Functionalized DNA Dendrimers For Gene Delivery To Cells
Compositions are disclosed which comprise a DNA dendrimer having one or more DNA sequences linked thereto, the DNA sequences comprising DNA sequences encoding a polypeptide or regulatory RNA linked to DNA sequences that regulate expression of the DNA sequences encoding a polypeptide or regulatory RNA to produce an RNA coding for the polypeptide or to produce the regulatory RNA. Also disclosed are methods for treating diseases and conditions of cells by delivering the dendrimers to the cells, with subsequent expression of the encoded polypeptide or regulatory RNA.
1 . A composition comprising a DNA dendrimer, wherein one or more linear DNA sequences encoding a cytotoxin operably linked to a promoter having specific transcriptional activity in pancreatic cancer cells or prostate cancer cells are linked to the DNA dendrimer.
2 . The composition of claim 1 , wherein the promoter is MSLN or K5.
3 . The composition of claim 1 , wherein the cytotoxin is Diphtheria toxin A chain.
4 . The composition of claim 1 , comprising a plurality of linked DNA sequences, wherein the linked DNA sequences encode a plurality of cytotoxins.
5 . The composition of claim 1 , further comprising a cellular internalization moiety linked to the DNA dendrimer.
6 . The composition of claim 5 , wherein the internalization moiety is an antibody that binds to a cell surface protein, a peptide that binds to a cell surface protein or a ligand that binds to a cell surface receptor.
7 . The composition of claim 6 , wherein the internalization moiety is an antibody or peptide that binds to the transferrin receptor.
8 . The composition of claim 1 , wherein the DNA dendrimer is constructed from DNA monomers, each of which comprises two DNA strands having a sequence of complementarity located in a central portion of each strand forming a central double-stranded waist bordered by four single-stranded arms, and the DNA dendrimer is assembled by base pairing between single-stranded arms of complementary monomers in sequential layers, and wherein the one or more linear DNA sequences encoding the cytotoxin are hybridized to available single-stranded arms on a surface of the DNA dendrimer via oligonucleotides complementary to the single-stranded arms ligated to the DNA sequences.
9 . A pharmaceutical composition comprising a DNA dendrimer according to claim 1 and a pharmaceutically acceptable carrier.
10 . The pharmaceutical composition of claim 9 , wherein the pharmaceutically acceptable carrier is a dextran aldehyde polymer mucoadhesive which is a topical delivery vehicle for facilitating uptake of the DNA dendrimer by cells.
11 . The pharmaceutical composition of claim 10 , wherein the mucoadhesive composite is a PEG:dextran mucoadhesive composite.
12 . The pharmaceutical composition of claim 9 , wherein the pharmaceutically acceptable carrier is a carrier for systemic administration.
13 . The pharmaceutical composition of claim 9 , wherein the DNA dendrimer is linked to a cellular internalization moiety.
14 . A method for treating pancreatic cancer or prostate cancer in a patient comprising administering to the patient an amount of the DNA dendrimer of claim 1 effective to treat the pancreatic cancer or prostate cancer.