Lung cancer is the leading cause of cancer death in the United States and is responsible for more deaths each year than breast, prostate, colon, hepatic, renal and skin cancers combined. Viewed in economic terms, in the United States, the value of life lost from lung cancer deaths in the year 2000 was more than $240 billion, which is estimated to rise to more than $433 billion in 2020. Because of the immense health care and economic burden imposed by lung cancer, new therapy strategies that improve patient outcomes will lead to very a significant impact. Collaborators at UCLA (Laboratory of Lenny Rome) have identified and characterized a unique ribonucleoprotein nanoparticle structure that is highly stable, and is found ubiquitously in all higher eukaryotes. Major Vault Protein (MVP) is the core nanoparticle component, and it is readily engineered to permit attachment of other agents, including promising cancer therapeutics like chemokine ligand 21 (CCL21). CCL21 is a lymphoid chemokine that is chemoattractant for mature dendritic cells (DCs), naive and memory T cells. Preclinical studies have demonstrated that intratumoral administration of CCL21 gene-modified dendritic cells led to tumor eradication. Vaults have been expressed at Chesapeake PERL, Inc. (C-PERL) to very high levels using the PERLXpress protein expression platform. The unique and powerful system uses recombinant baculovirus expression in whole insects in an automated platform to generate high protein yields cost effectively. Preliminary data shows that MVP is readily expressed and correctly assembled to form nanoparticles. Further, packaging of a model protein (pCherry) has been demonstrated. . We expect that continued development will show that CCL21-Vaults can be readily prepared in high quality by expression, and packaging of the constituent proteins, followed by stringent purification methods to generate high yields of high quality nanocapsule. We hypothesize that intratumoral administration of recombinant CCL21-vaults derived from baculovirus infection of whole insects will circumvent autologous DC preparation, minimize batch to batch variability and allow for comparability and standardization so that the particle can be used as an off-the-shelf reagent for advanced NSCLC.
Specific Aim 1 : Engineer, express and purify Vault-CCL21 nanocapsule protein complexes for initial biochemical characterization using standard protein analytical tools as well as electron microscopy. Continue purification development to optimize yield and ensure highest quality and purity.
Specific Aim 2 : Perform in vitro analyses, including chemotactic and related assays to assess and compare the biochemical profile for CCL21-vault and compare to previously tested CCL21-vault expressed in SF9 cells.
This project is relevant to treatment of common causes of death and disability due to lung cancer. The research will support continued development of a candidate therapy using a recombinant version of naturally occurring human vault protein in combination with CCL21 antitumor agent. The combination technologies may result in a drug that targets lung cancer, demonstrating a large scale, low cost, reproducible therapy. This is a potentially disruptive approach that can revolutionize the lung cancer field, and may be applied to other cancers.
