CONTEXT: Understanding phenotypic connection between type II diabetes (T2D) mellitus and amyotrophic lateral sclerosis (ALS) can offer valuable sight into shared disease etiology and have important implication in drug repositioning and therapeutic intervention. OBJECTIVE: This work aims to disentangle the nature of the inverse relationship between T2D mellitus and ALS. METHODS: Depending on summary statistics of T2D (n = 898 130) and ALS (n = 80 610), we estimated the genetic correlation between them and prioritized pleiotropic genes through a multiple-tissue expression quantitative trait loci-weighted integrative analysis and the conjunction conditional false discovery rate (ccFDR) method. We implemented mendelian randomization (MR) analyses to evaluate the causal relationship between the 2 diseases. A mediation analysis was performed to assess the mediating role of T2D in the pathway from T2D-related glycemic/anthropometric traits to ALS. RESULTS: We found supportive evidence of a common genetic foundation between T2D and ALS (rg = -0.223, P = .004) and identified 8 pleiotropic genes (ccFDR < 0.10). The MR analyses confirmed that T2D exhibited a neuroprotective effect on ALS, leading to an approximately 5% (95% CI, 0% ~ 9.6%, P = .038) reduction in disease risk. In contrast, no substantial evidence was observed that supported the causal influence of ALS on T2D. The mediation analysis revealed T2D can also serve as an active mediator for several glycemic/anthropometric traits, including high-density lipoprotein cholesterol, overweight, body mass index, obesity class 1, and obesity class 2, with the mediation effect estimated to be 0.024, -0.022, -0.041, -0.016, and -0.012, respectively. CONCLUSION: We provide new evidence supporting the observed inverse link between T2D and ALS, and revealed that a shared genetic component and causal association commonly drove such a relationship. We also demonstrate the mediating role of T2D standing in the pathway from T2D-related glycemic/anthropometric traits to ALS.