Anti-interleukin-6 therapy for treatment of high platelet counts in cGMP-dependent protein kinase I...
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POSTER PRESENTATION Open Access Anti-interleukin-6 therapy for treatment of high platelet counts in cGMP-dependent protein kinase I gene-targeted mice Lin Zhang 1 , Robert Lukowski 2,3* , Florian Gaertner 1 , Michael Lorenz 1 , Kyle R Legate 1,4 , Katrin Domes 2 , Elisabeth Angermeier 2 , Franz Hofmann 2 , Steffen Massberg 1,2 From 6th International Conference on cGMP: Generators, Effectors and Therapeutic Implications Erfurt, Germany. 28-30 June 2013 Background The cyclic guanosine-3’ ,5’ -monophosphate (cGMP)/ cGMP-dependent protein kinase type I (cGKI) pathway is a potent negative regulator of platelet adhesion and aggre- gation [1]; however, the role of cGMP/cGKI for platelet biogenesis in vivo is unclear. Results Here we report thrombocytosis in conventional cGKI null mutants (cGKI L1/L1 ) and gene-targeted cGKI a/b rescue mice (referred to as cGKI-SM) with cGKI expres- sion specifically restored in smooth muscle (SM), but not in other cell types [2-4]. In contrast, conditional knockouts lacking the cGKI protein specifically in the megakaryocyte (MK)/platelet lineage (Pf4-Cre tg/+ ; cGKI L2/L2 ) did not display a related thrombocytosis phe- notype, indicating that the high platelet count of cGKI L1/L1 and cGKI-SM mutants is rather a reactive response than an intrinsic defect in megakaryopoiesis. In line with these findings, wild-type (WT) mice engrafted with cGKI-deficient bone-marrow (BM) cells showed full reconstitution of haematopoiesis and normal platelet counts upon myeloablative radiotherapy. Stimulation of BM-derived WT MKs using serum preparations from cGKI-SM mutants strongly accelerated megakaryopoiesis, suggesting that their high platelet counts develop in response to soluble factors. Indeed, we confirm elevated Interleukin-6 (IL-6) serum levels [5,6], a known cause for reactive thrombocytosis, in cGKI-SM mutants, whereas IL-6 was unaltered in Pf4-Cre tg/+ ; cGKI L2/L2 mice and cGKI-deficient BM chimaeras. Vice versa, antibody- mediated blockage of IL-6 reduced platelet counts in cGKI-SM mice, but not in WT mice. Conclusion We conclude that abnormal signalling of cGMP/cGKI in non-hematopoietic cells affects thrombopoiesis via IL-6 resulting in a reactive thrombocytosis in vivo. Authors’ details 1 Medizinische Klinik und Poliklinik I, Klinikum der Universität, Ludwig- Maximilians-Universität, München, Germany. 2 Forschergruppe 923, Institut für Pharmakologie und Toxikologie, Technische Universität München, München, Germany. 3 Pharmakologie, Toxikologie und Klinische Pharmazie, Institut für Pharmazie, Universität Tübingen, Tübingen, Germany. 4 Center for NanoScience, Department of Applied Physics, Ludwig-Maximilians- Universität, München, München, Germany. Published: 29 August 2013 References 1. Massberg S, Sausbier M, Klatt P, Bauer M, Pfeifer A, Siess W, Fassler R, Ruth P, Krombach F, Hofmann F: Increased adhesion and aggregation of platelets lacking cyclic guanosine 3’,5’-monophosphate kinase I. J Expt Med 1999, 189:1255-1264. 2. Weber S, Bernhard D, Lukowski R, Weinmeister P, Worner R, Wegener JW, Valtcheva N, Feil S, Schlossmann J, Hofmann F, Feil R: Rescue of cGMP kinase I knockout mice by smooth muscle specific expression of either isozyme. Circ Res 2007, 101:1096-1103. 3. Lukowski R, Rybalkin SD, Loga F, Leiss V, Beavo JA, Hofmann F: Cardiac hypertrophy is not amplified by deletion of cGMP-dependent protein kinase I in cardiomyocytes. Proc Natl Acadf Sci USA 2010, 107:5646-5651. 4. Leiss V, Friebe A, Welling A, Hofmann F, Lukowski R: Cyclic GMP kinase I modulates glucagon release from pancreatic alpha-cells. Diabetes 2011, 60:148-156. 5. Lut SZZ, Hennige AM, Feil S, Peter A, Gerling A, Machann J, Krober SM, Rath M, Schurmann A, Weigert C, Haring HU, Feil R: Genetic ablation of cGMP-dependent protein kinase type I causes liver inflammation and fasting hyperglycemia. Diabetes 2011, 60:1566-1576. 6. Mitschke MM, Hoffmann LS, Gnad T, Scholz D, Kruithoff K, Mayer P, Haas B, Sassmann A, Pfeifer A, Kilic A: Increased cGMP promotes healthy * Correspondence: [email protected] 2 Forschergruppe 923, Institut für Pharmakologie und Toxikologie, Technische Universität München, München, Germany Full list of author information is available at the end of the article Zhang et al. BMC Pharmacology and Toxicology 2013, 14(Suppl 1):P80 http://www.biomedcentral.com/2050-6511/14/S1/P80 © 2013 Zhang et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative Commons Attribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction in any medium, provided the original work is properly cited.
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Transcript of Anti-interleukin-6 therapy for treatment of high platelet counts in cGMP-dependent protein kinase I...
POSTER PRESENTATION Open Access
Anti-interleukin-6 therapy for treatment of highplatelet counts in cGMP-dependent proteinkinase I gene-targeted miceLin Zhang1, Robert Lukowski2,3*, Florian Gaertner1, Michael Lorenz1, Kyle R Legate1,4, Katrin Domes2,Elisabeth Angermeier2, Franz Hofmann2, Steffen Massberg1,2
From 6th International Conference on cGMP: Generators, Effectors and Therapeutic ImplicationsErfurt, Germany. 28-30 June 2013
BackgroundThe cyclic guanosine-3’,5’-monophosphate (cGMP)/cGMP-dependent protein kinase type I (cGKI) pathway isa potent negative regulator of platelet adhesion and aggre-gation [1]; however, the role of cGMP/cGKI for plateletbiogenesis in vivo is unclear.
ResultsHere we report thrombocytosis in conventional cGKInull mutants (cGKIL1/L1) and gene-targeted cGKIa/brescue mice (referred to as cGKI-SM) with cGKI expres-sion specifically restored in smooth muscle (SM), butnot in other cell types [2-4]. In contrast, conditionalknockouts lacking the cGKI protein specifically in themegakaryocyte (MK)/platelet lineage (Pf4-Cretg/+;cGKIL2/L2) did not display a related thrombocytosis phe-notype, indicating that the high platelet count ofcGKIL1/L1 and cGKI-SM mutants is rather a reactiveresponse than an intrinsic defect in megakaryopoiesis. Inline with these findings, wild-type (WT) mice engraftedwith cGKI-deficient bone-marrow (BM) cells showed fullreconstitution of haematopoiesis and normal plateletcounts upon myeloablative radiotherapy. Stimulation ofBM-derived WT MKs using serum preparations fromcGKI-SM mutants strongly accelerated megakaryopoiesis,suggesting that their high platelet counts develop inresponse to soluble factors. Indeed, we confirm elevatedInterleukin-6 (IL-6) serum levels [5,6], a known cause forreactive thrombocytosis, in cGKI-SM mutants, whereasIL-6 was unaltered in Pf4-Cretg/+; cGKIL2/L2 mice and
cGKI-deficient BM chimaeras. Vice versa, antibody-mediated blockage of IL-6 reduced platelet counts incGKI-SM mice, but not in WT mice.
ConclusionWe conclude that abnormal signalling of cGMP/cGKI innon-hematopoietic cells affects thrombopoiesis via IL-6resulting in a reactive thrombocytosis in vivo.
Authors’ details1Medizinische Klinik und Poliklinik I, Klinikum der Universität, Ludwig-Maximilians-Universität, München, Germany. 2Forschergruppe 923, Institut fürPharmakologie und Toxikologie, Technische Universität München, München,Germany. 3Pharmakologie, Toxikologie und Klinische Pharmazie, Institut fürPharmazie, Universität Tübingen, Tübingen, Germany. 4Center forNanoScience, Department of Applied Physics, Ludwig-Maximilians-Universität, München, München, Germany.
Published: 29 August 2013
References1. Massberg S, Sausbier M, Klatt P, Bauer M, Pfeifer A, Siess W, Fassler R,
Ruth P, Krombach F, Hofmann F: Increased adhesion and aggregation ofplatelets lacking cyclic guanosine 3’,5’-monophosphate kinase I. J ExptMed 1999, 189:1255-1264.
2. Weber S, Bernhard D, Lukowski R, Weinmeister P, Worner R, Wegener JW,Valtcheva N, Feil S, Schlossmann J, Hofmann F, Feil R: Rescue of cGMPkinase I knockout mice by smooth muscle specific expression of eitherisozyme. Circ Res 2007, 101:1096-1103.
3. Lukowski R, Rybalkin SD, Loga F, Leiss V, Beavo JA, Hofmann F: Cardiachypertrophy is not amplified by deletion of cGMP-dependent proteinkinase I in cardiomyocytes. Proc Natl Acadf Sci USA 2010, 107:5646-5651.
4. Leiss V, Friebe A, Welling A, Hofmann F, Lukowski R: Cyclic GMP kinase Imodulates glucagon release from pancreatic alpha-cells. Diabetes 2011,60:148-156.
5. Lut SZZ, Hennige AM, Feil S, Peter A, Gerling A, Machann J, Krober SM,Rath M, Schurmann A, Weigert C, Haring HU, Feil R: Genetic ablation ofcGMP-dependent protein kinase type I causes liver inflammation andfasting hyperglycemia. Diabetes 2011, 60:1566-1576.
6. Mitschke MM, Hoffmann LS, Gnad T, Scholz D, Kruithoff K, Mayer P, Haas B,Sassmann A, Pfeifer A, Kilic A: Increased cGMP promotes healthy
* Correspondence: [email protected] 923, Institut für Pharmakologie und Toxikologie, TechnischeUniversität München, München, GermanyFull list of author information is available at the end of the article
Zhang et al. BMC Pharmacology and Toxicology 2013, 14(Suppl 1):P80http://www.biomedcentral.com/2050-6511/14/S1/P80
© 2013 Zhang et al; licensee BioMed Central Ltd. This is an Open Access article distributed under the terms of the Creative CommonsAttribution License (http://creativecommons.org/licenses/by/2.0), which permits unrestricted use, distribution, and reproduction inany medium, provided the original work is properly cited.
expansion and browning of white adipose tissue. FASEB J 2013,27:1621-1630.
doi:10.1186/2050-6511-14-S1-P80Cite this article as: Zhang et al.: Anti-interleukin-6 therapy for treatmentof high platelet counts in cGMP-dependent protein kinase I gene-targeted mice. BMC Pharmacology and Toxicology 2013 14(Suppl 1):P80.
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Zhang et al. BMC Pharmacology and Toxicology 2013, 14(Suppl 1):P80http://www.biomedcentral.com/2050-6511/14/S1/P80
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