Native cell derived vesicles containing tumor suppressor proteins for therapy
A method of obtaining cell derived vesicles comprising an active wild-type p53 is disclosed. The method comprising: (i) isolating cell derived vesicles from a biological sample comprising cells; and (ii) treating the cell derived vesicles with a DNA damaging agent, or the method comprising: (i) treating cells with a DNA damaging agent; and (ii) isolating cell derived vesicles from a biological sample comprising the cells. A proteinaceous preparation comprising cell derived vesicles and a pharmaceutical composition comprising the proteinaceous preparation are also disclosed. Methods of treating a disease, disorder or condition associated with a mutant or a nonfunctional p53 protein and methods of inducing apoptosis of a target cell comprising a mutant or a nonfunctional p53 protein are also disclosed.
1 . A proteinaceous preparation comprising cell derived vesicles from corneal epithelium cells, said cell derived vesicles comprising phosphorylated wild-type p53 protein and being devoid of recombinant p53, wherein the preparation is devoid of intact cells and wherein said cell derived vesicles have been isolated from corneal epithelium cells treated with a DNA damaging agent.
2 . The preparation of claim 1 , wherein said corneal epithelium cells are non-human corneal epithelium cells.
3 . The preparation of claim 1 , wherein said non-human corneal epithelium cell are rat corneal epithelium cells or chicken corneal epithelium cells.
4 . A pharmaceutical composition comprising the proteinaceous preparation of claim 1 and a pharmaceutically acceptable carrier.
5 . The proteinaceous preparation of claim 1 , wherein said cell derived vesicles comprise cell secreted vesicles.
6 . The proteinaceous preparation of claim 1 , wherein said cell derived vesicles have a mean particle diameter of about 20 to about 200 nm.
7 . The proteinaceous preparation of claim 1 , wherein said cell derived vesicles comprise exosomes.
8 . The proteinaceous preparation of claim 1 , wherein an outer surface of said cell derived vesicles comprise a heterologous moiety for targeted delivery of said cell derived vesicles to a target cell.
9 . The proteinaceous preparation of claim 1 , wherein said heterologous moiety is selected from the group consisting of a protein, a peptide and a glycolipid molecule.
10 . The proteinaceous preparation of claim 1 , wherein said corneal epithelium cells are genetically non-modified.
11 . A method of treating cancer associated with a mutant or a nonfunctional p53 protein in a human subject in need thereof, the method comprising administering to the subject a therapeutically effective amount of the pharmaceutical composition of claim 4 , thereby treating the cancer in the human subject.
12 . The method of claim 11 , wherein said cancer is a solid tumor or metastasis.
13 . The method of claim 12 , wherein the solid tumor or metastasis is selected from the group consisting of an ovarian cancer, a cervical cancer, a vaginal cancer, a vulvar cancer, an anal cancer, a penile cancer, a breast cancer, an endometrial cancer, a head and neck cancer, a colon cancer, a colorectal cancer, a prostate cancer, a lung cancer, a melanoma, a pancreatic cancer, a liver cancer and a splenic cancer.
14 . The method of claim 12 , wherein said solid tumor is selected from the group consisting of a colon cancer, a lung cancer, an ovarian cancer and a head and neck cancer.
15 . The method of claim 11 , wherein said cell derived vesicles comprise cell secreted vesicles.
16 . The method of claim 11 , wherein said cell derived vesicles have a mean particle diameter of about 20 to about 200 nm.
17 . The method of claim 11 , wherein said cell derived vesicles comprise exosomes.
18 . The method of claim 11 , wherein said corneal epithelium cells are genetically non-modified.
19 . The proteinaceous preparation of claim 1 , wherein said corneal epithelium cells are genetically non-modified.
20 . The proteinaceous preparation of claim 1 , wherein said corneal epithelium cells are genetically modified.
21 . The proteinaceous preparation of claim 1 , wherein said corneal epithelium cells are human corneal epithelium cells.