SBIR Phase II: Developing a Platform for Multiplexed Drug Profiling Using Yeast Synthetic Agglutination

这个小企业创新研究（SBIR）第二阶段项目的更广泛的影响和商业潜力是开发一个发现分子胶的平台技术。许多疾病，包括癌症、自身免疫性疾病和神经系统疾病，都可以通过控制细胞中特定蛋白质的活性或丰度来治疗。然而，许多蛋白质不能被传统药物靶向。相反，制药公司现在正在使用一种新的策略来劫持细胞的天然质量控制途径，并降解蛋白质来控制它们的丰度，而不是它们的活性。这种方法已被证实是一种强大的治疗策略，但发现分子胶仍面临重大挑战。该分子胶发现平台有望通过高通量筛选产生重大的商业和社会影响。这项小型企业创新研究（SBIR）二期项目旨在推进一种新型平台的开发，用于发现通过蛋白质-蛋白质相互作用发挥作用的分子胶或药物。该平台通过将酵母单倍体交配效率与细胞表面显示的蛋白质亲和力联系起来，将基于细胞的测定的通量与生物分析技术的准确性相结合。初步结果表明，二倍体细胞的下一代测序可以同时高精度地测量许多蛋白质-蛋白质相互作用的亲和力，并正确确定具有良好特征的抑制或增强特定蛋白质-蛋白质相互作用的小分子的作用。此外，该平台支持96孔板格式，这对于标准高通量筛选工作流程的兼容性非常重要。该项目的主要目标是提高平台检测诱导弱蛋白-蛋白相互作用的分子胶的灵敏度，通过提高检测效率降低每孔筛选成本，并将新蛋白纳入平台并与现有小分子验证其功能。该奖项反映了美国国家科学基金会的法定使命，并通过使用基金会的知识价值和更广泛的影响审查标准进行评估，被认为值得支持。

The broader impact/commercial potential of this Small Business Innovation Research (SBIR) Phase II project is to develop a platform technology for discovering molecular glues. Many illnesses, including cancers, autoimmune diseases, and neurological diseases, can be treated by controlling the activity or abundance of specific proteins in the cell. However, many proteins cannot be targeted by traditional drugs. Instead, pharmaceutical companies are now using a new strategy to hijack the cell’s native quality control pathways and degrade proteins to control their abundance rather than their activity. This approach has been validated as a powerful therapeutic strategy, but significant challenges remain for discovering molecular glues. The proposed platform for molecular glue discovery is expected to have a major commercial and societal impact by conducting high-throughput screening. This Small Business Innovation Research (SBIR) Phase II project proposes to advance the development of a novel platform for discovering molecular glues, or drugs that function by agonizing protein-protein interactions. This platform combines the throughput of a cell-based assay with the accuracy of a bioanalytical technique by linking yeast haploid mating efficiency to the affinity of proteins displayed on the cells' surfaces. Initial results demonstrate that next generation sequencing of diploid cells can be used to simultaneously measure the affinity of many protein-protein interactions with high accuracy and correctly determine the effect of well-characterized small molecules that inhibit or enhance particular protein-protein interactions. Additionally, the platform is functional in a 96-well plate format, which is important for compatibility with standard high-throughput screening workflows. The primary goals of this project are to improve the sensitivity of the platform for the detection of molecular glues that induce a weak protein-protein interaction, reduce the per-well screening cost by improving assay efficiency, and incorporate new proteins into the platform and validate their function with existing small-molecules.This award reflects NSF's statutory mission and has been deemed worthy of support through evaluation using the Foundation's intellectual merit and broader impacts review criteria.