Journal: Acta Pharmaceutica Sinica. B

Article Title: CCL9/CCR1 axis-driven chemotactic nanovesicles for attenuating metastasis of SMAD4-deficient colorectal cancer by trapping TGF- β

doi: 10.1016/j.apsb.2024.05.009

Figure Lengend Snippet: CCR1 + -G-MDSCs accumulate to SMAD4-deficient CRC tissues via the CCL15/CCR1 axis and CCL9/CCR1 axis in clinical specimens and mouse models. (A) Clinical specimens of human CRC were examined using immunohistochemistry staining for SMAD4, CCR1 and CD33, and (B) representative images of indicated specimens. Scale bar = 100 μm. (C) Immunohistochemistry analysis of human CRC specimens showing the correlations of SMAD4 and CCR1 expression. Pearson correlation analysis was used to indicate correlation. (D) Fluorescence Activated Cell Sorter (FACS) analysis of the percent of MDSCs in PBMC from healthy subjects and clinical patients with CRC. (E) Representative FACS plots of CD33 + CCR1 + in CD11b + HLA-DR – cells. (F) Quantification of CCL15 concentration in plasma from healthy subjects and clinical patients with CRC. (G) Representative FACS plots and (H) quantification of the percent of CD14 – CD15 + (G-MDSCs) in MDSCs, respectively. G: G-MDSCs, M: MDSCs. (I) MC38 or CT26 cells were injected into spleens of C57BL/6 or BALB/c mice respectively. Mice were reared for 14 days and sacrificed for analysis of liver metastasis, n = 4. (J) Representative FACS plots of MDSCs in CD11b + , CCR1 + in MDSCs from MC38 and CT26 metastasis tissues, respectively. (K) Liver metastasis from mice was stained for CD11b (green) and Ly6G/Ly6C (red). Inset shows invasion front of liver metastasis. Scale bar = 100 μm, scale bar for zoom out = 30 μm. (L) Quantification of the percent of G-MDSCs in CD11b + , n = 4, from MC38 and CT26 metastasis tissues, respectively. (M) Immunohistochemistry staining for CCL9 and CCR1 from MC38 and CT26 metastasis tissues, Scale bar = 100 μm, scale bar for zoom out = 20 μm. (N) Quantification of CCL9 concentration in tumors, n = 4. (O) Schematic summary showing SMAD4-deficient CRC recruited CCR1 + -G-MDSC via the CCL9/CCR1 axis. Data are presented as mean ± SD of at least two independent experiments. Two-tailed unpaired Student's t -test (H, L, N), and Brown-Forsythe and Welch ANOVA tests were used (D, F) for statistical analysis with calculated P values shown.

Article Snippet: The concentrations of CCL15 in the plasma of patients or healthy subjects were measured by Human CCL15 ELISA KIT (Solarbio, Cat# SEKH-0248).

Techniques: Immunohistochemistry, Staining, Expressing, Fluorescence, Concentration Assay, Clinical Proteomics, Injection, Two Tailed Test

Journal: Clinical Epigenetics

Article Title: Abnormal DNA methylation analysis of leucine-rich glioma-inactivated 1 antibody encephalitis reveals novel methylation-driven genes related to prognostic and clinical features

doi: 10.1186/s13148-023-01550-5

Figure Lengend Snippet: A A cluster heatmap showing methylation changes in promoter of 13 microRNAs between LGI1 encephalitis cohort and normal donors. B Morphological characteristics of apoptotic body, microvesicles, and exosome isolated from serum of one LGI1 encephalitis patient by TEM. C Particle size of microvesicles and exosome using NTA. D Protein markers of apoptotic body (C3B, C1QC), microvesicles (ARF6), exosome (CD63, TSG101) and serum supernatant by western blot. E Volcano plot of differentially expressive exosome-microRNAs between LGI1 encephalitis and healthy control. F A scatter plot assessing the expression variation of exosome microRNAs between LGI1 encephalitis patients and healthy donors. G Venn diagram showing the overlap of 71 differential exosome microRNAs and 13 methylated-driven microRNAs. H The methylation changes in promoter of hsa-miR-2467-5p between LGI1 encephalitis cases and healthy donors. I PCR analysis of miR-2467-5p expression in PBMCs between LGI1 encephalitis patients and healthy donors. J PCR analysis of miR-2467-5p expression in apoptotic bodies isolated from LGI1 encephalitis patients and healthy donors. K PCR analysis of miR-2467-5p in microvesicles isolated from LGI1 encephalitis patients and healthy donors. L PCR analysis of miR-2467-5p in exosomes isolated from LGI1 encephalitis patients and healthy donors. M Schematic representation of the complementary binding sites of CSF3 and PDCD1 with miR-2467-5p. Relative luciferase activity of wild-type and 3`UTR mutant constructs of CSF3 N and PDCD1 O cotransfected with miR-2467-5p mimics and miRNA-NC. PCR analysis of PDCD1 P , CSF3 Q and CCL15 R expression in PBMCs after transfection of miR-2467-5p mimics, inhibitor or scramble control into PBMCs. ELISA analysis of PDCD1 S , CSF3 T, and CCL15 U expression in cell supernatants after transfection of miR-2467-5p mimics, inhibitor or scramble control into PBMCs. Scatterplots showing the association of miR-2467-5p in PBMCs with the expression of PDCD1 V , CSF3 W , and CCL15 X in serum in LGI1 encephalitis patients ( n = 6) and healthy donors ( n = 4). LGI1-E LGI1 encephalitis; ND normal donor; AE autoimmune encephalitis; MV microvesicles; ABs apoptotic bodies; Sup supernatant; NC negative control. Data are means ± SD of three experiments. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: ELISA kits were used to measure CSF3, PDCD1, and CCL15 according to the manufacturer's instructions (Boster, Wuhan, China).

Techniques: Methylation, Isolation, Western Blot, Control, Expressing, Binding Assay, Luciferase, Activity Assay, Mutagenesis, Construct, Transfection, Enzyme-linked Immunosorbent Assay, Negative Control

Journal: Clinical Epigenetics

Article Title: Abnormal DNA methylation analysis of leucine-rich glioma-inactivated 1 antibody encephalitis reveals novel methylation-driven genes related to prognostic and clinical features

doi: 10.1186/s13148-023-01550-5

Figure Lengend Snippet: PDCD1 and CSF3 expression level in thymoma. A and B PDCD1 and CSF3 expression were compared between thymoma and normal tissues. C and D PDCD1 and CSF3 expression in thymoma were indicated in Masaoka stage I–IV. E and F PDCD1 and CSF3 expression in histological type A–C of thymoma. G and H PDCD1 and CSF3 expression in different tumor sites. All expression value (TPM value) were acquired using the TCGA dataset on the Xiantao platform. ns no significant. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: ELISA kits were used to measure CSF3, PDCD1, and CCL15 according to the manufacturer's instructions (Boster, Wuhan, China).

Techniques: Expressing

Journal: Clinical Epigenetics

Article Title: Abnormal DNA methylation analysis of leucine-rich glioma-inactivated 1 antibody encephalitis reveals novel methylation-driven genes related to prognostic and clinical features

doi: 10.1186/s13148-023-01550-5

Figure Lengend Snippet: The association of PDCD1 and CSF3 expression with thymoma immune infiltration. A The expression of IFNγ, ICAM1, TNF and CCL2 were compared between thymoma and normal tissues using the TCGA dataset on the Xiantao platform. B Spearman correlation were shown between PDCD1 and CSF3 expression in thymoma. C and D The correlations of PDCD1 and CSF3 with all 24 immune cell types. iDC immature DC; Tem T effector memory; TFH T follicular helper; Tgd T gamma delta; pDC plasmacytoid DC; aDC activated DC; Tcm T central memory. * p < 0.05, ** p < 0.01, *** p < 0.001

Article Snippet: ELISA kits were used to measure CSF3, PDCD1, and CCL15 according to the manufacturer's instructions (Boster, Wuhan, China).

Techniques: Expressing