Journal: bioRxiv

Article Title: Scalable microvascular networks-on-chip enable 8-week unidirectional perfusion for long-term vascular toxicity screening

doi: 10.64898/2026.06.10.731320

Figure Lengend Snippet: a, Schematic of the experimental design illustrating acute (72-hour) exposure of microvascular networks to six compounds administered exclusively through the perfusion channel. Vascular toxicity was characterised across five readouts: nuclear coverage, αSMA and ICAM-1 expression levels, perfusable area fraction, and dextran extravasation fraction. b, Nuclear coverage as a measure of cytotoxicity following exposure to six compounds at four concentrations at day 15 (N=2 independent experiments across 12 plates, n=9 chips for VC n=6–7 chips per compound). Data are presented as mean ± SD. Two-way ANOVA revealed significant effects of compound (F(5,139)=41.30, P<0.0001), concentration (F(3,139)=47.49, P<0.0001), and a significant compound x concentration interaction (F(15,139)=10.70, P<0.0001). Dunnett’s post hoc test was applied relative to VC; exact P values for all pairwise comparisons are provided in Supplementary Table 10. Dashed line indicates the 70% viability threshold. c, Multiparametric bubble plot summarising compound-induced changes across all five readouts. Bubble colour represents log2 fold change relative to VC; bubble size represents statistical significance. All readouts were assessed per compound using Welch one-way ANOVA with Dunnett’s T3 post hoc test relative to VC (N=2 independent experiments, n=9 chips for VC, n=6–7 chips per compound). Exact P values for all comparisons are provided in Supplementary Table 11. d, Representative maximum intensity projection images of network perfusability for VC and each compound at low, medium, and high concentrations. Images represent 4-site acquisitions using a 10X objective. e, Representative immunofluorescent images of αSMA and ICAM-1 expression in VC, sorafenib (10 µM), and LY2090314 (0.1 µM) conditions. Images were acquired as single-site maximum intensity projections using a 10X objective; scale bar, 100 µm. Lower panels show digitally magnified insets from the corresponding upper images; scale bar, 100 µm (left) 25 µm (right). Yellow arrows indicate cytoplasmic vesicle accumulation in αSMA-positive cells following sorafenib exposure. f, Comparative quantification of total nuclear area following acute exposure to cabozantinib and doxorubicin in day 15 versus day 57 microvascular cultures (N=2, n=9 chips for VC, n=6–7 chips per compound). Data are presented as mean ± SD. Two-way ANOVA with Dunnett’s post hoc test relative to VC; exact P values are provided in Supplementary Table 12. Images in d and e are representative of the stated N and n. Significance thresholds: *P<0.05, **P<0.01, ***P<0.001, *** P<0.0001 .

Article Snippet: Lenvatinib (TargetMol, T0520), sorafenib (MedChem Express, HY-10201), cabozantinib (Selleck Chemicals, S1119), LY2090314 (Selleck Chemicals, S7063), Chloroquine diphosphate (Selleck Chemicals, S4157), and Doxorubicin hydrochloride (MedChem Express, HY-15142) were prepared according to the manufacturer’s instructions and diluted in EGM-2 medium immediately prior to use.

Techniques: Toxicity, Expressing, Concentration Assay, Significance Assay, Immunofluorescence

Journal: Cancer Discovery

Article Title: A Targetable Developmental Program Coregulates Angiogenesis and Immune Evasion in Melanoma

doi: 10.1158/2159-8290.CD-24-1853

Figure Lengend Snippet: CD73 is a key mediator of the HOXD13-dependent T-cell exclusion phenotype. A, The NT5E locus displays different regulatory elements contacting each other and bound by HOXD13. Silencing of HOXD13 reduces the contacts around the NT5E promoter. B, Bar dot plot representing CD73 protein quantification in tumor (left) and blood vessel (right) areas of a TMA in which cores were segregated into HOXD13 -expressing ( n = 61) and HOXD13 -negative ( n = 39). An unpaired t test was conducted, and P values are displayed. C, Representative HOXD13 low and HOXD13 high cores (RNAScope, top) and CD73, SOX10, and CD31/CD34 (mIF, bottom). The white scale bar measures 200 μm. D, qRT-PCR of Nt5e (Cd73) mRNA expression in B16F10 OVA cells that are EMPTY or Cd73 overexpressing, incubated with siSCR or siHoxd13. A paired t test was performed, and P values are shown in the graphs. E, Relative cell growth of B16F10 OVA carrying Cd73 ectopic expression cocultured with OT-I T cells upon Hoxd13 KD. A two-way ANOVA was conducted between the conditions. P values are shown. F, YUMM1.7 EMPTY or HOXD13 OE tumor growth expressing an shSCR (control) or shCd73. Mixed-effect analysis with Tukey comparison was conducted. The P value of the comparison YUMM1.7 HOXD13 shSCR vs. shCd73 is displayed. G, Tumor growth measurements of YUMM1.7 HOXD13 syngeneic grafts upon vehicle (control), lenvatinib, etrumadenant, or lenvatinib + etrumadenant (combo). Mixed-effects analysis through Tukey multiple comparison was performed, and P values are shown in the curves’ graph. H, Representative mIF staining for DAPI, CD45, CD3, CD11b, CD31, and α-SMA in YUMM1.7 HOXD13 tumors upon vehicle, etrumadenant, lenvatinib, and combo treatments. Combo dramatically decreased intratumor vasculature (CD31, α-SMA) and increased immune cells, particularly macrophage infiltration (CD45, CD11b). White bars reflect 500 mm size. I, B16F10 tumor growth in syngeneic mice upon shNTC, shHoxd13, or treated with vehicle or combo ( n = 9 mice per condition). Mixed-effect Tukey comparison was used as a statistical study; P values are shown. J, mIF opal staining for fibrin, α-SMA, and CD31 in representative tumor areas of B16F10 tumors shNTC, shHoxd13, vehicle, and combo treated. The scale bar is a reference of 50 μm.

Article Snippet: For the etrumadenant and lenvatinib experiment, 8-week-old male mice C57BL/6J ( n = 10 per group) were injected intradermally in the flanks with 1 × 10 5 YUMM1.7 HOXD13 or B16F10, and after 3 days of grafting, mice were daily administered by oral gavage with 10 μg/g of lenvatinib (MedChemExpress, HY-10981) and/or 100 μg/g etrumadenant (MedChemExpress, HY-129393) or vehicle (95% corn oil + 5% DMSO).

Techniques: Expressing, RNAscope, Quantitative RT-PCR, Incubation, Control, Comparison, Staining