Human ITGAV/ITGB3 Stable Cell Line - CHO-K1

Researchers performed a CRISPR screen in multiple cancer models using human cell surface proteome and integrin family libraries and identified ITGAV (integrin αV) and its heterodimeric partner ITGB5 (integrin β5) as cancer cell-expanded Key integrin pairs. High-density CRISPR gene splicing further pinpointed key pockets within the ITGAV β-propeller domain necessary for integrin αVβ5 dimerization. Combined with computer simulation compound docking, the researchers developed the CRISPR-Tiling-Instructed Computer-Aided (CRISPR-TICA) drug discovery process and identified Cpd_AV2 as a candidate inhibitor targeting the central pocket of the ITGAV β-propeller. Cpd_AV2 treatment rapidly leads to integrin αVβ5 uncoupling and apoptosis, providing a novel therapeutic mechanism to eliminate integrin signaling through heterodimer dissociation. Researchers foresee that the CRISPR-TICA method will become a powerful tool for future drug discovery research.

Figure 1. ITGAV supports cancer cell expansion through RAC1. Gene ranking, RNA sequencing, Western blot, and growth competition assays showed that inhibition of RAC1 or ITGAV resulted in reduced apoptosis and proliferation. (Mattson, N.M., et al., 2024)

Researchers can verify the above experimental results using Creative Biogene's Human ITGAV/ITGB3 Stable Cell Line - CHO cell line. Although the original experiment focused on the ITGAV/ITGB5 heterodimer, ITGAV/ITGB3 (integrin αV/β3) also belongs to the integrin family and has a similar structure and function. Using the ITGAV/ITGB3 cell line can help confirm the uncoupling effect of Cpd_AV2 on integrin αV heterodimer and its resulting apoptotic effect. Specific verification methods may include:

1. Integrin αVβ3 dimerization detection: Detect the uncoupling of integrin αVβ3 after Cpd_AV2 treatment through co-immunoprecipitation or fluorescence co-localization experiments.

2. Cell apoptosis detection: Use methods such as flow cytometry or TUNEL staining to detect cell apoptosis levels and evaluate the impact of Cpd_AV2 on cell survival.

3. Signaling pathway analysis: Detect the phosphorylation status of downstream signaling molecules such as FAK and Akt to evaluate changes in the integrin signaling pathway.

Through these experiments, the researchers can verify whether Cpd_AV2 has an effect on ITGAV/ITGB5 similar to that observed in the original experiment, while also further understanding its general effect on different integrin heterodimers.