Journal: Alzheimer's & Dementia

Article Title: Plasma extracellular vesicles from APOE3 Christchurch carriers display a protective phenotype in early stages of autosomal dominant Alzheimer's disease

doi: 10.1002/alz.71148

Figure Lengend Snippet: Reduction of endothelial and leukocyte EV markers with concomitant platelet marker elevation in APOE3 Ch carriers. A, Percentage of EVs positive for CD45 (leukocytes), CD105 (endothelial cells), CD235a (erythrocytes), CD41a (platelets), and double‐positive CD45‐CD105. B, Normalized MFI of EVs positive for markers in (A). C, Heatmap showing EV positivity for the marker panel in (A), comparing PSEN1 E280A ‐APOE3 carriers with and without MCI. D, Heatmap of MFI for PSEN1 E280A ‐APOE3 carriers with and without MCI. E, Violin plots comparing positivity of markers in non‐MCI PSEN1 E280A carriers to APOE3 and APOE3 Ch . F, Violin plots comparing the MFI in non‐MCI PSEN1 E280A carriers with APOE3 and APOE3 Ch . In (C) and (D), each column in the heatmap is labeled with a number, in which each number represents an individual patient. Representative data from CNT, n = 10 (C1, C2, C3, C4, C5, C6, C7, C8, C9, C10); APOE3 Ch , n = 10 (Ch1, Ch2, Ch3, Ch4, Ch5, Ch6, Ch7, Ch8, Ch9, Ch10); MCI PSEN1 E280A ‐APOE3 , n = 10 (MCI1, MCI2, MCI3, MCI4, MCI5, MCI6, MCI7, MCI8, MCI9, MCI10); non‐MCI PSEN1 E280A ‐APOE3 , n = 10 (non‐MCI‐1, non‐MCI‐2, non‐MCI‐3, non‐MCI‐4, non‐MCI‐5, non‐MCI‐6, non‐MCI‐7, non‐MCI‐8, non‐MCI‐9, non‐MCI‐10) and non‐MCI PSEN1 E280A ‐APOE3 Ch , n = 9 (E‐Ch1, E‐Ch2, E‐Ch3, E‐Ch4, E‐Ch5, E‐Ch6, E‐Ch7, E‐Ch8, E‐Ch9). Data are presented as boxplots or violin plots. Boxplots display the median, interquartile range, and whiskers representing minimum and maximum values. Violin plots illustrate the full data distribution, with individual data points overlaid and a line indicating the median. The data for the homozygous APOE3 Ch patient is indicated by a red dot. One‐way analysis of variance, Tukey multiple comparison test; unpaired t test, Mann–Whitney test were performed. Comparison to CNT is denoted by number sign (#); with non‐MCI PSEN1 E280A ‐ APOE3 by asterisk (*). */# indicates P < 0.05, **/## P < 0.01, and ***/### P < 0.001. APOE , apolipoprotein E; CD, cluster of differentiation; CNT, cognitively unimpaired control; EV, extracellular vesicle; MCI, mild cognitive impairment; MFI, mean fluorescence intensity; PSEN, presenilin

Article Snippet: Cells were then stimulated with 50 ng/mL vascular endothelial growth factor (VEGF, 293‐VE, R&D Systems) in combination with either 1 μM wild‐type APOE3 or 1 μM APOE3Ch, diluted in experimental medium consisting of EBM‐2 supplemented with 5% FBS (30‐2020, ATCC), 1% GlutaMAX (35050061, Thermo Fisher Scientific), 0.2% Normocin, and 10 mM LiCl.

Techniques: Marker, Labeling, Comparison, MANN-WHITNEY, Control, Fluorescence

Journal: Alzheimer's & Dementia

Article Title: Plasma extracellular vesicles from APOE3 Christchurch carriers display a protective phenotype in early stages of autosomal dominant Alzheimer's disease

doi: 10.1002/alz.71148

Figure Lengend Snippet: Wnt signaling expression in APOE3 Ch across vascular cells and conditions of AD, and its role in promoting vascular integrity. A, UMAP visualization of 81,440 cells from post mortem human prefrontal cortex. B, Heatmap of relative expression of canonical Wnt signaling mediators across vascular cell types in APOE3 Ch and non‐carries with PSEN1 E280A , showing a significant increase WNT TF in endothelial cells. Significance showed for each gene and cell type (* P < 0.05). C, Violin plot of lymphoid enhancer‐binding factor‐1 expression in endothelial for non‐disease (Control), FAD ( PSEN1 E280A ), APOE3Ch ( PSEN1 E280A APOE3 Ch ). D, TEER measurements in HBMECs showed that treatment with VEGF (50 ng/mL) led to a progressive reduction in TEER over a 48‐hour period, reflecting increased endothelial permeability. However, this VEGF‐induced decrease in barrier integrity was prevented when cells were co‐treated with either wild‐type ApoE3 or the ApoE3Ch variant at 1 µM concentration. E, On the left, representative images show β‐catenin (green), CD31 (red), and DAPI (blue) in HBMECs co‐cultured with astrocytes, treated with ApoE3Ch (1 µM) in combination with ApoE4 (1 µM). On the right, quantification of nuclear β‐catenin demonstrates that ApoE3Ch enhances β‐catenin nuclear translocation and restores adherens junction integrity disrupted by ApoE4. AD, Alzheimer's disease; APOE , apolipoprotein E; FAD, familial Alzheimer's disease; HBMEC, human brain microvascular endothelial cell; MCI, mild cognitive impairment; PSEN, presenilin; TEER, transendothelial electrical resistance; UMAP, Uniform Manifold Approximation and Projection; VEGF, vascular endothelial growth factor

Article Snippet: Cells were then stimulated with 50 ng/mL vascular endothelial growth factor (VEGF, 293‐VE, R&D Systems) in combination with either 1 μM wild‐type APOE3 or 1 μM APOE3Ch, diluted in experimental medium consisting of EBM‐2 supplemented with 5% FBS (30‐2020, ATCC), 1% GlutaMAX (35050061, Thermo Fisher Scientific), 0.2% Normocin, and 10 mM LiCl.

Techniques: Expressing, Binding Assay, Control, Permeability, Variant Assay, Concentration Assay, Cell Culture, Translocation Assay

Journal: Alzheimer's & Dementia

Article Title: Plasma extracellular vesicles from APOE3 Christchurch carriers display a protective phenotype in early stages of autosomal dominant Alzheimer's disease

doi: 10.1002/alz.71148

Figure Lengend Snippet: PSEN1 E280A ‐APOE3 Ch EVs preserve gliovascular integrity in an in vitro coculture model. A, Representative images of endothelial cells treated with EVs from PSEN1 E280A carriers with APOE3 and APOE3 Ch . B, Quantification shows the number and area of intercellular gaps within the endothelial monolayer, as well as the fraction of nuclear β‐catenin. C, Representative images of astrocytes treated with EVs. D, Quantification (right) includes GFAP intensity (red), reflecting astrocyte reactivity, and the fraction of nuclear β‐catenin (green). E, Representative images showing ApoE signal localization in endothelial cells and astrocytes. F, Quantification depicts the number of ApoE‐positive particles normalized to the total number of cells, highlighting differential ApoE uptake. Representative data from; non‐MCI PSEN1 E280A ‐APOE3 , n = 4 (non‐MCI‐2, non‐MCI‐3, non‐MCI‐6, non‐MCI‐7) and non‐MCI PSEN1 E280A ‐APOE3 Ch , n = 4 (E‐Ch6, E‐Ch7, E‐Ch8, E‐Ch9). Magnification 20x and 40x, Scale bars: 50 µm. Violin plots illustrate the full data distribution, with individual data points overlaid and a line indicating the median. Unpaired t test, Mann Whitney test were performed. * indicates P < 0.05; ** P < 0.01; *** P < 0.001. APOE , apolipoprotein E; EV, extracellular vesicle; GFAP, glial fibrillary acidic protein; MCI, mild cognitive impairment; PSEN, presenilin

Article Snippet: Cells were then stimulated with 50 ng/mL vascular endothelial growth factor (VEGF, 293‐VE, R&D Systems) in combination with either 1 μM wild‐type APOE3 or 1 μM APOE3Ch, diluted in experimental medium consisting of EBM‐2 supplemented with 5% FBS (30‐2020, ATCC), 1% GlutaMAX (35050061, Thermo Fisher Scientific), 0.2% Normocin, and 10 mM LiCl.

Techniques: In Vitro, MANN-WHITNEY

Journal: Journal of Translational Medicine

Article Title: The ANGPTL3-integrin α5 axis drives retinal vascular leakage in diabetic retinopathy

doi: 10.1186/s12967-026-07710-4

Figure Lengend Snippet: ANGPTL3 knockout preserves retinal endothelial barrier function by upregulating tight junction proteins. ( A ) Representative TEM images showing tight junction structures in retinal capillary endothelial cells of control, diabetic, and Angptl3 knockout mice. ( B ) Immunofluorescence staining of retinal sections for tight junction proteins ZO-1 and VE-cadherin. Scale bars = 10 μm. ( C and D ) Quantification of ZO-1 and VE-cadherin fluorescence intensity from ( B ). ( E ) Proliferation assay of HUVECs treated with recombinant human ANGPTL3. ( F ) Western blot analysis of ZO-1 and VE-cadherin protein levels in HUVECs following ANGPTL3 treatment. ( G - H ) Quantification of protein expression from ( F ). * P < 0.05, ** P < 0.01

Article Snippet: The cells were equilibrated in ECM containing 1% FBS (Gibco, Cat# 26140079) for 12 h before treatment with recombinant ANGPTL3 (0.25, 0.50, 0.75 μg/mL) or recombinant vascular endothelial growth factor (VEGFA; 50 ng/mL, R&D Systems, Cat# 293-VE/CF) as a positive control.

Techniques: Knock-Out, Control, Immunofluorescence, Staining, Fluorescence, Proliferation Assay, Recombinant, Western Blot, Expressing