Compositions and methods for multiplexed tumor vaccination with endogenous gene activation
The present invention includes compositions and methods for treating or preventing cancer. Embodiments include cell-based and viral vector-based vaccines that utilize gene expression activation systems to augment the product of endogenous genes to treat or prevent cancer.
1 . A composition comprising a cancer vaccine, comprising a modified cell comprising a CRISPRa system capable of increasing expression of a plurality of endogenous human antigen presentation genes resulting in the modified cell expressing at least one tumor associated antigen (TAA),
wherein the CRISPRa system comprises an sgRNA library,
wherein the plurality of endogenous human antigen presentation genes are selected from the group consisting of CD70, CD80, CD86, IFNA4, IFNB1, and IFNG; and
wherein the sgRNA library comprises every nucleic acid sequence of SEQ ID NOs: 1, 2, 5, 6, 9, 11-13, 16, 18, 21, 22, 24, 25, 28, 30, 31, and 33-35.
2 . A method of vaccinating against cancer in a subject in need thereof, the method comprising
administering to the subject a therapeutically effective amount of the composition of claim 1 .
3 . The method of claim 2 , wherein the CRISPRa system is in an AAV vector, a lentiviral vector, or an adenoviral vector, wherein the AAV vector is selected from the group consisting of AAV2, AAV8, AAV9, or AAV-DJ.
4 . The method of claim 2 , wherein the CRISPRa system comprises:
(a) a nucleic acid encoding dCas9-VP64, a nucleic acid encoding MS2-p65-HSF 1, and a genome-scale lentiviral synergistic activation mediator (SAM) CRISPRa sgRNA library; or
(b) a nucleic acid encoding MS2-p65-HSF 1, and a nucleic acid encoding a genome-scale lentiviral SAM CRISPRa sgRNA library.
5 . The method of claim 2 , wherein the subject is a human.
6 . The method of claim 2 , wherein administering the therapeutically effective amount of the composition comprises a one dose, a two dose, a three dose, a four dose, or a multi-dose treatment.
7 . The method of claim 2 , further comprising contacting the cell with a substance that induces senescence in the cell prior to administering to the subject, wherein the substance that induces senescence in the cell is mitomycin.