Key Takeaways: Repressor element-1 silencing transcription factor (REST) protein and mRNA levels were associated with Alzheimer’s Disease (AD). REST expression was regulated by miR-153-3p (miR153). We identified that miR153, as a master regulator, reduced levels of a group of neurodegeneration-related proteins: amyloid-beta precursor protein (APP), alpha-synuclein (SNCA), and REST. The elevation of miR153 levels is associated with a reduced probability of AD in the posterior cingulate cortex, whereas REST is associated with a greater probability of AD. Further, modulation of the miR153–REST axis influences neuronal differentiation and cell survival in human iPSC-derived neuronal cells, highlighting this regulatory pathway as a promising therapeutic target for AD.