Neural Circuits, Kinetics and Energetics HTS of Human iPSC-Neurons, -Microglia, and -Astrocytes: AI-Enabled Platform for Target ID, and Drug Discovery and Toxicity (e.g., Cancer Chemo & HIV ARTs)

点击翻译按钮获取中文摘要

Neurotoxicity is among the most common reasons for failure of drugs during clinical trials, illustrating that current preclinical test models, which feature whole animals or animal cells are extremely poor at predicting human neurotoxicity. Additionally, many currently FDA-approved therapeutics, such as anti-cancer chemotherapeutics, are neurotoxic, which limits their therapeutic dosage and often results in impaired cognition during and after treatment. FDA “black box” warnings, describing neurological side-effects, are common and currently prescribed therapeutics for AIDs may contribute to HIV-Associated Neurocognitive Disorder (HAND). Accordingly, Vala Sciences Inc., proposes a Fast-Track SBIR project to develop an assay system utilizing neurons, astrocytes, and microglia, derived from human induced pluripotent stem cells (hiPSC-neurons/astrocytes/microglia) to test candidate therapeutics for neurotoxicity. The cells will be cultured in 384-well dishes to enable high throughput multiplexed assay of compound effects on neuronal activity (via Kinetic Image Cytometry [KIC] – acquisition/analysis of videos of cells labeled with calcium and voltage indicators), and synapses, mitochondria, endoplasmic reticulum, and cell viability. Since the assay will feature human cells, it will likely be more predictive of human neurotoxicity vs. animal test systems currently used in preclinical research. Phase I activities will develop the multiplexed system, to enable testing of compounds for toxic effects on mechanisms of action (MOA) including alteration of plasma membrane ion channels, ER function and integrity, mitochondrial function, and neuronal circuitry (correlated activity of neurons within a culture well). A total of 30 compounds with known MOA and effects of humans, will be tested in Phase I, these data will be used to train artificial intelligence (AI) clustering algorithms to recognize the MoAs, particularly for low doses of the test agents. The compounds will also be tested on hiPSC-neurons/astrocytes/microglia representing APOE-3/4 and APOE-4/4 genotypes, as the APOE-4 is associated increased neurotoxicity. In Phase II a total of 350 probes/drugs (of which 275 are known to have clinical effects, and 75 will be expected to have no effect) will be screened, with the results used to further train the AI and improve the overall clinical predictivity. Additional versions of the assay will be developed featuring mutations/genotypes relevant to Alzheimer’s Disease and Bipolar Disease. In addition to recognition of neurotoxicity, the assay system also will increase our understanding of neuronal circuits, which is responsive to the BRAIN Initiative program. The neurotoxicity assay will be marketed to pharmaceutical companies developing therapeutics across a broad spectrum of diseases/afflictions, including chemotherapeutics and potential therapeutics for neurodegenerative diseases.