Cell targeting compositions and methods
AssigneeThe Trustees of Indiana University
InventorsMilan Radovich, Jeffrey Solzak, Chao Wang
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Abstract
The present disclosure provides cell targeting constructs that specifically target cells of interest through the use of binding agents to cell surface nucleic acids, including without limitation extracellular DNA (exDNA). The cell targeting constructs can be engineered to deliver desired payload, including without limitation therapeutic agents and/or detectable labels, to the target cells.
Claims107
A cell targeting construct comprising a binding agent to a target nucleic acid on the surface of a target cell, wherein the binding agent comprises a protein, and wherein the binding agent is attached covalently or non-covalently to at least one payload.
The cell targeting construct of any preceding claim, wherein the binding agent is encoded by a nucleic acid sequence that has at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO. 3, SEQ ID NO. 5, SEQ ID NO. 9, or SEQ ID NO. 11, or the binding agent is encoded by a nucleic acid sequence that encodes a protein sequence that has at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 10, or SEQ ID NO. 12.
The cell targeting construct of any preceding claim, wherein the binding agent comprises a protein sequence that has at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO. 4, SEQ ID NO. 6, SEQ ID NO. 10, or SEQ ID NO. 12.
The cell targeting construct of any preceding claim, wherein the at least one payload comprises a small molecule, peptide, protein, nucleic acid, toxin, therapeutic agent, drug, chemotherapeutic agent, liposome, nanoparticle, dendrimer, detectable label, or any useful combination thereof.
The cell targeting construct of any preceding claim, wherein the target nucleic acid comprises DNA or RNA, optionally wherein the RNA comprises messenger RNA (mRNA) or microRNA (miRNA).
The cell targeting construct of any preceding claim, wherein the target nucleic acid originated within the target cell or within the target cell microenvironment.
The cell targeting construct of any preceding claim, wherein the target nucleic acid comprises genomic DNA (gDNA).
The cell targeting construct of any preceding claim, wherein the target nucleic acid has a wild-type (WT) sequence or a sequence comprising one or more mutations, optionally wherein the one or more mutations comprise at least one single nucleotide variant (whether pathogenic or not), an insertion, a deletion, a substitution, inversion, translocation, fusion, break, loss, duplication, amplification, or repeat;
- and optionally wherein the one or more mutation comprises one or more cancer mutation.
The cell targeting construct of any preceding claim, wherein the target nucleic acid comprises a KRAS sequence, optionally wherein the KRAS sequence comprises a Q61H, G12D and/or G13D mutation.
The cell targeting construct of any preceding claim, wherein the target nucleic acid comprises at least a portion of SEQ ID NO. 1, SEQ ID NO. 2, SEQ ID NO. 7, or SEQ ID NO. 8.
The cell targeting construct of any preceding claim, wherein the target nucleic acid comprises a sequence that as at least 80, 85, 90, 91, 92, 93, 94, 95, 96, 97, 98, 99, or 100% identity to SEQ ID NO. 2 or SEQ ID NO. 8.
The cell targeting construct of any preceding claim, wherein the target cell comprises a diseased cell, optionally wherein the disease comprises a cancer, a premalignant condition, an inflammatory disease, an immune disease, an autoimmune disease or disorder, a cardiovascular disease or disorder, a neurological disease or disorder, an infectious disease or pain.
A nucleic acid polymer encoding some or all of the cell targeting construct of any preceding claim, optionally wherein the nucleic acid polymer encodes the binding agent.
A protein encoded by a nucleic acid sequence that has at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO. 3 or SEQ ID NO. 5.
A protein having a sequence that has at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO. 4 or SEQ ID NO. 6.
A protein encoded by a nucleic acid sequence that has at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO. 9 or SEQ ID NO. 11.
A protein having a sequence that has at least 50%, 60%, 70%, 75%, 80%, 85%, 90%, 91%, 92%, 93%, 94%, 95%, 96%, 97%, 98%, 99%, or 100% identity to SEQ ID NO. 10 or SEQ ID NO. 12.
A method of treating cancer in a patient comprising the administration of a zinc finger protein to the patient in need thereof wherein
- i) the zinc finger protein binds to a cell surface extracellular DNA (exDNA) that is specific for a cancer cell and the zinc finger protein forms a zinc finger protein/exDNA complex with the exDNA;
- ii) the zinc finger protein/exDNA complex is internalized into the cancer cell;
- and,
- iii) the zinc finger protein/exDNA complex binds to an intracellular target.
A method of targeting or identifying cancer cells comprising using a zinc finger protein wherein
- i) the zinc finger protein binds to a cell surface extracellular DNA (exDNA) that is specific for a cancer cell and the zinc finger protein forms a zinc finger protein/exDNA complex with the exDNA;
- ii) the zinc finger protein/exDNA complex is internalized into the cancer cell;
- and,
- iii) the zinc finger protein/exDNA complex binds to an intracellular target.
A method of inhibiting the growth of a cancer cell in a patient comprising the administration of a zinc finger protein to the patient in need thereof wherein
- i) the zinc finger protein binds to a cell surface extracellular DNA (exDNA) that is specific for a cancer cell and the zinc finger protein forms a zinc finger protein/exDNA complex with the exDNA;
- ii) the zinc finger protein/exDNA complex is internalized into the cancer cell;
- and,
- iii) the zinc finger protein/exDNA complex binds to an intracellular target.
A method of diagnosing cancer in a patient comprising administration of a zinc finger protein to the patient suspected of having cancer, wherein the zinc finger protein has a biomarker attached and wherein the zinc finger protein binds to a cell surface extracellular DNA (exDNA) that is specific for a cancer cell, the zinc finger protein forms a zinc finger protein/exDNA complex with the exDNA, and wherein the the biomarker is visualized or quantified to diagnose the cancer.
A composition to treat cancer in a patient in need thereof comprising a zinc finger protein wherein
- i) the zinc finger protein binds to a cell surface extracellular DNA (exDNA) that is specific for a cancer cell and the zinc finger protein forms a zinc finger protein/exDNA complex with the exDNA;
- ii) the zinc finger protein/exDNA complex is internalized into the cancer cell;
- and,
- iii) the zinc finger protein/exDNA complex binds to an intracellular target.
A composition for diagnosing cancer in a patient suspected of having cancer comprising a zinc finger protein, wherein the zinc finger protein has a biomarker attached and wherein the zinc finger protein binds to a cell surface extracellular DNA (exDNA) that is specific for a cancer cell, the zinc finger protein forms a zinc finger protein/exDNA complex with the exDNA, and wherein the biomarker is visualized or quantified to diagnose the cancer.
Dependencies are inferred from the claim text.