Journal: Cancer Research

Article Title: Integrative Multiomic Classification Reveals Distinct Origins and Evolutionary Trajectories of Thymic Epithelial Tumors

doi: 10.1158/0008-5472.CAN-24-4977

Figure Lengend Snippet: CN-type thymoma–specific IRS4 activation. A, Volcano plot showing differentially expressed genes between GTF2I-type and CN-type thymomas. The P values were calculated by a Student t test and adjusted with the Benjamini–Hochberg procedure. Known oncogenes (reported in COSMIC Cancer Gene Census) are highlighted in red. B, Box plot showing exclusive IRS4 expression in CN-type thymomas. The unit of the y -axis is TPM. The P values were calculated by the Wilcoxon test. C, Relationship between IRS4 expressions and IGF/PI3K/AKT cascade in CN-type tumors. IRS4 showed strong positive correlation with PI3K cascade score (REACT_976; top right), IGF receptor signaling pathway score (GO:0048009; top left), and their representative individual genes, such as IGFR1 (bottom left) and PIK3R1 (bottom right). Pearson correlation tests were conducted. D, Western blot results comparing IRS4 -overexpressed cells and control cells. Overexpression of IRS4 in HEK293T cells results in an increase of AKT and phosphorylated AKT (pAKT), implying activation of the PI3K/AKT pathway. The activation of the PI3K/AKT pathway was sustained in a serum-free condition, which further supports the role of IRS4 in the pathway activation. E, Cell proliferation assay comparing IRS4 -overexpressed cells and control cells. Overexpression of IRS4 increases cell proliferation rate of HEK293T cells. F, Cell viability assay comparing IRS4 -overexpressed cells and control cells. Overexpression of IRS4 decreases response to OSI-906 (IGF1R inhibitor). G, Kaplan–Meier curves of recurrence-free survival showing worse prognosis of IRS4 -high CN-type thymomas (red) than GTF2I-type (blue) or IRS 4-low CN-type thymomas (orange). The median value of IRS4 expression (TPM = 30) separates CN-type thymomas into IRS4 -high or -low subgroups. The P values were calculated by the log-rank test. H, Percentage of TET cases harboring driver mutations ( HRAS , GTF2I , and CYLD ) or showing IRS4 activation in each histologic type.

Article Snippet: An insulin-like growth factor 1 receptor (IGF1R) inhibitor, linsitinib (OSI-906), was purchased from Selleckchem.

Techniques: Activation Assay, Expressing, Western Blot, Control, Over Expression, Proliferation Assay, Viability Assay

Journal: Stem Cell Reports

Article Title: An innovative in vitro system unveils IGF1R signaling regulating Merkel cell generation

doi: 10.1016/j.stemcr.2025.102756

Figure Lengend Snippet: IGF2-IGF1R signaling pathways regulate MC development (A) Uniform manifold approximation and projection visualization of cells from integrated published scRNA-seq data ( GSE154579 ) collected from E14.5, E16.5, and P0 skin of mice. The MC cluster is indicated in red ( n = 81 cells) with dashed box enlarged. IFE, interfollicular epidermis. (B) Dot plot shows the expression of selected marker genes of different types of cells. (C) Circle plot shows the significant ligand-receptor interactions identified by cell-cell communication analysis by iTALK. Line thickness and arrowheads are scaled to represent interaction strength between ligands and receptors. IGF1/2-IGF1R interaction was highlighted in red. (D) Violin plots show the normalized expression levels of Igf1/2 and Igf1r in the indicated cells in scRNA-seq data. (E and F) (E) Spatial transcriptomics analysis of P0 skin by Stereo-seq. Image of single-stranded DNA staining section is shown. The blue box highlights the touch dome region. Spatial distribution of representative cell clusters in the region is shown on the right. The dashed lines indicate the edges of hair follicle. IGF1/2-IGF1R-mediated adjacent fibroblast (FB)-MC communications are indicated by arrows. (F) Spatial expression of indicated genes in the highlighted region of (E) (see also ).

Article Snippet: The experimental groups were treated with a PRC inhibitor cocktail (PRCi), the FGFR2i infigratinib, the IGF1R inhibitor (BMS-754807), or with the AKT agonist SC79 (HY-18749) (all from MCE).

Techniques: Protein-Protein interactions, Expressing, Marker, Spatial Transcriptomics, Staining

Journal: Stem Cell Reports

Article Title: An innovative in vitro system unveils IGF1R signaling regulating Merkel cell generation

doi: 10.1016/j.stemcr.2025.102756

Figure Lengend Snippet: FGF20-FGFR2 and IGF2-IGF1R are involved in MC development (A and G) Whole-mount IF staining for SOX2 (magenta) and K8 (yellow) was performed on E14.5 mouse vibrissae following 6 days’ treatment with 10 μM IGF1R inhibitor (IGF1Ri, BMS-754807). The quantification of MC number per vibrissa in (G) was shown as box blots (each data point represents a vibrissa explant, n = 9 vibrissae per group from 1 independent experiment). (B and H) Whole-mount IF staining for SOX2 (magenta) and K8 (yellow) was performed on E14.5 mouse vibrissae following 6 days’ treatment with 5 nM FGFR2 inhibitor (FGFR2i, infigratinib). The quantification of MC number per vibrissa was shown in (H) (each data point represents a vibrissa explant, n = 8 vibrissa per group from 2 independent experiments). (C–F and I–J) Whole-mount IF staining for K20 + MC (magenta) and DAPI (blue) was performed on hSKO following 4 weeks’ treatment with vehicle, 10 μM IGF1Ri, or 10 μM FGFR2i. The quantification of MC number per field was shown in (I and J) as box blots (each data point represents a hSKO sample, n = 7 hSKOs per group from 2 independent experiments) (4 fields averaged per hSKO, field area = 0.3364 mm 2 ). (K–M) Western blot (K) and statistical analysis were performed to assess the protein expression of total AKT (L) and phospho-AKT (pAKT-Ser473) (M) in vibrissae following 6 days’ treatment with vehicle or IGF1Ri. Data are presented as mean ± SEM; each data point represents an independent experiment, ( n = 5). (N) The quantification of MC number per vibrissa after 6 days’ treatment with DMSO (vehicle), 10 μM IGF1Ri, 10 μM IGF1Ri and 10 μM AKT agonist SC79 (each data point represents a vibrissa explant, n = 11 vibrissa per group from 3 independent experiment). Statistical significance was assessed using the t test (G, H, I, L, and M) or non-parametric t test (J and N), with ns, p > 0.05; ∗ p < 0.5; ∗∗∗ p < 0.001. Scale bars: 100 μm in (A and B) and 50 μm in (I–L).

Article Snippet: The experimental groups were treated with a PRC inhibitor cocktail (PRCi), the FGFR2i infigratinib, the IGF1R inhibitor (BMS-754807), or with the AKT agonist SC79 (HY-18749) (all from MCE).

Techniques: Staining, Western Blot, Expressing

Journal: Journal of Nanobiotechnology

Article Title: RVG-targeted extracellular vesicles loaded with echinatin attenuate dopaminergic neurodegeneration via the IGF-2/PI3K/Akt pathway in Parkinson’s disease mice

doi: 10.1186/s12951-025-03997-5

Figure Lengend Snippet: RVG-EVs@Echi attenuate oxidative stress in the in vitro PD models. ( a ) Representative blots and ( b ) quantification of the expression levels of IGF2, P-PI3K, PI3K, P-Akt, Akt, and Nrf2 in MN9D cells. ( c ) Representative blots and ( d ) quantification of Bcl-2, Bax, and the resulting Bcl-2/Bax ratio in MN9D cells. ( e , g ) Representative blots and ( f , h ) quantification showing the effects of the IGF-1R/IR inhibitor NVP-AEW541 in MN9D cells. ( i ) Representative graphs of ROS generation in control or MPP+-treated cells upon RVG-EVs@Echi treatment. Scale bars, 50 μm and 10 μm for the original and magnified images, respectively. ( k ) MitoSOX was used to stain live control or MPP+-treated cells upon treatment with RVG-EVs@Echi. Scale bars, 50 μm and 10 μm for the original and magnified images, respectively. ( j , l ) Quantification of the relative ROS and MitoSOX fluorescence intensity. n = 6 per group. ( m ) Representative flow cytometry histograms and ( n ) quantification of mean fluorescence intensity (MFI) showing intracellular ROS levels in MN9D cells. n = 3 per group. Statistical Analysis: For (a-d): Groups are Control, MPP+, and MPP + + RVG-EVs@Echi. n = 3 per group. * p < 0.05, ** p < 0.01 vs. Control group; # p < 0.05, ## p < 0.01 vs. MPP + group. For (e-h): Groups are Control, MPP + + RVG-EVs@Echi, and MPP + + RVG-EVs@Echi + NVP-AEW541. n = 3 per group. * p < 0.05, ** p < 0.01 vs. Control group; # p < 0.05, ## p < 0.01 vs. MPP + + RVG-EVs@Echi group. For (i-n): Groups are Control, RVG-EVs@Echi, MPP+, and MPP + + RVG-EVs@Echi. * p < 0.05, ** p < 0.01 vs. Control group; # p < 0.05, ## p < 0.01 vs. RVG-EVs@Echi group; & p < 0.05, && p < 0.01 vs. MPP + group

Article Snippet: To assess the effects of RVG-EVs@Echi, MN9D cells were treated with MPP+ (600 μM) or RVG-EVs@Echi (1 μM) for 24 h. For the IGF-1R inhibition experiment, cells were pre-treated with the IGF-1R/IR inhibitor NVP-AEW541 (0.2 μM; MCE, HY-50866) [ ] for 2 h, followed by co-incubation with MPP + and RVG-EVs@Echi for 24 h.

Techniques: In Vitro, Expressing, Control, Staining, Fluorescence, Flow Cytometry