A pesticide and iPSC dopaminergic neuron screen identifies and classifies Parkinson-relevant pesticides.

Parkinson's disease (PD) is a complex neurodegenerative disease with etiology rooted in genetic vulnerability and environmental factors. Here we combine quantitative epidemiologic study of pesticide exposures and PD with toxicity screening in dopaminergic neurons derived from PD patient induced pluripotent stem cells (iPSCs) to identify Parkinson's-relevant pesticides. Agricultural records enable investigation of 288 specific pesticides and PD risk in a comprehensive, pesticide-wide association study. We associate long-term exposure to 53 pesticides with PD and identify co-exposure profiles. We then employ a live-cell imaging screening paradigm exposing dopaminergic neurons to 39 PD-associated pesticides. We find that 10 pesticides are directly toxic to these neurons. Further, we analyze pesticides typically used in combinations in cotton farming, demonstrating that co-exposures result in greater toxicity than any single pesticide. We find trifluralin is a driver of toxicity to dopaminergic neurons and leads to mitochondrial dysfunction. Our paradigm may prove useful to mechanistically dissect pesticide exposures implicated in PD risk and guide agricultural policy.

Small-molecule screen reveals pathways that regulate C4 secretion in stem cell-derived astrocytes.

In the brain, the complement system plays a crucial role in the immune response and in synaptic elimination during normal development and disease. Here, we sought to identify pathways that modulate the production of complement component 4 (C4), recently associated with an increased risk of schizophrenia. To design a disease-relevant assay, we first developed a rapid and robust 3D protocol capable of producing large numbers of astrocytes from pluripotent cells. Transcriptional profiling of these astrocytes confirmed the homogeneity of this population of dorsal fetal-like astrocytes. Using a novel ELISA-based small-molecule screen, we identified epigenetic regulators, as well as inhibitors of intracellular signaling pathways, able to modulate C4 secretion from astrocytes. We then built a connectivity map to predict and validate additional key regulatory pathways, including one involving c-Jun-kinase. This work provides a foundation for developing therapies for CNS diseases involving the complement cascade.

Simulation of proximal catheter occlusion and design of a shunt tap aspiration system.

PURPOSE: Total and partial proximal catheter occlusions are well-known complications of ventriculoperitoneal shunts (VPS). When this occurs, surgeons often attempt to perform a shunt tap. However, the degree of obstruction in a proximal catheter that ultimately leads to shunt malfunction is unknown. METHODS: We developed a benchtop model to simulate proximal catheter occlusion with two hydrostatic reservoirs connected by a VPS catheter system. The Centurion compass device was used to measure pressure across the valve digitally. Wires of varying diameters (equalling different occlusion percentages) were inserted into the catheter's proximal end to stimulate obstruction. A mock shunt tap aspiration was then performed by incorporating a pressure transducer. RESULTS: As a general trend, pressure reading on the device decreases as occlusion increases. At higher levels of occlusion (> 45%), the blockage begins to significantly impede the flow through the catheter, and the pressure drops at a faster rate compared with lower occlusion percentages. The pressure reading converges quickly to 0 with increasing blockage after about 70%. The Centurion compass is able to detect large changes in pressure as evidenced by the major differences in pressure readings between no occlusion, 45%, and 84%. The shunt will not function at 84%. In order to determine the threshold for occlusion beyond which fluid cannot be withdrawn, we tested five levels of occlusion (0%, 33%, 63%, 84%, and 100%) at various aspiration pressures and determined that fluid can still be produced with 0-84% occlusion, but no fluid could be produced at 100% occlusion. CONCLUSIONS: We developed a model of proximal shunt obstruction and found that cerebrospinal fluid (CSF) flow through a VPS is unaffected up to 33% occlusion, begins to become impaired at 45% occlusion, and is miniscule at 84% occlusion. Shunt aspiration was not possible at 84% occlusion. Pressure measured at the reservoir is accurate and correlates with intracranial pressure (ICP) up to approximately 60% proximal occlusion. With partial occlusion up to 70%, ventricular pressure will dictate shunt function.