Biological characterization of the therapeutic role of Rab10 modulation in the development of Alzheimer’s disease

Abstract

AbstractBackgroundAlzheimer’s disease (AD) is the leading cause of dementia in the elderly. The continuous genetic research done on this disease demonstrates the vast complexity of the genetic architecture. The most recent whole‐genome sequence analysis (GWAS)included 1.1 million individuals and identified 38 loci associated with AD status with seven novel loci. However, there is no reliable therapeutics to combat the late stages of AD as of late. The identified rare variants in the RAB10 gene show protective effects against developing AD. This work demonstrates that inhibition of the expression of RAB10 yields better outcomes in terms of the amyloid‐beta ratio and cognition. In addition, we demonstrated that RAB10 modulates the NLRC4 inflammasome complex and their derived products IL‐1‐beta, IL‐18, and TNF‐alpha.MethodIn‐vitro experiments were carried out using human neuroblastoma (N2A) cell lines transfected with a mammalian plasmid to overexpress the APP gene and the immortalized microglia HMC3 cell line. We used the compound reference MLi2 and house compounds HT4253 and HT4403 to inhibit the expression of the RAB10. We used ELISA protocols to test the amyloid‐beta 40 and 42 ratios, IL‐1‐beta, IL‐18, and TNF‐alpha secretion. Western blots immunocytochemistry allowed us to determine the phosphorylation of RAB10 and LRRK2, LAMP1, and ASC. In‐vivo behavioral experiments were done using APP/PS1 mice. Hippocampus tissue was collected to run RNA sequence and total proteomics analysis.ResultA higher dosage of compound HT4253 and HT4403 inhibits phosphorylation of RAB10 with an IC50 of 82 nM and 63 nM, respectably. The inhibition of RAB10 and LRRK2 activates a higher expression of the LAMP1 protein. The incubation with a high compound dosage improved the secretion of the amyloid‐beta 42/40 ratio. The additional experiments and results will be presented at the conference.ConclusionThis work provides insights into the protective role of RAB10 against AD and neuroinflammation. RAB10 could be a promising therapeutic target for AD prevention.