Journal: bioRxiv

Article Title: Development of Integrin α5β1-targeted PET/NIR imaging probes for glioblastoma intraoperative navigation and intracavity targeted radionuclide therapy

doi: 10.64898/2026.01.09.698741

Figure Lengend Snippet: (A) Strategies for modification of GS, including amino acid mutation, covalent modification, cyclization, and multivalency. (B) SPR sensorgrams demonstrating the binding affinity of GS and GR for human integrin α5β1 in a concentration-dependent manner. The equilibrium dissociation constant (K D ) of each peptide was calculated based on SPR measurements. The K D values of each precursor are shown. (C) Molecular docking of GR and GS binding integrin a5b1 protein (grey; PDB: 7NWL) showing the selected possible ligation residues. (D-F) Analysis of integrin α5β1 expression in U87MG cells and tumor tissues by western blot ( D ), and immunohistochemistry ( E-F ) analysis. The band for integrin α5 was approximately 150 kDa. M, marker. C, cell. T, tumor. For immunofluorescence images, green is for integrin α5, red for integrin β1, and blue for nucleus. Scale bar, 50□μm ( E-F ). (G) In vitro cellular uptake of [ 68 Ga]GS and [ 68 Ga]GR in U87MG cell lines. All results are expressed as means ± SEM, as indicated in at least three independent experiments. “*” represents differences compared with the [ 68 Ga]GS. * p < 0.05.

Article Snippet: Recombinant human integrin α5β1 (alpha 5 beta 1, HY-P77718, MCE, NJ, USA) was immobilized on a CM5 sensor chip at 25 □ following a standard amine coupling kit.

Techniques: Modification, Mutagenesis, Binding Assay, Concentration Assay, Ligation, Expressing, Western Blot, Immunohistochemistry, Marker, Immunofluorescence, In Vitro

Journal: bioRxiv

Article Title: Development of Integrin α5β1-targeted PET/NIR imaging probes for glioblastoma intraoperative navigation and intracavity targeted radionuclide therapy

doi: 10.64898/2026.01.09.698741

Figure Lengend Snippet: (A) Immunofluorescence images show the expression of integrin α5β1, GFAP, and NEUN of U87MG tumors in orthotopic glioblastoma tumor-bearing mice. Green indicates integrin α5, Red for NEUN, yellow for GFAP, and blue for nucleus. Scale bar, 1 mm. (B-C) Representative fluorescence imaging of mice bearing in orthotopic U87MG tumors ( B ) and ex vivo brains ( C ). Cy5-GS and Cy5-GR were intravenously injected with a dose of 5 mg/kg. (D) Quantification of fluorescence intensity in tumors corresponding to ( C ). (E) Immunohistochemical analysis of integrin α5β1 expression in brain tissue sections from orthotopic glioblastoma tumor-bearing mice. Scale bar, 1 mm. (F-G) Immunofluorescence images of brain tissue sections from orthotopic glioblastoma tumor-bearing mice treated with Cy5-GS ( F ) and Cy5-GR ( G ). Green indicates integrin α5, Red for GS or GR, and blue for nucleus. Scale bar, 1 mm. (H-I) Magnific imaging of brain tissue sections from orthotopic glioblastoma tumor-bearing mice treated with GS-Cy5. Scale bar, 50 μm. All results are expressed as means ± SEM, as indicated in at least three independent experiments. A multiple t-test was used when two groups were compared. The symbol “*” represents differences compared with the Cy5-GS. *** p < 0.001.

Article Snippet: Recombinant human integrin α5β1 (alpha 5 beta 1, HY-P77718, MCE, NJ, USA) was immobilized on a CM5 sensor chip at 25 □ following a standard amine coupling kit.

Techniques: Immunofluorescence, Expressing, Fluorescence, Imaging, Ex Vivo, Injection, Immunohistochemical staining

Journal: PLoS ONE

Article Title: An innovative strategy to identify new targets for delivering antibodies to the brain has led to the exploration of the integrin family

doi: 10.1371/journal.pone.0274667

Figure Lengend Snippet: Anti-integrin antibodies apparent affinities to different integrin subunits investigated by ELISA.

Article Snippet: Recombinant integrin proteins were purchased from OriGene for human monomer α3, α5 and β1 (tp320975, tp301151 and tp303818), from GeneTex for human monomer α4 (GTX48181), from R&D Systems for human and mouse dimer α3β1, α4β1 and α5β1 (2840-a3, 3230-a5, 5668-a4, 7728-a5, 9374-a3) for human ALCAM 656-AL, from Sino Biological for human α5β1 (CT-014-H2508H), from Abcam for Striatin3 (ab162295) Antibodies were purchased from antibodies-online (natalizumab ABIN5668196), from Abcam (Anti-VE Cadherin ab33168), from BD Biosciences (553715), from BioLegend (343802 and MFR5 103801), from Interchim (DCABH-8217), from Invitrogen (14-0299-82, MA5-17103, MA1-25298), from Millipore (MABT409, MABT199, MAB2079Z) from Novus Biological (NBP2-52708), from Proteintech (66070-1-Ig), from R&DSystems (MAB1345), from Sigma-Aldrich (MAB1965), from ThermoFisher Scientific (Anti-ZO-1 #61–7300; Anti-Occludin #33–1500; all other antibodies were produced in-house by Sanofi Biological Research. hCMEC/D3 cells were obtained from Cedarlane.

Techniques: Enzyme-linked Immunosorbent Assay

Journal: The Journal of biological chemistry

Article Title: The cell surface hyaluronidase TMEM2 regulates cell adhesion and migration via degradation of hyaluronan at focal adhesion sites.

doi: 10.1016/j.jbc.2021.100481

Figure Lengend Snippet: Figure 5. Association of transmembrane protein 2 (TMEM2) with integrins via interactions between the extracellular domains. A and B, targeting of TMEM2 to focal adhesions (FAs) does not require the cyto- plasmic domain of TMEM2. In this experiment, mCherry-mTMEM2 (full length) and mCherry-mTMEM2/Δcyto (Δcyto) cells were analyzed for their in situ hyaluronan (HA) degradation activities. To allow specific analysis of the activity of the full-length mouse TMEM2 and its Δcyto deletion mutant, expression of endogenous human TMEM2 was silenced by siRNA treatment prior to the assay. A, in situ HA degradation assays were performed on substrate immobilized with FA-HA, as described in Experimental procedures section. Note that the pattern of in situ HA degradation is indistinguishable between mCherry-mTMEM2/Δcyto and mCherry-mTMEM2 cells. The scale bar represents 10 μm. B, immunostaining for vinculin in mCherry-mTMEM2 and mCherry-mTMEM2/Δcyto cells on the FA-HA substrate. Note that the sites of HA degradation colocalize with vinculin-immunoreactive puncta in both mCherry-mTMEM2/Δcyto and mCherry-mTMEM2 cells. The scale bar represents 2 μm. C–E, TMEM2 associates with integrins via extracellular interactions. C, cell surface–expressed TMEM2 is coimmunoprecipitated with integrin α5β1. mCherry-mTMEM2 cells were treated with the membrane-impermeable crosslinker 3’,3’-dithiobis(sulfosuccinimidyl

Article Snippet: Two micrograms of recombinant heterodimer of integrin ECD (α5β1; R&D Systems: 3230-A5-050; αLβ2; R&D Systems: 3868-AV-050) were applied to the TMEM2–ECD–bound and control unbound resin and incubated in HBSS++ overnight at 4 C. After extensive washing, bound materials were eluted by boiling in SDS-PAGE sample buffer, and eluents were analyzed by SDSPAGE and immunoblotting with rabbit polyclonal antiintegrin α5 (Proteintech; 10569-1-AP), rabbit monoclonal anti-integrin β1 (Abcam; ab52971), rabbit polyclonal antiintegrin β2 (Proteintech; 10544-1-AP), or mouse monoclonal anti-polyhistidine (Sigma; A7058; clone: HIS-1, peroxidase conjugated).

Techniques: In Situ, Activity Assay, Mutagenesis, Expressing, Immunostaining, Membrane

Journal: The Journal of biological chemistry

Article Title: The cell surface hyaluronidase TMEM2 regulates cell adhesion and migration via degradation of hyaluronan at focal adhesion sites.

doi: 10.1016/j.jbc.2021.100481

Figure Lengend Snippet: Figure 6. A model for the role of transmembrane protein 2 (TMEM2) in integrin-mediated cell adhesion and migration. Our results suggest that TMEM2-dependent degradation of hyaluronan (HA) is critical for cells to form strong cell–matrix adhesion on HA-rich extracellular matrix (ECM). A, high levels of HA in the ECM are inhibitory to the direct engagement of integrins to their ECM ligands. B, in the presence of TMEM2, HA in the ECM is locally removed, which generates a microenvironment that is permissible to the direct integrin–ECM engagement. C, the association between TMEM2 and integrins promotes the FA formation and maturation via further removal of HA in the vicinity of the integrin–ECM engagement. D, this in turn facilitates integrin clustering, integrin-mediated downstream signaling, and cellular responses. See the text for further discussion.

Article Snippet: Two micrograms of recombinant heterodimer of integrin ECD (α5β1; R&D Systems: 3230-A5-050; αLβ2; R&D Systems: 3868-AV-050) were applied to the TMEM2–ECD–bound and control unbound resin and incubated in HBSS++ overnight at 4 C. After extensive washing, bound materials were eluted by boiling in SDS-PAGE sample buffer, and eluents were analyzed by SDSPAGE and immunoblotting with rabbit polyclonal antiintegrin α5 (Proteintech; 10569-1-AP), rabbit monoclonal anti-integrin β1 (Abcam; ab52971), rabbit polyclonal antiintegrin β2 (Proteintech; 10544-1-AP), or mouse monoclonal anti-polyhistidine (Sigma; A7058; clone: HIS-1, peroxidase conjugated).

Techniques: Migration

Journal: Cellular and molecular life sciences : CMLS

Article Title: Low density lipoprotein receptor-related protein 1 couples β1 integrin activation to degradation.

doi: 10.1007/s00018-017-2707-6

Figure Lengend Snippet: Fig. 7 Regulation of β1 integrin activation, subcellular traffick- ing and degradation by LRP1. a Inactive α5β1 integrin is present in the complex with LRP1 on the cell surface. Upon cell contact with fibronectin, kindlin2 is sequentially recruited to LRP1 (b) and to β1 integrin (c) which results in integrin activation. d Following endocy- tosis, LRP1 employs myosins to direct ubiquitinated integrins toward lysosomal and proteasomal degradation. The molecular mechanism by which PKC mediates LRP1-induced activation and subsequent trafficking of α5β1 integrin and whether LRP1 remains bound to α5β1 integrin during endocytosis, transport and degradation are cur- rently unknown

Article Snippet: To this end, cells were transfected with GFP-kindlin2-encoding plasmid [3] and lysed after 24 h. A 12-fold titration series of purified recombinant α5β1 integrin (1750 nM to 0.854 nM, R&D Systems) diluted 1:1 with PBS containing 0.05% Tween-20 was prepared, while the lysate concentration was kept constant at 7.25 mg/ ml.

Techniques: Activation Assay

Journal: Cell Adhesion & Migration

Article Title: F4, a collagen XIX-derived peptide, inhibits tumor angiogenesis through αvβ3 and α5β1 integrin interaction

doi: 10.1080/19336918.2021.1951425

Figure Lengend Snippet: F4 binds to αvβ3 and α5β1 integrins on HUVEC . (a) HUVECs were analyzed by flow cytometry for the expression of αvβ3 and α5β1 integrins. (b) HUVEC extracts were submitted to F4 affinity chromatography. Bound proteins were eluted with increasing concentrations of NaCl (0.15, 0.6 and 1 M) and the elution profile was checked by recording the absorbance at 280 nm. (c) Eluted samples were then analyzed by SDS-PAGE and western blot HUVEC total extracts and recombinant αvβ3 and α5β1 integrins were used as positive controls. The 0.6 M eluted sample revealed bands which matched the molecular weight of the α5, β1, αv and β3 recombinant integrin subunits. The experiment was repeated twice (N = 2)

Article Snippet: HUVEC extracts and recombinant human αvβ3 and α5β1 integrins (3230-A5-050 and 3050-AV-050, R&D systems) were used as positive controls.

Techniques: Flow Cytometry, Expressing, Affinity Chromatography, SDS Page, Western Blot, Recombinant, Molecular Weight

Journal: Cell Adhesion & Migration

Article Title: F4, a collagen XIX-derived peptide, inhibits tumor angiogenesis through αvβ3 and α5β1 integrin interaction

doi: 10.1080/19336918.2021.1951425

Figure Lengend Snippet: F4 directly binds to αvβ3 and α5β1 integrins . Direct interaction between αvβ3 or α5β1 integrin and F4 was studied using solid phase assays as described in the material and method section. The F4 peptide was biotinylated to allow its detection using the streptavidin-peroxidase complex. Different conditions were used. (a-b) The amounts of coated αvβ3 (a) or α5β1 integrin (b) were increased while biotinylated F4 peptide was kept constant. (c-d) The concentration of biotinylated F4 peptide was increased while coated amounts of αvβ3 (c) or α5β1 (d) integrins were kept constant. (e-f) Competitive assays were performed using increasing concentrations of F4 peptide while biotinylated-F4 peptide and αvβ3 (e) or α5β1 (f) integrins were kept constant. The solid phase assays were repeated twice (N = 2; n = 2)

Article Snippet: HUVEC extracts and recombinant human αvβ3 and α5β1 integrins (3230-A5-050 and 3050-AV-050, R&D systems) were used as positive controls.

Techniques: Concentration Assay

Journal: Signal Transduction and Targeted Therapy

Article Title: Targeting integrin pathways: mechanisms and advances in therapy

doi: 10.1038/s41392-022-01259-6

Figure Lengend Snippet: Integrin-targeting therapies in clinical trials

Article Snippet: Pegylated recombinant human endostatin , NCT01527864 , Protgen Ltd , Peptide , 2012-02-07 , α5β1 , Non-small cell lung cancer , 10 mg/m 2 QW , IV , Phase II.

Techniques: Recombinant, Methylation, Injection, Imaging, Biomarker Discovery, Cream, Ointment, Synthesized