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Article ; Online: Visualising and understanding cGMP signals in the cardiovascular system.

Feil, Robert / Lehners, Moritz / Stehle, Daniel / Feil, Susanne

2021 Volume 179, Issue 11, Page(s) 2394–2412

Abstract: cGMP is an important signalling molecule in humans. Fluorescent cGMP biosensors have emerged as powerful tools for the sensitive analysis of cGMP pathways at the single-cell level. Here, we briefly outline cGMP's multifaceted role in (patho)physiology ... ...

Abstract	cGMP is an important signalling molecule in humans. Fluorescent cGMP biosensors have emerged as powerful tools for the sensitive analysis of cGMP pathways at the single-cell level. Here, we briefly outline cGMP's multifaceted role in (patho)physiology and pharmacotherapy. Then we summarise what new insights cGMP imaging has provided into endogenous cGMP signalling and drug action, with a focus on the cardiovascular system. Indeed, the use of cGMP biosensors has led to several conceptual advances, such as the discovery of local, intercellular and mechanosensitive cGMP signals. Importantly, single-cell imaging can provide valuable information about the heterogeneity of cGMP signals within and between individual cells of an isolated cell population or tissue. We also discuss current challenges and future directions of cGMP imaging, such as the direct visualisation of cGMP microdomains, simultaneous monitoring of cGMP and other signalling molecules and, ultimately, cGMP imaging in tissues and animals under close-to-native conditions. LINKED ARTICLES: This article is part of a themed issue on cGMP Signalling in Cell Growth and Survival. To view the other articles in this section visit http://onlinelibrary.wiley.com/doi/10.1111/bph.v179.11/issuetoc.
MeSH term(s)	Animals ; Cardiovascular System/metabolism ; Cyclic GMP/metabolism ; Signal Transduction
Chemical Substances	Cyclic GMP (H2D2X058MU)
Language	English
Publishing date	2021-05-22
Publishing country	England
Document type	Journal Article ; Review ; Research Support, Non-U.S. Gov't
ZDB-ID	80081-8
ISSN	1476-5381 ; 0007-1188
ISSN (online)	1476-5381
ISSN	0007-1188
DOI	10.1111/bph.15500
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Article ; Online: Noninvasive Detection of Smooth Muscle Cell-Derived Hot Spots to Study Atherosclerosis by PET/MRI in Mice.

Feil, Susanne / Stowbur, Dimitri / Schörg, Barbara F / Ehrlichmann, Walter / Reischl, Gerald / Kneilling, Manfred / Pichler, Bernd J / Feil, Robert

Circulation research

2023 Volume 132, Issue 6, Page(s) 747–750

MeSH term(s)	Animals ; Mice ; Positron-Emission Tomography ; Atherosclerosis/diagnostic imaging ; Magnetic Resonance Imaging ; Fluorodeoxyglucose F18
Chemical Substances	Fluorodeoxyglucose F18 (0Z5B2CJX4D)
Language	English
Publishing date	2023-02-16
Publishing country	United States
Document type	Research Support, Non-U.S. Gov't ; Journal Article
ZDB-ID	80100-8
ISSN	1524-4571 ; 0009-7330 ; 0931-6876
ISSN (online)	1524-4571
ISSN	0009-7330 ; 0931-6876
DOI	10.1161/CIRCRESAHA.122.322296
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Article ; Online: Role of the NO-GC/cGMP signaling pathway in platelet biomechanics.

Balmes, Aylin / Rodríguez, Johanna G / Seifert, Jan / Pinto-Quintero, Daniel / Khawaja, Akif A / Boffito, Marta / Frye, Maike / Friebe, Andreas / Emerson, Michael / Seta, Francesca / Feil, Robert / Feil, Susanne / Schäffer, Tilman E

Platelets

2024 Volume 35, Issue 1, Page(s) 2313359

Abstract: Cyclic guanosine monophosphate (cGMP) is a second messenger produced by the NO-sensitive guanylyl cyclase (NO-GC). The NO-GC/cGMP pathway in platelets has been extensively studied. However, its role in regulating the biomechanical properties of platelets ...

Abstract	Cyclic guanosine monophosphate (cGMP) is a second messenger produced by the NO-sensitive guanylyl cyclase (NO-GC). The NO-GC/cGMP pathway in platelets has been extensively studied. However, its role in regulating the biomechanical properties of platelets has not yet been addressed and remains unknown. We therefore investigated the stiffness of living platelets after treatment with the NO-GC stimulator riociguat or the NO-GC activator cinaciguat using scanning ion conductance microscopy (SICM). Stimulation of human and murine platelets with cGMP-modulating drugs decreased cellular stiffness and downregulated P-selectin, a marker for platelet activation. We also quantified changes in platelet shape using deep learning-based platelet morphometry, finding that platelets become more circular upon treatment with cGMP-modulating drugs. To test for clinical applicability of NO-GC stimulators in the context of increased thrombogenicity risk, we investigated the effect of riociguat on platelets from human immunodeficiency virus (HIV)-positive patients taking abacavir sulfate (ABC)-containing regimens. Our results corroborate a functional role of the NO-GC/cGMP pathway in platelet biomechanics, indicating that biomechanical properties such as stiffness or shape could be used as novel biomarkers in clinical research.
MeSH term(s)	Humans ; Mice ; Animals ; Biomechanical Phenomena ; Blood Platelets/metabolism ; Signal Transduction ; Guanylate Cyclase/metabolism ; Guanylate Cyclase/pharmacology ; Platelet Activation ; Cyclic GMP/metabolism ; Cyclic GMP/pharmacology ; Nitric Oxide/metabolism ; Platelet Aggregation
Chemical Substances	Guanylate Cyclase (EC 4.6.1.2) ; Cyclic GMP (H2D2X058MU) ; Nitric Oxide (31C4KY9ESH)
Language	English
Publishing date	2024-02-14
Publishing country	England
Document type	Journal Article
ZDB-ID	1034283-7
ISSN	1369-1635 ; 0953-7104
ISSN (online)	1369-1635
ISSN	0953-7104
DOI	10.1080/09537104.2024.2313359
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Article ; Online: cGMP Signaling and Vascular Smooth Muscle Cell Plasticity.

Lehners, Moritz / Dobrowinski, Hyazinth / Feil, Susanne / Feil, Robert

Journal of cardiovascular development and disease

2018 Volume 5, Issue 2

Abstract: Cyclic GMP regulates multiple cell types and functions of the cardiovascular system. This review summarizes the effects of cGMP on the growth and survival of vascular smooth muscle cells (VSMCs), which display remarkable phenotypic plasticity during the ... ...

Abstract	Cyclic GMP regulates multiple cell types and functions of the cardiovascular system. This review summarizes the effects of cGMP on the growth and survival of vascular smooth muscle cells (VSMCs), which display remarkable phenotypic plasticity during the development of vascular diseases, such as atherosclerosis. Recent studies have shown that VSMCs contribute to the development of atherosclerotic plaques by clonal expansion and transdifferentiation to macrophage-like cells. VSMCs express a variety of cGMP generators and effectors, including NO-sensitive guanylyl cyclase (NO-GC) and cGMP-dependent protein kinase type I (cGKI), respectively. According to the traditional view, cGMP inhibits VSMC proliferation, but this concept has been challenged by recent findings supporting a stimulatory effect of the NO-cGMP-cGKI axis on VSMC growth. Here, we summarize the relevant studies with a focus on VSMC growth regulation by the NO-cGMP-cGKI pathway in cultured VSMCs and mouse models of atherosclerosis, restenosis, and angiogenesis. We discuss potential reasons for inconsistent results, such as the use of genetic versus pharmacological approaches and primary versus subcultured cells. We also explore how modern methods for cGMP imaging and cell tracking could help to improve our understanding of cGMP’s role in vascular plasticity. We present a revised model proposing that cGMP promotes phenotypic switching of contractile VSMCs to VSMC-derived plaque cells in atherosclerotic lesions. Regulation of vascular remodeling by cGMP is not only an interesting new therapeutic strategy, but could also result in side effects of clinically used cGMP-elevating drugs.
Language	English
Publishing date	2018-04-19
Publishing country	Switzerland
Document type	Journal Article ; Review
ZDB-ID	2777082-5
ISSN	2308-3425 ; 2308-3425
ISSN (online)	2308-3425
ISSN	2308-3425
DOI	10.3390/jcdd5020020
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Article ; Online: Protocol to visualize CD11c

Sauter, Manuela / Sauter, Reinhard J / Olbrich, Marcus / Thunemann, Martin / Feil, Susanne / Feil, Robert / Langer, Harald F

STAR protocols

2022 Volume 3, Issue 3, Page(s) 101645

Abstract: Here, we describe ... ...

Abstract	Here, we describe an
MeSH term(s)	Animals ; Atherosclerosis/genetics ; CD11c Antigen/genetics ; Lac Operon/genetics ; Mice ; Mice, Knockout ; Plaque, Atherosclerotic/genetics
Chemical Substances	CD11c Antigen
Language	English
Publishing date	2022-08-19
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	2666-1667
ISSN (online)	2666-1667
DOI	10.1016/j.xpro.2022.101645
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Article ; Online: Optogenetic manipulation of cyclic guanosine monophosphate to probe phosphodiesterase activities in megakaryocytes.

Zhang, Yujing / Benz, Pascal / Stehle, Daniel / Yang, Shang / Kurz, Hendrikje / Feil, Susanne / Nagel, Georg / Feil, Robert / Gao, Shiqiang / Bender, Markus

Open biology

2022 Volume 12, Issue 8, Page(s) 220058

Abstract: Cyclic guanosine monophosphate (cGMP) signalling plays a fundamental role in many cell types, including platelets. cGMP has been implicated in platelet formation, but mechanistic detail about its spatio-temporal regulation in megakaryocytes (MKs) is ... ...

Abstract	Cyclic guanosine monophosphate (cGMP) signalling plays a fundamental role in many cell types, including platelets. cGMP has been implicated in platelet formation, but mechanistic detail about its spatio-temporal regulation in megakaryocytes (MKs) is lacking. Optogenetics is a technique which allows spatio-temporal manipulation of molecular events in living cells or organisms. We took advantage of this method and expressed a photo-activated guanylyl cyclase,
MeSH term(s)	Blastocladiella ; Cyclic GMP/metabolism ; Guanosine Monophosphate ; Megakaryocytes/metabolism ; Nitric Oxide/metabolism ; Optogenetics ; Phosphoric Diester Hydrolases
Chemical Substances	Nitric Oxide (31C4KY9ESH) ; Guanosine Monophosphate (85-32-5) ; Phosphoric Diester Hydrolases (EC 3.1.4.-) ; Cyclic GMP (H2D2X058MU)
Language	English
Publishing date	2022-08-17
Publishing country	England
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ZDB-ID	2630944-0
ISSN	2046-2441 ; 2046-2441
ISSN (online)	2046-2441
ISSN	2046-2441
DOI	10.1098/rsob.220058
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Article ; Online: Near-Infrared II Photobiomodulation Preconditioning Ameliorates Stroke Injury via Phosphorylation of eNOS.

Yokomizo, Shinya / Kopp, Timo / Roessing, Malte / Morita, Atsuyo / Lee, Seeun / Cho, Suin / Ogawa, Emiyu / Komai, Eri / Inoue, Kazumasa / Fukushi, Masahiro / Feil, Susanne / Kim, Hyung-Hwan / Bragin, Denis E / Gerashchenko, Dmitry / Huang, Paul L / Kashiwagi, Satoshi / Atochin, Dmitriy N

Stroke

2024

Abstract: Background: The current management of patients with stroke with intravenous thrombolysis and endovascular thrombectomy is effective only when it is timely performed on an appropriately selected but minor fraction of patients. The development of novel ... ...

Abstract	Background: The current management of patients with stroke with intravenous thrombolysis and endovascular thrombectomy is effective only when it is timely performed on an appropriately selected but minor fraction of patients. The development of novel adjunctive therapy is highly desired to reduce morbidity and mortality with stroke. Since endothelial dysfunction is implicated in the pathogenesis of stroke and is featured with suppressed endothelial nitric oxide synthase (eNOS) with concomitant nitric oxide deficiency, restoring endothelial nitric oxide represents a promising approach to treating stroke injury. Methods: This is a preclinical proof-of-concept study to determine the therapeutic effect of transcranial treatment with a low-power near-infrared laser in a mouse model of ischemic stroke. The laser treatment was performed before the middle cerebral artery occlusion with a filament. To determine the involvement of eNOS phosphorylation, unphosphorylatable eNOS S1176A knock-in mice were used. Each measurement was analyzed by a 2-way ANOVA to assess the effect of the treatment on cerebral blood flow with laser Doppler flowmetry, eNOS phosphorylation by immunoblot analysis, and stroke outcomes by infarct volumes and neurological deficits. Results: Pretreatment with a 1064-nm laser at an irradiance of 50 mW/cm Conclusions: Near-infrared II photobiomodulation could offer a noninvasive and low-risk adjunctive therapy for stroke injury. This new modality using a physical parameter merits further consideration to develop innovative therapies to prevent and treat a wide array of cardiovascular diseases.
Language	English
Publishing date	2024-04-04
Publishing country	United States
Document type	Journal Article
ZDB-ID	80381-9
ISSN	1524-4628 ; 0039-2499 ; 0749-7954
ISSN (online)	1524-4628
ISSN	0039-2499 ; 0749-7954
DOI	10.1161/STROKEAHA.123.045358
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Article ; Online: Near-infrared II photobiomodulation augments nitric oxide bioavailability via phosphorylation of endothelial nitric oxide synthase.

Yokomizo, Shinya / Roessing, Malte / Morita, Atsuyo / Kopp, Timo / Ogawa, Emiyu / Katagiri, Wataru / Feil, Susanne / Huang, Paul L / Atochin, Dmitriy N / Kashiwagi, Satoshi

FASEB journal : official publication of the Federation of American Societies for Experimental Biology

2022 Volume 36, Issue 9, Page(s) e22490

Abstract: There is solid evidence of the beneficial effect of photobiomodulation (PBM) with low-power near-infrared (NIR) light in the NIR-I window in increasing bioavailable nitric oxide (NO). However, it is not established whether this effect can be extended to ... ...

Abstract	There is solid evidence of the beneficial effect of photobiomodulation (PBM) with low-power near-infrared (NIR) light in the NIR-I window in increasing bioavailable nitric oxide (NO). However, it is not established whether this effect can be extended to NIR-II light, limiting broader applications of this therapeutic modality. Since we have demonstrated PBM with NIR laser in the NIR-II window, we determined the causal relationship between NIR-II irradiation and its specific biological effects on NO bioavailability. We analyzed the impact of NIR-II irradiation on NO release in cultured human endothelial cells using a NO-sensitive fluorescence probe and single-cell live imaging. Two distinct wavelengths of NIR-II laser (1064 and 1270 nm) and NIR-I (808 nm) at an irradiance of 10 mW/cm
MeSH term(s)	Cells, Cultured ; Endothelial Cells/metabolism ; Humans ; Nitric Oxide/metabolism ; Nitric Oxide Synthase Type III/metabolism ; Phosphorylation ; Proto-Oncogene Proteins c-akt/metabolism
Chemical Substances	Nitric Oxide (31C4KY9ESH) ; Nitric Oxide Synthase Type III (EC 1.14.13.39) ; Proto-Oncogene Proteins c-akt (EC 2.7.11.1)
Language	English
Publishing date	2022-08-05
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
ZDB-ID	639186-2
ISSN	1530-6860 ; 0892-6638
ISSN (online)	1530-6860
ISSN	0892-6638
DOI	10.1096/fj.202101890R
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Article ; Online: Genetic inducible fate mapping in adult mice using tamoxifen-dependent Cre recombinases.

Feil, Susanne / Krauss, Jana / Thunemann, Martin / Feil, Robert

Methods in molecular biology (Clifton, N.J.)

2014 Volume 1194, Page(s) 113–139

Abstract: The Cre/lox site-specific recombination system allows the control of gene activity in space and time in almost any tissue of the mouse. A major technical advance was the development of tamoxifen-dependent Cre recombinases, such as CreER(T2), that can be ... ...

Abstract	The Cre/lox site-specific recombination system allows the control of gene activity in space and time in almost any tissue of the mouse. A major technical advance was the development of tamoxifen-dependent Cre recombinases, such as CreER(T2), that can be activated by administration of tamoxifen to the animal. This powerful tool greatly facilitates the study of gene functions and the generation of more realistic animal models of sporadic human diseases. Another important application of tamoxifen-dependent Cre recombinases is genetic inducible fate mapping (GIFM). In GIFM studies, the inducible Cre/lox system is used to genetically label a defined cell population at a selected time by irreversible activation of the expression of a Cre-responsive reporter transgene. Then, marked cells are detected at later time points to determine how the originally labeled progenitors contribute to specific structures and cell types during pre- and postnatal development. GIFM was initially applied during mouse embryogenesis, but is now increasingly used for cell lineage tracing in adult mice under physiological and pathophysiological conditions. Here we describe the design of GIFM experiments in adult mice as exemplified by CreER(T2)-assisted tracing of vascular smooth muscle cells during the development of atherosclerotic lesions. First, we give an overview of reporter transgenes available for genetic cell marking that are expressed from the Rosa26 locus, such as β-galactosidase and fluorescent proteins. Then we present detailed protocols for the generation of experimental mice for GIFM studies, the induction of cell labeling by tamoxifen treatment, and the detection of marked cells in fixed and live tissues. Each section also provides a discussion of limitations and common pitfalls of GIFM experiments. Most of the protocols can be easily adapted to other developmental stages, cell types, Cre recombinases, and reporter transgenes and, thus, can be used as general guidelines for GIFM studies in mice.
MeSH term(s)	Animals ; Breeding ; Cell Differentiation/drug effects ; Cell Differentiation/genetics ; Cell Lineage/genetics ; Cell Tracking/methods ; Female ; Genes, Reporter/genetics ; Genetic Techniques ; Integrases/metabolism ; Male ; Mice ; Paraffin Embedding ; Plaque, Atherosclerotic/genetics ; Polymerase Chain Reaction ; Recombination, Genetic/drug effects ; Staining and Labeling ; Tamoxifen/pharmacology ; Transgenes/genetics
Chemical Substances	Tamoxifen (094ZI81Y45) ; Cre recombinase (EC 2.7.7.-) ; Integrases (EC 2.7.7.-)
Language	English
Publishing date	2014-06-28
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't
ISSN	1940-6029
ISSN (online)	1940-6029
DOI	10.1007/978-1-4939-1215-5_6
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Article ; Online: Dual near-infrared II laser modulates the cellular redox state of T cells and augments the efficacy of cancer immunotherapy.

FASEB journal : official publication of the Federation of American Societies for Experimental Biology

2022 Volume 36, Issue 10, Page(s) e22521

Abstract: Immunotherapy, including immune checkpoint inhibitors, has revolutionized cancer treatment, but only a minor fraction of patients shows durable responses. A new approach to overcome this limitation is yet to be identified. Recently, we have shown that ... ...

Abstract	Immunotherapy, including immune checkpoint inhibitors, has revolutionized cancer treatment, but only a minor fraction of patients shows durable responses. A new approach to overcome this limitation is yet to be identified. Recently, we have shown that photobiomodulation (PBM) with near-infrared (NIR) light in the NIR-II window reduces oxidative stress and supports the proliferation of CD8
MeSH term(s)	Animals ; CD8-Positive T-Lymphocytes ; Immunotherapy ; Lasers ; Mice ; Neoplasms/therapy ; Oxidation-Reduction
Language	English
Publishing date	2022-09-02
Publishing country	United States
Document type	Journal Article ; Research Support, Non-U.S. Gov't ; Research Support, N.I.H., Extramural
ZDB-ID	639186-2
ISSN	1530-6860 ; 0892-6638
ISSN (online)	1530-6860
ISSN	0892-6638
DOI	10.1096/fj.202200033R
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