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Rainer M. Bohle - One of the best experts on this subject based on the ideXlab platform.

  • 3-Deazaadenosine Prevents Smooth Muscle Cell Proliferation and Neointima Formation by Interfering With Ras Signaling
    Circulation research, 2009
    Co-Authors: Daniel Sedding, Gerhard Walker, Ludger Fink, Werner Haberbosch, Harald Tillmanns, Monique Tröbs, Fabian Reich, Wigbert S. Rau, Klaus T. Preissner, Rainer M. Bohle
    Abstract:

    3-Deazaadenosine (c3Ado) is a potent inhibitor of S -adenosylhomocysteine hydrolase, which regulates cellular methyltransferase activity. In the present study, we sought to determine the effect of c3Ado on vascular smooth muscle cell (VSMC) function and neointima formation in vivo. c3Ado dose-dependently prevented the proliferation and migration of human coronary VSMCs in vitro. This was accompanied by an increased expression of the cyclin-dependent kinase inhibitors p21 WAF1/Cip1 , p27 Kip1 , a decreased expression of G 1 /S phase cyclins, and a lack of retinoblastoma protein hyperphosphorylation. In accordance with these findings, fluorescence-activated cell-sorting analysis of propidium iodide–stained cells indicated a cell cycle arrest in the G 0 /G 1 phase. Importantly, c3Ado did not affect the number of viable (trypan blue exclusion) or apoptotic cells (TUNEL). Mechanistically, c3Ado prevented FCS-induced Ras carboxyl methylation and membrane translocation and activity by inhibiting isoprenylcysteine carboxyl methyltransferase and reduced FCS-induced extracellular signal-regulated kinase (ERK)1/2 and Akt phosphorylation in a dose-dependent manner. Conversely, rescuing signal transduction by overexpression of a constitutive active Ras mutant abrogated c3Ado’s effect on proliferation. For in vivo studies, the femoral artery of C57BL/6 mice was dilated and mice were fed a diet containing 150 μg of c3Ado per day. c3Ado prevented dilation-induced Ras activation, as well as ERK1/2 and Akt phosphorylation in vivo. At day 21, VSMC proliferation (proliferating-cell nuclear antigen [PCNA]-positive cells), as well as the neointima/media ratio (0.7±0.2 versus 1.6±0.4; P S -adenosylhomocysteine hydrolase by c3Ado may represent a save and effective novel approach to prevent vascular proliferative disease.

  • 3 Deazaadenosine inhibits vasa vasorum neovascularization in aortas of apoe ldl double knockout mice
    Atherosclerosis, 2009
    Co-Authors: A C Langheinrich, Daniel Sedding, Werner Haberbosch, Marian Kampschulte, Regina Moritz, Jochen Wilhelm, Erik L Ritman, Rainer M. Bohle
    Abstract:

    Abstract Background Atherosclerosis and inflammation/angiogenesis are strongly associated including growth of vasa vasorum (VV) and plaque neovascularization, but a causative role for neovascularization has still not been established. Hence, we investigated the effect of 3-Deazaadenosine (c 3 Ado), an anti-inflammatory and anti-proliferative drug, on plaque progression and VV neovascularization in apoE −/− /LDL −/− double knockout mice. Methods The arterial trees from apoE −/− /LDL −/− mice with, or without c 3 Ado at the age of 16 weeks ( n =10), 18 weeks ( n =8) and 20 weeks ( n =7) were infused in situ with Microfil, and the aortas harvested and scanned with micro-CT (12μm cubic voxel). We characterized plaque volume and VV luminal volume along the descending aorta using Analyze 6.0 software. Cellular effects of c 3 Ado on human endothelial and vascular smooth muscle cells were investigated in cell cultures and on nylon cDNA expression arrays. Results Lesions spatially connected to VV increased from 16 to 20 weeks significantly ( p 3 Ado ( p p 3 Ado. Moreover, c 3 Ado dose-dependently prevented the proliferation and migration of human coronary artery endothelial cells in vitro. Conclusion The smaller lesion volume in animals treated with c 3 Ado was closely associated with a reduced VV neovascularization, suggesting a direct relationship between lesion growth and VV development.

  • 3-Deazaadenosine inhibits vasa vasorum neovascularization in aortas of ApoE(-/-)/LDL(-/-) double knockout mice.
    Atherosclerosis, 2008
    Co-Authors: Alexander C. Langheinrich, Daniel Sedding, Werner Haberbosch, Marian Kampschulte, Regina Moritz, Jochen Wilhelm, Erik L Ritman, Rainer M. Bohle
    Abstract:

    Atherosclerosis and inflammation/angiogenesis are strongly associated including growth of vasa vasorum (VV) and plaque neovascularization, but a causative role for neovascularization has still not been established. Hence, we investigated the effect of 3-Deazaadenosine (c(3)Ado), an anti-inflammatory and anti-proliferative drug, on plaque progression and VV neovascularization in apoE(-/-)/LDL(-/-) double knockout mice. The arterial trees from apoE(-/-)/LDL(-/-) mice with, or without c(3)Ado at the age of 16 weeks (n=10), 18 weeks (n=8) and 20 weeks (n=7) were infused in situ with Microfil, and the aortas harvested and scanned with micro-CT (12mum cubic voxel). We characterized plaque volume and VV luminal volume along the descending aorta using Analyze 6.0 software. Cellular effects of c(3)Ado on human endothelial and vascular smooth muscle cells were investigated in cell cultures and on nylon cDNA expression arrays. Lesions spatially connected to VV increased from 16 to 20 weeks significantly (p<0.001). The volume of atherosclerotic lesions was significantly reduced in animals treated with c(3)Ado (p<0.01). This was accompanied by a significant decrease of vasa vasorum neovascularization along the descending aorta (p<0.01). Using nylon cDNA expression arrays, we identified the regulation of anti-proliferative, anti-inflammatory genes in human smooth muscle cells which might be involved in the anti-angiogenic effects of c(3)Ado. Moreover, c(3)Ado dose-dependently prevented the proliferation and migration of human coronary artery endothelial cells in vitro. The smaller lesion volume in animals treated with c(3)Ado was closely associated with a reduced VV neovascularization, suggesting a direct relationship between lesion growth and VV development.

  • Effects of 3-Deazaadenosine on homocysteine and atherosclerosis in apolipoprotein E-deficient mice.
    Atherosclerosis, 2003
    Co-Authors: Alexander C. Langheinrich, Ruediger C. Braun-dullaeus, Gerhard Walker, Ina Jeide, Ralph Schilling, Kai Tammoscheit, Thomas Dreyer, Ludger Fink, Rainer M. Bohle, Werner Haberbosch
    Abstract:

    Abstract Objective: In the past decade, elevated homocysteine concentration has achieved widespread recognition as an independent risk factor in the development of atherosclerosis. 3-Deazaadenosine (c 3 Ado) is a potent inhibitor and substrate for S -adenosylhomocysteine hydrolase and therefore may reduce homocysteine concentrations. The current study investigated the effect of c 3 Ado on serum homocysteine, atherosclerotic lesions, and the expression of adhesion molecules in apoE-knockout mice. Methods and results: Animals were placed on an atherogenic diet with or without c 3 Ado for 12 and 24 weeks. Frozen cross-sections of the aortic sinus and the proximal aorta were analyzed by computer-aided planimetry for fatty plaque formation. Macrophages, VCAM-1 and ICAM-1 were quantified by immunhistochemistry and oligo-cell reverse transcription polymerase chain reaction after laser microdissection. Application of c 3 Ado resulted in significant reduction of homocysteine levels by 35.9 and 45.3% after 12 and 24 weeks, respectively ( P 3 Ado ( P P P Conclusion: Our results demonstrate that c 3 Ado induces a marked reduction of homocysteine concentrations which might explain in part the anti-atherogenic effect of the drug.

  • 3 Deazaadenosine prevents adhesion molecule expression and atherosclerotic lesion formation in the aortas of c57bl 6j mice
    Arteriosclerosis Thrombosis and Vascular Biology, 1999
    Co-Authors: Gerhard Walker, Jörg Kreuzer, Ruediger C Braundullaeus, Thomas Dreyer, Rainer M. Bohle, Elisabeth Dennhauser, Harald Tillmanns, A C Langheinrich, Werner Haberbosch
    Abstract:

    Abstract —Adhesion molecules such as vascular cell adhesion molecule-1 (VCAM-1) and intercellular adhesion molecule-1 (ICAM-1) play an important role during the development of atherosclerosis. 3-Deazaadenosine (c3Ado), an adenosine analogue, inhibits endothelial-leukocyte adhesion and ICAM-1-expression in vitro. We hypothesized that c3Ado is able to prevent the expression of adhesion molecules and atherosclerotic lesion formation in female C57BL/6J mice. The animals were placed on an atherogenic diet with or without c3Ado for 9 weeks. Frozen cross sections of the proximal ascending aorta just beyond the aortic sinus were stained with oil red O, hematoxylin, and elastic van Gieson’s stains and were analyzed by computer-aided planimetry for fatty plaque formation and neointimal proliferation. Monoclonal antibodies against CD11b (macrophages), VCAM-1, and ICAM-1 were used for immunohistochemistry. Mice on the atherogenic diet demonstrated multiple (5.4±1.6 per animal) lesions covering 3.4±2.8% of the endothelium and a marked neointima when compared with control mice (4501±775 versus 160±38 μm2, P <0.001). Mice on the cholesterol-rich diet without c3Ado showed strong endothelial coexpression of ICAM-1 and VCAM-1. Moreover, there was a 10-fold increase in monocyte accumulation on the endothelial surface (33.3±4.9 versus 3.8±1.2, P <0.004). In contrast, in mice treated with c3Ado, expression of ICAM-1 and VCAM-1 as well as monocyte adhesion and infiltration were almost completely inhibited. Furthermore, these mice did not show any fatty streak formation or neointima formation (125±32 μm2). Our results demonstrate that c3Ado can inhibit diet-induced fatty streak formation and the expression of endothelial ICAM-1 and VCAM-1 in C57BL/6J mice. This may provide a novel pharmacological approach in the prevention and treatment of atherosclerosis.

Noriaki Minakawa - One of the best experts on this subject based on the ideXlab platform.

  • Groove modification of siRNA duplexes to elucidate siRNA–protein interactions using 7-bromo-7-Deazaadenosine and 3-bromo-3-Deazaadenosine as chemical probes
    Organic & biomolecular chemistry, 2016
    Co-Authors: Noriko Saito-tarashima, Akira Matsuda, Hirotaka Kira, Tomoya Wada, Kazuya Miki, Shiho Ide, Naoshi Yamazaki, Noriaki Minakawa
    Abstract:

    Elucidation of dynamic interactions between RNA and proteins is essential for understanding the biological processes regulated by RNA, such as RNA interference (RNAi). In this study, the logical chemical probes, comprising 7-bromo-7-Deazaadenosine (Br7C7A) and 3-bromo-3-Deazaadenosine (Br3C3A), to investigate small interfering RNA (siRNA)-RNAi related protein interactions, were developed. The bromo substituents of Br7C7A and Br3C3A are expected to be located in the major and the minor grooves, respectively, and to act as a steric hindrance in each groove when these chemical probes are incorporated into siRNAs. A comprehensive investigation using siRNAs containing these chemical probes revealed that (i) Br3C3A(s) at the 5'-end of the passenger strand enhanced their RNAi activity, and (ii) the direction of RISC assembly is determined by the interaction between Argonaute2, which is the main component of RISC, and siRNA in the minor groove near the 5'-end of the passenger strand. Utilization of these chemical probes enables the investigation of the dynamic interactions between RNA and proteins.

  • groove modification of sirna duplexes to elucidate sirna protein interactions using 7 bromo 7 Deazaadenosine and 3 bromo 3 Deazaadenosine as chemical probes
    Organic and Biomolecular Chemistry, 2016
    Co-Authors: Noriko Saitotarashima, Akira Matsuda, Hirotaka Kira, Tomoya Wada, Kazuya Miki, Shiho Ide, Naoshi Yamazaki, Noriaki Minakawa
    Abstract:

    Elucidation of dynamic interactions between RNA and proteins is essential for understanding the biological processes regulated by RNA, such as RNA interference (RNAi). In this study, the logical chemical probes, comprising 7-bromo-7-Deazaadenosine (Br7C7A) and 3-bromo-3-Deazaadenosine (Br3C3A), to investigate small interfering RNA (siRNA)–RNAi related protein interactions, were developed. The bromo substituents of Br7C7A and Br3C3A are expected to be located in the major and the minor grooves, respectively, and to act as a steric hindrance in each groove when these chemical probes are incorporated into siRNAs. A comprehensive investigation using siRNAs containing these chemical probes revealed that (i) Br3C3A(s) at the 5′-end of the passenger strand enhanced their RNAi activity, and (ii) the direction of RISC assembly is determined by the interaction between Argonaute2, which is the main component of RISC, and siRNA in the minor groove near the 5′-end of the passenger strand. Utilization of these chemical probes enables the investigation of the dynamic interactions between RNA and proteins.

  • Identification of A-minor tertiary interactions within a bacterial group I intron active site by 3-Deazaadenosine interference mapping.
    Biochemistry, 2002
    Co-Authors: Juliane K. Soukup, Noriaki Minakawa, And Akira Matsuda, Scott A. Strobel
    Abstract:

    The A-minor motifs appear to be the most ubiquitous helix packing elements within RNA tertiary structures. These motifs have been identified throughout the ribosome and almost every other tertiary-folded RNA for which structural information is available. These motifs utilize the packing of the donor adenosine's N1, N3, and/or 2'-OH against the 2'-OHs and minor groove edge of the acceptor base pair. The ability to identify biochemically which adenosines form A-minor motifs and which base pairs they contact is an important experimental objective. Toward this goal, we report the synthesis and transcriptional incorporation of 5'-O-(1-thio)-3-Deazaadenosine triphosphate and its use in Nucleotide Analogue Interference Mapping (NAIM) and Nucleotide Analogue Interference Suppression (NAIS). This analogue makes it possible for the first time to explore the functional importance of the N3 imino group of adenosine in RNA polymers. Interference analysis of the group I self-splicing introns from Tetrahymena and Azoarcus indicates that A-minor motifs are integral to the helix packing interactions that define the 5'-splice site of the intron. Specifically, Azoarcus A58 in the J4/5 region contacts the G.U wobble pair at the cleavage site in the P1 helix, and Azoarcus A167 in the J8/7 region contacts the C13-G37 base pair in the P2 helix. Both of these structural features are conserved between the eukaryotic and bacterial introns. These results suggest that nucleotide analogue interference patterns can identify and distinguish A-minor interactions in RNA tertiary structure, particularly the most prevalent type I and type II varieties. Furthermore, clustering of 3-Deazaadenosine interferences is suggestive of A patches, in which a series of consecutive A-minor motifs mediate helix packing. Biochemical identification of these interactions may provide valuable constraints for RNA structure prediction.

  • nucleosides and nucleotides 116 convenient syntheses of 3 Deazaadenosine 3 deazaguanosine and 3 deazainosine via ring closure of 5 ethynyl 1 β d ribofuranosylimidazole 4 carboxamide or carbonitrile
    Tetrahedron, 1993
    Co-Authors: Noriaki Minakawa, Akira Matsuda
    Abstract:

    Abstract An easy chemical synthesis of 3-deazapurine nucleosides, 3-deazainosine [1-β- D -ribofuranosylimidazo[4,5-c]pyridin-4(5H)-one (8)], 3-deazaguanosine [6-amino-1-β- D -ribofuranosylimidazo[4,5-c]pyridin-4(5H)-one (23)], and 3-Deazaadenosine [4-amino-1-β- D -ribofuranosylimidazo[4,5-c]pyridine (29)] is described. The approach consists of ring closure between substituents at 4- and 5-positions of the imidazole ring. Treatment of 5-ethynyl-1-β- D -ribofuranosylimidazole-4-carboxamide (2), which was readily obtained from AICA-riboside (1), with aqueous dimethylamine, followed by aqueous acetic acid gave 8 in 64% yield. 5-(2-Hydroxyiminoethyl)-1-(2,3,5-tri-O-tert-butyldimethylsilyl-β- D -ribofuranosyl)imidazole-4-carboxamide (19) was synthesized from 3 by treatment of aqueous dimethylamine, followed by hydroxylamine hydrochloride. Dehydration of 19 was achieved by phenyl isocyanate to give 5-cyanomethyl derivative 21, from which 3-deazaguanosine (23) was easily obtained. 3-Deazaadenosine (29) was synthesized from 5-ethynyl-1-(2,3,5-tri-O-tert-butyldimethylsilyl-β- D -ribofuranosyl)imidazole-4-carbonitrile (25).

K Müller - One of the best experts on this subject based on the ideXlab platform.

Werner Haberbosch - One of the best experts on this subject based on the ideXlab platform.

  • 3-Deazaadenosine Prevents Smooth Muscle Cell Proliferation and Neointima Formation by Interfering With Ras Signaling
    Circulation research, 2009
    Co-Authors: Daniel Sedding, Gerhard Walker, Ludger Fink, Werner Haberbosch, Harald Tillmanns, Monique Tröbs, Fabian Reich, Wigbert S. Rau, Klaus T. Preissner, Rainer M. Bohle
    Abstract:

    3-Deazaadenosine (c3Ado) is a potent inhibitor of S -adenosylhomocysteine hydrolase, which regulates cellular methyltransferase activity. In the present study, we sought to determine the effect of c3Ado on vascular smooth muscle cell (VSMC) function and neointima formation in vivo. c3Ado dose-dependently prevented the proliferation and migration of human coronary VSMCs in vitro. This was accompanied by an increased expression of the cyclin-dependent kinase inhibitors p21 WAF1/Cip1 , p27 Kip1 , a decreased expression of G 1 /S phase cyclins, and a lack of retinoblastoma protein hyperphosphorylation. In accordance with these findings, fluorescence-activated cell-sorting analysis of propidium iodide–stained cells indicated a cell cycle arrest in the G 0 /G 1 phase. Importantly, c3Ado did not affect the number of viable (trypan blue exclusion) or apoptotic cells (TUNEL). Mechanistically, c3Ado prevented FCS-induced Ras carboxyl methylation and membrane translocation and activity by inhibiting isoprenylcysteine carboxyl methyltransferase and reduced FCS-induced extracellular signal-regulated kinase (ERK)1/2 and Akt phosphorylation in a dose-dependent manner. Conversely, rescuing signal transduction by overexpression of a constitutive active Ras mutant abrogated c3Ado’s effect on proliferation. For in vivo studies, the femoral artery of C57BL/6 mice was dilated and mice were fed a diet containing 150 μg of c3Ado per day. c3Ado prevented dilation-induced Ras activation, as well as ERK1/2 and Akt phosphorylation in vivo. At day 21, VSMC proliferation (proliferating-cell nuclear antigen [PCNA]-positive cells), as well as the neointima/media ratio (0.7±0.2 versus 1.6±0.4; P S -adenosylhomocysteine hydrolase by c3Ado may represent a save and effective novel approach to prevent vascular proliferative disease.

  • 3 Deazaadenosine inhibits vasa vasorum neovascularization in aortas of apoe ldl double knockout mice
    Atherosclerosis, 2009
    Co-Authors: A C Langheinrich, Daniel Sedding, Werner Haberbosch, Marian Kampschulte, Regina Moritz, Jochen Wilhelm, Erik L Ritman, Rainer M. Bohle
    Abstract:

    Abstract Background Atherosclerosis and inflammation/angiogenesis are strongly associated including growth of vasa vasorum (VV) and plaque neovascularization, but a causative role for neovascularization has still not been established. Hence, we investigated the effect of 3-Deazaadenosine (c 3 Ado), an anti-inflammatory and anti-proliferative drug, on plaque progression and VV neovascularization in apoE −/− /LDL −/− double knockout mice. Methods The arterial trees from apoE −/− /LDL −/− mice with, or without c 3 Ado at the age of 16 weeks ( n =10), 18 weeks ( n =8) and 20 weeks ( n =7) were infused in situ with Microfil, and the aortas harvested and scanned with micro-CT (12μm cubic voxel). We characterized plaque volume and VV luminal volume along the descending aorta using Analyze 6.0 software. Cellular effects of c 3 Ado on human endothelial and vascular smooth muscle cells were investigated in cell cultures and on nylon cDNA expression arrays. Results Lesions spatially connected to VV increased from 16 to 20 weeks significantly ( p 3 Ado ( p p 3 Ado. Moreover, c 3 Ado dose-dependently prevented the proliferation and migration of human coronary artery endothelial cells in vitro. Conclusion The smaller lesion volume in animals treated with c 3 Ado was closely associated with a reduced VV neovascularization, suggesting a direct relationship between lesion growth and VV development.

  • 3-Deazaadenosine inhibits vasa vasorum neovascularization in aortas of ApoE(-/-)/LDL(-/-) double knockout mice.
    Atherosclerosis, 2008
    Co-Authors: Alexander C. Langheinrich, Daniel Sedding, Werner Haberbosch, Marian Kampschulte, Regina Moritz, Jochen Wilhelm, Erik L Ritman, Rainer M. Bohle
    Abstract:

    Atherosclerosis and inflammation/angiogenesis are strongly associated including growth of vasa vasorum (VV) and plaque neovascularization, but a causative role for neovascularization has still not been established. Hence, we investigated the effect of 3-Deazaadenosine (c(3)Ado), an anti-inflammatory and anti-proliferative drug, on plaque progression and VV neovascularization in apoE(-/-)/LDL(-/-) double knockout mice. The arterial trees from apoE(-/-)/LDL(-/-) mice with, or without c(3)Ado at the age of 16 weeks (n=10), 18 weeks (n=8) and 20 weeks (n=7) were infused in situ with Microfil, and the aortas harvested and scanned with micro-CT (12mum cubic voxel). We characterized plaque volume and VV luminal volume along the descending aorta using Analyze 6.0 software. Cellular effects of c(3)Ado on human endothelial and vascular smooth muscle cells were investigated in cell cultures and on nylon cDNA expression arrays. Lesions spatially connected to VV increased from 16 to 20 weeks significantly (p<0.001). The volume of atherosclerotic lesions was significantly reduced in animals treated with c(3)Ado (p<0.01). This was accompanied by a significant decrease of vasa vasorum neovascularization along the descending aorta (p<0.01). Using nylon cDNA expression arrays, we identified the regulation of anti-proliferative, anti-inflammatory genes in human smooth muscle cells which might be involved in the anti-angiogenic effects of c(3)Ado. Moreover, c(3)Ado dose-dependently prevented the proliferation and migration of human coronary artery endothelial cells in vitro. The smaller lesion volume in animals treated with c(3)Ado was closely associated with a reduced VV neovascularization, suggesting a direct relationship between lesion growth and VV development.

  • 3-Deazaadenosine prevents leukocyte invasion by suppression of adhesion molecule expression during acute cardiac allograft rejection: involvement of apoptotic cell death.
    The Journal of heart and lung transplantation : the official publication of the International Society for Heart Transplantation, 2004
    Co-Authors: Horst Fingerhuth, Ruediger C. Braun-dullaeus, Werner Haberbosch, Harald Tillmanns, Hans Hölschermann, H. Grimm, T.h.w Stadlbauer
    Abstract:

    Abstract Background In the initial phase after cardiac transplantation, mononuclear cells infiltrate the graft, initiating a relevant impulse for rejection. 3-Deazaadenosine (c3Ado), an analog of adenosine, has proven anti-inflammatory properties both in vitro and in vivo. We hypothesized that c3Ado can serve as a therapeutic tool to reduce cellular infiltration in cardiac allograft transplantation. Methods Using the Wistar-Furth-to-Lewis rat cardiac allograft model, animals were treated with 5 mg c3Ado subcutaneously twice per day. Allografts of untreated animals served as controls. Grafts were harvested on Days 1, 3 and 6 after transplantation for further examination (n = 4 per group and timepoint). Results Immunohistochemical examination of c3Ado-treated grafts revealed up to 80% reduction of infiltrating major histocompatability complex (MHC) II-positive cells and T-cell-receptor-positive cells (R73) as well as ED1-positive monocytes and macrophages at Days 3 and 6 after transplantation. Adhesion molecule (ICAM-1 and VCAM-1) expression at Days 1 and 3 was almost completely abolished in c3Ado-treated grafts. However, c3Ado treatment did not prevent apoptotic cell death (TUNEL assay, DNA laddering) at Day 6, nor did it prolong allograft survival. As in controls, grafts were rejected at Day 7. Conclusion c3Ado significantly reduces graft infiltration by preventing leukocyte invasion, most likely through suppression of adhesion molecule expression. Although graft survival was not prolonged, treatment with c3Ado may still serve as a strategy to protect hearts from early damage after transplantation. Further studies will show whether peri-operative use of c3Ado can bridge the critical phase after transplantation when standard immunosuppression is not yet completely efficacious.

  • Effects of 3-Deazaadenosine on homocysteine and atherosclerosis in apolipoprotein E-deficient mice.
    Atherosclerosis, 2003
    Co-Authors: Alexander C. Langheinrich, Ruediger C. Braun-dullaeus, Gerhard Walker, Ina Jeide, Ralph Schilling, Kai Tammoscheit, Thomas Dreyer, Ludger Fink, Rainer M. Bohle, Werner Haberbosch
    Abstract:

    Abstract Objective: In the past decade, elevated homocysteine concentration has achieved widespread recognition as an independent risk factor in the development of atherosclerosis. 3-Deazaadenosine (c 3 Ado) is a potent inhibitor and substrate for S -adenosylhomocysteine hydrolase and therefore may reduce homocysteine concentrations. The current study investigated the effect of c 3 Ado on serum homocysteine, atherosclerotic lesions, and the expression of adhesion molecules in apoE-knockout mice. Methods and results: Animals were placed on an atherogenic diet with or without c 3 Ado for 12 and 24 weeks. Frozen cross-sections of the aortic sinus and the proximal aorta were analyzed by computer-aided planimetry for fatty plaque formation. Macrophages, VCAM-1 and ICAM-1 were quantified by immunhistochemistry and oligo-cell reverse transcription polymerase chain reaction after laser microdissection. Application of c 3 Ado resulted in significant reduction of homocysteine levels by 35.9 and 45.3% after 12 and 24 weeks, respectively ( P 3 Ado ( P P P Conclusion: Our results demonstrate that c 3 Ado induces a marked reduction of homocysteine concentrations which might explain in part the anti-atherogenic effect of the drug.

Daniel Sedding - One of the best experts on this subject based on the ideXlab platform.

  • 3-Deazaadenosine Prevents Smooth Muscle Cell Proliferation and Neointima Formation by Interfering With Ras Signaling
    Circulation research, 2009
    Co-Authors: Daniel Sedding, Gerhard Walker, Ludger Fink, Werner Haberbosch, Harald Tillmanns, Monique Tröbs, Fabian Reich, Wigbert S. Rau, Klaus T. Preissner, Rainer M. Bohle
    Abstract:

    3-Deazaadenosine (c3Ado) is a potent inhibitor of S -adenosylhomocysteine hydrolase, which regulates cellular methyltransferase activity. In the present study, we sought to determine the effect of c3Ado on vascular smooth muscle cell (VSMC) function and neointima formation in vivo. c3Ado dose-dependently prevented the proliferation and migration of human coronary VSMCs in vitro. This was accompanied by an increased expression of the cyclin-dependent kinase inhibitors p21 WAF1/Cip1 , p27 Kip1 , a decreased expression of G 1 /S phase cyclins, and a lack of retinoblastoma protein hyperphosphorylation. In accordance with these findings, fluorescence-activated cell-sorting analysis of propidium iodide–stained cells indicated a cell cycle arrest in the G 0 /G 1 phase. Importantly, c3Ado did not affect the number of viable (trypan blue exclusion) or apoptotic cells (TUNEL). Mechanistically, c3Ado prevented FCS-induced Ras carboxyl methylation and membrane translocation and activity by inhibiting isoprenylcysteine carboxyl methyltransferase and reduced FCS-induced extracellular signal-regulated kinase (ERK)1/2 and Akt phosphorylation in a dose-dependent manner. Conversely, rescuing signal transduction by overexpression of a constitutive active Ras mutant abrogated c3Ado’s effect on proliferation. For in vivo studies, the femoral artery of C57BL/6 mice was dilated and mice were fed a diet containing 150 μg of c3Ado per day. c3Ado prevented dilation-induced Ras activation, as well as ERK1/2 and Akt phosphorylation in vivo. At day 21, VSMC proliferation (proliferating-cell nuclear antigen [PCNA]-positive cells), as well as the neointima/media ratio (0.7±0.2 versus 1.6±0.4; P S -adenosylhomocysteine hydrolase by c3Ado may represent a save and effective novel approach to prevent vascular proliferative disease.

  • the nucleotide analogue 3 Deazaadenosine prevents neointima formation after balloon injury
    Biochemical and Biophysical Research Communications, 2009
    Co-Authors: Florian H. Seeger, Wolfram Hess, Daniel Sedding, Gunter Becker, Ralf Kinscherf, Christiane Viedt, Ruediger C Braundullaeus, Jörg Kreuzer
    Abstract:

    We have recently shown that 3-Deazaadenosine (c3Ado) inhibits atherogenesis in mice. We studied whether its anti-inflammatory capacity would also affect neointima-formation after balloon injury. Sprague Dawley rats underwent balloon angioplasty. C3Ado was administered orally, starting 5 days prior to the balloon injury and continued for 2 weeks. Fourteen days after balloon injury the intima/media ratio in the c3Ado-treated group was reduced by 67% (p < 0.001) and luminal stenosis by 50% (p < 0.001). Neointimal cellular density was decreased by 25% (p < 0.001) and the induction of c-Jun and ki67 was markedly lower. The reduction of the intima/media ratio was still observed 3 months after balloon injury. Furthermore, a c3Ado-dependent inhibition of PDGF-mediated ERK-activation and proliferation could be demonstrated. Short-term administration of C3Ado inhibits neointima-formation in rats for at least 3 months after injury. The present findings implicate that c3Ado may be useful as an inhibitor of restenosis-formation after balloon angioplasty in humans.

  • 3 Deazaadenosine inhibits vasa vasorum neovascularization in aortas of apoe ldl double knockout mice
    Atherosclerosis, 2009
    Co-Authors: A C Langheinrich, Daniel Sedding, Werner Haberbosch, Marian Kampschulte, Regina Moritz, Jochen Wilhelm, Erik L Ritman, Rainer M. Bohle
    Abstract:

    Abstract Background Atherosclerosis and inflammation/angiogenesis are strongly associated including growth of vasa vasorum (VV) and plaque neovascularization, but a causative role for neovascularization has still not been established. Hence, we investigated the effect of 3-Deazaadenosine (c 3 Ado), an anti-inflammatory and anti-proliferative drug, on plaque progression and VV neovascularization in apoE −/− /LDL −/− double knockout mice. Methods The arterial trees from apoE −/− /LDL −/− mice with, or without c 3 Ado at the age of 16 weeks ( n =10), 18 weeks ( n =8) and 20 weeks ( n =7) were infused in situ with Microfil, and the aortas harvested and scanned with micro-CT (12μm cubic voxel). We characterized plaque volume and VV luminal volume along the descending aorta using Analyze 6.0 software. Cellular effects of c 3 Ado on human endothelial and vascular smooth muscle cells were investigated in cell cultures and on nylon cDNA expression arrays. Results Lesions spatially connected to VV increased from 16 to 20 weeks significantly ( p 3 Ado ( p p 3 Ado. Moreover, c 3 Ado dose-dependently prevented the proliferation and migration of human coronary artery endothelial cells in vitro. Conclusion The smaller lesion volume in animals treated with c 3 Ado was closely associated with a reduced VV neovascularization, suggesting a direct relationship between lesion growth and VV development.

  • The nucleotide analogue 3-Deazaadenosine prevents neointima-formation after balloon injury.
    Biochemical and biophysical research communications, 2008
    Co-Authors: Florian H. Seeger, Wolfram Hess, Daniel Sedding, Gunter Becker, Ralf Kinscherf, Ruediger C. Braun-dullaeus, Christiane Viedt, Jörg Kreuzer
    Abstract:

    We have recently shown that 3-Deazaadenosine (c3Ado) inhibits atherogenesis in mice. We studied whether its anti-inflammatory capacity would also affect neointima-formation after balloon injury. Sprague Dawley rats underwent balloon angioplasty. C3Ado was administered orally, starting 5 days prior to the balloon injury and continued for 2 weeks. Fourteen days after balloon injury the intima/media ratio in the c3Ado-treated group was reduced by 67% (p

  • 3-Deazaadenosine inhibits vasa vasorum neovascularization in aortas of ApoE(-/-)/LDL(-/-) double knockout mice.
    Atherosclerosis, 2008
    Co-Authors: Alexander C. Langheinrich, Daniel Sedding, Werner Haberbosch, Marian Kampschulte, Regina Moritz, Jochen Wilhelm, Erik L Ritman, Rainer M. Bohle
    Abstract:

    Atherosclerosis and inflammation/angiogenesis are strongly associated including growth of vasa vasorum (VV) and plaque neovascularization, but a causative role for neovascularization has still not been established. Hence, we investigated the effect of 3-Deazaadenosine (c(3)Ado), an anti-inflammatory and anti-proliferative drug, on plaque progression and VV neovascularization in apoE(-/-)/LDL(-/-) double knockout mice. The arterial trees from apoE(-/-)/LDL(-/-) mice with, or without c(3)Ado at the age of 16 weeks (n=10), 18 weeks (n=8) and 20 weeks (n=7) were infused in situ with Microfil, and the aortas harvested and scanned with micro-CT (12mum cubic voxel). We characterized plaque volume and VV luminal volume along the descending aorta using Analyze 6.0 software. Cellular effects of c(3)Ado on human endothelial and vascular smooth muscle cells were investigated in cell cultures and on nylon cDNA expression arrays. Lesions spatially connected to VV increased from 16 to 20 weeks significantly (p<0.001). The volume of atherosclerotic lesions was significantly reduced in animals treated with c(3)Ado (p<0.01). This was accompanied by a significant decrease of vasa vasorum neovascularization along the descending aorta (p<0.01). Using nylon cDNA expression arrays, we identified the regulation of anti-proliferative, anti-inflammatory genes in human smooth muscle cells which might be involved in the anti-angiogenic effects of c(3)Ado. Moreover, c(3)Ado dose-dependently prevented the proliferation and migration of human coronary artery endothelial cells in vitro. The smaller lesion volume in animals treated with c(3)Ado was closely associated with a reduced VV neovascularization, suggesting a direct relationship between lesion growth and VV development.