## 3 Results
### PPI and Pathway Analysis  
To investigate ADGRG3's role in cancer and immune pathways, performed PPI analysis using STRING (Figure 10A) and supplemented it with data from the Human Protein Atlas, gathering 29 interacting genes (Supplementary File S1: Information on PPI genes). The gene enrichment analysis via GO, KEGG, and Reactome identified key pathways that may link ADGRG3 to cancer and immune functions. The Reactome analysis (Figure 10B) highlighted pathways such as GPCR ligand binding and G-alpha (i) signaling, involving proteins such as GNB1, GNG2, CXCR1, and FPR2, which are central to cell signaling in cancer and immune regulation. Additionally, neutrophil degranulation and Ca\({}^{2+}\) signaling pathways suggest ADGRG3's involvement in innate immune responses and inflammation, which are processes critical to both cancer \begin{table}
\begin{tabular}{l c c c c c c c c} \hline \hline
**Complex** & **AVdwaals** & **\(\Delta\)EEL** & **\(\Delta\)EPB** & **\(\Delta\)ENPOLAR** & **\(\Delta\)EDISPER** & **AGGas** & **AGSolv** & **\(\Delta\)**GTotal** \\  & **(kcal/mol)** & **(kcal/mol)** & **(kcal/mol)** & **(kcal/mol)** & **(kcal/mol)** & **(kcal/mol)** & **(kcal/mol)** & **(kcal/mol)** \\ \hline
**ADGRG3–** & \(-52.8947\) & \(-14.1053\) & \(35\) & \(-9\) & \(0.9\) & \(-67\) & \(26.9\) & \(-40.1\) \\
**Ginsenoside Rg3** & \(-43.3333\) & \(-8.66667\) & \(20\) & \(-5\) & \(0.5\) & \(-52\) & \(15.5\) & \(-36.5\) \\
**ADGRG3–** & \(-60.0481\) & \(-17.4519\) & \(52.7\) & \(-12\) & \(1.2\) & \(-77.5\) & \(41.9\) & \(-35.6\) \\
**Ginsenoside F5** & \(-46.2609\) & \(-9.73913\) & \(25\) & \(-6\) & \(0.6\) & \(-56\) & \(19.6\) & \(-36.4\) \\
**ADGRG3–** & \(-58.0645\) & \(-13.9355\) & \(40\) & \(-8\) & \(1\) & \(-72\) & \(33\) & \(-39\) \\
**Ginsenoside F1** & \(-45.6522\) & \(-10.3478\) & \(21.5\) & \(-6\) & \(0.6\) & \(-56\) & \(16.1\) & \(-39.9\) \\
**ADGRG3–** & \(-45.8333\) & \(-9.16667\) & \(20\) & \(-5\) & \(0.5\) & \(-55\) & \(15.5\) & \(-39.5\) \\ \hline
**Abbreviation:** & \(\Delta\)Vdwaals = van der Waals energy; \(\Delta\)EEL = electrostatic molecular energy; \(\Delta\)EPB = polar contribution to the solvation energy; \(\Delta\)ENPOLAR = nonpolar contribution of repulsive solute–solvent interactions to the solvation energy; \(\Delta\)EDISPER = nonpolar contribution of attractive solute–solvent interactions to the solvation energy; \(\Delta\)GGas = total gas-phase molecular energy; \(\Delta\)GSolv = total solvation energy; and \(\Delta\)GGTotal = total binding energy. \\ \hline \hline \end{tabular}
\end{table} 
Table 2: Summarized data of the MMPBSA-based free energy calculation of ligand–receptor interactions. Figure 10: (**A**) PPI network; (**B**) Reactome enrichment analysis. progression and immune activity. These findings predict that ADGRG3 could possibly influence cancer and immune pathways through GPCR and cytokine signaling. Figure 11 presents the top 10 enriched Gene Ontology (GO) terms for ADGRG3 interactors across the following three categories: biological process (BP), cellular component (CC), and molecular function [67]. Detailed GO terms are available in Supplementary File S1. For BP, the most enriched terms included adenylate cyclase-modulating GPCR signaling and dopamine receptor signaling, involving genes such as ADGRG3, GNB1, and GNG2. These pathways suggest ADGRG3's involvement in GPCR-related signaling, which is important in cancer progression and immune modulation. In the CC category, enriched terms such as the heterotrimeric G-protein complex and specific granules highlight the localization of ADGRG3 interactors in structures relevant to signal transduction and immune responses, particularly those involving GNB1, GNG2, and GNAO1. For MF, terms such as G-protein beta-subunit binding and G-protein-coupled peptide receptor activity were prominent, underscoring the functional role of ADGRG3 and its interactors in GPCR binding and receptor activity regulation.