Abstract

The development of therapies to treat patients with neurodegeneration is hampered by the use of animal models, less than 10% of findings derived from these pre-clinical models translate to humans. Patient-derived induced pluripotent stem cells can generate in vitro systems to model neurological diseases that recapitulate relevant human disease phenotypes. Conventional human iPSC (hiPSCs) differentiation protocols are lengthy, inconsistent, and difficult to scale.

 

To overcome these problems, bit.bio’ precision reprogramming technology, opti-oxTM enables highly controlled expression of transcription factors to rapidly reprogram hiPSCs into pure somatic cell types in a scalable manner.

 

We have introduced a 50 ‘CAG’ trinucleotide repeat mutation in the Huntingtin (HTT) gene in wild type ioGlutamatergic Neurons. Mutant HTT proteins containing elongated polyglutamine stretches are aggregation-prone and affect a range of neuronal subtypes, including cortical glutamatergic neurons. Characterisation of these neurons by ICC and RT-qPCR showed the expression profile of pan-neuronal (MAP2 and TUBB3) and glutamatergic (VGLUT1 and VGLUT2) marker genes as well as of the HTT transcript itself are highly similar between ioGlutamatergic Neurons HTT 50CAG and the isogenic control. We are currently performing an in-depth phenotypic characterisation of this HD disease model and the isogenic control to determine the differences in their transcriptome, neuronal activity and mitochondrial functions. Beside the 50 CAG mutation in HTT, we have generated mutations in MAPT, TARDBP, GBA and PRKN to provide isogenic disease models for FTD, FTD/ALS and Parkinson’s disease.Our novel strategy to use the opti-oxTM technology for the scalable and consistent production of hiPSC-derived isogenic disease models, offers new avenues into drug discovery and can accelerate research and the development of new therapeutics.