Causal mediation analysis with multiple causally non-ordered and ordered mediators based on summarized genetic data.

Causal mediation analysis investigates the mechanism linking exposure and outcome. Dealing with the impact of unobserved confounders among exposure, mediator and outcome is an issue of great concern. Moreover, when multiple mediators exist, this causal pathway intertwines with other causal pathways, rendering it difficult to estimate the path-specific effects. In this study, we propose a method (PSE-MR) to identify and estimate path-specific effects of an exposure (e.g. education) on an outcome (e.g. osteoarthritis risk) through multiple causally ordered and non-ordered mediators (e.g. body mass index and pack-years of smoking) using summarized genetic data, when the sequential ignorability assumption is violated. Specifically, PSE-MR requires a specific rank condition in which the number of instrumental variables is larger than the number of mediators. Furthermore, we illustrate the utility of PSE-MR by providing guidance for practitioners and exploring the mediation effects of body mass index and pack-years of smoking in the causal pathways from education to osteoarthritis risk. Additionally, the results of simulation reveal that the causal estimates of path-specific effects are almost unbiased with good coverage and Type I error properties. Also, we summarize the least number of instrumental variables for the specific number of mediators to achieve 80% power.