Engineered APP C-terminus Alters Its Native Transcript Dynamics in Differentiated SH-SY5Y-Derived Neurons

Alzheimer's disease (AD) pathology is deeply connected with the processing of amyloid precursor protein (APP) and its cleavage products. APP processing generates functional fragments, such as the APP intracellular domain (AICD) and the APP carboxy-terminal fragment (CTF), which have been implicated in transcriptional regulation. However, their potential role in modulating endogenous APP splicing and expression remains unclear. This study investigates the self-regulatory potential of APP through its C-terminal fragments. Stable transgenic SH-SY5Y lines overexpressing APP695, its Swedish mutant form, and truncated and engineered C-terminal constructs were generated. They were differentiated over 24 days to get mature neuron-like cells and analyzed for APP transcript variants. The overexpression of APP fragments altered total APP levels and specific transcript variants, especially APPv3 and APPv11, predominantly in differentiated neurons. Differential splicing patterns were further confirmed through specific transcript fragment-size analysis. These findings highlight that APP derivatives actively influence transcriptional regulation and alternative splicing of native APP. This research advances our understanding of APP biology by revealing its self-regulatory complexity, suggesting that APP fragments could serve as transcription modulators of APP itself, providing insights into disease pathogenesis and novel therapeutic strategies in AD.