Journal: Pharmaceutics

Article Title: Targeted Therapy of Hepatocellular Carcinoma Using Gemcitabine-Incorporated GPC3 Aptamer

doi: 10.3390/pharmaceutics12100985

Figure Lengend Snippet: In vitro affinity and specificity of G12msi aptamers. ( a ) The specific binding capacity of G12msi to HCC cells was assessed by using flow cytometry. GPC3-positive HepG2, Hep3B, and Huh7 cell lines, and GPC3-negative A431 cells were stained with Cy5-labeled G12msi or scrambled G12msi. ( b ) Confocal microscopy analysis with Cy5-labeled G12msi or scrambled G12msi aptamers (Red) on GPC3 positive HepG2, Hep3B, Huh7 cells, and GPC3 negative A431 cells at 4 °C. The nuclei were stained with 4′,6-diamidino-2-phenylindole (DAPI, blue). Scale bars, 20 μm.

Article Snippet: G12msi, scrambled G12msi, and Cy5-labeled G12msi aptamers were provided by Aptamer Sciences.

Techniques: In Vitro, Binding Assay, Flow Cytometry, Staining, Labeling, Confocal Microscopy

Journal: Pharmaceutics

Article Title: Targeted Therapy of Hepatocellular Carcinoma Using Gemcitabine-Incorporated GPC3 Aptamer

doi: 10.3390/pharmaceutics12100985

Figure Lengend Snippet: Internalization and intracellular localization of G12msi aptamers. ( a ) Representative z-stack images with orthogonal side-views of HepG2, Hep3B, Huh7, and A431 cells treated with Cy5-labeled G12msi or scrambled G12msi aptamers (red) at 37 °C. Nuclei were stained with DAPI (blue). ( b ) The cellular trafficking of G12ms and G12msi aptamers in HepG2 cells. Representative images showing the intracellular colocalization of the Cy5-labeled G12ms and G12msi aptamers (red) with a lysosomal marker (LysoTracker™ Green DND-26; green) by using confocal microscopy. The nuclei were counterstained with DAPI (blue). Scale bars, 20 μm.

Article Snippet: G12msi, scrambled G12msi, and Cy5-labeled G12msi aptamers were provided by Aptamer Sciences.

Techniques: Labeling, Staining, Marker, Confocal Microscopy

Journal: Advanced Healthcare Materials

Article Title: Urine‐Stable Aptamer‐Conjugated Gold Nanorods for the Early Detection of High‐Grade Bladder Cancer Residual Disease

doi: 10.1002/adhm.202403314

Figure Lengend Snippet: Features of the aptamer and tested bladder cancer cell lines. A) The secondary structure of H02 full‐length and Apt‐Itg (nucleotides from 18 to 51, boxed); structures of the H02 full‐length and Apt‐Itg were predicted by using Mfold web server, and the calculated change in Gibbs free energy (ΔG) values are reported. B) AlphaFold 3 molecular docking result of integrin α5β1‐Apt‐Itg binding complex; Apt‐Itg, integrin α5 and integrin β1 predicted structures are shown in yellow, light blue and red, respectively. The predicted interaction of Apt‐Itg with integrin α5, involving D154 and Y208 residues, is framed on the side. C) The expression of integrin α5β1 by human and murine bladder cancer cell lines evaluated by western blot analysis of the α5 and β1 chain of the integrin. The expression of the housekeeping protein α‐tubulin was used to estimate the relative fold of expression of integrin chains D). MB49‐Luc; murine bladder cancer cell line.

Article Snippet: The SH‐terminated 2′Fluoro‐Pyrimidine (2′F‐Py)‐RNA aptamer targeting integrin α5β1 (Apt‐Itg) and unrelated aptamer were purchased from Metabion (Martinsried, Germany).

Techniques: Binding Assay, Expressing, Western Blot

Journal: Advanced Healthcare Materials

Article Title: Urine‐Stable Aptamer‐Conjugated Gold Nanorods for the Early Detection of High‐Grade Bladder Cancer Residual Disease

doi: 10.1002/adhm.202403314

Figure Lengend Snippet: Binding assay of Alexa 647‐Apt‐Itg to human and murine bladder cancer cells using flow cytometry. A–C) Representative binding profiles of increasing concentrations of Alexa 647‐Apt‐Itg and Unrelated aptamer incubated with T24 (A), RT112 (B) and MB49‐Luc (C) cells. Apparent dissociation curves of aptamer‐cell interaction and Kd value of Apt‐Itg is shown. Data are presented as the mean ± SD. D) Competition binding assay by flow cytometry (left) and quantification of the gMFI (right) using 1 µ m Alexa 647‐Apt‐Itg in the absence (blue) or in the presence (red) of 30‐fold excess of unlabeled unrelated aptamer.

Article Snippet: The SH‐terminated 2′Fluoro‐Pyrimidine (2′F‐Py)‐RNA aptamer targeting integrin α5β1 (Apt‐Itg) and unrelated aptamer were purchased from Metabion (Martinsried, Germany).

Techniques: Binding Assay, Flow Cytometry, Incubation

Journal: Advanced Healthcare Materials

Article Title: Urine‐Stable Aptamer‐Conjugated Gold Nanorods for the Early Detection of High‐Grade Bladder Cancer Residual Disease

doi: 10.1002/adhm.202403314

Figure Lengend Snippet: Apt‐Itg actively internalizes into T24 bladder cancer cells. A) Representative confocal images of T24 cells incubated for indicated times at 4 °C or 37 °C with 4 µm Alexa 647‐Apt‐Itg or Alexa 647‐Unrelated aptamer as a negative control. Aptamer, WGA‐488 (cell surface), LysoTracker (lysosomes), and DAPI (nuclei) are visualized in red, green, blue, and gray, respectively. All digital images were captured at the same setting to allow direct comparison of staining patterns. Magnification 63x, 1.0x digital zoom, scale bar = 10 µm. B) Plot of MFI of Alexa 647‐Apt‐Itg normalized to cell number. Bars depict mean ± SD. *** p < 0.001.

Article Snippet: The SH‐terminated 2′Fluoro‐Pyrimidine (2′F‐Py)‐RNA aptamer targeting integrin α5β1 (Apt‐Itg) and unrelated aptamer were purchased from Metabion (Martinsried, Germany).

Techniques: Incubation, Negative Control, Comparison, Staining

Journal: Advanced Healthcare Materials

Article Title: Urine‐Stable Aptamer‐Conjugated Gold Nanorods for the Early Detection of High‐Grade Bladder Cancer Residual Disease

doi: 10.1002/adhm.202403314

Figure Lengend Snippet: Integrin α5 silencing results in reduced Apt‐Itg binding. A) T24, RT112, and MB49‐Luc cells were left untreated (NT) or transfected with the indicated Integrin α5 siRNAs or siRNA ctrl. At 24 h post‐transfection, cells were harvested, and cell lysates prepared and immunoblotted with anti‐integrin α5 or anti‐integrin β1 antibody. Equal loading was confirmed by immunoblot with anti‐α‐tubulin antibody. The histograms indicate the integrin chains/α‐tubulin ratio of the densitometric signals. Values are shown relative to siRNA ctrl, arbitrarily set to 1. B) Binding of 1 µ m Alexa647‐Apt‐Itg to human and murine bladder cancer cells following 24 h‐transfection with si‐ITGα5 (blue) and siRNA ctrl (red). C) Binding of Alexa488‐Ab‐Itg to T24 cells 24 h post‐transfection. (B, C) The histogram indicates gMFI of aptamer‐ or antibody‐treated cells normalized to the gMFI of siRNA ctrl untreated cells (black), arbitrarily set to 1. Bars depict mean ± SD of two independent experiments. * p < 0.005, ** p < 0.001, and *** p < 0.0001.

Article Snippet: The SH‐terminated 2′Fluoro‐Pyrimidine (2′F‐Py)‐RNA aptamer targeting integrin α5β1 (Apt‐Itg) and unrelated aptamer were purchased from Metabion (Martinsried, Germany).

Techniques: Binding Assay, Transfection, Western Blot

Journal: Advanced Healthcare Materials

Article Title: Urine‐Stable Aptamer‐Conjugated Gold Nanorods for the Early Detection of High‐Grade Bladder Cancer Residual Disease

doi: 10.1002/adhm.202403314

Figure Lengend Snippet: Characterization of the final nanosystem, GNRs@Chit‐Apt‐Itg. A) Scheme representative of the aptamer Apt‐Itg conjugated to the GNRs@Chit via PEG 12 linker. B) TEM images of nanoparticles. C) Left , representative PCR calibration curve with unconjugated Apt‐Itg (red); samples from TOP, 1/2 and 1/10 GNRs@Chit‐Apt‐Itg preparations are shown in blue, green, and black, respectively. Right , quantification of the Apt‐Itg and unrelated aptamer loaded onto GNRs@Chitosan; data shown mean ± SD of three independent experiments. D) UV–vis spectra of GNRs@CTAB, GNRs@Chit, and GNRs@Chit‐Apt‐Itg/SCR showing no significant differences in the absorption spectrum of GNRs after surface modification; FAAS analysis of GNRs@Chit‐Apt‐Itg with calibration line; thermogravimetric analysis of GNRs@Chit in N 2 and air atmosphere (above 600 °C). GNRs@Chit‐Apt‐Itg stability in human urine was investigated by E) VIS‐NIR spectra of GNRs@Chit‐Apt‐Itg compared to the VIS‐NIR spectra of GNRs@CTAB and GNRs@Chit, and F) VIS‐NIR spectra and absorption intensity of GNRs@Chit‐Apt‐Itg in the presence of human urine over time. G) Binding of GNRs@Chit‐Apt‐Itg to MB49 cells after 15 min of incubation, followed by cell internalization of GNRs and accumulation into lysosome after 4 h. SCR: unrelated aptamer.

Article Snippet: The SH‐terminated 2′Fluoro‐Pyrimidine (2′F‐Py)‐RNA aptamer targeting integrin α5β1 (Apt‐Itg) and unrelated aptamer were purchased from Metabion (Martinsried, Germany).

Techniques: Modification, Binding Assay, Incubation

Journal: Cancers

Article Title: Are Integrins Still Practicable Targets for Anti-Cancer Therapy?

doi: 10.3390/cancers11070978

Figure Lengend Snippet: Selected clinical trials of agents targeting integrins. Non-exhaustive listing of the recent most important clinical studies with integrin inhibitors and their salient features and results).

Article Snippet: Volociximab (MEDI-522, anti α5β1 integrin antibody) (AbbVie): total trials 7 (+ 3 discontinued early) , , , , , , .

Techniques: Clinical Proteomics, Control, Activity Assay, Comparison