Noninvasive assessment of the lung inflammation-fibrosis axis by targeted imaging of CMKLR1.

Precision management of fibrotic lung diseases is challenging due to their diverse clinical trajectories and lack of reliable biomarkers for risk stratification and therapeutic monitoring. Here, we validated the accuracy of CMKLR1 as an imaging biomarker of the lung inflammation-fibrosis axis. By analyzing single-cell RNA sequencing datasets, we demonstrated expression as a transient signature of monocyte-derived macrophages (MDMφ) enriched in patients with idiopathic pulmonary fibrosis (IPF).

Targeted imaging of very late antigen-4 for noninvasive assessment of lung inflammation-fibrosis axis.

Background: The lack of noninvasive methods for assessment of dysregulated inflammation as a major driver of fibrosis (i.e., inflammation-fibrosis axis) has been a major challenge to precision management of fibrotic lung diseases.

2-deoxy-2-[F]fluoro-D-glucose Positron Emission Tomography to Monitor Lung Inflammation and Therapeutic Response to Dexamethasone in a Murine Model of Acute Lung Injury.

Purpose: To image inflammation and monitor therapeutic response to anti-inflammatory intervention using 2-deoxy-2-[F]fluoro-D-glucose ([F]FDG) positron emission tomography (PET) in a preclinical model of acute lung injury (ALI).

Procedures: Mice were intratracheally administered lipopolysaccharide (LPS, 2.5 mg/kg) to induce ALI or phosphate-buffered saline as the vehicle control.

Molecular imaging of chemokine-like receptor 1 (CMKLR1) in experimental acute lung injury.

The lack of techniques for noninvasive imaging of inflammation has challenged precision medicine management of acute respiratory distress syndrome (ARDS). Here, we determined the potential of positron emission tomography (PET) of chemokine-like receptor-1 (CMKLR1) to monitor lung inflammation in a murine model of lipopolysaccharide-induced injury. Lung uptake of a CMKLR1-targeting radiotracer, [Cu]NODAGA-CG34, was significantly increased in lipopolysaccharide-induced injury, correlated with the expression of multiple inflammatory markers, and reduced by dexamethasone treatment.

Student Substance Use Policies in US Allopathic Medical Schools: A National Study.

This quality improvement study examines US allopathic medical schools’ student handbook policies on substance use and assesses their adherence to current Association of American Medical Colleges guidelines.

Imaging Immunometabolism in Atherosclerosis.

Over the past decade, there has been a growing recognition of the links between intracellular metabolism and immune cell activation, that is, immunometabolism, and its consequences in atherogenesis. However, most immunometabolic investigations have been conducted in cultured cells through pharmacologic or genetic manipulations of selected immunologic or metabolic pathways, limiting their extrapolation to the complex microenvironment of plaques. In vivo metabolic imaging is ideally situated to address this gap and to determine the clinical implications of immunometabolic alterations for diagnosis and management of patients.

Divergence of acetate uptake in proinflammatory and inflammation-resolving macrophages: implications for imaging atherosclerosis.

Background: Metabolic divergence of macrophages polarized into different phenotypes represents a mechanistically relevant target for non-invasive characterization of atherosclerotic plaques using positron emission tomography (PET). Carbon-11 (C)-labeled acetate is a clinically available tracer which accumulates in atherosclerotic plaques, but its biological and clinical correlates in atherosclerosis are undefined.

Methods And Results: Histological correlates of C-acetate uptake were determined in brachiocephalic arteries of western diet-fed apoE mice.

Engineering bromodomains with a photoactive amino acid by engaging 'Privileged' tRNA synthetases.

Site-specific placement of unnatural amino acids, particularly those responsive to light, offers an elegant approach to control protein function and capture their fleeting 'interactome'. Herein, we have resurrected 4-(trifluoromethyldiazirinyl)-phenylalanine, an underutilized photo-crosslinker, by introducing several key features including easy synthetic access, site-specific incorporation by 'privileged' synthetases and superior crosslinking efficiency, to develop photo-crosslinkable bromodomains suitable for 'interactome' profiling.

Design and in Vivo Characterization of A Adenosine Receptor Agonists in the Native Ribose and Conformationally Constrained (N)-Methanocarba Series.

(N)-Methanocarba ([3.1.0]bicyclohexyl) adenosines and corresponding ribosides were synthesized to identify novel A adenosine receptor (AAR) agonists for CNS or peripheral applications.

Structure activity relationship of 2-arylalkynyl-adenine derivatives as human A adenosine receptor antagonists.

Recognition of nucleosides at adenosine receptors (ARs) is supported by multiple X-ray structures, but the structure of an adenine complex is unknown. We examined the selectivity of predicted AAR and AAR adenine antagonists that incorporated known agonist affinity-enhancing and C2 substituents. Adenines with AAR-favoring -alkyl, cycloalkyl and arylalkyl substitutions combined with an AAR-favoring 2-((5-chlorothiophen-2-yl)ethynyl) group were human (h) AAR-selective, MRS7497 (∼1000-fold over AAR).

Repurposing of a Nucleoside Scaffold from Adenosine Receptor Agonists to Opioid Receptor Antagonists.

While screening off-target effects of rigid ()-methanocarba-adenosine 5'-methylamides as A adenosine receptor (AR) agonists, we discovered μM binding hits at the δ-opioid receptor (DOR) and translocator protein (TSPO). In an effort to increase OR and decrease AR affinity by structure activity analysis of this series, antagonist activity at κ-(K)OR appeared in 5'-esters (ethyl and propyl ), which retained TSPO interaction (μM). 7-Deaza modification of C2-(arylethynyl)-5'-esters but not 4'-truncation enhanced KOR affinity (MRS7299 and , ≈ 40 nM), revealed μ-OR and DOR binding, and reduced AR affinity.

Extrinsic Tryptophans as NMR Probes of Allosteric Coupling in Membrane Proteins: Application to the A Adenosine Receptor.

Tryptophan indole N-H signals are well separated in nuclear magnetic resonance (NMR) spectra of proteins. Assignment of the indole N-H signals therefore enables one to obtain site-specific information on complex proteins in supramacromolecular systems, even when extensive assignment of backbone N-H resonances is challenging. Here we exploit the unique indole N-H chemical shift by introducing extrinsic tryptophan reporter residues at judiciously chosen locations in a membrane protein for increased coverage of structure and function by NMR.

Bitopic fluorescent antagonists of the A adenosine receptor based on pyrazolo[4,3-][1,2,4]triazolo[1,5-]pyrimidin-5-amine functionalized congeners.

A pyrazolo[4,3-][1,2,4]triazolo[1,5-]pyrimidin-5-amine antagonist of the A adenosine receptor (AR) was functionalized as amine congeners, fluorescent conjugates and a sulfonate, and the AAR binding modes were predicted computationally. The optimal -butyl spacer was incorporated into the following AAR-selective (, nM) conjugates: BODIPY630/650 derivative (MRS7396, 24.6) and AlexaFluor488 derivative (MRS7416, 30.

Identification of a novel spinal nociceptive-motor gate control for Aδ pain stimuli in rats.

Physiological responses to nociceptive stimuli are initiated within tens of milliseconds, but the corresponding sub-second behavioral responses have not been adequately explored in awake, unrestrained animals. A detailed understanding of these responses is crucial for progress in pain neurobiology. Here, high-speed videography during nociceptive Aδ fiber stimulation demonstrated engagement of a multi-segmental motor program coincident with, or even preceding, withdrawal of the stimulated paw.

Synthesis of a Masked 2,3-Diaminoindole.

A three-step synthesis of masked 2,3-diaminoindole 1 from 2-iodo-3-nitro-1-(phenylsulfonyl)indole (2) has been developed. Treatment of 1 with trifluoroacetic acid generates the unstable 2,3-diamino-1-(phenylsulfonyl)indole (3), which can be trapped with α-dicarbonyl compounds to afford 5H-pyrazino[2,3-b]indoles 7-10.

Photo-degradation of 2,4-dinitroanisole (DNAN): An emerging munitions compound.

The US military is developing insensitive munitions (IM) that are less sensitive to shock and high temperatures to minimize unintentional detonations. DNAN (2,4-dinitroanisole) is one of the main ingredients of these IM formulations. During live-fire training, chunks of IM formulations are scattered by partial detonations and, once on the soil, they weather and dissolve.

Triple Benzannulation of Naphthalene via a 1,3,6-Naphthotriyne Synthetic Equivalent. Synthesis of Dibenz[a,c]anthracene.

A new synthesis of dibenzo[a,c]anthracene (4) is described that features the generation, from tetrabromo-bis-triflate 1 and phenyllithium, of a 1,3,6-naphthotriyne (2) synthetic equivalent that is trapped with 3 equiv of furan to form Diels-Alder tris-adduct 3. A subsequent two-step deoxygenation of 3 represents the first synthesis of dibenz[a,c]anthracene (4) that involves a tandem aryne Diels-Alder cycloaddition-deoxygenation strategy.

Tonantzitlolone cytotoxicity toward renal cancer cells is PKCθ- and HSF1-dependent.

Elucidating the targets and mechanism of action of natural products is strategically important prior to drug development and assessment of potential clinical applications. In this report, we elucidated the main targets and mechanism of action of the natural product tonantzitlolone (TZL) in clear cell renal cell carcinoma (CCRCC). We identified TZL as a dual PKCα and PKCθ activator in vitro, although in CCRCC cells its activity was mostly PKCθ-dependent.

Targeting ABL1-mediated oxidative stress adaptation in fumarate hydratase-deficient cancer.

Patients with germline fumarate hydratase (FH) mutation are predisposed to develop aggressive kidney cancer with few treatment options and poor therapeutic outcomes. Activity of the proto-oncogene ABL1 is upregulated in FH-deficient kidney tumors and drives a metabolic and survival signaling network necessary to cope with impaired mitochondrial function and abnormal accumulation of intracellular fumarate. Excess fumarate indirectly stimulates ABL1 activity, while restoration of wild-type FH abrogates both ABL1 activation and the cytotoxicity caused by ABL1 inhibition or knockdown.