Tunning the micro-electric field at the metal-support interface offers great opportunities for developing heterogeneous catalysts with high stability, selectivity, and activity. However, the role of carbon support defects plays in manipulating the charge transfer process and activating the interfacial perimeter sites is still obscure due to the absence of direct experimental evidence. Herein, we develop a top-down method to synthesize a metal-defective-support model via both heavy ion irradiation and chemical impregnation. The interface electric field distribution, together with the magnitude of charge density distribution were reconstructed and visualized in the real space via four-dimension scanning transmission electron microscopy (4D-STEM).Combining the density functional theory (DFT) calculations, we further reveal the carbon topological defects induced atomic scale polarization mechanism is responsible for the reversed charge transfer behavior and demonstrate that the metal-support interaction could be precisely manipulated through controlling support defects density. This work offers new insights for understanding defects’ function of elevating the catalyst stability and performance, and facilitating the future development of advanced catalysts with high activity.