The Guanylyl Cyclase Receptor B and Cyclic GMP Activating Actions of a Unique Molecular Form of C-Type Natriuretic Peptide In Vitro and In Vivo

Abstract C-type natriuretic peptide (CNP), the third member of natriuretic peptides, has recently been discovered from the porcine brain. Using a polyclonal antiserum to CNP, we demonstrated that CNP-like immunoreactivity (CNP-LI) is present mainly in the central nervous system. Recently, however, we have discovered the production and secretion of CNP in vascular endothelial cells. These observations suggested that CNP may act not only as a neuropeptide but also as a local regulator of vascular tone or growth. In order to further clarify the pathophysiological significance of CNP, we aimed at the preparation of a monoclonal antibody to CNP. A monoclonal antibody to CNP, KY-CNP-I, has been produced. This monoclonal antibody belongs to the immunogloblin G1 subclass and has high affinity for CNP. Using this monoclonal antibody, we established a specific radioimmunoassay (RIA) for CNP. The RIA detected CNP-LI in rat brain extracts and culture media conditioned with bovine endothelial cells. In addition, the pretreatment of cultured aortic smooth muscle cells with KY-CNP-I attenuated cyclic GMP production induced by CNP in vitro. The preadministration of KY-CNP-I to rats also attenuated plasma cyclic GMP increase after intravenous injection of CNP in vivo. These results indicate that this monoclonal antibody is a useful tool to clarify the pathophysiological role of CNP as a neuropeptide and as a local vascular regulator. Journal of Endocrinology (1994) 141, 473–479

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Abstract 12951: Human C-Type Natriuretic Peptide-53: A Distinct Molecular Form With in vitro and in vivo Biological Actions Through cGMP Generation

Circulation ◽

10.1161/circ.130.suppl_2.12951 ◽

2014 ◽

Vol 130 (suppl_2) ◽

Author(s):

S. J Sangaralingham ◽

Brenda K Huntley ◽

Tomoko Ichiki ◽

Sharon M Sandberg ◽

John C Burnett

Keyword(s):

Natriuretic Peptide ◽

Molecular Form ◽

Cardiac Fibroblasts ◽

Active Form ◽

Biologically Active ◽

Hek 293 ◽

Hek Cells ◽

Biological Actions

Introduction: ANP and BNP are cardiac hormones that have a natriuretic and diuretic role in heart failure (HF) through guanylyl cyclase receptor A (GC-A) and cGMP generation. C-type natriuretic peptide (CNP) is part of this hormone family and is produced in the heart and endothelium. The biologically active form, CNP-22, is known to have potent anti-remodeling properties through the activation of GC-B and cGMP, but lacks natriuretic actions. Recently we have identified that another larger CNP form, CNP-53, is elevated in the plasma of acute decompensated HF patients, however it unclear if CNP-53 possesses biological actions. Hypothesis: We hypothesized that CNP-53 would be biologically active in vivo and in vitro, and would stimulate GC-B specific cGMP production. Methods: HEK 293 cells over-expressing GC-A and GC-B and human cardiac fibroblasts (hCFs), in which GC-B are in abundance, were stimulated with CNP-53 for 10 minutes at a dose of (10-8M). Two groups of anesthetized rats (n=8) received a 75-minute infusion of normal saline vehicle (V) or CNP-53 (1 ug/kg/min) and then mean arterial pressure (MAP), sodium excretion and urinary and plasma cGMP were assessed. Results: CNP-53 significantly activated cGMP in GC-B HEK cells (75±9 vs. 0.4±0.1 pmol/well, p<0.05) and in hCFs (0.17±0.03 vs. 0.00±0.00 pmol/well, p<0.05) compared to no treatment. In contrast CNP-53 failed to generate cGMP in GC-A HEK cells. In vivo, CNP-53 decreased MAP compared to vehicle (V: 101±2, CNP-53: 94±2 mmHg, p<0.05). CNP-53 had a greater natriuretic effect compared vehicle (V: 0.86±0.11, CNP-53: 1.39±0.21 uEq/min, p<0.05). Urinary cGMP excretion (V: 33±6, CNP-53: 177±28 pmol/min, p<0.05) and plasma cGMP generation was also significantly higher with CNP-53 compared to vehicle (V: 9±2, CNP-53: 91±3 pmol/ml, p<0.05). Conclusions: CNP-53, a higher molecular form of CNP that was reported to be increased human HF plasma, is bioactive with GC-B specific cGMP generation in vitro. Moreover, infusion of CNP-53 generates significant urinary and plasma cGMP and has natriuretic actions with minimal hypotensive effects in vivo. This study advances our understanding of CNP biology and provides new insights into potential CNP therapeutic opportunities targeting the progression of HF.

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Effects of natriuretic peptides and nitroprusside on venous function in trout

AJP Regulatory Integrative and Comparative Physiology ◽

10.1152/ajpregu.1997.273.2.r527 ◽

1997 ◽

Vol 273 (2) ◽

pp. R527-R539 ◽

Cited By ~ 19

Author(s):

K. R. Olson ◽

D. J. Conklin ◽

A. P. Farrell ◽

J. E. Keen ◽

Y. Takei ◽

...

Keyword(s):

Natriuretic Peptide ◽

Natriuretic Peptides ◽

Aortic Pressure ◽

Systemic Resistance ◽

Venous Compliance ◽

Venous Capacitance ◽

Unstressed Volume ◽

Ventral Aorta

Active venous regulation of cardiovascular function is well known in mammals but has not been demonstrated in fish. In the present studies, the natriuretic peptides (NP) rat atrial natriuretic peptide (ANP) and trout ventricular natriuretic peptide (VNP), clearance receptor inhibitor SC-46542, and sodium nitroprusside (SNP) were infused into unanesthetized trout fitted with pressure cannulas in the ventral aorta, dorsal aorta, and ductus Cuvier, and a ventral aorta (VA) flow probe was used to measure cardiac output (CO). In another group, in vivo vascular (venous) capacitance curves were obtained during ANP or SNP infusion. The in vitro effects of NP on vessels and the heart were also examined. ANP, VNP, and SC-46542 decreased central venous pressure (PVen), CO, stroke volume (SV), and gill resistance (RG), whereas systemic resistance (RS) and heart rate (HR) increased. Dorsal aortic pressure (PDA) transiently increased and then fell even though RS remained elevated. ANP decreased mean circulatory filling pressure (MCFP), increased vascular compliance at all blood volumes, and increased unstressed volume in hypovolemic fish. ANP had no direct effect on the heart. ANP responses in vivo were not altered in trout made hypotensive by prior treatment with the angiotensin-converting enzyme inhibitor lisinopril. SNP reduced ventral aortic pressure (PVA), PDA, and RS, increased CO and HR, but did not affect PVen, SV, or RG. SNP slightly decreased MCFP but did not affect compliance or unstressed volume. In vitro, large systemic arteries were more responsive than veins to NP, whereas SNP relaxed both. These results show that, in vivo, NP decrease venous compliance, thereby decreasing venous return, CO, and arterial pressure. Conversely, SNP hypotension is due to decreased RS. This is the first evidence for active regulation of venous capacitance in fish, which probably occurs in small veins or venules. The presence of venous baroreceptors is also suggested.

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Development of Small-Molecule STING Activators for Cancer Immunotherapy

Biomedicines ◽

10.3390/biomedicines10010033 ◽

2021 ◽

Vol 10 (1) ◽

pp. 33

Author(s):

Hee Ra Jung ◽

Seongman Jo ◽

Min Jae Jeon ◽

Hyelim Lee ◽

Yeonjeong Chu ◽

...

Keyword(s):

Cancer Immunotherapy ◽

Cyclic Gmp ◽

Therapeutic Potential ◽

Interferon Response ◽

Type I ◽

Anti Cancer ◽

Cancer Agent ◽

Sting Pathway

In cancer immunotherapy, the cyclic GMP–AMP synthase–stimulator of interferon genes (STING) pathway is an attractive target for switching the tumor immunophenotype from ‘cold’ to ‘hot’ through the activation of the type I interferon response. To develop a new chemical entity for STING activator to improve cyclic GMP-AMP (cGAMP)-induced innate immune response, we identified KAS-08 via the structural modification of DW2282, which was previously reported as an anti-cancer agent with an unknown mechanism. Further investigation revealed that direct STING binding or the enhanced phosphorylation of STING and downstream effectors were responsible for DW2282-or KAS-08-mediated STING activity. Furthermore, KAS-08 was validated as an effective STING pathway activator in vitro and in vivo. The synergistic effect of cGAMP-mediated immunity and efficient anti-cancer effects successfully demonstrated the therapeutic potential of KAS-08 for combination therapy in cancer treatment.

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Differential effects of doxorubicin on atrial natriuretic peptide expression in vivo and in vitro

Biological Research ◽

10.4067/s0716-97602001000300007 ◽

2001 ◽

Vol 34 (3-4) ◽

Cited By ~ 4

Author(s):

ASIM RAHMAN ◽

MAHMOOD ALAM ◽

SUDHA RAO ◽

LIN CAI ◽

CLARK LUTHER T. ◽

...

Keyword(s):

Atrial Natriuretic Peptide ◽

Natriuretic Peptide ◽

Differential Effects ◽

Peptide Expression ◽

Atrial Natriuretic

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Possible role of cyclic GMP in stimulus-secretion coupling by salt gland of the duck

AJP Cell Physiology ◽

10.1152/ajpcell.1979.237.5.c200 ◽

1979 ◽

Vol 237 (5) ◽

pp. C200-C204 ◽

Cited By ~ 11

Author(s):

D. J. Stewart ◽

J. Sax ◽

R. Funk ◽

A. K. Sen

Keyword(s):

Cyclic Amp ◽

Guanylate Cyclase ◽

Cyclic Gmp ◽

Salt Gland ◽

Domestic Ducks ◽

Stimulus Secretion Coupling ◽

Stimulation Of

Stimulation of salt galnd secretion in domestic ducks in vivo increased the cyclic GMP concentration of the tissue, but had no effect on cyclic AMP levels. Methacholine, which is known to stimulate sodium transport by the glands both in vivo and in vitro, stimulated ouabain-sensitive respiration in salt gland slices. Cyclic GMP stimulated ouabain-sensitive respiration to the same extent as methacholine. Guanylate cyclase stimulators, hydroxylamine and sodium azide, also stimulated ouabain-sensitive respiration. The stimulation of ouabain-sensitive respiration by methacholine was blocked either by atropine or by removal of calcium from the incubation medium. The stimulation of ouabain-sensitive respiration by cyclic GMP still occurred in the absence of calcium. The above observations seem to indicate that cyclic GMP acts as a tertiary link in the process of stimulus-secretion coupling in the tissue.

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Direct effect of α-human atrial natriuretic peptide on human vasculature in vivo and in vitro

Clinical Science ◽

10.1042/cs0740207 ◽

1988 ◽

Vol 74 (2) ◽

pp. 207-211 ◽

Cited By ~ 28

Author(s):

A. Hughes ◽

S. Thom ◽

P. Goldberg ◽

G. Martin ◽

P. Sever

Keyword(s):

Blood Flow ◽

Atrial Natriuretic Peptide ◽

Natriuretic Peptide ◽

Forearm Blood Flow ◽

Renin Angiotensin System ◽

Organ Bath ◽

Human Atrial Natriuretic Peptide ◽

Atrial Natriuretic

1. The effect of a α-human atrial natriuretic peptide (1–28) (ANP) on human vasculature was investigated in vivo and in vitro. Possible involvement of vascular dopamine receptors and the renin-angiotensin system in the response to ANP was also studied in vivo. 2. Forearm blood blow was measured by venous occlusion plethysmography. Isolated human blood vessels were studied using conventional organ bath techniques. 3. ANP (0.1–1 μg/min, intra-arterially) produced a dose-dependent increase in forearm blood flow, corresponding to a 163% increase in net forearm blood flow in the study arm. This action of ANP was not antagonized by (R)-sulpiride (100 μg/min, intra-arterially), a selective vascular dopamine receptor antagonist, or 50 mg of oral captopril, an inhibitor of angiotensin-converting enzyme. 4. ANP (1 nmol/l–1 μmol/l) produced concentration-dependent relaxation of isolated human arteries, including brachial artery, but was without effect on isolated human saphenous vein. 5. ANP produces vasodilatation in vivo and relaxes isolated human arterial smooth muscle. This action of ANP may contribute to its reported hypotensive effects in vivo.

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Cleaved CD31 as a target for in vivo molecular imaging of inflammation

Scientific Reports ◽

10.1038/s41598-019-56163-x ◽

2019 ◽

Vol 9 (1) ◽

Cited By ~ 1

Author(s):

Jonathan Vigne ◽

Sylvie Bay ◽

Rachida Aid-Launais ◽

Guillaume Pariscoat ◽

Guillaume Rucher ◽

...

Keyword(s):

Molecular Imaging ◽

Molecular Form ◽

Positive Control ◽

Negative Control ◽

Stability Study ◽

Biodistribution Studies ◽

Wide Range ◽

Significant Difference

AbstractThere is a need for new targets to specifically localize inflammatory foci, usable in a wide range of organs. Here, we hypothesized that the cleaved molecular form of CD31 is a suitable target for molecular imaging of inflammation. We evaluated a bioconjugate of D-P8RI, a synthetic peptide that binds all cells with cleaved CD31, in an experimental rat model of sterile acute inflammation. Male Wistar rats were injected with turpentine oil into the gastrocnemius muscle two days before 99mTc-HYNIC-D-P8RI (or its analogue with L-Proline) SPECT/CT or [18F]FDG PET/MRI. Biodistribution, stability study, histology, imaging and autoradiography of 99mTc-HYNIC-D-P8RI were further performed. Biodistribution studies revealed rapid elimination of 99mTc-HYNIC-D-P8RI through renal excretion with almost no uptake from most organs and excellent in vitro and in vivo stability were observed. SPECT/CT imaging showed a significant higher 99mTc-HYNIC-D-P8RI uptake compared with its analogue with L-Proline (negative control) and no significant difference compared with [18F]FDG (positive control). Moreover, autoradiography and histology revealed a co-localization between 99mTc-HYNIC-D-P8RI uptake and inflammatory cell infiltration. 99mTc-HYNIC-D-P8RI constitutes a new tool for the detection and localization of inflammatory sites. Our work suggests that targeting cleaved CD31 is an attractive strategy for the specific in vivo imaging of inflammatory processes.

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Renal hemodynamic and natriuretic effects of manganese and interactions with atrial natriuretic peptide

AJP Renal Physiology ◽

10.1152/ajprenal.1990.258.4.f998 ◽

1990 ◽

Vol 258 (4) ◽

pp. F998-F1004 ◽

Cited By ~ 1

Author(s):

H. M. Lafferty ◽

M. Gunning ◽

H. R. Brady ◽

B. M. Brenner ◽

S. Anderson

Keyword(s):

Atrial Natriuretic Peptide ◽

Natriuretic Peptide ◽

Flow Rate ◽

Plasma Flow ◽

Collecting Duct ◽

Hydraulic Pressure ◽

Cgmp Production ◽

Atrial Natriuretic

Manganese (Mn2+) is a cofactor for guanylate cyclase (GC), which is involved in the generation of guanosine 3',5'-cyclic monophosphate (cGMP), a second messenger for atrial natriuretic peptide (ANP) action. Mn2+ is also, however, a nonselective calcium-channel blocker. We examined the effects of infusion of MnCl2 into normal rats and its interaction in vivo and in vitro with GC and ANP. MnCl2 significantly increased glomerular filtration rate (GFR) and effective renal plasma flow rate (RPF). These effects were caused by selective afferent arteriolar vasodilatation, which allowed the glomerular capillary plasma flow rate and hydraulic pressure to rise, thus elevating single-nephron GFR. Urinary Na+ excretion (UNaV) also increased with MnCl2. The natriuresis was, unlike ANP, not mediated by GC activation and cGMP production, as MnCl2 had no effect on either urinary cGMP excretion or cGMP accumulation in intact inner medullary collecting duct cell (IMCD) suspensions, nor did it affect Na(+)-dependent oxygen consumption in these cells. When superimposed on an infusion of ANP, MnCl2 resulted in significant increases in UNaV, GFR, and RPF. These effects were associated with small but significant increments in urinary cGMP excretion. However, MnCl2 did not affect in vitro cGMP production in intact IMCDs or glomeruli in response to ANP stimulation. It is uncertain therefore whether the in vivo augmentation of the natriuretic effect of ANP by MnCl2 is related to GC activation and cGMP production.

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