Parkinson’s disease (PD) is a progressive neurodegenerative disease that is currently incurable with significant unmet medical needs. In the US, over a million people suffer from this disease with an estimated cost of $27 billion per year. The mainstay of PD management is symptomatic treatment with drugs that increase dopamine in the striatum. However, the utility of these drugs is significantly curtailed by waning efficacy and debilitating side effects. Since PD stems from the degeneration of a single cell type, dopaminergic (DA) neurons that supply dopamine to a defined location the striatum, PD patients have been viewed as optimal candidate for cell transplantation therapy. Preclinical and clinical studies have demonstrated that transplantation of primary fetal midbrain DA neurons into the striatum, provide significant and sustained restoration of function. However ethical, practical and safety issues with using fetal aborted tissue and the recent progress in induced pluripotent stem cells (iPSCs), grafting trials in PD are beginning to re-emerge world-wide with a new focus on pluripotent stem cell technologies. However, clinical studies to date suggest that critical refinements in the cellular product, in the delivery method, and in the clinical protocol by considering the incorporation of adjunctive therapies, are essential to realize the potential of cell therapy in PD. NeoNeuron LLC has developed intellectual property on scalable and effective technologies for generating an unlimited supply of DA neurons from iPSCs and an image-guided methodology for delivering the cells into the target area of the brain enabling functional recovery in the rat and in the nonhuman primate models of PD. In preparation for our Investigational New Drug (IND) submission, NeoNeuron met with the U.S. Food and Drug Administration (FDA) and received recommendations to conduct optimal dose range for the cellular product, iPSC-DA neurons, in the immunocompromised rat model of PD. The company has established standard operating procedures to expand this product under current good manufacturing practice (cGMP), and to produce DA neurons that will be available as an off-the-shelf product. This feature is desirable for the development of the intended product for clinical use and for commercialization. In Aim 1 we will show evidence of dose-response and adequate levels of DA cell engraftment and survival for a 6-month duration in the hemi- parkinsonian rat model of PD. Aim 2 will assess the impact of adjunctive physical and cognitive training on motor and cognitive functions in the grafted animals. We will leverage single cell spatial transcriptomics profiling of the grafted cells to gain insights into the mechanisms of actions of our product and of the adjunctive training intervention in enhancing functional recovery. This Phase 1 will enable us to respond to the FDA recommendations by identifying the optimal dose level of iPSC-DA cell engraftment and durable effects in t...