IP Library › Granted Patent US 10,329,543
Granted Patent B2
US 10,329,543 · App. 15/790,762 · Granted Jun 25, 2019

Modified stem cell memory T cells, methods of making and methods of using same

Inventors: Eric Ostertag (San Diego, CA); Devon Shedlock (San Diego, CA)
Assignee: Poseida Therapeutics, Inc.
C12N9/1241C07K14/7051C07K14/70564C07K14/70589C12N5/0636C12N9/644C12N15/85C12N2800/90
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Quick Facts
Patent No.
US 10,329,543
App. No.
15/790,762
Granted
Jun 25, 2019
Kind
B2
Abstract

The disclosure provides a method of producing modified stem memory T cells (e.g. CAR-T cells) for administration to a subject as, for example an adoptive cell therapy.

Claims (31)

1. A method of producing a plurality of expanded modified T cells, comprising

introducing into a plurality of primary human T cells (a) a transposon composition comprising a transposon comprising an antigen receptor, a therapeutic protein or a sequence encoding the same and (b) a transposase composition comprising a transposase or a sequence encoding the transposase to produce a plurality of modified T cells, under conditions that preserve desirable stem-like properties of the plurality of modified T cells;

wherein the conditions comprise culturing the plurality of modified T cells in vitro in a liquid culture media comprising a sterol; an alkane; phosphorus and one or more of an octanoic acid, a palmitic acid, a linoleic acid, and an oleic acid;

wherein at least 25% of the plurality of modified T cells expresses one or more cell-surface marker(s) of a stem memory T cell (T SCM ) or a T SCM -like cell and wherein the one or more cell-surface marker(s) comprises CD62L and CD45RA.

2. The method of claim 1 , wherein at least 60% of the plurality of modified T cells expresses one or more cell-surface marker(s) of a stem memory T cell (T SCM ) or a T SCM -like cell.

3. The method of claim 1 , wherein the transposon is a plasmid DNA transposon with a sequence encoding the antigen receptor or the therapeutic protein flanked by two cis-regulatory insulator elements.

4. The method of claim 1 , wherein the transposon is a piggyBac transposon.

5. The method of claim 4 , wherein the transposase is a piggyBac transposase.

6. The method of claim 5 , wherein the piggyBac transposase comprises an amino acid sequence comprising SEQ ID NO: 4.

7. The method of claim 6 , wherein the piggyBac transposase is a hyperactive variant and wherein the hyperactive variant comprises an amino acid substitution at one or more of positions 30, 165, 282 and 538 of SEQ ID NO: 4.

8. The method of claim 7 , wherein the amino acid substitution at position 30 of SEQ ID NO: 4 is a substitution of a valine (V) for an isoleucine (I) (I30V).

9. The method of claim 7 , wherein the amino acid substitution at position 165 of SEQ ID NO: 4 is a substitution of a serine (S) for a glycine (G) (G165S).

10. The method of claim 7 , wherein the amino acid substitution at position 282 of SEQ ID NO: 4 is a substitution of a valine (V) for a methionine (M) (M282V).

11. The method of claim 7 , wherein the amino acid substitution at position 538 of SEQ ID NO: 4 is a substitution of a lysine (K) for an asparagine (N) (N538K).

12. The method of claim 4 , wherein the transposase is a Super piggyBac (SPB) transposase.

13. The method of claim 12 , wherein the Super piggyBac (SPB) transposase comprises an amino acid sequence comprising SEQ ID NO: 5.

14. The method of claim 1 , wherein the sequence encoding the transposase is an mRNA sequence.

15. The method of claim 1 , wherein the transposon is a Sleeping Beauty transposon.

16. The method of claim 15 , wherein the transposase is a Sleeping Beauty transposase or a hyperactive Sleeping Beauty transposase (SB100X).

17. The method of claim 1 , wherein the transposon is a Helraiser transposon.

18. The method of claim 17 , wherein the transposase is a Helitron transposase.

19. The method of claim 1 , wherein the transposon is a Tol2 transposon.

20. The method of claim 19 , wherein the transposase is a Tol2 transposase.

21. The method of claim 1 , wherein the transposon is derived or recombined from any species.

22. The method of claim 1 , wherein the transposon is synthetic.

23. The method of claim 1 , wherein the antigen receptor is a T-cell receptor.

24. The method of claim 23 , wherein the T-cell receptor is not naturally-occurring.

25. The method of claim 1 , wherein the antigen receptor is a Chimeric Antigen Receptor (CAR).

26. The method of claim 25 , wherein the CAR is a CARTyrin.

27. The method of claim 1 , wherein the therapeutic protein is a secreted or secretable protein.

28. The method of claim 25 , wherein the CAR is a VCAR.

Assignments (1)
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Nov 27, 2017
From: OSTERTAG, ERIC; SHEDLOCK, DEVON
To: POSEIDA THERAPEUTICS, INC.
Reel/Frame 044223/0380 →
Continuity (1)
Related Publication 20190119658A1 · Apr 25, 2019
Cited By (3)
US 12,275,685 US 12,421,284 US 12,686,726