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.