Background: The Zucker Diabetic Fatty (ZDF) rat is a popular model of type 2 diabetes as it develops pathologic changes in the kidney similar to human diabetic nephropathy. To identify metabolic ...pathways linked to pathologic features MALDI mass spectrometry imaging (MSI) can be a powerful platform. However, quantification of metabolites from MALDI-MSI is very challenging. Our MSI-DeepPath computational platform enables spatial quantitation of metabolites from any tissue section size.
Methods: Control and obese ZDF male rats (age 6mo, n=5/group) were studied. Spatial metabolomics analysis of kidney tissue sections was performed using MALDI-MSI at 20 µm resolution. Metabolites were detected at mass accuracy <2 ppm using Q-Exactive orbitrap MS followed by annotations using METASPACE. SygnaMap’s MSI-DeepPath computational platform was used to quantify the annotated metabolites.
Results: Of 400 metabolites annotated at m/z range 70-500 Da the top features were selected using t-test, p value and fold change analysis. The top 20 biomarkers that separated ZDF from controls included glucose, serine, histidine, and glutamate. The most significantly upregulated metabolite was tyrosinamide (8.2-fold, p=8.02 x 10-9 ). The most significant downregulated metabolite was L-threonine (0.3-fold, p=2.03 x 10-9). Pathway analysis revealed that aminoacyl-tRNA biosynthesis, galactose, glyoxylate and dicarboxylate metabolic pathways can discriminate healthy from diabetic kidney samples.
Conclusion: MSIDeepPath enables quantification of spatial metabolomics of tissue sections using MALDI-MSI data. This new tool can be applied across normal and diseased samples allowing for robust statistical analysis of spatial omics data. Using MSI-DeepPath on kidney sections from the ZDF rat we identified regulation of metabolites linked to key pathways of diabetic kidney disease.
Disclosure
L.Hejazi: None. S.Sharma: None. A.Ruiz: None. G.Zhang: None. F.C.Tucci: None. K.Sharma: Advisory Panel; Reata Pharmaceuticals, Inc., Otsuka America Pharmaceutical, Inc.
Funding
National Institutes of Health (1R43DK130732-01A1)
Diabetic kidney disease (DKD) can lead to end-stage kidney disease (ESKD) and mortality; however, few mechanistic biomarkers are available for high-risk patients, especially those without ...macroalbuminuria. Urine from participants with diabetes from the Chronic Renal Insufficiency Cohort (CRIC) study, the Singapore Study of Macro-angiopathy and Micro-vascular Reactivity in Type 2 Diabetes (SMART2D), and the American Indian Study determined whether urine adenine/creatinine ratio (UAdCR) could be a mechanistic biomarker for ESKD. ESKD and mortality were associated with the highest UAdCR tertile in the CRIC study and SMART2D. ESKD was associated with the highest UAdCR tertile in patients without macroalbuminuria in the CRIC study, SMART2D, and the American Indian study. Empagliflozin lowered UAdCR in nonmacroalbuminuric participants. Spatial metabolomics localized adenine to kidney pathology, and single-cell transcriptomics identified ribonucleoprotein biogenesis as a top pathway in proximal tubules of patients without macroalbuminuria, implicating mTOR. Adenine stimulated matrix in tubular cells via mTOR and stimulated mTOR in mouse kidneys. A specific inhibitor of adenine production was found to reduce kidney hypertrophy and kidney injury in diabetic mice. We propose that endogenous adenine may be a causative factor in DKD.
Background and Purpose
Targeting α7 nicotinic ACh receptors (nAChRs) in neuroinflammatory disorders including acute ischaemic stroke holds significant therapeutic promise. However, therapeutically ...relevant signalling mechanisms remain unidentified. Activation of neuronal α7 nAChRs triggers ionotropic signalling, but there is limited evidence for it in immunoglial tissues. The α7 ligands which are effective in reducing acute ischaemic stroke damage promote α7 ionotropic activity, suggesting a link between their therapeutic effects for treating acute ischaemic stroke and activation of α7 conductive states.
Experimental Approach
This hypothesis was tested using a transient middle cerebral artery occlusion (MCAO) model of acute ischaemic stroke, NS6740, a known selective non‐ionotropic agonist of α7 nAChRs and 4OH‐GTS‐21, a partial α7 agonist. NS6740‐like ligands exhibiting low efficacy/potency for ionotropic activity will be referred to as non‐ionotropic agonists or “metagonists”.
Key Results
4OH‐GTS‐21, used as a positive control, significantly reduced neurological deficits and brain injury after MCAO as compared to vehicle and NS6740. By contrast, NS6740 was ineffective in identical assays and reversed the effects of 4OH‐GTS‐21 when these compounds were co‐applied. Electrophysiological recordings from acute hippocampal slices obtained from NS6740‐injected animals demonstrated its remarkable brain availability and protracted effects on α7 nAChRs as evidenced by sustained (>8 h) alterations in α7 ionotropic responsiveness.
Conclusion and Implications
These results suggest that α7 ionotropic activity may be obligatory for therapeutic efficacy of α7 ligands after acute ischaemic stroke yet, highlight the potential for selective application of α7 ligands to disease states based on their mode of receptor activation.
Highlights • TAK-242 prevents LPS evoked TNF release from RAW267.4 cells and male and female macrophages. • IV, IT, and IP TAK-242 administration prevents LPS-induced allodynia in males. • TLR4 ...deficiency and TAK-242 treatment prevents the delayed onset of formalin-induced tactile allodynia in both sexes. • TAK-242 does not reverse established tactile allodynia in the current experiments.
Oral ferrous salts are standard treatment for children with iron deficiency anemia (IDA). The objective of our study was to monitor oral iron therapy in children, aged 3 months–12 years, with IDA. We ...prospectively collected clinical and hematological data of children with IDA, from 15 AIEOP (Associazione Italiana di Ematologia ed. Oncologia Pediatrica) centers. Response was measured by the increase of Hb from baseline. Of the 107 analyzed patients, 18 received ferrous gluconate/sulfate 2 mg/kg (ferrous 2), 7 ferrous gluconate/sulfate 4 mg/kg (ferrous 4), 7 ferric iron salts 2 mg/kg (ferric), 62 bis-glycinate iron 0.45 mg/kg (glycinate), and 13 liposomal iron 0.7–1.4 mg/kg (liposomal). Increase in reticulocytes was evident at 3 days, while Hb increase appeared at 2 weeks. Gain of Hb at 2 and 8 weeks revealed a higher median increase in both ferrous 2 and ferrous 4 groups. Gastro-intestinal side effects were reported in 16% (ferrous 2), 14% (ferrous 4), 6% (glycinate), and 0 (ferric and liposomal) patients. The reticulocyte counts significantly increased after 3 days from the start of oral iron supplementation. Bis-glycinate iron formulation had a good efficacy/safety profile and offers an acceptable alternative to ferrous iron preparations.
Drug-eluting embolic transarterial chemoembolization (DEE-TACE) improves the overall survival of hepatocellular carcinoma (HCC), but the agents used are not tailored to HCC. Our patented liposomal ...formulation enables the loading and elution of targeted therapies onto DEEs. This study aimed to establish the safety, feasibility, and pharmacokinetics of sorafenib or regorafenib DEE-TACE in a VX2 model. DEE-TACE was performed in VX2 hepatic tumors in a selective manner until stasis using liposomal sorafenib- or regorafenib-loaded DEEs. The animals were euthanized at 1, 24, and 72 h timepoints post embolization. Blood samples were taken for pharmacokinetics at 5 and 20 min and at 1, 24, and 72 h. Measurements of sorafenib or regorafenib were performed in all tissue samples on explanted hepatic tissue using the same mass spectrometry method. Histopathological examinations were carried out on tumor tissues and non-embolized hepatic specimens. DEE-TACE was performed on 23 rabbits. The plasma concentrations of sorafenib and regorafenib were statistically significantly several folds lower than the embolized liver at all examined timepoints. This study demonstrates the feasibility of loading sorafenib or regorafenib onto commercially available DEEs for use in TACE. The drugs eluted locally without release into systemic circulation.
The accurate assessment of tree crowns is important for agriculture, for example, to adjust spraying rates, to adjust irrigation rates or even to estimate biomass. Among the available methodologies, ...there are the traditional methods that estimate with a three-dimensional approximation figure, the HDS (High Definition Survey), or TLS (Terrestrial Laser Scanning) based on LiDAR technology, the aerial photogrammetry that has re-emerged with unmanned aerial vehicles (UAVs), as they are considered low cost. There are situations where either the cost or location does not allow for modern methods and prices such as HDS or the use of UAVs. This study proposes, as an alternative methodology, the evaluation of images extracted from Google Maps (GM) for the calculation of tree crown volume. For this purpose, measurements were taken on orange trees in the south of Spain using the four methods mentioned above to evaluate the suitability, accuracy, and limitations of GM. Using the HDS method as a reference, the photogrammetric method with UAV images has shown an average error of 10%, GM has obtained approximately 50%, while the traditional methods, in our case considering ellipsoids, have obtained 100% error. Therefore, the results with GM are encouraging and open new perspectives for the estimation of tree crown volumes at low cost compared to HDS, and without geographical flight restrictions like those of UAVs.
Neoantigens (nAgs) are promising tumor antigens for cancer vaccination with the potential of inducing robust and selective T cell responses. Genetic vaccines based on Adenoviruses derived from ...non-human Great Apes (GAd) elicit strong and effective T cell-mediated immunity in humans. Here, we investigate for the first time the potency and efficacy of a novel GAd encoding multiple neoantigens. Prophylactic or early therapeutic vaccination with GAd efficiently control tumor growth in mice. In contrast, combination of the vaccine with checkpoint inhibitors is required to eradicate large tumors. Gene expression profile of tumors in regression shows abundance of activated tumor infiltrating T cells with a more diversified TCR repertoire in animals treated with GAd and anti-PD1 compared to anti-PD1. Data suggest that effectiveness of vaccination in the presence of high tumor burden correlates with the breadth of nAgs-specific T cells and requires concomitant reversal of tumor suppression by checkpoint blockade.
Acquired aplastic anemia (AA) is a rare heterogeneous disease characterized by pancytopenia and hypoplastic bone marrow. The incidence is 2–3/million inhabitants/year, in Europe, but higher in East ...Asia. Survival in severe aplastic anemia (SAA) has markedly improved in the past 2decades because of advances in hematopoietic stem cell transplantation, immunosuppressive and biologic drugs, and supportive care. In SAA hematopoietic stem cell transplant (HSCT) from a matched sibling donor (MSD) is the treatment of choice. If a MSD is not available, the options include immunosuppressive therapy (IST) or unrelated donor HSCT. The objective of this guideline is to provide healthcare professionals with clear guidance on the diagnosis and management of pediatric patients with AA. A preliminary, evidence-based document issued by a group of pediatric hematologists was discussed, modified and approved during a series of “Consensus Conferences” according to procedures previously validated by the AIEOP Board. The guidelines highlight the importance of referring pediatric patients with AA to pediatric centers with long experience in diagnosis, differential diagnosis, management, supportive care and follow-up of AA.
NASH causes a tremendous health care burden in the United States. A glucagon-like peptide-1 agonist, semaglutide (Sema), treatment resulted in hepatic steatosis reduction in clinical trials of NASH. ...Lysophosphatidic acid receptor 1 antagonists are known to have antifibrotic effects in several organs. We tested Sema and a novel lysophosphatidic acid receptor 1 antagonist, EPGN2154, individually and in combination to evaluate their efficacy for NASH remission in preclinical models.
In the present study, we used (1) C57Bl6/J wild-type mice fed on a high-fat, high-carbohydrate (HFHC) diet for 16 weeks and (2) leptin-deficient mice (ob/ob) fed on an Amylin liver NASH diet for 16 weeks. After 16 weeks, the mice were randomly distributed in equal numbers in (1) no-drug, (2) EPGN2154, (3) Sema, and (4) EPGN2154+Sema treatment groups for 8 additional weeks at a dosage of 10 mg/kg body weight for EPGN2154 (oral gavage, 5 days a week) and 6.17 μg/kg body weight of Sema (subcutaneous injection every alternate day, 3 days a week).
In the wild-type-high-fat, high-carbohydrate model, we observed the most body weight loss in the EPGN2154+Sema combination group compared to the other treatment groups. All groups led to a significant reduction in alanine transaminase levels when compared to high-fat, high-carbohydrate-fed wild type. However, no significant difference in alanine transaminase levels was observed among the treatment groups. In the ob/ob mice study, Sema did not cause body weight loss. Moreover, the EPGN2154 and the combination groups had a lower NAFLD Activity Score and incidence of advanced-stage hepatic fibrosis than the Sema group.
EPGN2154 demonstrated a hepato-protective effect independent of body weight loss in preclinical NASH models.
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