IP Library › Granted Patent US 12,222,347
Granted Patent B2
US 12,222,347 · App. 18/458,657 · Granted Feb 11, 2025

Ligand discovery and gene delivery via retroviral surface display

Inventors: Michael Birnbaum (Arlington, MA); Connor Dobson (Somerville, MA)
Assignee: Massachusetts Institute of Technology
G01N33/505C07K14/005C07K14/4748C07K14/5437C07K14/70532C07K14/70539C07K16/2803C12N15/86C07K2317/622C07K2319/02C07K2319/035C07K2319/60C12N2740/15045
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Quick Facts
Patent No.
US 12,222,347
App. No.
18/458,657
Granted
Feb 11, 2025
Kind
B2
Abstract

Compositions of retroviruses and methods of using the same for gene delivery are disclosed, wherein the retroviruses comprise a viral envelope protein comprising at least one mutation that diminishes its native function, a non-viral membrane-bound protein comprising a membrane-bound domain and an extracellular targeting domain.

Claims (39)

1. A lentivirus comprising:

(a) a nucleic acid encoding a gene of interest; and

(b) a viral envelope that comprises at least two different proteins: (i) a vesicular stomatitis virus (VSV-G) envelope protein comprising an amino acid sequence at least 97% identical to SEQ ID NO: 13, wherein the VSV-G envelope protein comprises an amino acid substitution at position 47 (lysine, K) and an amino acid substitution at position 354 (arginine, R) to alanine (A) or glutamine (Q); and

(ii) a non-viral membrane-bound protein comprising a transmembrane domain and an extracellular targeting domain that binds to a T cell or B cell.

2. The lentivirus of claim 1 , wherein:

(a) the VSV-G envelope protein comprises a K47A or K47Q amino acid substitution and an R354A or R354Q amino acid substitution;

(b) the VSV-G envelope protein comprises a K47A amino acid substitution and an R354A amino acid substitution;

(c) the VSV-G envelope protein comprises a K47A amino acid substitution and an R354Q amino acid substitution;

(d) the VSV-G envelope protein comprises a K47Q amino acid substitution and an R354A amino acid substitution; or

(e) the VSV-G envelope protein comprises a K47Q amino acid substitution and an R354Q amino acid substitution.

3. The lentivirus of claim 1 , wherein the extracellular targeting domain comprises a protein, a peptide, or an antibody.

4. The lentivirus of claim 1 , wherein the extracellular targeting domain comprises an interleukin-13 protein domain, a CD80 protein domain, a full-length antibody, an antibody fragment, a nanobody, a single chain antibody (scFv), an anti-CD19 antibody, an anti-TCR antibody, or an anti-CD3 antibody.

5. The lentivirus of claim 1 , wherein the extracellular targeting domain binds a T cell receptor (TCR), a cytokine receptor, a cytokine, a T cell surface marker, CD3, CD19, or CD20.

6. A lentivirus, comprising

(a) a nucleic acid encoding a gene of interest; and

(b) a viral envelope that comprises at least two different proteins: (i) a CD80 protein comprising a CD80 extracellular domain and a transmembrane domain; and

(ii) a VSV-G envelope protein comprising an amino acid sequence at least 97% identical to SEQ ID NO: 13, wherein the VSV-G protein comprises an amino acid substitution at position K47 and an R354A or R354Q amino acid substitution.

7. The lentivirus of claim 6 , wherein:

(a) the VSV-G envelope protein comprises a K47A or K47Q amino acid substitution and an R354A or R354Q amino acid substitution;

(b) the VSV-G envelope protein comprises a K47A amino acid substitution and an R354A amino acid substitution;

(c) the VSV-G envelope protein comprises a K47A amino acid substitution and an R354Q amino acid substitution;

(d) the VSV-G envelope protein comprises a K47Q amino acid substitution and an R354A amino acid substitution; or

(e) the VSV-G envelope protein comprises a K47Q amino acid substitution and an R354Q amino acid substitution.

8. The lentivirus of claim 6 , further comprising a non-viral membrane-bound protein comprising an extracellular targeting domain that binds to a cognate ligand of a cell.

9. The lentivirus of claim 8 , wherein the non-viral membrane-bound protein comprises an extracellular targeting domain extracellular targeting domain that comprises an interleukin-13 protein domain, a full-length antibody, an antibody fragment, a nanobody, a single chain antibody (scFv), an anti-CD19 antibody, an anti-TCR antibody, or an anti-CD3 antibody.

10. The lentivirus of claim 8 , wherein the cognate ligand is a T cell receptor (TCR), a cytokine receptor, a cytokine, a T cell surface marker, CD3, CD19, or CD20.

11. A lentivirus, comprising

(a) a nucleic acid encoding a gene of interest; and

(b) a viral envelope that comprises at least two different proteins: (i) a non-viral membrane-bound protein comprising a transmembrane domain and an extracellular targeting domain that comprises an anti-CD3 antibody; and

(ii) a VSV-G envelope protein comprising an amino acid sequence at least 97% identical to SEQ ID NO: 13, wherein the VSV-G protein comprises an amino acid substitution at position K47 and an R354A or R354Q amino acid substitution.

12. The lentivirus of claim 11 , wherein:

(a) the VSV-G envelope protein comprises a K47A or K47Q amino acid substitution and an R354A or R354Q amino acid substitution;

(b) the VSV-G envelope protein comprises a K47A amino acid substitution and an R354A amino acid substitution;

(c) the VSV-G envelope protein comprises a K47A amino acid substitution and an R354Q amino acid substitution;

(d) the VSV-G envelope protein comprises a K47Q amino acid substitution and an R354A amino acid substitution; or

(e) the VSV-G envelope protein comprises a K47Q amino acid substitution and an R354Q amino acid substitution.

13. The lentivirus of claim 11 , further comprising a non-viral membrane-bound protein comprising an extracellular targeting domain that binds to a cognate ligand of a cell.

14. The lentivirus of claim 13 , wherein the non-viral membrane-bound protein comprises an extracellular targeting domain extracellular targeting domain that comprises an interleukin-13 protein domain, a CD80 protein domain, a full-length antibody, an antibody fragment, a nanobody, a single chain antibody (scFv), an anti-CD19 antibody, or an anti-TCR antibody.

15. The lentivirus of claim 13 , wherein the cognate ligand is a T cell receptor (TCR), a cytokine receptor, a cytokine, a T cell surface marker, CD19, or CD20.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Feb 20, 2024
From: BIRNBAUM, MICHAEL; DOBSON, CONNOR
To: MASSACHUSETTS INSTITUTE OF TECHNOLOGY
Reel/Frame 066499/0263 →
Continuity (3)
Continuation 16826665 · Mar 23, 2020
Provisional Application 62851889 · May 23, 2019
Related Publication 20240230627A1 · Jul 11, 2024
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