Journal: Acta Pharmaceutica Sinica. B

Article Title: AI-powered therapeutic aptamer drug discovery: Targeting the CT-domain of CTGF for duchenne muscular dystrophy

doi: 10.1016/j.apsb.2026.02.016

Figure Lengend Snippet: CT-domain is an optimal domain without elevating compensatory TGF- β 1 level rather than VWC-domain. (a) The levels of supernatant fibrosis markers (fibronectin and collagen III) in CTGF ko (CTGF knock out) fibroblast expressing full-length CTGF (FL-CTGF), CTGF with VWC-domain deficiency (CTGF-ΔVWC), and CTGF with CT-domain deficiency (CTGF-ΔCT), respectively (upper). Statistical analysis of ratios of fibronectin and collagen III to GAPDH (lower). (b) The levels of supernatant TGF- β 1 in CTGF ko fibroblast expressing FL-CTGF, CTGF-ΔVWC and CTGF-ΔCT, respectively (upper). Statistical analysis of ratios of TGF- β 1 to GAPDH (lower). (c) The ELISA analysis of TGF- β 1 levels in fibroblast treated with CTGF (1.5 μg/mL), CTGF + FG-3019 (0.01 μg/mL), CTGF + IgG (0.05 μg/mL), CTGF + FG-3019 (0.1 μg/mL), CTGF + IgG (0.01 μg/mL), CTGF + FG-3019 (0.05 μg/mL), CTGF + IgG (0.1 μg/mL) for 48 h, respectively. (d) The Western blot analysis of fibronectin and collagen III levels in fibroblast treated with CTGF, CTGF + FG-3019, CTGF + TGF- β 1 antibody (CTGF + TGF- β 1 ab), CTGF + FG-3019 + TGF- β 1 antibody (CTGF + F + T), and CTGF + IgG for 48 h, respectively (left). Statistical analysis of ratios of fibronectin and collagen III levels to GAPDH, respectively (right). (e) The Western blot analysis of fibronectin and collagen III levels in diagram muscle of WT mice treated with PBS, Mdx mice treated with PBS, FG-3019, TGF- β 1 antibody (TGF- β 1 ab), FG-3019 + TGF- β 1 antibody (CTGF + F + T), and IgG for 6 weeks, respectively (left). Statistical analysis of ratios of fibronectin and collagen III levels to GAPDH, respectively (right). Data are expressed as mean ± SD followed by one-way ANOVA with Tukey’s post hoc test, n = 3 per group. ∗ P < 0.05. ∗∗ P < 0.01. ∗∗∗ P < 0.001, ∗∗∗∗ P < 0.0001.

Article Snippet: Mice were randomly assigned to one of four experimental groups: (1) Vehicle Control Group: Received subcutaneous (s.c.) injections of the formulation buffer alone ( e.g., PBS or vehicle used for Apc003OA), once weekly for 12 weeks. (2) Apc003OA Treatment Group: Received s.c. injections of Apc003OA at a dose of 100 mg/kg body weight, dissolved in the appropriate vehicle, once weekly for 12 weeks. (3) FG-3019 Treatment Group: Received s.c. injections of the CTGF VWC-domain specific antibody FG-3019 (purchased from Med Chem Express) administered according to an equivalent schedule and dose regimen as a comparator. (4) Apc003OA + BP Pre-treatment Group: Received an s.c. injection of the blocking peptide (BP) designed against the CT-domain of CTGF binding site for Apc003 ( e.g ., at a dose and timepoint defined in pilot studies, such as 1 h prior to each Apc003OA dose), followed by s.c. injection of Apc003OA (100 mg/kg), once weekly for 12 weeks.

Techniques: Knock-Out, Expressing, Enzyme-linked Immunosorbent Assay, Western Blot

Journal: Acta Pharmaceutica Sinica. B

Article Title: AI-powered therapeutic aptamer drug discovery: Targeting the CT-domain of CTGF for duchenne muscular dystrophy

doi: 10.1016/j.apsb.2026.02.016

Figure Lengend Snippet: Apc003OA could distribute and remain in muscle tissues for an extended period, whereas FG-3019 could not do so. (a) The in vivo distribution of Cy3-Apc003OA and Cy3-FG-3019 in muscles (diaphragm and gastrocnemius) and major organs (heart, liver, spleen, lung, and kidney) 2, 4, 12 and 24 h after subcutaneous injection, respectively. (b) Fluorescence intensities of Cy3-Apc003OA and Cy3-FG-3019 in muscles and major organs 2, 4, 12 and 24 h after subcutaneous injection, respectively. Data are expressed as mean ± standard deviation followed by multiple unpaired t tests, n = 3 per group. ∗ P < 0.05. ∗∗ P < 0.01. ∗∗∗ P < 0.001. ∗∗∗∗ P < 0.0001. The P values for the respective comparisons are indicated on the graph. Note: The data were presented as the means ± standard deviation.

Article Snippet: Mice were randomly assigned to one of four experimental groups: (1) Vehicle Control Group: Received subcutaneous (s.c.) injections of the formulation buffer alone ( e.g., PBS or vehicle used for Apc003OA), once weekly for 12 weeks. (2) Apc003OA Treatment Group: Received s.c. injections of Apc003OA at a dose of 100 mg/kg body weight, dissolved in the appropriate vehicle, once weekly for 12 weeks. (3) FG-3019 Treatment Group: Received s.c. injections of the CTGF VWC-domain specific antibody FG-3019 (purchased from Med Chem Express) administered according to an equivalent schedule and dose regimen as a comparator. (4) Apc003OA + BP Pre-treatment Group: Received an s.c. injection of the blocking peptide (BP) designed against the CT-domain of CTGF binding site for Apc003 ( e.g ., at a dose and timepoint defined in pilot studies, such as 1 h prior to each Apc003OA dose), followed by s.c. injection of Apc003OA (100 mg/kg), once weekly for 12 weeks.

Techniques: In Vivo, Muscles, Injection, Fluorescence, Standard Deviation

Journal: Acta Pharmaceutica Sinica. B

Article Title: AI-powered therapeutic aptamer drug discovery: Targeting the CT-domain of CTGF for duchenne muscular dystrophy

doi: 10.1016/j.apsb.2026.02.016

Figure Lengend Snippet: Apc003OA exerted better fibrosis inhibitory activity without elevating compensatory TGF- β 1 levels than FG-3019 in mdx mice. (a) The specific force against stimulation frequency of soleus muscles in WT mice treated with PBS, and mdx mice treated with PBS, Apc003OA, FG-3019, Apc003OA + BP, and BP for 12 weeks, respectively. (b) The forelimb grip strengths of mdx mice treated with PBS, and mdx mice treated with PBS, Apc003OA, FG-3019, Apc003OA + BP and BP for 12 weeks, respectively. (c) The fibronectin and collagen III levels in diaphragm muscle of WT mice treated with PBS, mdx mice treated with PBS, Apc003OA, FG-3019, Apc003OA + BP and BP for 12 weeks, respectively (left). Statistical analysis of fibronectin and collagen III intensity ratios to GAPDH (right). (d) The diaphragm muscle using Masson's staining from WT mice treated with PBS, and mdx mice treated with PBS, Apc003OA, FG-3019, Apc003OA + BP, and BP for 12 weeks, respectively (left). The statistical analysis of fibrosis index (right). (e) The serum levels of TGF- β 1 in WT mice treated with PBS, and mdx mice treated with PBS, Apc003OA, FG-3019, Apc003OA + BP and BP for 12 weeks, respectively. Data are expressed as mean ± standard deviation followed by one-way ANOVA with Tukey’s post hoc test, n = 5 per group. ∗ P < 0.05. ∗∗ P < 0.01. ∗∗∗ P < 0.001. ∗∗∗∗ P < 0.0001. The P values for the respective comparisons are indicated on the graph. Note: BP: Blocking peptide. WT: wild type (C57BL/10ScSn mice). Administration route: subcutaneous injection. Dosing frequency: once a week.

Article Snippet: Mice were randomly assigned to one of four experimental groups: (1) Vehicle Control Group: Received subcutaneous (s.c.) injections of the formulation buffer alone ( e.g., PBS or vehicle used for Apc003OA), once weekly for 12 weeks. (2) Apc003OA Treatment Group: Received s.c. injections of Apc003OA at a dose of 100 mg/kg body weight, dissolved in the appropriate vehicle, once weekly for 12 weeks. (3) FG-3019 Treatment Group: Received s.c. injections of the CTGF VWC-domain specific antibody FG-3019 (purchased from Med Chem Express) administered according to an equivalent schedule and dose regimen as a comparator. (4) Apc003OA + BP Pre-treatment Group: Received an s.c. injection of the blocking peptide (BP) designed against the CT-domain of CTGF binding site for Apc003 ( e.g ., at a dose and timepoint defined in pilot studies, such as 1 h prior to each Apc003OA dose), followed by s.c. injection of Apc003OA (100 mg/kg), once weekly for 12 weeks.

Techniques: Activity Assay, Muscles, Staining, Standard Deviation, Blocking Assay, Injection

Journal: Oncotarget

Article Title: Epithelial derived CTGF promotes breast tumor progression via inducing EMT and collagen I fibers deposition

doi:

Figure Lengend Snippet: Kaplan–Meier curves showing the overall survival and recurrence-free survival of patients with low or high tumor epithelial or stroma CTGF expression in the combined cohort of 103 breast cancer patients, P values were determined by log rank test.

Article Snippet: Overexpression of full-length CTGF gene was achieved by transfection of cDNA for CTGF (OriGene Technologies), which clones the open reading frame (ORF) of this gene into the pCMV6 Entry vector.

Techniques: Expressing

Journal: Oncotarget

Article Title: Epithelial derived CTGF promotes breast tumor progression via inducing EMT and collagen I fibers deposition

doi:

Figure Lengend Snippet: A–B. Quantitative graphs and representative CTGF immunohistochemistry staining of the breast tumor PDX tissue microarray. H score was calculated as described in Methods. P values were determined by student t -test. C. Representative images showing the Masson's trichrome staining and CTGF immunohistochemistry staining of the triple-negative (TNBC) and HER2-positive (HRE2+) breast tumor PDX tissue microarray. D. Correlation between the H scores of CTGF in tumor cells and % of stroma area in the TNBC tissue microarrays. R 2 = 0.66. E. Correlation between the H scores of CTGF in stromal cells and % of stroma area in the TNBC tissue microarrays. R 2 = 0.39. F. Correlation between the H scores of CTGF in stromal cells and % of stroma area in the HER2+ tissue microarrays. R 2 = 0.43.

Article Snippet: Overexpression of full-length CTGF gene was achieved by transfection of cDNA for CTGF (OriGene Technologies), which clones the open reading frame (ORF) of this gene into the pCMV6 Entry vector.

Techniques: Immunohistochemistry, Staining, Microarray

Journal: Oncotarget

Article Title: Epithelial derived CTGF promotes breast tumor progression via inducing EMT and collagen I fibers deposition

doi:

Figure Lengend Snippet: A–B. The co-expression analysis between CTGF and FN1, VIM, CDH1 in the n = 22 clinical breast tumor cohort . All gene probes were listed for FN1, VIM and CDH1 . The correlation co-efficient (cc) were calculated based on the log2 median-centered intensity of FN1, VIM, CDH1 and CTGF. C–D. The co-expression analysis between CTGF and FN1, VIM, CDH1 in the n = 66 clinical breast tumor cohort . E–H. CTGF gene expression in a series EMT-induced human mammary epithelial cell (HMLER) cell lines, including over-expression of EMT transcription factor Goosecoid (GSC), Snail, or Twist, and induction of EMT by TGFB1 transfection. P values were determined by student t -test. Fold change was log 2 scale.

Article Snippet: Overexpression of full-length CTGF gene was achieved by transfection of cDNA for CTGF (OriGene Technologies), which clones the open reading frame (ORF) of this gene into the pCMV6 Entry vector.

Techniques: Expressing, Gene Expression, Over Expression, Transfection

Journal: Oncotarget

Article Title: Epithelial derived CTGF promotes breast tumor progression via inducing EMT and collagen I fibers deposition

doi:

Figure Lengend Snippet: A. Representative phase-contrast images showing the elongated morphology of over-CTGF-HMLER cells which is typically associated with mesenchymal phenotype. B. RT-PCR analysis of the EMT genes and CTGF in the control and over-CTGF-HMLER cells. Relative gene expression level to GAPDH was plotted. C. Western blot analysis of the EMT markers and CTGF in the control and over-CTGF-HMLER cells. D–E. Quantitative graphs and representative membrane images showing the migration ability of the control and over-CTGF-HMLER cells measured by the Boyden Chamber assay. F. Quantitative graphs showing the invasion ability of the control and over-CTGF-HMLER cells measured by coating Matrigel in the Boyden Chamber assay. G–H. Enforced expression of CTGF in the HMLER cells promoted the number and size of mammospheres. I. Western blot analysis of enriched CD44 expression in the mammospheres. ** P < 0.01, vs control. P values were determined by student t -test.

Article Snippet: Overexpression of full-length CTGF gene was achieved by transfection of cDNA for CTGF (OriGene Technologies), which clones the open reading frame (ORF) of this gene into the pCMV6 Entry vector.

Techniques: Reverse Transcription Polymerase Chain Reaction, Control, Gene Expression, Western Blot, Membrane, Migration, Boyden Chamber Assay, Expressing

Journal: Oncotarget

Article Title: Epithelial derived CTGF promotes breast tumor progression via inducing EMT and collagen I fibers deposition

doi:

Figure Lengend Snippet: A. Representative phase-contrast images showing the epithelial morphology of the shCTGFs knockdown cells. B. Western blot analysis of the CTGF and N-cadherin, E-cadherin in the control and shCTGFs cells. C–D. Quantitative graphs and representative membrane images showing the migration ability of the control and shCTGFs cells measured by the Boyden Chamber assay. E. Quantitative graphs showing the invasion ability of the control and shCTGFs cells measured by coating Matrigel in the Boyden Chamber assay. F–G. CTGF depletion in the HMLER-snail cells reduced the number and size of mammospheres. H. Western blot analysis of CD44 in the mammospheres. * P < 0.05, vs control; ** P < 0.01, vs control. P values were determined by student t -test.

Article Snippet: Overexpression of full-length CTGF gene was achieved by transfection of cDNA for CTGF (OriGene Technologies), which clones the open reading frame (ORF) of this gene into the pCMV6 Entry vector.

Techniques: Knockdown, Western Blot, Control, Membrane, Migration, Boyden Chamber Assay

Journal: Oncotarget

Article Title: Epithelial derived CTGF promotes breast tumor progression via inducing EMT and collagen I fibers deposition

doi:

Figure Lengend Snippet: A–B. In vivo tumor growth of the over-CTGF-HMLER cells and control HMLER cells mixed with Matrigel (A) and PBS (B) C. Representative images showing the Masson's trichrome staining of the tumor connective tissue, and second harmonic generation (SHG) imaging of the intra-tumoral collagen I fibers, of the Matrigel-mixed over-CTGF-HMLER tumors. D–E. In vivo tumor growth of the shCTGF-HMLER cells (D), and representative Masson's trichrome staining and SHG imaging of the intra-tumoral collagen I fibers. F. Quantification of the percentage of collagen fiber and perpendicular collagen fiber to the overall connective tissue area. Mouse tumor sections (10 μm thick) were cut serially, and one section from every 100 μm was stained by Masson's trichrome staining, and the adjacent section was imaged by SHG imaging. In each section 5–10 fields were quantified. N = 4 mouse tumors in each group. * P < 0.05, vs control HMLER; ^ P < 0.05, vs HMLER-snail. P values were determined by student t -test.

Article Snippet: Overexpression of full-length CTGF gene was achieved by transfection of cDNA for CTGF (OriGene Technologies), which clones the open reading frame (ORF) of this gene into the pCMV6 Entry vector.

Techniques: In Vivo, Control, Staining, Imaging

Journal: Oncotarget

Article Title: Epithelial derived CTGF promotes breast tumor progression via inducing EMT and collagen I fibers deposition

doi:

Figure Lengend Snippet: A–B. A precedent treatment with anti-TNFR1 monoclonal antibody (10 μg/ml) more profoundly prevented the increase of CTGF-induced TNFR1 expression in the CTGF-highly expressed cells (over-CTGF-HMLER and HMLER-snail) than that in the CTGF-lowly expressed cells (control-HMLER and shCTGF2). C. Representative phase-contrast images showing the cell morphology changes in the indicated cell lines and treatments. D–E. The activation of IKBKA and IKBKB by CTGF could be specifically attenuated by the anti-TNFR1 antibody in the CTGF-high tumor cells. F. JUK but not ERK1/2 and p38 in the MAPK signaling pathway was up-regulated by exogenous CTGF in the CTGF-low tumor cells. G. Knockdown CTGF in the HMLER-snail cells down-regulated the IKBKA and IKBKB, and such down-regulation could be rescued by exogenous CTGF. All experimental data are representative of at least two independent experiments performed in triplicate. Data are expressed as mean ± s.e.m.

Article Snippet: Overexpression of full-length CTGF gene was achieved by transfection of cDNA for CTGF (OriGene Technologies), which clones the open reading frame (ORF) of this gene into the pCMV6 Entry vector.

Techniques: Expressing, Control, Activation Assay, Knockdown

Journal: Oncotarget

Article Title: Epithelial derived CTGF promotes breast tumor progression via inducing EMT and collagen I fibers deposition

doi:

Figure Lengend Snippet: A–B. Cell proliferation and mammosphere formation were significantly inhibited by the anti-TNFR1 treatment in over-CTGF-HMLER and HMLE-snail cells but not control-HMLER cells or shCTGF-HMLE-snail. C–D. Cell proliferation and mammosphere formation were significantly inhibited by resveratrol treatment in over-CTGF-HMLER and HMLE-snail cells but not control-HMLER cells or shCTGF-HMLE-snail. * P < 0.05, ** P < 0.01, vs anti-TNFR1(−) or resveratrol (−) group of the corresponding cell lines. P values were determined by student t -test.

Article Snippet: Overexpression of full-length CTGF gene was achieved by transfection of cDNA for CTGF (OriGene Technologies), which clones the open reading frame (ORF) of this gene into the pCMV6 Entry vector.

Techniques: Control

Journal: Oncotarget

Article Title: Epithelial derived CTGF promotes breast tumor progression via inducing EMT and collagen I fibers deposition

doi:

Figure Lengend Snippet: CTGF is mainly derived from epithelial tumor cells. Through the CTGF-TNFR1-IκB autocrine signaling, CTGF promotes tumor EMT which contributes to the stemness and metastasis. Through promoting the deposition and orientation of collagen I fibers in the primary tumor stroma, CTGF promotes stroma-tumor interaction which facilitates tumor progression.

Article Snippet: Overexpression of full-length CTGF gene was achieved by transfection of cDNA for CTGF (OriGene Technologies), which clones the open reading frame (ORF) of this gene into the pCMV6 Entry vector.

Techniques: Derivative Assay