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: Investigative Ophthalmology & Visual Science

Article Title: Hepatocyte Growth Factor Confers Trabecular Meshwork Stem Cell Resilience and Paracrine Protection of Trabecular Meshwork Cells in Glaucoma

doi: 10.1167/iovs.67.4.1

Figure Lengend Snippet: TMSCs exhibit attenuated fibrotic and oxidative stress responses compared with TM cells under TGF-β2 stimulation, and NAC mitigates these effects. ( A , B ) MTT assays in TMSCs ( A ) and TM cells ( B ) used to determine the NAC concentration applied in subsequent experiments. ( C–E ) Immunofluorescent staining of CTGF ( C ), α-SMA ( D ), and FN ( E ) in TMSCs and TM cells after a 5-day treatment with control, TGF-β2 (3 ng/mL), or TGF-β2 combined with NAC (40 µM). ( F–H ) Quantification of fluorescence intensities for CTGF ( F ), α-SMA ( G ), and FN ( H ). ( I ) Western blotting analyses of CTGF, α-SMA, and FN expression in TMSCs and TM cells across the same treatment groups. ( J–L ) Densitometric quantification of CTGF ( J ), α-SMA ( K ), and FN ( L ) protein levels. ( M–O ) Measurements of total ROS levels ( M ), lipid peroxidation assessed by C11-BODIPY ( N ), and mitochondrial membrane potential measured by TMRM ( O ) in TMSCs and TM cells across treatment groups. Data are presented as mean ± SD ( n ≥ 3). Statistical analyses were conducted using two-way ANOVA, followed by Tukey's multiple comparisons test. * P < 0.05; ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns, not significant.

Article Snippet: After blocking with 1% bovine serum albumin (MilliporeSigma) for 30 minutes, samples were incubated overnight at 4°C with primary antibodies against CHI3L1 (R&D Systems, 1:100), MYOC (a gift from Dr. Stamer, 1:300), OCT4 (Cell Signaling Technology, 1:200), Ki67 (Abcam, 1:200), 8F1-3 (a kind gift from Dr. Nirmala SundarRaj, University of Pittsburgh, 1:200), CTGF (Santa Cruz Biotechnology, 1:200), α-SMA (R&D Systems, 1:200), and FN (Abcam, 1:200).

Techniques: Concentration Assay, Staining, Control, Fluorescence, Western Blot, Expressing, Membrane

Journal: Investigative Ophthalmology & Visual Science

Article Title: Hepatocyte Growth Factor Confers Trabecular Meshwork Stem Cell Resilience and Paracrine Protection of Trabecular Meshwork Cells in Glaucoma

doi: 10.1167/iovs.67.4.1

Figure Lengend Snippet: HGF and TMSC-derived secretome alleviate TGF-β2–induced fibrotic and oxidative stress responses in TM cells. ( A , B ) MTT assays in TM cells assessing the effects of HGF ( A ) and HGFI ( B ) on cell viability. ( C ) Bright-field images showing TM cell morphology after 5-day treatment with control, TGF-β2 (3 ng/mL), HGF (10 ng/mL), HGFI (100 nM), TMSC-derived secretome (TMSC-Scr), or TMSC-Scr combined with HGFI. ( D ) Immunofluorescent staining of CTGF, α-SMA, and FN. ( E–G ) Quantification of fluorescence intensities for CTGF ( E ), α-SMA ( F ), and FN ( G ). ( H ) Western blotting analyses of CTGF, α-SMA, and FN expression under the same treatment conditions. ( I–K ) Densitometric quantification of CTGF ( I ), α-SMA ( J ), and FN ( K ) protein levels. ( L–N ) Oxidative stress analyses including total ROS levels ( L ), lipid peroxidation measured by C11-BODIPY ( M ), and mitochondrial membrane potential assessed by TMRM ( N ). Data are presented as mean ± SD ( n ≥ 3). Statistical analyses were performed using one-way ANOVA, followed by Tukey's multiple comparisons test. * P < 0.05, ** P < 0.01, *** P < 0.001, **** P < 0.0001; ns, not significant.

Article Snippet: After blocking with 1% bovine serum albumin (MilliporeSigma) for 30 minutes, samples were incubated overnight at 4°C with primary antibodies against CHI3L1 (R&D Systems, 1:100), MYOC (a gift from Dr. Stamer, 1:300), OCT4 (Cell Signaling Technology, 1:200), Ki67 (Abcam, 1:200), 8F1-3 (a kind gift from Dr. Nirmala SundarRaj, University of Pittsburgh, 1:200), CTGF (Santa Cruz Biotechnology, 1:200), α-SMA (R&D Systems, 1:200), and FN (Abcam, 1:200).

Techniques: Derivative Assay, Control, Staining, Fluorescence, Western Blot, Expressing, Membrane

Journal: Investigative Ophthalmology & Visual Science

Article Title: Hepatocyte Growth Factor Confers Trabecular Meshwork Stem Cell Resilience and Paracrine Protection of Trabecular Meshwork Cells in Glaucoma

doi: 10.1167/iovs.67.4.1

Figure Lengend Snippet: Proposed mechanism by which TMSCs and TMSC-derived factors protect TM cells from TGF-β2–induced injury. This schematic summarizes the integrated model derived from our experimental findings. In TM cells, TGF-β2 activates oxidative stress pathways, leading to increased ROS production, lipid peroxidation, mitochondrial depolarization, and induction of profibrotic markers, including CTGF, α-SMA, and FN, ultimately promoting cytoskeletal remodeling and extracellular matrix accumulation, thereby increasing aqueous humor outflow resistance and elevating IOP. TMSCs exhibit intrinsic resistance to these deleterious pathways, maintaining low oxidative burden, preserved mitochondrial membrane potential, and attenuated fibrotic activation. The TMSC-Scr, enriched in HGF and additional trophic factors, reduces ROS and lipid peroxidation, stabilizes mitochondrial function, and suppresses α-SMA and FN expression while partially modulating CTGF.

Article Snippet: After blocking with 1% bovine serum albumin (MilliporeSigma) for 30 minutes, samples were incubated overnight at 4°C with primary antibodies against CHI3L1 (R&D Systems, 1:100), MYOC (a gift from Dr. Stamer, 1:300), OCT4 (Cell Signaling Technology, 1:200), Ki67 (Abcam, 1:200), 8F1-3 (a kind gift from Dr. Nirmala SundarRaj, University of Pittsburgh, 1:200), CTGF (Santa Cruz Biotechnology, 1:200), α-SMA (R&D Systems, 1:200), and FN (Abcam, 1:200).

Techniques: Derivative Assay, Membrane, Activation Assay, Expressing