Novel approach to modulate the TME on-demand with therapeutic peptides and proteins. - Abstract We have developed a transformable nanoparticle (TNP) platform capable of transforming its morphology from nanoparticles to nanofibrils when interact with receptors (e.g., EGFR or α3β1 integrin) over-expressed on the cell surface of many tumor types, including non-small cell lung cancer (NSCLC). TNPs displaying peptide ligands against these receptors, will transform into nanofibrillar networks surrounding the tumor cells, in vitro or in vivo. Through the above work, we found that the nanofibrillar network generated was able to retain at the tumor microenvironment (TME) for a week, whereas nanostructures retained at normal organs such as liver and lung disappeared after 2 days. Here, we want to exploit this unique tumor retention property of TNP by utilizing the highly specific bioorthogonal chemistry to introduce, on- demand, desired therapeutic payloads, to modulate the tumor microenvironment (TME), while sparing normal tissues. This novel two-component two-step (TCTS) nanotherapeutic strategy is based on the use of (i) TNP for pre-targeting, and (ii) the rapid and highly specific bioorthogonal click reaction for on- demand targeted delivery of peptides, small molecules and proteins that can modulate the TME. Our hypothesis is that this highly versatile TNP/TCTS therapeutic platform will allow effective on- demand sequential or combination delivery to the TME orthogonal immunomodulatory molecules such as immune cell capturing ligands (e.g., LLP2A), small molecule immunostimulants (e.g., resiquimod), and sialidase (to shave the immunoinhibitory oligo-siliac acids off the tumor cell membrane). As a consequence, a robust anti-tumor immune response will be elicited, resulting in durable regression of the cancer. The 3 specific aims of this project are: Aim 1. To design, synthesize and characterize EGFR and α3β1 integrin targeting transformable nanoparticle (TNP) constructs. Aim 2. To use in vivo and ex vivo optical imaging to optimize TNP/TCTS platform for cancer immunotherapy. Aim 3. To determine the in vivo toxicity and efficacy of the novel TNP/TCTS immunotherapeutic approach using syngeneic murine Lewis lung cancer model.
