Journal: Advanced Science

Article Title: PARPi Combining Nanoparticle LIN28B siRNA for the Management of Malignant Ascites

doi: 10.1002/advs.202510547

Figure Lengend Snippet: Aberrant LIN28B expression and DNA repair pathway are associate with MSE. (a) Integrated analysis combining scRNA‐seq data from ascites in a constructed preclinical OC model; scRNA‐seq data from MA in OC patients ( n = 13), and gene mutation profiles from MSE in cancer patients ( n = 442) to explore novel dual‐targeting therapeutic strategies (created at BioRender.com). (b) Venn diagram analysis of DEG targets between the preclinical OC model and OC patients (Nat Cancer. 2023). GO‐BP enrichment analysis for (c) SNV gene mutations and (d) CNV gene mutations in MSE from cancer patients. (e) Waterfall plot for top 45 SNV genes and relative pathways. Genotype Quality (GQ): Minimum threshold of 20 (99% confidence in the genotype call). Read Depth (DP): Site‐specific depth filters (DP ≥ 10 for high‐confidence calls, adjusted for cohort mean depth). lternate Allele Support: Minimum alternative allele reads (≥ 3) and fraction (AF ≥ 0.25 for heterozygous germline calls). opulation Frequency: Filtering against population databases (gnomAD allele frequency < 0.01 for rare variants, < 0.001 for ultra‐rare). (f) Heatmap of CNV genes in MSE from cancer patients ( n = 442). Quality Metrics: Segment‐wise log2 ratio standard deviation or confidence score (e.g., CNVkit quality score > 0.5). Size Threshold: Typically, > 1 kb for targeted sequencing and > 10–50 kb for whole‐genome sequencing to exclude small, noisy calls. Copy State Threshold: Log2 ratio thresholds define copy states (e.g., deletion: log2 ratio < −0.4, duplication: log2 ratio > 0.2).

Article Snippet: The following antibodies were used: anti‐human LIN28B (Cell Signaling Technology, 4196S, 1:1000), anti‐mouse Lin28b (Proteintech, 11724‐1‐AP, 1:500), anti‐β‐actin (ABclonal, AC026, 1:10000), StarBright Blue 520 goat anti‐mouse IgG (Bio‐Rad, 12005867, 1:10000), Alexa Fluor Plus 800 goat anti‐rabbit IgG (H + L) (Invitrogen, A32735, 1:10000), Anti‐Mouse CD8a‐Purified In vivo (C375‐25 mg), Anti‐Mouse NK1.1‐Purified In vivo (N123‐25 mg), Human/Primate IL‐6 Mab(Clone 6708, MAB206‐SP, R&D), Human TNF‐alpha Mab(Clone28401, MAB610‐SP, R&D), anti‐CD31 antibody (Thermo Fisher Scientific, MA3105, 1:200), and anti‐TER‐119 antibody (BD Biosciences, 557915, 1:100).

Techniques: Expressing, Construct, Mutagenesis, Standard Deviation, Targeted Sequencing, Sequencing

Journal: Advanced Science

Article Title: PARPi Combining Nanoparticle LIN28B siRNA for the Management of Malignant Ascites

doi: 10.1002/advs.202510547

Figure Lengend Snippet: Synthesis and characterization of DSSP@lip‐PEG‐FA targeting LIN28B.(a) Schematic illustration of the synthesis process for siRNA/DSSP@lip‐PEG‐FA. Representative TEM images and size distribution profiles of siRNA/DSSP NPs (b) and siRNA/DSSP@lip‐PEG‐FA (c). (d) hydrogen peroxide (H 2 O 2 )‐ and glutathione (GSH)‐responsive siRNA release profiles under different conditions (pH 7.4, pH7.4& 1 mM H 2 O 2 , and pH 7.4 & 10 mM GSH) over 24 h. (e) Confocal microscopy images (scale bars: 50 µm) and quantitative analysis of cellular uptake efficiency for free siRNA, siRNA/DSSP@lip‐PEG, and siRNA/DSSP@lip‐PEG‐FA. (f) Flow cytometric analysis quantifying Cy3‐labeled siRNA fluorescence intensity in treated cells. (g) Flow cytometry evaluation of folate receptor (FA)‐targeting specificity. (h) Mean fluorescence intensity (MFI) comparison across different treatment groups at 6 h post‐incubation. (i) Confocal images showing intracellular distribution of siRNA/DSSP@lip‐PEG‐FA (scale bar: 10 µm). (j) Quantitative colocalization analysis performed using ImageJ software. (k) and (l) Western blot analysis of Lin28b protein expression in A2780 and ID8 ovarian cancer cells following various treatments. Data are presented as mean ± SD ( n = 3, unpaired t‐test). n = independent biological replicates. * p < 0.05, ** p < 0.01, *** p < 0.001; ns, not significant.

Article Snippet: The following antibodies were used: anti‐human LIN28B (Cell Signaling Technology, 4196S, 1:1000), anti‐mouse Lin28b (Proteintech, 11724‐1‐AP, 1:500), anti‐β‐actin (ABclonal, AC026, 1:10000), StarBright Blue 520 goat anti‐mouse IgG (Bio‐Rad, 12005867, 1:10000), Alexa Fluor Plus 800 goat anti‐rabbit IgG (H + L) (Invitrogen, A32735, 1:10000), Anti‐Mouse CD8a‐Purified In vivo (C375‐25 mg), Anti‐Mouse NK1.1‐Purified In vivo (N123‐25 mg), Human/Primate IL‐6 Mab(Clone 6708, MAB206‐SP, R&D), Human TNF‐alpha Mab(Clone28401, MAB610‐SP, R&D), anti‐CD31 antibody (Thermo Fisher Scientific, MA3105, 1:200), and anti‐TER‐119 antibody (BD Biosciences, 557915, 1:100).

Techniques: Confocal Microscopy, Labeling, Fluorescence, Flow Cytometry, Comparison, Incubation, Software, Western Blot, Expressing