IP Library › Granted Patent US 10,036,040
Granted Patent B2
US 10,036,040 · App. 15/849,062 · Granted Jul 31, 2018

Methods and compositions for the activation of gamma-delta T-cells

Inventors: Charles David Pauza (Baltimore, MD); Haishan Li (North Potomac, MD); Tyler Lahusen (Frederick, MD); Mei-Ling Liou (Germantown, MD)
Assignee: American Gene Technologies International Inc.
C12N15/86A61K38/16C12N5/0636C12N15/113A61K48/00A61K2039/5158C12N5/00C12N2310/14C12N2310/141C12N2310/531
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Quick Facts
Patent No.
US 10,036,040
App. No.
15/849,062
Granted
Jul 31, 2018
Kind
B2
Abstract

The present invention relates generally to methods and compositions for gene therapy and immunotherapy that activate gamma delta T-cells, and in particular, can be used in the treatment of various cancers and infectious diseases.

Claims (326)

1. A viral vector for treating a condition associated with the mevalonate pathway, the viral vector comprising:

a. at least one encoded shRNA capable of inhibiting production of an enzyme of the mevalonate pathway, wherein the at least one encoded shRNA comprises a sequence having at least 80% percent identity with:

(SEQ ID NO: 1)

  i.

GTCCTGGAGTACAATGCCATTCTCGAGAATGGCATTGTACTCCA

GGACTTTTT;

(SEQ ID NO: 2)

 ii.

GCAGGATTTCGTTCAGCACTTCTCGAGAAGTGCTGAACGAAATC

CTGCTTTTT;

(SEQ ID NO: 3)

iii.

GCCATGTACATGGCAGGAATTCTCGAGAATTCCTGCCATGTACA

TGGCTTTTT;

or

(SEQ ID NO: 4)

 iv.

GCAGAAGGAGGCTGAGAAAGTCTCGAGACTTTCTCAGCCTCCTT

CTGCTTTTT;

 or

b. at least one encoded microRNA capable of inhibiting production of an enzyme of the mevalonate pathway, wherein the at least one encoded microRNA comprises a sequence having at least 80% percent identity with:

(SEQ ID NO: 5)

i.

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCCT

TCTGCGTGAAGCCACAGATGGCAGAAGGAGGCTGAGAAAGTGCTG

CCTACTGCCTCGGACTTCAAGGGGCT;

(SEQ ID NO: 6)

ii.

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCCT

TCTGCGTGAAGCCACAGATGGCAGAAGGGCTGAGAAAGTGCTGCC

TACTGCCTCGGACTTCAAGGGGCT;

(SEQ ID NO: 7)

iii.

TGCTGTTGACAGTGAGCGACTTTCTCAGCCTCCTTCTGCGTGAAG

CCACAGATGGCAGAAGGAGGCTGAGAAAGTTGCCTACTGCCTCGG

A;

(SEQ ID NO: 8)

iv.

CCTGGAGGCTTGCTGAAGGCTGTATGCTGACTTTCTCAGCCTCCT

TCTGCTTTTGGCCACTGACTGAGCAGAAGGGCTGAGAAAGTCAGG

ACACAAGGCCTGTTACTAGCACTCA;

(SEQ ID NO: 9)

v.

CATCTCCATGGCTGTACCACCTTGTCGGGACTTTCTCAGCCTCCT

TCTGCCTGTTGAATCTCATGGCAGAAGGAGGCGAGAAAGTCTGAC

ATTTTGGTATCTTTCATCTGACCA;

or

(SEQ ID NO: 10)

vi.

GGGCCTGGCTCGAGCAGGGGGCGAGGGATACTTTCTCAGCCTCCT

TCTGCTGGTCCCCTCCCCGCAGAAGGAGGCTGAGAAAGTCCTTCC

CTCCCAATGACCGCGTCTTCGTCG.

2. The viral vector of claim 1 , wherein:

a. the at least one encoded shRNA comprises a sequence having at least 85% percent identity with:

(SEQ ID NO: 1)

  i.

GTCCTGGAGTACAATGCCATTCTCGAGAATGGCATTGTACTCCA

GGACTTTTT;

(SEQ ID NO: 2)

 ii.

GCAGGATTTCGTTCAGCACTTCTCGAGAAGTGCTGAACGAAATC

CTGCTTTTT;

(SEQ ID NO: 3)

iii.

GCCATGTACATGGCAGGAATTCTCGAGAATTCCTGCCATGTACA

TGGCTTTTT;

or

(SEQ ID NO: 4)

 iv.

GCAGAAGGAGGCTGAGAAAGTCTCGAGACTTTCTCAGCCTCCTT

CTGCTTTTT;

 or

b. the at least one encoded microRNA comprises a sequence having at least 85% percent identity with:

(SEQ ID NO: 5)

i.

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCCT

TCTGCGTGAAGCCACAGATGGCAGAAGGAGGCTGAGAAAGTGCTG

CCTACTGCCTCGGACTTCAAGGGGCT;

(SEQ ID NO: 6)

ii.

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCCT

TCTGCGTGAAGCCACAGATGGCAGAAGGGCTGAGAAAGTGCTGCC

TACTGCCTCGGACTTCAAGGGGCT;

(SEQ ID NO: 7)

iii.

TGCTGTTGACAGTGAGCGACTTTCTCAGCCTCCTTCTGCGTGAAG

CCACAGATGGCAGAAGGAGGCTGAGAAAGTTGCCTACTGCCTCGG

A;

(SEQ ID NO: 8)

iv.

CCTGGAGGCTTGCTGAAGGCTGTATGCTGACTTTCTCAGCCTCCT

TCTGCTTTTGGCCACTGACTGAGCAGAAGGGCTGAGAAAGTCAGG

ACACAAGGCCTGTTACTAGCACTCA;

(SEQ ID NO: 9)

v.

CATCTCCATGGCTGTACCACCTTGTCGGGACTTTCTCAGCCTCCT

TCTGCCTGTTGAATCTCATGGCAGAAGGAGGCGAGAAAGTCTGAC

ATTTTGGTATCTTTCATCTGACCA;

or

(SEQ ID NO: 10)

vi.

GGGCCTGGCTCGAGCAGGGGGCGAGGGATACTTTCTCAGCCTCCT

TCTGCTGGTCCCCTCCCCGCAGAAGGAGGCTGAGAAAGTCCTTCC

CTCCCAATGACCGCGTCTTCGTCG.

3. The viral vector of claim 1 , wherein the enzyme is farnesyl diphosphate synthase (FDPS).

4. The viral vector of claim 1 , wherein the viral vector is a lentiviral vector or an adeno-associated virus vector.

5. The viral vector of claim 4 , wherein the viral vector is a lentiviral vector.

6. A lentiviral particle produced by a packaging cell and capable of infecting a target cell, the lentiviral particle comprising an envelope protein capable of infecting a target cell, and:

a. at least one encoded shRNA capable of inhibiting production of an enzyme of the mevalonate pathway, wherein the at least one encoded shRNA comprises a sequence having at least 80% percent identity with:

(SEQ ID NO: 1)

  i.

GTCCTGGAGTACAATGCCATTCTCGAGAATGGCATTGTACTCCA

GGACTTTTT;

(SEQ ID NO: 2)

 ii.

GCAGGATTTCGTTCAGCACTTCTCGAGAAGTGCTGAACGAAATC

CTGCTTTTT;

(SEQ ID NO: 3)

iii.

GCCATGTACATGGCAGGAATTCTCGAGAATTCCTGCCATGTACA

TGGCTTTTT;

or

(SEQ ID NO: 4)

 iv.

GCAGAAGGAGGCTGAGAAAGTCTCGAGACTTTCTCAGCCTCCTT

CTGCTTTTT;

 or

b. at least one encoded microRNA capable of inhibiting production of an enzyme of the mevalonate pathway, wherein the at least one encoded microRNA comprises a sequence having at least 80% percent identity with:

(SEQ ID NO: 5)

i.

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCCT

TCTGCGTGAAGCCACAGATGGCAGAAGGAGGCTGAGAAAGTGCTG

CCTACTGCCTCGGACTTCAAGGGGCT;

(SEQ ID NO: 6)

ii.

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCCT

TCTGCGTGAAGCCACAGATGGCAGAAGGGCTGAGAAAGTGCTGCC

TACTGCCTCGGACTTCAAGGGGCT;

(SEQ ID N0: 7)

iii.

TGCTGTTGACAGTGAGCGACTTTCTCAGCCTCCTTCTGCGTGAAG

CCACAGATGGCAGAAGGAGGCTGAGAAAGTTGCCTACTGCCTCGG

A;

(SEQ ID NO: 8)

iv.

CCTGGAGGCTTGCTGAAGGCTGTATGCTGACTTTCTCAGCCTCCT

TCTGCTTTTGGCCACTGACTGAGCAGAAGGGCTGAGAAAGTCAGG

ACACAAGGCCTGTTACTAGCACTCA;

(SEQ ID NO: 9)

v.

CATCTCCATGGCTGTACCACCTTGTCGGGACTTTCTCAGCCTCCT

TCTGCCTGTTGAATCTCATGGCAGAAGGAGGCGAGAAAGTCTGAC

ATTTTGGTATCTTTCATCTGACCA;

or

(SEQ ID NO: 10)

vi.

GGGCCTGGCTCGAGCAGGGGGCGAGGGATACTTTCTCAGCCTCCT

TCTGCTGGTCCCCTCCCCGCAGAAGGAGGCTGAGAAAGTCCTTCC

CTCCCAATGACCGCGTCTTCGTCG.

7. The lentiviral particle of claim 6 , wherein:

a. the at least one encoded shRNA comprises a sequence having at least 85% percent identity with:

(SEQ ID NO: 1)

  i.

GTCCTGGAGTACAATGCCATTCTCGAGAATGGCATTGTACTCCA

GGACTTTTT;

(SEQ ID NO: 2)

 ii.

GCAGGATTTCGTTCAGCACTTCTCGAGAAGTGCTGAACGAAATC

CTGCTTTTT;

(SEQ ID NO: 3)

iii.

GCCATGTACATGGCAGGAATTCTCGAGAATTCCTGCCATGTACA

TGGCTTTTT;

or

(SEQ ID NO: 4)

 iv.

GCAGAAGGAGGCTGAGAAAGTCTCGAGACTTTCTCAGCCTCCTT

CTGCTTTTT;

 or

b. the at least one encoded microRNA comprises a sequence having at least 85% percent identity with:

(SEQ ID NO: 5)

i.

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCCT

TCTGCGTGAAGCCACAGATGGCAGAAGGAGGCTGAGAAAGTGCTG

CCTACTGCCTCGGACTTCAAGGGGCT;

(SEQ ID NO: 6)

ii.

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCCT

TCTGCGTGAAGCCACAGATGGCAGAAGGGCTGAGAAAGTGCTGCC

TACTGCCTCGGACTTCAAGGGGCT;

(SEQ ID NO: 7)

iii.

TGCTGTTGACAGTGAGCGACTTTCTCAGCCTCCTTCTGCGTGAAG

CCACAGATGGCAGAAGGAGGCTGAGAAAGTTGCCTACTGCCTCGG

A;

(SEQ ID NO: 8)

iv.

CCTGGAGGCTTGCTGAAGGCTGTATGCTGACTTTCTCAGCCTCCT

TCTGCTTTTGGCCACTGACTGAGCAGAAGGGCTGAGAAAGTCAGG

ACACAAGGCCTGTTACTAGCACTCA;

(SEQ ID NO: 9)

v.

CATCTCCATGGCTGTACCACCTTGTCGGGACTTTCTCAGCCTCCT

TCTGCCTGTTGAATCTCATGGCAGAAGGAGGCGAGAAAGTCTGAC

ATTTTGGTATCTTTCATCTGACCA;

or

(SEQ ID NO: 10)

vi.

GGGCCTGGCTCGAGCAGGGGGCGAGGGATACTTTCTCAGCCTCCT

TCTGCTGGTCCCCTCCCCGCAGAAGGAGGCTGAGAAAGTCCTTCC

CTCCCAATGACCGCGTCTTCGTCG.

8. The lentiviral particle of claim 6 , wherein the envelope protein targets an endocytic compartment of the target cell.

9. The lentiviral particle of claim 6 , wherein the target cell is one or more cancer cells that are present in a cancer selected from one or more of a carcinoma, a leukemia, a lymphoma, a sarcoma, a myeloma, a mesothelioma, a mixed type, or mixtures thereof.

10. The lentiviral particle of claim 6 , wherein the target cell is one or more cancer cells that that are present in a hepatocellular carcinoma.

11. The lentiviral particle of claim 6 , wherein the target cell is capable of activating a gamma delta T cell following infection with the lentiviral particle.

12. The lentiviral particle of claim 6 , wherein the enzyme is FDPS.

13. A pharmaceutical combination for treating a condition associated with the mevalonate pathway, the pharmaceutical combination comprising: a lentiviral particle that is produced by a packaging cell and is capable of infecting a target cell, wherein the lentiviral particle comprises an envelope protein capable of infecting a target cell, and

a. at least one encoded shRNA capable of inhibiting production of an enzyme of the mevalonate pathway, wherein the at least one encoded shRNA comprises a sequence having at least 80% percent identity with:

(SEQ ID NO: 1)

  i.

GTCCTGGAGTACAATGCCATTCTCGAGAATGGCATTGTACTCCA

GGACTTTTT;

(SEQ ID NO: 2)

 ii.

GCAGGATTTCGTTCAGCACTTCTCGAGAAGTGCTGAACGAAATC

CTGCTTTTT;

(SEQ ID NO: 3)

iii.

GCCATGTACATGGCAGGAATTCTCGAGAATTCCTGCCATGTACA

TGGCTTTTT;

or

(SEQ ID NO: 4)

 iv.

GCAGAAGGAGGCTGAGAAAGTCTCGAGACTTTCTCAGCCTCCTT

CTGCTTTTT;

 or

b. at least one encoded microRNA capable of inhibiting production of an enzyme of the mevalonate pathway, wherein the at least one encoded microRNA comprises a sequence having at least 80% percent identity with:

(SEQ ID NO: 5)

  i.

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCC

TTCTGCGTGAAGCCACAGATGGCAGAAGGAGGCTGAGAAAGTGC

TGCCTACTGCCTCGGACTTCAAGGGGCT;

(SEQ ID NO: 6)

 ii.

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCC

TTCTGCGTGAAGCCACAGATGGCAGAAGGGCTGAGAAAGTGCTG

CCTACTGCCTCGGACTTCAAGGGGCT;

(SEQ ID NO: 7)

iii.

TGCTGTTGACAGTGAGCGACTTTCTCAGCCTCCTTCTGCGTGAA

GCCACAGATGGCAGAAGGAGGCTGAGAAAGTTGCCTACTGCCTC

GGA;

(SEQ ID NO: 8)

 iv.

CCTGGAGGCTTGCTGAAGGCTGTATGCTGACTTTCTCAGCCTCC

TTCTGCTTTTGGCCACTGACTGAGCAGAAGGGCTGAGAAAGTCA

GGACACAAGGCCTGTTACTAGCACTCA;

(SEQ ID NO: 9)

  v.

CATCTCCATGGCTGTACCACCTTGTCGGGACTTTCTCAGCCTCC

TTCTGCCTGTTGAATCTCATGGCAGAAGGAGGCGAGAAAGTCTG

ACATTTTGGTATCTTTCATCTGACCA;

or

(SEQ ID NO: 10)

 vi.

GGGCCTGGCTCGAGCAGGGGGCGAGGGATACTTTCTCAGCCTCC

TTCTGCTGGTCCCCTCCCCGCAGAAGGAGGCTGAGAAAGTCCTT

CCCTCCCAATGACCGCGTCTTCGTCG;

and

c. an aminobisphosphonate compound; separately or together.

14. The pharmaceutical combination of claim 13 , wherein the at least one encoded shRNA comprises a sequence having at least 85% percent identity with:

(SEQ ID NO: 1)

  i.

GTCCTGGAGTACAATGCCATTCTCGAGAATGGCATTGTACTCCA

GGACTTTTT;

(SEQ ID NO: 2)

 ii.

GCAGGATTTCGTTCAGCACTTCTCGAGAAGTGCTGAACGAAATC

CTGCTTTTT;

(SEQ ID NO: 3)

iii.

GCCATGTACATGGCAGGAATTCTCGAGAATTCCTGCCATGTACA

TGGCTTTTT;

or

(SEQ ID NO: 4)

 iv.

GCAGAAGGAGGCTGAGAAAGTCTCGAGACTTTCTCAGCCTCCTT

CTGCTTTTT;

or

b. the at least one encoded microRNA comprises a sequence having at least 85% percent identity with:

(SEQ ID NO: 5)

i.

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCCT

TCTGCGTGAAGCCACAGATGGCAGAAGGAGGCTGAGAAAGTGCTG

CCTACTGCCTCGGACTTCAAGGGGCT;

(SEQ ID NO: 6)

ii.

AAGGTATATTGCTGTTGACAGTGAGCGACACTTTCTCAGCCTCCT

TCTGCGTGAAGCCACAGATGGCAGAAGGGCTGAGAAAGTGCTGCC

TACTGCCTCGGACTTCAAGGGGCT;

(SEQ ID NO: 7)

iii.

TGCTGTTGACAGTGAGCGACTTTCTCAGCCTCCTTCTGCGTGAAG

CCACAGATGGCAGAAGGAGGCTGAGAAAGTTGCCTACTGCCTCGG

A;

(SEQ ID NO: 8)

iv.

CCTGGAGGCTTGCTGAAGGCTGTATGCTGACTTTCTCAGCCTCCT

TCTGCTTTTGGCCACTGACTGAGCAGAAGGGCTGAGAAAGTCAGG

ACACAAGGCCTGTTACTAGCACTCA;

(SEQ ID NO: 9)

v.

CATCTCCATGGCTGTACCACCTTGTCGGGACTTTCTCAGCCTCCT

TCTGCCTGTTGAATCTCATGGCAGAAGGAGGCGAGAAAGTCTGAC

ATTTTGGTATCTTTCATCTGACCA;

or

(SEQ ID NO: 10)

vi.

GGGCCTGGCTCGAGCAGGGGGCGAGGGATACTTTCTCAGCCTCCT

TCTGCTGGTCCCCTCCCCGCAGAAGGAGGCTGAGAAAGTCCTTCC

CTCCCAATGACCGCGTCTTCGTCG.

15. The pharmaceutical combination of claim 13 , wherein the aminobisphosphonate compound comprises zoledronic acid.

16. The pharmaceutic combination of claim 13 , wherein the target cell is one or more cancer cells that are present in a cancer selected from one or more of a carcinoma, a leukemia, a lymphoma, a sarcoma, a myeloma, a mesothelioma, a mixed type, or mixtures thereof.

17. The pharmaceutical combination of claim 13 , wherein the target cell is one or more cancer cells that are present in a hepatocellular carcinoma.

18. The pharmaceutical combination of claim 13 , wherein the target cell is capable of activating a gamma delta T cell following infection with the lentiviral particle.

19. The pharmaceutical combination of claim 13 , wherein the enzyme is FDPS.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Jun 20, 2018
From: PAUZA, CHARLES DAVID; LI, HAISHAN; LAHUSEN, TYLER; LIOU, MEI-LING
To: AMERICAN GENE TECHNOLOGIES INTERNATIONAL INC.
Reel/Frame 046147/0870 →
Continuity (4)
Continuation 15652080 · Jul 17, 2017
Continuation PCTUS2017013399 · Jan 13, 2017
Provisional Application 62279474 · Jan 15, 2016
Related Publication 20180142257A1 · May 24, 2018
Cited By (8)
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