Engineering Saffron Plant Cell Culture to Produce Neuroprotective Bioactives

ABSTRACT The saffron plant (Crocus sativa L.) contains a variety of bioactive compounds with demonstrated clinical benefit, including the terpenoids crocin, crocetin, safranal, and picrocrocin. The clinical benefits of these molecules have been demonstrated in depression, anxiety, obsessive-compulsive disorder, attention deficit hyperactivity disorder, ischemic stroke, multiple sclerosis, Alzheimer’s disease, Parkinson’s disease, and Huntington’s disease. Data from randomized controlled clinical trials in individuals with neurocognitive disorders provide compelling evidence to further develop pharmaceutical, supplement, and food-based treatments. These approaches hold commercial and therapeutic promise. And customer demand is high. For example, saffron supplements comprise a current market of more than $1 billion (2022), which is expected to grow (reaching nearly $1.7 billion by 2029). But the paucity of high-quality supplies limits market access to saffron bioactives. Ayana Bio is developing a platform approach called ASCENT (Analyze to Scale Cellular ENhancement Technology) to realize the power of plant cell culture technology for scalable, consistent, and cost-effective commercial scale production of plant-specific bioactives. In this proposal, we will demonstrate proof-of-concept for ASCENT by generating the first-ever bioactive-enriched saffron cell line and demonstrate preliminary safety and efficacy in a C. elegans model of Huntington’s disease. In Aim 1, top-of-the-line screening, sequencing, and metabolomics approaches will be used to simultaneously optimize cell culture growth, increase production of Crocus sativa L. bioactives, and elucidate the genes, enzymes, and pathways that can be targeted to further optimize the strain. In Aim 2, lead lines will be engineered to overexpress key biosynthetic enzymes that drive saffron bioactive production. In Aim 3, we will determine toxicology and efficacy of an engineered Crocus sativa L. cell culture extract in a C. elegans model of Huntington’s disease. Our preliminary data using a large-scale microfluidics approach to screen the C. elegans poly-glutamine (PolyQ) aggregation model discovered four compounds that significantly reduced protein aggregation and mitigated disease phenotypes. These results demonstrate the utility of C. elegans as a model of polyQ disease.