Journal: PLOS Biology

Article Title: Angptl5 restricts primitive hematopoiesis by promoting retinoic acid signaling in zebrafish

doi: 10.1371/journal.pbio.3003858

Figure Lengend Snippet: (A) WISH of cyp26a1 in WT embryos. Embryos were treated with ERK activator (C16-PAF, ERKa) or ERK inhibitor (Mirdametinib, ERKi) from the shield stage, or injected with angptl5 + itga6l + itgb5 mRNA at the 1-cell stage. Untreated embryos were used as control. (B–D) Schematic diagram of experimental setup (B) for WISH of dhrs9 and aldh1a2 in WT embryos. caERK2 (C) or angptl5 + itga6l + itgb5 (D) mRNA injected into one blastomere on the animal pole at the 128-cell stage and then detected at 6 hpf. (E–G) Schematic diagram of experimental setup (E) for WISH of dhrs9 . WT embryos were first injected with angptl5 mRNA at the 1-cell stage. Subsequently, itga6l ± itgb5 mRNA was injected into one blastomere on the animal pole at the 128-cell stage. Embryos were then continuously treated with or without ERK inhibitor until the shield stage (G) . Uninjected embryos and only angptl5 mRNA injected embryos (F) were used as control. (H) WISH of dhrs9 in WT embryos. caERK2 mRNA ± junba/bb MO injected into one blastomere on the animal pole at the 128-cell stage and then detected at 6 hpf. (I) ChIP-qPCR analysis of Junb binding to upstream regions of dhrs9. Embryos injected with HA-Junba mRNA were subjected to chromatin immunoprecipitation with HA-agarose, using IgG-agarose as a control. Enrichment at three predicted binding sites upstream of the dhrs9 gene was quantified by qPCR. Data are presented as the mean ± SD from three independent biological replicates. *** P < 0.001 (Unpaired t t est). (J and K) WISH of spi1b in angptl5 Δ10/Δ10 embryos. Embryos were injected with itga6l + itgb5 mRNA at the 1-cell stage and treated with or without ERK inhibitor from the shield stage to the 18-somite stage (J), or injected with caERK 2 mRNA at the 1-cell stage and treated with or without RA receptor antagonist AGN 193109 (K). Uninjected embryos were used as control. Statistics are shown below. Data presented as the mean ± SD, n( angptl5 Δ10/Δ10 ) = 22, n( angptl5 Δ10/Δ10 + itga6l + itgb5 ) = 26, n( angptl5 Δ10/Δ10 + itga6l + itgb5 + ERKi) = 28 (J); n( angptl5 Δ10/Δ10 ) = 22, n( angptl5 Δ10/Δ10 + caERK ) = 22, and n( angptl5 Δ10/Δ10 + caERKi + AGN) = 26 (K). Statistical significance: ** P < 0.01, *** P < 0.001 (One-way ANOVA). LV, lateral view; AV, animal view (C–H), anterior view (J and K). The data for this figure can be found in .

Article Snippet: Detailed pharmacological parameters and experimental concentrations are tabulated below: RA (sigma), Aldehyde dehydrogenase inhibitors 4-diethylaminobenzaldehyde (DEAB) (MCE, 10 μM), RA receptor (RARs) antagonists AGN 193109 (MCE, 20 μM), FAK inhibitor Defactinib (MCE, 2 μM/8 μM), MAPK inhibitor Adezmapimod (MCE, 10 μM), ERK inhibitor Mirdametinib (MCE, 10 μM), ERK activator C16-PAF (MCE, 1 μM/5 μM), NF-κB inhibitor BAY 11-7082 (MCE, 0.1 μg/mL, 0.4 μg/mL).

Techniques: Injection, Control, ChIP-qPCR, Binding Assay, Chromatin Immunoprecipitation

Journal: International Journal of Pharmaceutics: X

Article Title: Brain-targeted delivery of siRNA via non-viral delivery systems, the therapeutic strategy for Alzheimer's disease—Unveiling challenges and prospects

doi: 10.1016/j.ijpx.2026.100503

Figure Lengend Snippet: (A) Unconjugated or C16-modified Sod1-targeting siRNA was administered as a single intrathecal bolus injection to rats at 0.9 mg, and siRNA biodistribution was assessed in whole brain at 24 h post-dose using IHC with anti-siRNA antibody.The C16-siRNA demonstrated superior brain biodistribution compared to its unconjugated version after a single intrathecal injection in rats. (B) Robust neuronal and glial cell drug accumulation (magenta) in cerebral cortex (left panel). Dual IHC for the detection of siRNA and cell-type-specific targets in neurons (Map2), astrocytes (Gfap) and microglia (Iba1). (C) APP-targeting siRNA was administered as a single intrathecal bolus injection to cynomolgus monkeys at 60 mg, and, 3 months post-dose, tissue samples were analyzed for APP mRNA levels by RT–qPCR after 85 days. NHP, non-human primates; Images reproduced from Ref. . (D) Co-administration of differentially targeting siRNA and anti-siRNAs can be adapted as a strategy to achieve tissue selectivity in different organ combinations. Images reproduced from Ref. ( , ). (E) The dual-targeting divalent siRNA is highly programmable, enabling simultaneous modulation of two different disease-relevant gene pairs (e.g. Huntington's disease: MSH3 and HTT; Alzheimer's disease: APOE and JAK1) with similar potency to a mixture of single-targeting divalent siRNAs against each gene. Images reproduced from Ref. . (For interpretation of the references to colour in this figure legend, the reader is referred to the web version of this article.)

Article Snippet: Early results from this phase I trial of ALN-APP ( NCT05231785 ) demonstrate that Alnylam's proprietary C16-siRNA conjugate platform has achieved the first human clinical transformation in CNS delivery and is the first clinical demonstration of RNA therapy silencing disease-causing genes in the human brain.

Techniques: Modification, Injection, Quantitative RT-PCR