Blimp-1 and c-Maf regulate immune gene networks to protect against distinct pathways of pathobiont-induced colitis.

Intestinal immune responses to microbes are controlled by the cytokine IL-10 to avoid immune pathology. Here, we use single-cell RNA sequencing of colon lamina propria leukocytes (LPLs) along with RNA-seq and ATAC-seq of purified CD4+ T cells to show that the transcription factors Blimp-1 (encoded by Prdm1) and c-Maf co-dominantly regulate Il10 while negatively regulating proinflammatory cytokines in effector T cells. Double-deficient Prdm1fl/flMaffl/flCd4Cre mice infected with Helicobacter hepaticus developed severe colitis with an increase in TH1/NK/ILC1 effector genes in LPLs, while Prdm1fl/flCd4Cre and Maffl/flCd4Cre mice exhibited moderate pathology and a less-marked type 1 effector response. LPLs from infected Maffl/flCd4Cre mice had increased type 17 responses with increased Il17a and Il22 expression and an increase in granulocytes and myeloid cell numbers, resulting in increased T cell-myeloid-neutrophil interactions. Genes over-expressed in human inflammatory bowel disease showed differential expression in LPLs from infected mice in the absence of Prdm1 or Maf, revealing potential mechanisms of human disease.

Deep Learning Enables Spatial Mapping of the Mosaic Microenvironment of Myeloma Bone Marrow Trephine Biopsies.

Bone marrow trephine biopsy is crucial for the diagnosis of multiple myeloma. However, the complexity of bone marrow cellular, morphologic, and spatial architecture preserved in trephine samples hinders comprehensive evaluation. To dissect the diverse cellular communities and mosaic tissue habitats, we developed a superpixel-inspired deep learning method (MoSaicNet) that adapts to complex tissue architectures and a cell imbalance aware deep learning pipeline (AwareNet) to enable accurate detection and classification of rare cell types in multiplex immunohistochemistry images. MoSaicNet and AwareNet achieved an AUC of >0.98 for tissue and cellular classification on separate test datasets. Application of MoSaicNet and AwareNet enabled investigation of bone heterogeneity and thickness as well as spatial histology analysis of bone marrow trephine samples from monoclonal gammopathies of undetermined significance (MGUS) and from paired newly diagnosed and posttreatment multiple myeloma. The most significant difference between MGUS and newly diagnosed multiple myeloma (NDMM) samples was not related to cell density but to spatial heterogeneity, with reduced spatial proximity of BLIMP1+ tumor cells to CD8+ cells in MGUS compared with NDMM samples. Following treatment of patients with multiple myeloma, there was a reduction in the density of BLIMP1+ tumor cells, effector CD8+ T cells, and regulatory T cells, indicative of an altered immune microenvironment. Finally, bone heterogeneity decreased following treatment of patients with multiple myeloma. In summary, deep learning-based spatial mapping of bone marrow trephine biopsies can provide insights into the cellular topography of the myeloma marrow microenvironment and complement aspirate-based techniques. SIGNIFICANCE: Spatial analysis of bone marrow trephine biopsies using histology, deep learning, and tailored algorithms reveals the bone marrow architectural heterogeneity and evolution during myeloma progression and treatment.

Accelerated developmental adipogenesis programs adipose tissue dysfunction and cardiometabolic risk in offspring born to dams with metabolic dysfunction.

This study determined if a perturbation in in utero adipogenesis leading to later life adipose tissue (AT) dysfunction underlies programming of cardiometabolic risk in offspring born to dams with metabolic dysfunction. Female mice heterozygous for the leptin receptor deficiency (Hetdb) had 2.4-fold higher prepregnancy fat mass and in late gestation had higher plasma insulin and triglycerides compared with wild-type (Wt) females (P < 0.05). To isolate the role of the intrauterine milieu, wild-type (Wt) offspring from each pregnancy were studied. Differentiation potential in isolated progenitors and cell size distribution analysis revealed accelerated adipogenesis in Wt pups born to Hetdb dams, accompanied by a higher accumulation of neonatal fat mass. In adulthood, whole body fat mass by NMR was higher in male (69%) and female (20%) Wt offspring born to Hetdb versus Wt pregnancies, along with adipocyte hypertrophy and hyperlipidemia (all P < 0.05). Lipidomic analyses by gas chromatography revealed an increased lipogenic index (16:0/18:2n6) after high-fat/fructose diet (HFFD). Postprandial insulin, ADIPO-IR, and ex vivo AT lipolytic responses to isoproterenol were all higher in Wt offspring born to Hetdb dams (P < 0.05). Intrauterine metabolic stimuli may direct a greater proportion of progenitors toward terminal differentiation, thereby predisposing to hypertrophy-induced adipocyte dysfunction.NEW & NOTEWORTHY This study reveals that accelerated adipogenesis during the perinatal window of adipose tissue development predisposes to later life hypertrophic adipocyte dysfunction, thereby compromising the buffering function of the subcutaneous depot.

The relationship between concentrations of magnesium and oxidized low density lipoprotein and the activity of platelet activating factor acetylhydrolase in the serum of patients with type 1 diabetes.

The study was aimed at comparing the concentration of metabolic parameters, the serum concentration of oxidized low density lipoproteins (oxLDL) and the activity of platelet activating factor acetylhydrolase (PAF-AH) in the relation to the serum concentration of magnesium (Mg) in patients with type 1 diabetes (DM1). DM1 patients (n=78) were divided into 2 groups: patients with low serum Mg concentration (<0.7 mmol/L, group 1, n=34) and patients with reference levels of Mg (>or=0.7 mmol/L, group 2, n=44). A control group (n=24) of healthy subjects was also recruited. Our results showed that DM1 patients had lower serum Mg concentrations than the control group. It was found that parameters of poor metabolic control and lipid profile are not related to the serum Mg concentration in DM1 patients. However, both the Mg concentration and the PAF-AH activity are independently related to the serum oxLDL concentration. In group 1 the oxLDL concentration and the PAF-AH activity were higher than in group 2, and the control group. Two groups of DM1 patients did not show any differences with regard to the metabolic control. Therefore, the oxidative modification of LDL and the higher activity of PAF-AH are related with the low Mg status; however, no relation has been observed between these parameters and the poor metabolic control in DM1 patients.

The evaluation of IL-12 concentration, PAF-AH, and PLA(2) activity in patients with type 1 diabetes treated with intensive insulin therapy.

OBJECTIVES: The aim of the study was to evaluate the concentration of interleukin 12 (IL-12), the activity of phospholipase A(2) (PLA(2)), and platelet-activating factor acetylhydrolase (PAF-AH) in type 1 diabetes (DM1) patients treated with intensive insulin therapy. DESIGN AND METHODS: Studied parameters were measured in 81 patients, who were subdivided according to the HbA(1)c value, hsCRP concentration, and presence or absence of late complications. RESULTS: PAF-AH activity was higher in the DM1 patients versus the control group (P=0.042). IL-12 concentration was the highest in subgroup with > or =3 mg/L hsCRP (P<0.05). Negative correlations were found for the IL-12 and age of patients and for apo A-I in the subgroup with poor metabolic control. In addition, positive correlation for hsCRP and PAF-AH activity in the subgroup with > or =3 mg/L CRP (P<0.05) was also found. CONCLUSIONS: PAF-AH and IL-12 appear to be implicated in the development of a chronic inflammation in DM1. In addition, our results emphasize a protective role of apo A-I against an increase in IL-12 production.

Acamprosate involvement in triacylglycerol hydrolysis and transacylation with cholesterol in chronically ethanol-drinking rats.

Acamprosate (AC) is used as a drug for treating alcoholism. We evaluated the effect of AC on serum triacylglycerol hydrolysis (GEH, glycerol ester hydrolysis), triacylglycerol transacylation with cholesterol (GECAT, glycerol ester:cholesterol acyltransferase), and acylcholesterol hydrolysis (Cease, cholesterol ester hydrolysis) in an experimental model of alcoholism. Ethanol-preferring (PRF), non-preferring (NPF), and control (CR) male Wistar rats were treated with AC (500 mg/kg, p.o.) for 21 consecutive days. The beneficial effect of AC on lipid parameters of PRF rats included decreased triacylglycerol, total cholesterol, and LDL-cholesterol, and increased HDL-cholesterol levels. Acamprosate-compensated changes associated with ethanol consumption were observed. Acamprosate treatment decreased GECAT and increased Cease control rats, but increased GECAT and decreased CEase in PRF animals. In all groups of rats, AC treatment did not influence GEH. In conclusion, our results suggest that AC can influence triacylglycerol metabolism by its action on the balance between hydrolysis and transacylation in rats.