Feasibility of Ex Vivo Ligandomics
We developed ligandomics for the in vivo profiling of vascular ligands in mice, discovering secretogranin III (Scg3) as a novel angiogenic factor that selectively binds to retinal vessels of diabetic but not healthy mice. This discovery led to the development of anti-Scg3 therapy for ocular vasculop...
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| Format: | Article |
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MDPI AG
2025-01-01
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| Series: | Biomolecules |
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| Online Access: | https://www.mdpi.com/2218-273X/15/1/145 |
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| author | Prabuddha Waduge Remya Ammassam Veettil Bojun Zhang Chengchi Huang Hong Tian Wei Li |
| author_facet | Prabuddha Waduge Remya Ammassam Veettil Bojun Zhang Chengchi Huang Hong Tian Wei Li |
| author_sort | Prabuddha Waduge |
| collection | DOAJ |
| description | We developed ligandomics for the in vivo profiling of vascular ligands in mice, discovering secretogranin III (Scg3) as a novel angiogenic factor that selectively binds to retinal vessels of diabetic but not healthy mice. This discovery led to the development of anti-Scg3 therapy for ocular vasculopathies. However, in vivo ligandomics requires intracardial perfusion to remove unbound phage clones, limiting its use to vascular endothelial cells (ECs). To extend ligandomics to non-vascular cells, we investigated ex vivo ligandomics. We isolated ECs and retinal ganglion cells (RGCs) from diabetic and healthy mouse retinas by immunopanning. We quantified the binding of clonal phages displaying Scg3 and vascular endothelial growth factor (VEGF), confirming that their binding patterns to isolated diabetic versus healthy ECs matched in vivo patterns. Additionally, Scg3 and VEGF binding to isolated RGCs reflected their in vivo activity. These results support the feasibility of ex vivo ligandomics. We further mapped ligands binding to immunopanned diabetic and healthy ECs and RGCs by ligandomics, confirming that Scg3 was enriched with selective binding to diabetic ECs but not healthy ECs or diabetic/healthy RGCs. These findings demonstrate the feasibility of ex vivo ligandomics, which can be broadly applied to various cell types, tissues, diseases, and species. |
| format | Article |
| id | doaj-art-24309ea1bd89451388bbce62bfcd8593 |
| institution | Kabale University |
| issn | 2218-273X |
| language | English |
| publishDate | 2025-01-01 |
| publisher | MDPI AG |
| record_format | Article |
| series | Biomolecules |
| spelling | doaj-art-24309ea1bd89451388bbce62bfcd85932025-01-24T13:25:21ZengMDPI AGBiomolecules2218-273X2025-01-0115114510.3390/biom15010145Feasibility of Ex Vivo LigandomicsPrabuddha Waduge0Remya Ammassam Veettil1Bojun Zhang2Chengchi Huang3Hong Tian4Wei Li5Cullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, TX 77030, USACullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, TX 77030, USACullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, TX 77030, USACullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, TX 77030, USALigandomicsRx, LLC, Houston, TX 77098, USACullen Eye Institute, Department of Ophthalmology, Baylor College of Medicine, Houston, TX 77030, USAWe developed ligandomics for the in vivo profiling of vascular ligands in mice, discovering secretogranin III (Scg3) as a novel angiogenic factor that selectively binds to retinal vessels of diabetic but not healthy mice. This discovery led to the development of anti-Scg3 therapy for ocular vasculopathies. However, in vivo ligandomics requires intracardial perfusion to remove unbound phage clones, limiting its use to vascular endothelial cells (ECs). To extend ligandomics to non-vascular cells, we investigated ex vivo ligandomics. We isolated ECs and retinal ganglion cells (RGCs) from diabetic and healthy mouse retinas by immunopanning. We quantified the binding of clonal phages displaying Scg3 and vascular endothelial growth factor (VEGF), confirming that their binding patterns to isolated diabetic versus healthy ECs matched in vivo patterns. Additionally, Scg3 and VEGF binding to isolated RGCs reflected their in vivo activity. These results support the feasibility of ex vivo ligandomics. We further mapped ligands binding to immunopanned diabetic and healthy ECs and RGCs by ligandomics, confirming that Scg3 was enriched with selective binding to diabetic ECs but not healthy ECs or diabetic/healthy RGCs. These findings demonstrate the feasibility of ex vivo ligandomics, which can be broadly applied to various cell types, tissues, diseases, and species.https://www.mdpi.com/2218-273X/15/1/145ligandomicsex vivo ligandomicsin vivo ligandomicsScg3VEGFex vivo ligand binding assay |
| spellingShingle | Prabuddha Waduge Remya Ammassam Veettil Bojun Zhang Chengchi Huang Hong Tian Wei Li Feasibility of Ex Vivo Ligandomics Biomolecules ligandomics ex vivo ligandomics in vivo ligandomics Scg3 VEGF ex vivo ligand binding assay |
| title | Feasibility of Ex Vivo Ligandomics |
| title_full | Feasibility of Ex Vivo Ligandomics |
| title_fullStr | Feasibility of Ex Vivo Ligandomics |
| title_full_unstemmed | Feasibility of Ex Vivo Ligandomics |
| title_short | Feasibility of Ex Vivo Ligandomics |
| title_sort | feasibility of ex vivo ligandomics |
| topic | ligandomics ex vivo ligandomics in vivo ligandomics Scg3 VEGF ex vivo ligand binding assay |
| url | https://www.mdpi.com/2218-273X/15/1/145 |
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