The determine the effect of tissue-type plasminogen activator and its derivative tenecteplase on neuronal signalling and injury

Project overview: The serine protease tissue-type plasminogen activator (tPA) is well known for its fibrinolytic properties to remove blood clots from patients with acute ischaemic stroke (AIS) and approved by the FDA over 30 years ago. However, tPA has a short plasma half-life and needs to be administered as an infusion over 1h that can be complicated in emergency settings. tPA can also disrupt blood-brain barrier (BBB) permeability and promote neuron excitotoxicity and these effects impact on the safety of this drug in treating patients with AIS. A variant of tPA called tenecteplase (TNK) that is 98.5% identical to tPA (6 amino acids altered) has a longer plasma half-life and is likely to become the front-line thrombolytic for patients with AIS. However, information on the effect of TNK on neurons is limited. In this project, human iPSC-derived neurons (iPSCs) will be used to investigate the stimulatory effects of tPA and TNK on neuronal signalling and at promoting excitotoxic injury. In this project, human iPSCs will be stimulated with tPA and TNK in the presence and absence of glutamate analogies (NMDA, kainic acid) and neuronal responses compared using markers of cell death and injury. In addition, neurons will be plated onto multielectrode array (MEA) plates to allow us to evaluate how tPA and TNK alters neuronal activity. We anticipate that important and novel information will be gained from this project.