The wildland-urban interface (WUI) represents landscapes where human settlements coexist with natural features. Trails within the WUI areas, valued for their ecological, recreational, and educational values, lack comprehensive research on landscape sensitivity influenced by both landscape and urban development. This paper addresses the gap by proposing a comprehensive landscape sensitivity index (CLSI) using multiple regression, cluster analysis, and correlation analysis. The Appalachian Trail (AT) serves as a case study to explore the characteristics of high sensitivity areas, considering various attributes and their connection with federal reserved land. Results show that eliminating covariance in landscape indices refines the landscape aggregation pattern, with Moran's I decreasing from 0.776 to 0.449, aligning with the observed fragmented landscape. In comparison to modified landscape indices (MLSI), the CLSI reveals that 85.6% of the area experiences changes in landscape sensitivity, with 42.5% of the AT region displaying significant landscape sensitivity, including 4.9% as having high landscape sensitivity (HLS), influenced by rock formations, wetlands, and biodiversity. A spatial mismatch is identified between HLS and current federal preservation efforts, with a correlation of only 0.011. The paper proposes tailored conservation strategies for HLS areas in urban, wilderness, and protected regions. Considering the combined impact of ecological and urbanization forces, this study assists in prioritizing land conservation objectives and finding a balance between wilderness protection and urban development. Display omitted •The comprehensive landscape sensitivity index is applicable in the WUI region.•Rock, wetlands, and biodiversity are the key factors of high landscape sensitivity.•Only 14.3% of the landscape sensitivity is agreed considering urban disturbances.•Elimination of co-linear landscape indices reveals fragmented landscape pattern.•Spatial mismatch exists between federal land and high landscape sensitivity areas.