Perinatal brain therapeutics: a high throughput biology approach to build next generation RNA precision medicines

Project Summary Early electrical patterns in the human brain guide development, and even slightly disrupted electrical activity can result in severe neurodevelopmental diseases. Epilepsy alone affects ~3.5 million individuals in the United States, including ~450,000 children (CDC). For children with severe epilepsy syndromes, few effective treatments are available, and clinicians rely on decades-old anticonvulsant drugs. Recently, our lab, in collaboration with Tim Yu’s lab at Boston Children’s Hospital, obtained Food and Drug Administration approval for a precision-guided antisense oligonucleotide (ASO) RNA therapy that we developed and validated for a child who, due to potassium-channel malfunction, had a treatment-resistant epilepsy. This safe and efficacious RNA therapeutic modality (see also FDA approved Nusinersen for SMA) has transformed the therapeutic landscape as a cross-cutting approach for children with untreatable diseases. However, while ASOs are a proven therapeutic strategy, the “personalized precision medicine” model for individual patients costs millions of dollars per child, and is therefore both limited in its treatment reach and highly inequitable for low resource/underserved patient populations. Moreover, developing a personalized therapy takes an average of 3 years, by which time irreversible brain damage has occurred. Instead, this DP2 proposal aims to provide an immediately available and broadly applicable RNA therapeutic to stabilize children early with severe neurodevelopmental diseases. Here, we propose a transformational approach that will enable us to reprogram brain cells, enhancing their resilience against a broad array of neurodevelopmental diseases by stabilizing electrical activity before permanent damage can occur. To this end, we combine our proven RNA therapeutics approach to stabilize excitotoxicity pathways with a high-throughput 3-dimensional (3D) neurobiology platform optimized for the evaluation of complex neocortical neurophysiology in 100,000+ cerebral organoids over the project duration. If successful, our results will build a new era of stabilizing therapeutics for a wide range of diseases in infants. To support our HTS 3D cell culture and RNA-approach, we employ leading in silico modeling techniques, including machine learning, to simulate neuronal activity responses to ASOs. In vitro models will be screened against computationally designed ASO cocktail combinations to systematically modulate levels of neuronal activity in specific cell-types, monitored with dual physiological calcium and voltage readouts. We are committed to share our longitudinal single-cell and 3D-neurophysiology datasets with colleagues to enhance therapeutic targeting in human brain development and power the switch from traditional monolayer neuron models to 3D models that best capture complex human neocortex neurophysiology. This DP2 will reveal the full potential of this new class of ASO, improving the lives of children with neurodevelopmental diseases and pave the way for our highly efficacious RNA chemistry to be utilized for prenatal therapies (akin to in utero spina bifida surgery).