Protein-based Self-assembly and Disassembly of Nanostructures

University Of Massachusetts Amherst, Amherst MA

Investigators

Linked publications & trials

Abstract

PROJECT SUMMARY / ABSTRACT Our research program is focused on developing robust platforms that enable protein-based biologics as therapeutics for intracellular targets. Biologics offer distinct advantages over small molecule drugs, because of their high specificity, including against targets that are considered undruggable with small molecules. However, large size and hydrophilic nature of proteins present the formidable challenge of transporting them to intracellular locations. Protein-based therapeutics have been, thus, largely limited to extracellular targets. Recognizing this gap, we have focused on developing a platform with the combination of characteristics, critical for transporting proteins inside the cells. Accordingly, we arrived at a new protein-encapsulating nanogel system that effectively and tracelessly delivers the proteins inside the cells in its functional form. Over the next five years, we seek to build on these accomplishments to explore new scientific directions. We will create a new antibody-directed antibody conjugates (ADACs) platform with the goal of engaging currently undruggable intracellular targets in specific cells. ADACs contain two distinct antibodies – one for targeting extracellular epitopes for cellular specificity and another for engaging a specific intracellular target. We leverage the combined capabilities of the protein-polymer self-assembly to generate nanogels with the bioorthogonal chemistries developed for decorating the nanogel surfaces with antibodies, to create ADACs with tunable physiochemical characteristics. We will investigate the potential for ADACs in targeted protein degradation (TPD) of model intracellular targets that are considered undruggable, because of a single missense mutation or single site post-translational modification. In this context, we will also fully test the potential of the TRIM21 E3 ligase pathway for proteasomal TPD using antibodies and compare its efficacy with the more classical inhibition pathway in eliciting downstream responses. Concurrently, we will also develop new strategies for increasing the efficacy and specificity of antibody-based biologics using the ADACs platform. Endosomal escape has been identified as the major bottleneck in the efficacy of intracellular biologics. We propose two new, mechanistically distinct strategies for functionalizing nanogel surfaces that promote endosomal escape. Similarly, to substantially enhance cellular specificities of therapeutics, we propose a high-risk, high-payoff strategy for in-cell assembly of antibody-like molecules using Boolean logic based convergence. We have chosen the components of our approaches to be independent, such that each of our scientific goals independently inform new strategies for solving critical and unsolved issues in the field of intracellular biologics. Simultaneously, these goals are also designed to be convergent such that when put together, the impact of our overall approach will be exponentially higher. Collectively, our program will inform and develop strategies for enabling biologics as the next generation therapeutics for intracellular targets.