Granted Patent B2

US 12,239,693 · App. 17/050,949 · Granted Mar 4, 2025

Inventors: Jeffrey A. Medin (Shorewood, WI); Daniel H. Fowler (Milwaukee, WI); Murtaza S. Nagree (Milwaukee, WI); Tania Felizardo (Milwaukee, WI)

Assignee: The Medical College of Wisconsin, Inc.

A61K38/443A61K38/47A61K39/4611A61K39/464A61P43/00C12N5/0636C12N15/86A61K2239/31A61K2239/38C12N15/79C12N2510/00C12Y101/01205C12Y302/01022

Abstract

Claims (18)

1. A method comprising:

(a) conditioning T-cells from a subject with rapamycin ex vivo to generate T-Rapa cells, wherein the subject is suffering from Fabry disease;

(b) transducing the T-Rapa cells in vitro with a vector comprising a transgene of interest that encodes an enzyme associated with a lysosomal storage disorder, wherein the enzyme associated with the lysosomal storage disorder is a-galactosidase A (α-gal A);

(c) expanding the vector-transduced T-Rapa cells by culturing in vitro; and

(d) administering the expanded transduced T-Rapa cells of step (c) derived from the subject to the subject by intravenous infusion, wherein the T-Rapa cells express the enzyme associated with the lysosomal storage disorder in the subject and reduce a level of globotriaosylceramide (Gb 3 ) in the subject.

2. The method of claim 1 further comprising cryopreserving a portion of the expanded transduced T-Rapa cells for future administration to the subject.

3. The method of claim 1 further comprising detecting and isolating CD4+ T-cells from the subject and culturing the CD4+ T-cells in vitro prior to step (a), wherein the purity of the isolated CD4+ T cells is at least 75%.

4. The method of claim 1 , wherein step (a) comprises culturing the T-cells in chemically defined medium comprising about 0.1 to about 2 micromolar rapamycin.

5. The method of claim 4 , wherein the chemically defined medium further comprises recombinant human interleukin 4 (IL-4) and recombinant human interleukin 2 (IL-2).

6. The method of claim 1 , wherein the vector is a lentiviral vector.

7. The method of claim 6 , wherein the lentiviral vector is a dual promoter lentivirus vector, and wherein the vector expresses the transgene of interest and a mutant form of inosine-5′-monophosphate dehydrogenase 2 (IMPDH2(IY)) when transduced into the T-Rapa cells.

8. The method of claim 7 , wherein the vector comprises SEQ ID NO:11.

9. The method of claim 8 , wherein the method further comprises administering to the subject an amount of mycophenolate mofetil (MMF) sufficient to enrich the population of transduced T-Rapa cells in the subject.

10. The method of claim 1 , wherein the transgene is:

a) an a-galactosidase A (AGA) comprising SEQ ID NO: 1 or a sequence with at least 75% sequence identity to SEQ ID NO:1.

11. The method of claim 10 , wherein the vector is a lentiviral vector that comprises:

SEQ ID NO:2 or a sequence with at least 75% sequence identity to SEQ ID NO:2.

12. The method of claim 1 , wherein the reduced level of globotriaosylceramide (Gb 3 ) is detected in the blood, liver, spleen, heart or kidneys of the subject.

Assignments (1)

Continuity (2)

References Cited (101)