Harnessing Nanoscale Presentation to Elucidate the Impact of Therapeutic Packaging on Innate Immunity - Nanomaterial delivery systems are critical in the packaging and delivery of immunostimulatory, immunoinhibitory, or immunomodulatory cues, which range from small molecules to biologics. However, an underappreciated factor in the delivery process is the interface of delivery systems with immune cells, which critically affects downstream signal propagation. A challenge in designing effective delivery systems across varied patient populations is elucidating how particular structures propagate distinct downstream functions. Understanding propagation due to cargo presentation at the immune interface would enable the design of delivery systems that can be programmed with specific biodistribution, uptake, and processing outcomes. Addressing this effectively requires nanosystems with high molecular precision to fully evaluate interfacial interactions on a molecular level. This gap is unsolved by current delivery systems, which are inherently polydisperse, heterogeneous, and lack molecular precision; they are unable to molecularly tune cargo stoichiometry, placement, and orientation. Thus, the structure-function relationships that have been obtained cannot reveal the intricacies of cargo packaging at the nano-immune interface. The overall goal of this program is to develop synthetic oligonucleotide nanostructures that molecularly program interfacial interactions at immune interfaces to uncover the impact of molecular cargo packaging on immunomodulation. We will explore two focus areas: Area 1: How does cargo component spacing modulate receptor interactions and affect propagation? Many nanomaterials deliver cargo that binds with immune cell receptors and multivalent interactions can enhance responses. However, precise control over avidity-based binding and an understanding of the molecular positioning requirements for optimal binding kinetics is underexplored due to structural polydispersity. Using our molecular and programmable oligonucleotide platform, we will explore these interactions and identify nanoscale features (positioning, stoichiometry, cargo type) that drive effective cargo binding across multiple receptors and signal propagation. Area 2: What is the role of anisotropy at the nano-immune interface? The anisometric nature of biologics and their preferred alignment can be harnessed in therapeutic design to access elevated potencies. However, programming this synthetically and molecularly with current delivery systems is a challenge. We will molecularly package cargo anisotropically to elucidate preferential interactions with cell-surface receptors. Holistically, this program will: 1) generate insights for how molecular component placement and anisotropy in nanoscale delivery systems alter nano-immune interactions and propagate distinct immunity; 2) uncover preferential receptor cooperation mediated by therapeutic co-delivery; and 3) reveal biophysical interactions at the nano-immune interface. This work will pave the way for our ability to elevate immunotherapeutic potency for various drugs (small molecules, biologics) against different diseases (cancer, autoimmunity, infectious disease).
