central regulator of antigen receptor signaling that is frequently mutated in human lymphoma. As a scaffold, it assembles protein complexes for NF-κB activation, and its proteolytic domain cleaves negative NF-κB regulators for signal enforcement. Still, the physiological functions of Malt1-protease are unknown. We demonstrate that targeted Malt1-paracaspase inactivation induces a lethal inflammatory syndrome with lymphocyte-dependent neurodegeneration in vivo. Paracaspase activity is essential for regulatory T cell (Treg) and innate-like B cell development, but it is largely dispensable for overcoming Malt1-dependent thresholds for lymphocyte activation. In addition to NF-κB inhibitors, Malt1 cleaves an entire set of mRNA stability regulators, including Roquin-1, Roquin-2, and Regnase-1, and paracaspase inactivation results in excessive interferon gamma (IFNγ) production by effector lymphocytes that drive pathology. Together, our results reveal distinct threshold and modulatory functions of Malt1 that differentially control lymphocyte differentiation and activation pathways and demonstrate that selective paracaspase blockage skews systemic immunity toward destructive autoinflammation., (Copyright © 2014 The Authors. Published by Elsevier Inc. All rights reserved.)
- Published
- 2014
- Full Text
- View/download PDF
48. Structural basis for RNA recognition in roquin-mediated post-transcriptional gene regulation.
- Author
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Schlundt A, Heinz GA, Janowski R, Geerlof A, Stehle R, Heissmeyer V, Niessing D, and Sattler M
- Subjects
- Amino Acid Substitution, Animals, Base Pairing, Base Sequence, Binding Sites, Consensus Sequence, Crystallography, X-Ray, Mice, Models, Molecular, Mutagenesis, Site-Directed, Nuclear Magnetic Resonance, Biomolecular, Protein Binding, Protein Structure, Secondary, Protein Structure, Tertiary, RNA Stability, Ubiquitin-Protein Ligases genetics, RNA Interference, RNA, Messenger chemistry, Ubiquitin-Protein Ligases chemistry
- Abstract
Roquin function in T cells is essential for the prevention of autoimmune disease. Roquin interacts with the 3' untranslated regions (UTRs) of co-stimulatory receptors and controls T-cell activation and differentiation. Here we show that the N-terminal ROQ domain from mouse roquin adopts an extended winged-helix (WH) fold, which is sufficient for binding to the constitutive decay element (CDE) in the Tnf 3' UTR. The crystal structure of the ROQ domain in complex with a prototypical CDE RNA stem-loop reveals tight recognition of the RNA stem and its triloop. Surprisingly, roquin uses mainly non-sequence-specific contacts to the RNA, thus suggesting a relaxed CDE consensus and implicating a broader spectrum of target mRNAs than previously anticipated. Consistently with this, NMR and binding experiments with CDE-like stem-loops together with cell-based assays confirm roquin-dependent regulation of relaxed CDE consensus motifs in natural 3' UTRs.
- Published
- 2014
- Full Text
- View/download PDF
49. Tfh cell differentiation: missing Stat3 uncovers interferons' interference.
- Author
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Edelmann SL and Heissmeyer V
- Subjects
- Animals, Cell Differentiation, Interferon Type I metabolism, STAT3 Transcription Factor metabolism, T-Lymphocytes, Helper-Inducer cytology, T-Lymphocytes, Helper-Inducer metabolism
- Abstract
Viral infection induces a number of cytokines that shape T cell responses. In this issue of Immunity, Ray et al. (2014) describe how CD4(+) T cells decide on T follicular helper (Tfh) or T helper 1 (Th1) cell skewed gene expression during acute viral infection., (Copyright © 2014 Elsevier Inc. All rights reserved.)
- Published
- 2014
- Full Text
- View/download PDF
50. Cyclin-dependent kinase 9 links RNA polymerase II transcription to processing of ribosomal RNA.
- Author
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Burger K, Mühl B, Rohrmoser M, Coordes B, Heidemann M, Kellner M, Gruber-Eber A, Heissmeyer V, Strässer K, and Eick D
- Subjects
- Animals, Cell Line, Tumor, Cell Nucleolus drug effects, Cell Nucleolus enzymology, Cyclin-Dependent Kinase 9 antagonists & inhibitors, DEAD-box RNA Helicases metabolism, Feedback, Physiological drug effects, Flavonoids pharmacology, Gene Knockdown Techniques, Humans, Mice, Mice, Knockout, Piperidines pharmacology, RNA 3' End Processing drug effects, RNA 3' End Processing genetics, RNA Polymerase II antagonists & inhibitors, RNA, Small Nucleolar metabolism, Ribonuclease III metabolism, Cyclin-Dependent Kinase 9 metabolism, RNA Polymerase II metabolism, RNA Processing, Post-Transcriptional drug effects, RNA, Ribosomal genetics, Transcription, Genetic drug effects
- Abstract
Ribosome biogenesis is a process required for cellular growth and proliferation. Processing of ribosomal RNA (rRNA) is highly sensitive to flavopiridol, a specific inhibitor of cyclin-dependent kinase 9 (Cdk9). Cdk9 has been characterized as the catalytic subunit of the positive transcription elongation factor b (P-TEFb) of RNA polymerase II (RNAPII). Here we studied the connection between RNAPII transcription and rRNA processing. We show that inhibition of RNAPII activity by α-amanitin specifically blocks processing of rRNA. The block is characterized by accumulation of 3' extended unprocessed 47 S rRNAs and the entire inhibition of other 47 S rRNA-specific processing steps. The transcription rate of rRNA is moderately reduced after inhibition of Cdk9, suggesting that defective 3' processing of rRNA negatively feeds back on RNAPI transcription. Knockdown of Cdk9 caused a strong reduction of the levels of RNAPII-transcribed U8 small nucleolar RNA, which is essential for 3' rRNA processing in mammalian cells. Our data demonstrate a pivotal role of Cdk9 activity for coupling of RNAPII transcription with small nucleolar RNA production and rRNA processing.
- Published
- 2013
- Full Text
- View/download PDF
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