Vault complexes for cytokine delivery
The invention relates to compositions of vault complexes containing recombinant cytokine fusion proteins that include a cytokine and a vault targeting domain, and methods of using the vault complexes to deliver the cytokines to a cell or subject, and methods for using the compositions to treat cancer, such as lung cancer.
1. A method of preparing a composition comprising a vault complex, said vault complex comprising a vault particle and a cytokine fusion protein, the method comprising
a) mixing the cytokine fusion protein with the vault particle to generate a mixture, wherein the cytokine fusion protein comprises a cytokine and a vault targeting domain, which said vault targeting domain is a vault poly ADP-ribose polymerase (VPARP) sequence or a portion thereof that interacts with the major vault protein, or a telomerase/vault associated protein 1 (TEP 1) sequence or a portion thereof that interacts with the major vault protein, and wherein the vault particle comprises a major vault protein (MVP); and
b) incubating the mixture for a sufficient period of time to allow non-covalent association of the cytokine fusion protein with the vault particle, thereby generating the vault complex.
2. The method of claim 1 , wherein the MVP is human MVP.
3. The method of claim 1 , wherein the cytokine fusion protein and the MVP are generated in Sf9 or insect larvae cells.
4. The method of claim 1 , wherein the vault targeting domain is a) a vault poly-ADP ribose polymerase (VPARP) sequence or a portion thereof that interacts with the MVP; or b) a telomerase/vault associated protein 1 (TEP 1) sequence or a portion thereof that interacts with the MVP.
5. The method of claim 1 , wherein the cytokine fusion protein is non-covalently associated with the vault particle.
6. The method of claim 1 , wherein the cytokine is a chemokine.
7. The method of claim 6 , wherein the chemokine is a cysteine-cysteine (CC) chemokine.
8. The method of claim 7 , wherein the CC chemokine is a CCL-21 chemokine.
9. The method of claim 1 , wherein the cytokine comprises the amino acid sequence of SEQ ID NO: 2.
10. The method of claim 4 , wherein the vault targeting domain is the portion of the VPARP that is a major vault protein interaction (mINT) domain.
11. The method of claim 10 , wherein the mINT domain comprises the amino acid sequence of SEQ ID NO: 8.
12. The method of claim 1 , wherein the cytokine fusion protein comprises the amino acid sequence of SEQ ID NO: 13.