Self-replicating cell selective gene delivery compositions, methods, and uses thereof
Described herein are cell-selective mRNA constructs that can contain a RNA of interest and one or more miRNA targets. The cell-selective mRNA constructs described herein can be used to express an RNA of interest to a cell in a cell-selective manner.
1. A polyribonucleotide comprising:
a RNA molecule of interest (ROI), wherein the ROI is an RNA capable of being translated into p27;
a microRNA (miRNA) target sequence, wherein the miRNA target sequence is a target for miR-126, and wherein the miRNA target sequence is operatively linked to the ROI; and
a RNA molecule capable of being translated into a viral RNA replicase, wherein the RNA molecule capable of being translated into a viral RNA replicase is operatively linked to the ROI, the miRNA target sequence, or both the ROI and miRNA target sequence.
2. The polyribonucleotide of claim 1 , wherein the viral replicase is a Venezuelan Equine Encephalitis viral replicase, a Sindbis viral replicase, or a Semliki Forest virus replicase.
3. The polyribonucleotide of claim 1 , further comprising a RNA molecule capable of being translated into one or more additional nonstructural viral proteins, wherein the RNA molecule capable of being translated into one or more additional nonstructural viral proteins is operatively linked to the ROI, the miRNA target sequence, or both the ROI and miRNA target sequence.
4. The polyribonucleotide of claim 1 , wherein the polyribonucleotide is a linear polyribonucleotide or a circular polyribonucleotide.
5. The polyribonucleotide of claim 1 , wherein one or more ribonucleotides of the polyribonucleotide is modified.
6. The polyribonucleotide of claim 5 , wherein the modification is a Pseudouridine, N-1-methylpseudouridineridine, 5-methoxy-Uridine, a 5-hydroxymethyl-C, a 5-methyl-C, or a combination thereof.
7. The polyribonucleotide of claim 1 , wherein p27 is differentially expressed.
8. The polyribonucleotide of claim 1 , wherein the ROI has a sequence that is 85-100% identical to or corresponds to a sequence that is 85% to 100% identical to SEQ ID NO: 2.
9. The polyribonucleotide of claim 1 , wherein the miRNA target has a sequence that is complementary to a sequence that is 20-100% SEQ ID NO: 10.
10. The polyribonucleotide of claim 1 , wherein the miRNA target has a sequence that is complementary to a sequence that is 90-100% identical to SEQ ID NO: 10, where the portion is 5 or more consecutive nucleotides.
11. A pharmaceutical formulation comprising:
a polyribonucleotide comprising:
a RNA molecule of interest (ROI), wherein the ROI is an RNA capable of being translated into p27;
a microRNA (miRNA) target sequence, wherein the miRNA target sequence is a target for miR-126, and wherein the miRNA target sequence is operatively linked to the ROI; and
a RNA molecule capable of being translated into a viral RNA replicase, wherein the RNA molecule capable of being translated into a viral RNA replicase is operatively linked to the ROI, the miRNA target sequence, or both the ROI and miRNA target sequence; and
a pharmaceutically acceptable carrier.
12. The pharmaceutical formulation of claim 11 , wherein the viral replicase is a Venezuelan Equine Encephalitis viral replicase, a Sindbis viral replicase, or a Semliki Forest virus replicase.
13. A method comprising:
administering a polyribonucleotide to subject, wherein the polyribonucleotide comprises
a RNA molecule of interest (ROI), wherein the ROI is an RNA capable of being translated into p27;
a microRNA (miRNA) target sequence, wherein the miRNA target sequence is a target for miR-126, and wherein the miRNA target sequence is operatively linked to the ROI; and
a RNA molecule capable of being translated into a viral RNA replicase, wherein the RNA molecule capable of being translated into a viral RNA replicase is operatively linked to the ROI, the miRNA target sequence, or both the ROI and miRNA target sequence.
14. The method of claim 13 , wherein the viral replicase is a Venezuelan Equine Encephalitis viral replicase, a Sindbis viral replicase, or a Semliki Forest virus replicase.
15. The pharmaceutical formulation of claim 11 , wherein the polynucleotide is present in the pharmaceutical formulation at an amount effective to inhibit neointimal hyperplasia, inhibit inflammatory cell infiltration to a site of vascular injury, or inhibit neointimal hyperplasia and inhibit inflammatory cell infiltration to a site of vascular injury while not adversely affecting endothelial cells.
16. The pharmaceutical formulation of claim 11 , wherein the polynucleotide is present in the pharmaceutical formulation at an amount effective to inhibit proliferation of vascular smooth muscle cells while not adversely affecting endothelial cells.
17. The pharmaceutical formulation of claim 11 , wherein the polynucleotide is present in the pharmaceutical formulation at an amount effective to overexpress p27 in a vascular smooth muscle cell but not overexpress p27 in a vascular endothelial cell.