Dual Messenger Research Articles

Realveolarization with inhalable mucus-penetrating lipid nanoparticles for the treatment of pulmonary fibrosis in mice.

The stemness loss-associated dysregeneration of impaired alveolar type 2 epithelial (AT2) cells abolishes the reversible therapy of idiopathic pulmonary fibrosis (IPF). We here report an inhalable mucus-penetrating lipid nanoparticle (LNP) for codelivering dual mRNAs, promoting realveolarization via restoring AT2 stemness for IPF treatment. Inhalable LNPs were first formulated with dipalmitoylphosphatidylcholine and our in-house-made ionizable lipids for high-efficiency pulmonary mucus penetration and codelivery of dual messenger RNAs (mRNAs), encoding cytochrome b5 reductase 3 and bone morphogenetic protein 4, respectively. After being inhaled in a bleomycin model, LNPs reverses the mitochondrial dysfunction through ameliorating nicotinamide adenine dinucleotide biosynthesis, which inhibits the accelerated senescence of AT2 cells. Concurrently, pathological epithelial remodeling and fibroblast activation induced by impaired AT2 cells are terminated, ultimately prompting alveolar regeneration. Our data demonstrated that the mRNA-LNP system exhibited high protein expression in lung epithelial cells, which markedly extricated the alveolar collapse and prolonged the survival of fibrosis mice, providing a clinically viable strategy against IPF.

Translational Pharmacokinetic/Pharmacodynamic Model for mRNA-3927, an Investigational Therapeutic for the Treatment of Propionic Acidemia.

Propionic acidemia (PA) is an ultrarare disorder caused by deficiency of the mitochondrial enzyme, propionyl-CoA carboxylase (PCC), composed of PCCA and PCCB subunits. An enzyme replacement therapy is being developed using dual messenger RNA (mRNA) therapy composed of lipid nanoparticles (LNPs) encapsulating mRNAs encoding PCCA and PCCB subunits of the PCC enzyme. We herein report on development of a translational semimechanistic pharmacokinetic (PK) and PK/pharmacodynamic (PD) model to quantify the relationship between the mRNA components of mRNA-3927 (an LNP encapsulating PCCA and PCCB mRNAs) and dose levels; PCCA/B mRNA PK and PD responses were assessed as circulating levels of primary disease markers 2-methyl citrate, 3-hydroxypropionate, and propionyl carnitine normalized to acetyl carnitine (C3/C2 ratio) to inform the first-in-human dose range and regimen selection. The translational PK/PD model was developed using preclinical data available in mice with PA, Sprague Dawley rats, and cynomolgus monkeys at dose levels ranging from 0.2 to 9 mg/kg. PCCA/B mRNA PK in mice, rats, and monkeys was adequately described using allometric scaling of volume and clearance parameters. The interspecies preclinical model was scaled allometrically to humans to predict the dose-response relationship in adult and pediatric patients with PA to guide selection of dose range and regimen for the Phase 1 clinical trial (ClinicalTrials.gov Identifier NCT04159103).

Adventitial matrix metalloproteinase production and distribution of immunoglobulin G4-related abdominal aortic aneurysms

ObjectiveIgG4-related diseases are systemic inflammatory fibrous lesions characterized by elevated serum IgG4 and infiltration of IgG4-positive plasmacytes. They can manifest in vascular lesions as frequently formed aneurysms with prominent thickening of the adventitia (IgG4-related abdominal aortic aneurysm; IgG4-AAA). Matrix metalloproteinases (MMPs) degrade the extracellular matrix, mainly in the tunica media, resulting in destruction of aortic structures to cause enlargement of the aneurysm. However, the expression of adventitial MMPs in IgG4-AAAs is poorly understood. MethodsMMPs and MMPs-presenting cells in the adventitia of IgG4-AAAs (n = 19) of human surgical specimens were evaluated by immunohistochemistry and dual messenger RNA in situ hybridization. The results were compared with those from control groups of non-IgG4-related inflammatory AAA (n = 18), atherosclerotic AAA (aAAA; n = 11), and autopsy cases (n = 11). Preoperative serum MMPs levels of these groups were compared with the histologic data. ResultsExpression of MMP-9, MMP-2, and MMP-14 at the protein and messenger RNA levels in the adventitia was significantly higher in IgG4-AAAs than in controls. Other MMPs were scarce. The total number of MMP-9-positive cells was positively correlated with the diameter of the aneurysm (R = 0.461; P = .031), the adventitial thickness (R = 0.688; P < .001), and the number of IgG4-positive cells (R = 0.764; P < .001). Within lymphoid follicles, MMP-9-presenting cells were predominantly detected in large follicular dendritic cells, followed by histiocytes, fibroblasts, and plasmacytic dendritic cells. Outside lymphoid follicles, fibroblasts, and histiocytes mainly expressed MMP-9, and tissue dendritic cells also produced MMP-9. The levels of MMP-9 derived from follicular dendritic cells and histiocytes and plasmacytic dendritic cells outside lymphoid follicles were significantly higher in IgG4-AAA group than in other groups. Expression of adventitial MMP-2 and MMP-14 by histiocytes and fibroblasts was predominantly detected outside lymphoid follicles. Serum MMP-9 levels were significantly higher in IgG4-AAAs (835 ng/mL) than in controls, and correlated with serum IgG4 levels and the total numbers of adventitial MMP-9-positive cells, whereas serum MMP-2 levels did not differ among the three aneurysmal groups. ConclusionsMMP-9 production in adventitial immune cells concerning lymphoid follicles was characteristic of IgG4-AAAs and might work in its activity with aneurysmal dilatation and adventitial thickening. Expressions of adventitial MMP-2 and MMP-14 were detected in histiocytes and fibroblasts outside lymphoid follicles, and were less concerned with the activity of IgG4-AAAs.

Wiener filtering and pure $\mathcal {E}/\mathcal {B}$ decomposition of CMB maps with anisotropic correlated noise

ABSTRACTWe present an augmented version of our dual messenger algorithm for spin field reconstruction on the sphere, while accounting for highly non-trivial and realistic noise models such as modulated correlated noise. We also describe an optimization method for the estimation of noise covariance from Monte Carlo simulations. Using simulated Planck polarized cosmic microwave background (CMB) maps as a showcase, we demonstrate the capabilities of the algorithm in reconstructing pure $\mathcal {E}$ and $\mathcal {B}$ maps, guaranteed to be free from ambiguous modes resulting from the leakage or coupling issue that plagues conventional methods of $\mathcal {E}/\mathcal {B}$ separation. Due to its high speed execution, coupled with lenient memory requirements, the algorithm can be optimized in exact global Bayesian analyses of state-of-the-art CMB data for a statistically optimal separation of pure $\mathcal {E}$ and $\mathcal {B}$ modes. Our algorithm, therefore, has a potentially key role in the data analysis of high-resolution and high-sensitivity CMB data, especially with the range of upcoming CMB experiments tailored for the detection of the elusive primordial $\mathcal {B}$-mode signal.

Optimal and fast $\mathcal {E}/\mathcal {B}$ separation with a dual messenger field

We adapt our recently proposed dual messenger algorithm for spin field reconstruction and showcase its efficiency and effectiveness in Wiener filtering polarized cosmic microwave background (CMB) maps. Unlike conventional preconditioned conjugate gradient (PCG) solvers, our preconditioner-free technique can deal with high-resolution joint temperature and polarization maps with inhomogeneous noise distributions and arbitrary mask geometries with relative ease. Various convergence diagnostics illustrate the high quality of the dual messenger reconstruction. In contrast, the PCG implementation fails to converge to a reasonable solution for the specific problem considered. The implementation of the dual messenger method is straightforward and guarantees numerical stability and convergence. We show how the algorithm can be modified to generate fluctuation maps, which, combined with the Wiener filter solution, yield unbiased constrained signal realizations, consistent with observed data. This algorithm presents a pathway to exact global analyses of high-resolution and high-sensitivity CMB data for a statistically optimal separation of E and B modes. It is therefore relevant for current and next-generation CMB experiments, in the quest for the elusive primordial B-mode signal.

Wiener filter reloaded: fast signal reconstruction without preconditioning

We present a high performance solution to the Wiener filtering problem via a formulation that is dual to the recently developed messenger technique. This new dual messenger algorithm, like its predecessor, efficiently calculates the Wiener filter solution of large and complex data sets without preconditioning and can account for inhomogeneous noise distributions and arbitrary mask geometries. We demonstrate the capabilities of this scheme in signal reconstruction by applying it on a simulated cosmic microwave background (CMB) temperature data set. The performance of this new method is compared to that of the standard messenger algorithm and the preconditioned conjugate gradient (PCG) approach, using a series of well-known convergence diagnostics and their processing times, for the particular problem under consideration. This variant of the messenger algorithm matches the performance of the PCG method in terms of the effectiveness of reconstruction of the input angular power spectrum and converges smoothly to the final solution. The dual messenger algorithm outperforms the standard messenger and PCG methods in terms of execution time, as it runs to completion around 2 and 3-4 times faster than the respective methods, for the specific problem considered.

Ceramide 1-Phosphate: First, Second, or Dual Messenger?

Ceramide 1-Phosphate: First, Second, or Dual Messenger?

Second messengers in neuronal signaling.

The past decade has witnessed an enormous increase in our knowledge of the variety and complexity of intracellular signaling events that follow receptor binding on the cell surface. This overview emphasizes the phosphoinositidase C-mediated dual messenger pathway in brain and in brain-derived cells, with special reference to possible significance for research on the dementias.

Neurotransmitter Receptors and Phosphoinositide Turnover

Neurotransmitters, neuromodulators, hormones and growth-promoting fac­ tors are now known to exhibit their diverse metabolic and physiological responses by interaction with their selective receptors located on the cell surface. These surface receptors transduce and amplify extracellular signals by the generation of so-called second messengers. Among these are inositol phosphates and 1 ,2-diacylglycerol (DG) which are formed by hydrolysis of membrane-bound phosphoinositides in response to receptor stimulation. This dual messenger system has attracted enormous attention in recent years because of its involvement in an array of physiological processes, including neuronal membrane excitability, synaptic transmission, cellular metabolism, muscle contraction, sensory transduction, motility, growth, DNA synthesis, and platelet aggregation. Several recent articles review various aspects of this receptor-signaling system (1-15). This review focuses on recent de­ velopments concerning the metabolism of second messengers , their mode of action, the regulation of neurotransmitter receptor signalling, and the interac­ tion with other receptor-coupled effectors . The explosive growth of the field of phosphoinositide turnover has touched virtually every major biological system, and a detailed review of each system is impractical . I therefore concentrate on a few systems, in particular the eNS and related neural tissues where the metabolism of phosphoinositides is particularly active. Phosphatidylinositol (PI) , phosphatidylinositol 4-phosphate (PIP) and phosphatidylinositol 4, 5-bisphosphate (PIP2) are collectively referred to as

Cholinergic-induced [ 3H] noradrenaline release in rat brain cortical slices is mediated via a pertussis toxin sensitive GTP binding protein and involves activation of protein kinase C

The involvement of a GTP-binding protein (G-protein) in the process of neurotransmitter release was examined using pertussis toxin and cholerra toxin. Cholinergic agonists are shown to mediate [ 3H]noradrenaline release in rat brain slices via a pertussis toxin (1.2 μg/ml) sensitive, and cholera toxin (0.5 μg/ml) insenstive G. protein. An indication for the involvement of a G-protein and phospholipase C activation in the release process was implied from the inhibitory effect of neomycin on K +-, veratridine, and carbachol-induced-norepinephrine release. Depolarizing agents mediate a neomycin-senstive release, which is not affected either by pertussis toxin or cholera toxin, suggesting a different mode of phospholipase C activation, unlike carbachol-induced release, which is both neomycin and pertussis toxin senstive. Similarly, a hormone-sensitive carrier activated by phenylephrine not via α 1-adrenergic receptors, mediates a non-exocytosis efflux which is not affected by neomycin and is shown to be pertussis toxin-insensitive. The inhibitory action of protein kinase C inhibitors polymyxin B, K252 a and H-7 [(1-(1-(5-isoquinolinesulphonyl)-2-methyl-piperazine] on release, strongly suggests its participation in the process. Polymyxin B, a relatively selective protein kinase C inhibitor, inhibted carbachol-induced release ( ic 50 = 0.53 μ M) as well as the K + and the veratridine induced [ 3H] noradrenaline release. K252 a, an inhibitor of various protein kinases at the ATP site, and H-7, another protein kinase C inhibitor, inhibited carbachol-induced noradrenaline released with ic = 35 nM and 3 μ M respectively. Consistent with its inability to activate phospholipase C, phenylephrine-induced noradrenaline efflux was unaffected by polymyxin B (> 70 μM). The results offer more supportive evidence for a major role played by the dual messengers inositol triphosphate and diacylglycerol (IP 3/DG) in the mechanisms of neuronal release.

Calcium and cAMP as Synarchic Messengers

ABSTRACT The topics addressed in this monograph are intracellular communication and the biochemical mechanisms used by cells for information flow from cell surface to cell interior.Howard Rasmussen has long been a leader in the field of cellular regulation by the second messengers, Ca++ and cAMP. Here, he surveys data gathered from nervous and endocrine systems during the past 15 years and concludes that Ca++ and cAMP, rather than operating independently of each other, almost always function as dual messengers in stimulus-response coupling. To define the interrelatedness of Ca++ and cAMP functions, he uses the term synarchic regulation, which he says originates from discussions with a friend in the Classics Department at Yale. In Greek mythology, archons were heralds entrusted to carry information of such great importance that they were employed in pairs to carry either the same message or, under other circumstances, only part of the same message. Thus, the