Induced pluripotent stem cells derived from patient cells have emerged as complementary tools to study human neurological disorders.  Neurxstem Inc has generated human iPSC-derived synthetic neural organoids (Neural Organoid Platform^TM) which exhibit a remarkable level of development that morphologically resembles a nearly complete human central nervous system at ~5 weeks in development in utero, but after ~12 weeks in culture in vitro.  These synthetic organoids express a complex milieu of markers characteristic of nearly all types of neurons in the human embryonic central nervous system, as well as cells that are astrocytic, oligodendritic, microglial, and vascular in lineage.  Our analysis supports the formation of all the major regions of the central nervous system including the retina, cortex, midbrain, brain stem and the spinal cord in a single complete neural organoid and is the first time this milestone in human brain organogenesis has been achieved. Using Neurxstem’s proprietary process, we have now engineered synthetic neural organoids from Alzheimer’s disease and tuberous sclerosis patient skin cells. Whole genome transcriptomic shows close genotype-clinical phenotype correlations.

This success strongly suggests that Neurxstem’s Neural Organoid Platform^TM has a multitude of future applications to study neurodegenerative diseases (Alzheimer’s and Parkinsons’s disease, ALS, and MS), neurodevelopmental disorders (autism), and neuropsychiatric disorders (Post Traumatic Stress Disorder (PTSD), bipolar and schizophrenia).

The Neural Organoid Platform^TM is also useful to study the effect of nutrition and environmental toxins on prenatal brain development.

Future exciting applications include assessing effects of changes in gravity and geomagnetic fields on prenatal development of the human brain for space explorations and colonization of new planets.

Methods: We characterize neural organoid responses to genetic or environmental perturbations at the molecular, cellular and structural level using transcriptomics, immunohisto-chemistry, and 3D whole brain imaging. 

Results: Neurxstem has generated human iPSC-derived synthetic Neural Organoid Platforms which exhibit a remarkable level of development that morphologically resembles a nearly complete human embryonic central nervous system at ~5 week in utero development, but after ~12 weeks in culture in vitro.  Using our proprietary process, we have engineered neural organoids from Alzheimer’s disease and tuberous sclerosis patient skin cells with identified genetic mutations in susceptibility genes.  Our transcriptomics results remarkably show comprehensive and accurate correlation of the dysregulated expression of hundreds of genes previously correlated with the clinical symptoms and/or pathologies of both of these diseases. For Alzheimer’s disease these include genes for lipid homeostasis, inflammation, metal ion homeostasis, and water homeostasis and amyloid peptide beta and tau metabolism. For tuberous sclerosis these include genes for tumor formation, autism, blood pressure regulation, Zn++ ion homeostasis, Pb++ ion toxicity, round worm infections, and cholesterol metabolism among others.

Conclusions: Neurxstem’s “NexGen” synthetic Neural Organoid Platform^TM allows the use of whole genome transcriptomic “digitization” of the gene expression response of the models to genetic or environmental perturbations of human brain physiology and thus accurately predict susceptibility to brain disease.
 

Cell Sciences Innovation, Merck Research Laboratories, 2015 Galloping Hill Road, Kenilworth, NJ USA
In vitro translational models play a critical role in the drug discovery process because they are often used to select potential new pharmaceutical entities and predict in vivo dosing.  Most, if not all, of those who take part in basic research and pharmacology have focused efforts on decreasing the attrition rate of clinical candidates by creating more biologically relevant systems.

These new platforms are instrumental in simulating a biological micro-environment and are typically comprised of primary or stem cell-derived tissue.  Current data suggests that three dimensional culture methods enable retention of native in vivo properties such as gene expression and biological function where more facile two dimensional tissue culture methods may have detrimental effects such as altered gene expression and decreased physiological activity. 

This represents a change in the paradigm where target-based genetically engineered recombinant cell lines cultured in monolayers in plastic vessels were the foundation of screening and profiling biological and chemical matter.  The move to more advanced tissue-based physiologically active models ushers in an era of complexity, but arrives with the promise that researchers will gain a more accurate understanding of clinical efficacy and toxicity much earlier in the process.  If successful, drug discovery will become more efficient and cost-effective for the industry, while leading to improved pharmaceutical treatments for disease.
 

The study of target-focused, disease relevant biology for drug discovery requires a comprehensive suite of tools that allow multiple aspects of the biology to be assessed.  Thermo Fisher Scientific has pioneered the development of such tools to study the role of the kinase LRRK2 in Parkinson’s Disease, from the first commercial production of full-length protein to enable biochemical assays, to tools that allow the generation and interrogation of dopaminergic neurons harboring disease-specific mutations.  This presentation will focus on how such tools can be used to better understand the disease, and accelerate the drug discovery process.