Charcot-Marie-Tooth disease type 4D (CMT4D) is a demyelinating peripheral neuropathy caused by mutations in NDRG1. While progressive myelin degeneration has been observed in Ndrg1 knockout (KO) mice, the early transcriptional events leading to demyelination and the accompanying proteomic changes remain inadequately understood. This study analyzed sciatic nerves from Ndrg1 KO mice at presymptomatic (2 weeks) and symptomatic (4 weeks) stages, employing transcriptomic profiling, quantitative proteomics, and integrated multi-omics analysis. At 2 weeks, despite normal myelin structure, KO sciatic nerves exhibited transcriptional shifts indicative of extracellular matrix remodeling and neurofilament gene downregulation. By 4 weeks, the molecular profile emphasized immune activation and cholesterol metabolism disruption. Proteomics revealed decreased myelin proteins and increased immune- and lipid-related proteins. Integration identified cholesterol metabolism as a key pathway, with APOE, HSD17B7, and HMGCR as central hubs. Lipid droplet accumulation was also observed. To evaluate therapeutic potential, we tested 2-hydroxypropyl-β-cyclodextrin (HPβCD), a cholesterol-mobilizing agent, from 4 to 12 weeks; however, no significant improvement in myelin integrity was detected via G-ratio analysis. Overall, the findings demonstrate a temporal progression of molecular pathology in Ndrg1-deficient nerves, evolving from extracellular matrix and cytoskeletal perturbations to immune activation and cholesterol metabolic dysregulation. These insights refine our understanding of NDRG1's role in myelin maintenance and may guide future therapeutic approaches for CMT4D.