Modelling Parkinson’s Disease Using Human iPSC-derived Neurons

Parkinson’s disease (PD) is characterized by progressive neurodegeneration that primarily depletes dopaminergic neurons in the substantia nigra pars compacta, resulting in the PD quintessential motor symptoms. Animal models for PD have replicated many features of the neurodegeneration and LB pathology; however, there is no model that fully reproduces the human disease. Therefore, a human model of PD neurons is needed to accurately investigate the underlying molecular mechanisms. Recent advancements in the field of stem cell research have opened up new possibilities for modelling of difficult-to-obtain cell types. In 2006, Yamanaka et al. were the first to dedifferentiate fibroblasts into induced pluripotent stem cells (iPSCs), and thereby identifying the necessary transcription factors to express that mediate dedifferentiation. With an improved delivery method these factors remain the basis for today’s iPSCs. Combined with a directed differentiation, it is possible to create a new replenishable source of human derived neurons. This is especially important for PD, as research suffers from lack of access and availability of affected human midbrain dopaminergic neurons. Research in this thesis applies these new techniques to set up models based on patients with familial PD and aims to find affected cellular mechanisms in PD

Telomerase reverse transcriptase promoter mutations in bladder cancer: High frequency across stages, detection in urine, and lack of association with outcome

Background Hotspot mutations in the promoter of the gene coding for telomerase reverse transcriptase (TERT) have been described and proposed to activate gene expression. Objectives To investigate TERT mutation frequency, spectrum, association with expression and clinical outcome, and potential for detection of recurrences in urine in patients with urothelial bladder cancer (UBC). D

The SAC1 domain in synaptojanin is required for autophagosome maturation at presynaptic terminals

Presynaptic terminals are metabolically active and accrue damage through continuous vesicle cycling. How synapses locally regulate protein homeostasis is poorly understood. We show that the presynaptic lipid phosphatase synaptojanin is required for macroautophagy, and this role is inhibited by the Parkinson's disease mutation R258Q. Synaptojanin drives synaptic endocytosis by dephosphorylating PI(4,5)P2, but this function appears normal in SynaptojaninRQ knock-in flies. Instead, R258Q affects the synaptojanin SAC1 domain that dephosphorylates PI(3)P and PI(3,5)P2, two lipids found in autophagosomal membranes. Using advanced imaging, we show that SynaptojaninRQ mutants accumulate the PI(3)P/PI(3,5)P2-binding protein Atg18a on nascent synaptic autophagosomes, blocking autophagosome maturation at fly synapses and in neurites of human patient induced pluripotent stem cell-derived neurons. Additionally, we observe neurodegeneration, including dopaminergic neuron loss, in SynaptojaninRQ flies. Thus, synaptojanin is essential for macroautophagy within presynaptic terminals, coupling protein turnover with synaptic vesicle cycling and linking presynaptic-specific autophagy defects to Parkinson's disease