DNA vector and transformed tumor cell vaccines
Inventors
Lawman, Michael J. P. • Lawman, Patricia D. • RAMIYA, VIJAY • GENTILINI, MEGHAN
Assignees
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Publication Number
US-10391158-B2
Publication Date
2019-08-27
Expiration Date
Abstract
Customized whole cell cancer vaccines can be produced from autologous (ex vivo or in situ) or allogeneic human or veterinary patient cell lines. Cells are transformed with S. pyogenes DNA that expresses an Emm protein on the cell surface and cytosol. Treatment of cancer patients with an Emm vector vaccine induces an immunologic response to the cancer by enhancing immunogenicity of a tumor. Emm vaccines can be used in patients where the cancer is not identified due to lower tumor burden or used to treat a specific cancer and subsequently treat for a second type that may have arisen through metastasis.
Core Innovation
The document describes a therapeutic DNA vector cancer vaccine based on an Emm-based approach using S. pyogenes emm genes, such as pAc/emm. The emm gene is used to express an Emm protein on a cell surface and in the cytosol, with the Emm protein functioning as a membrane-anchoring protein design that supports immune activation.
The disclosed vaccine strategy addresses a cancer immune response problem by promoting APC/DC activation and cross-presentation, and by engaging innate immunity as a danger signal/danger-associated innate activation. The approach is presented as being relevant to suppressive elements of the tumor microenvironment, including immune suppression mechanisms involving T regulatory (Treg) cells, myeloid-derived suppressor cells, and tumor-associated macrophages.
The document further includes designs for Emm55, including helical transmembrane and anchor features corresponding to predicted N-terminal and C-terminal regions, along with experimental confirmation of membrane and cytosolic expression using Western blot and flow cytometry. It also discusses use in both direct intratumoral/in situ DNA administration and ex vivo transformed whole-cell vaccine preparation, and it describes potential combination approaches with checkpoint inhibitors and other immune-modulating agents.
Claims Coverage
The independent claim set covers a membrane anchoring protein defined by the placement of SEQ ID NO: 5 within both an N-terminal signal sequence and a C-terminal anchor region, with additional dependents adding sequence-identity constraints, membrane topology/orientation, and a functional in vivo immune-response requirement. The inventive coverage is centered on the specific sequence location and topology/orientation features associated with membrane anchoring.
Membrane anchoring protein with SEQ ID NO: 5 located in N-terminal signal and C-terminal anchor
A membrane anchoring protein comprising the amino acid sequence SEQ ID NO: 5 located within the N-terminal signal sequence and the C-terminal anchor region of the protein.
N-terminal signal sequence identity defined by SEQ ID NO: 7
The membrane anchoring protein of claim 1 specified so that its N-terminal signal sequence contains the amino acid sequence of SEQ ID NO: 7.
C-terminal anchor region identity defined by SEQ ID NO: 9
The membrane anchoring protein of claim 1 includes a C-terminal anchor region whose amino acid sequence is SEQ ID NO: 9.
Helical C-terminal anchor orientation outside to inside
The membrane anchoring protein of claim 1 has a helical C-terminal anchor region oriented from the outside to the inside on the cell membrane.
Helical N-terminal signal peptide region inside to outside for egress to membrane surface
The membrane anchoring protein of claim 1 includes a helical N-terminal signal peptide region that orients from inside to outside to provide egress to the cell membrane surface.
Inducing an immune response in vivo
The membrane anchoring protein of claim 1 is described as inducing an immune response in vivo.
Overall, the claim coverage is focused on a membrane anchoring protein defined by SEQ ID NO: 5 positioned within an N-terminal signal sequence and a C-terminal anchor region, refined by specific sequence identities (SEQ ID NO: 7 and SEQ ID NO: 9), defined helical topology/orientation (inside-to-outside for the N-terminal signal peptide region and outside-to-inside for the C-terminal anchor region), and the functional requirement to induce an immune response in vivo.
Stated Advantages
Induces an immune response in vivo.
Documented Applications
Direct intratumoral/in situ DNA administration for whole-cell vaccine generation in a cancer setting.
Ex vivo preparation of autologous or allogeneic transformed whole-cell vaccines using tumor cells transformed to express Emm.
Cancer vaccine use discussed across solid, liquid, and metastatic cancers, including tumor contexts involving immune suppression mechanisms in the tumor microenvironment.
Supportive in-vivo examples in canine lymphoma model and mammary adenocarcinoma (Golden Retriever).
Supportive in-vivo examples in cutaneous melanoma model (equine).
Supportive in-vivo examples in transitional cell carcinoma model (Shetland Sheepdog).
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