Granted Patent B2

US 12,472,209 · App. 16/999,459 · Granted Nov 18, 2025

Inventors: Marcin Tomasz Kortylewski (Monrovia, CA); Karen Aboody (Arcadia, CA); Alexandra Jacqueline Annala (Pasadena, CA); Mohamed Hammad (Alhambra, CA)

Assignee: CITY OF HOPE

A61K35/30A61P35/00C12N5/0623C12N15/1138C12N2310/11C12N2310/17

Abstract

Claims (28)

1 . A neural stem cell comprising an exogenous nucleic acid, wherein the exogenous nucleic acid comprises a neural stem cell-loading sequence bonded to an immunostimulatory sequence via a linking group.

2 . The neural stem cell of claim 1 , wherein the neural stem cell is a human leukocyte antigen II-negative neural stem cell.

3 . The neural stem cell of claim 1 , wherein the exogenous nucleic acid comprises from about 10 base pairs to about 500 base pairs.

4 . The neural stem cell of claim 1 , wherein the neural stem cell-loading sequence comprises a Toll-like receptor 3-binding nucleic acid sequence, a Toll-like receptor 9-binding nucleic acid sequence, or a combination thereof.

5 . The neural stem cell of claim 4 , wherein the Toll-like receptor 9-binding nucleic acid sequence comprises a CpG oligodeoxynucleotide (ODN) selected from the group consisting of a CpG-A ODN, a CpG-B ODN, and a CpG-C ODN; and wherein the Toll-like receptor 3-binding nucleic acid sequence comprises dsRNA, polyinosine-polycytidylic acid, or a combination thereof.

6 . The neural stem cell of claim 1 , wherein the neural stem cell-loading sequence comprises CpG ODN 1585, CpG ODN 2216, CpG ODN 2336, CpG ODN 1668, CpG ODN 1826, CpG ODN 2006, CpG ODN 2007, CpG ODN BW006, CpG ODN D-SL01, CpG ODN 2395, CpG ODN M362, CpG ODN D-SL03, or CpG ODN D19.

7 . The neural stem cell of claim 1 , wherein the immunostimulatory sequence comprises a STAT-inhibiting nucleic acid sequence, a Toll like receptor 7/8-activating nucleic acid sequence, a STING receptor-activating nucleic acid sequence, or a RIG-1 receptor-activating nucleic acid sequence.

8 . The neural stem cell of claim 7 , wherein:

(i) the STAT-inhibiting nucleic acid sequence comprises a STAT3-inhibiting nucleic acid sequence, a STAT4-inhibiting nucleic acid sequence, a STAT5A-inhibiting nucleic acid sequence, a STAT5B-inhibiting nucleic acid sequence, a STAT6-inhibiting nucleic acid sequence, or a combination of two or more thereof;

(ii) the Toll like receptor 7/8-activating nucleic acid sequence comprises ssRNA, thymidine homopolymer oligodeoxynucleotide, guanosine, or an analog of any of the foregoing;

(iii) the STING receptor-activating nucleic acid sequence comprises a cGAMP, a c-di-CMP, a c-di-AMP, a c-AIMP, or an analog of any of the foregoing; and

(iv) the RIG-1 receptor-activating nucleic acid sequence comprises dsRNA, polyinosinic: polycytidylic acid, or an analog of any of the foregoing.

9 . The neural stem cell of claim 1 , wherein the immunostimulatory sequence comprises a STAT3-inhibiting nucleic acid sequence.

10 . The neural stem cell of claim 9 , wherein the STAT3-inhibiting nucleic acid sequence comprises a nucleic acid sequence having at least 90% sequence identity to SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, SEQ ID NO:35, or a combination of two or more thereof.

11 . The neural stem cell of claim 9 , wherein the exogenous nucleic acid comprises a Toll-like receptor 9-binding nucleic acid sequence covalently bonded to the STAT3-inhibiting nucleic acid sequence via the linking group.

12 . The neural stem cell of claim 1 , wherein the neural stem cell-loading sequence is a nucleic acid sequence having at least 90% sequence identity to CpG ODN 1585, CpG ODN 2216, CpG ODN 2336, CpG ODN 1668, CpG ODN 1826, CpG ODN 2006, CpG ODN 2007, CpG ODN BW006, CpG ODN D-SL01, CpG ODN 2395, CpG ODN M362, CpG ODN D-SL03, or CpG ODN D19; and wherein the immunostimulatory sequence is a nucleic acid sequence having at least 90% sequence identity to SEQ ID NO:17, SEQ ID NO:18, SEQ ID NO:19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO:24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO:30, SEQ ID NO:31, SEQ ID NO:32, or SEQ ID NO:35.

13 . The neural stem cell of claim 1 , wherein the linking group comprises a bond, a nucleic acid sequence, a DNA sequence, substituted or unsubstituted alkylene, substituted or unsubstituted heteroalkylene, substituted or unsubstituted cycloalkylene, substituted or unsubstituted heterocycloalkylene, substituted or unsubstituted arylene, substituted or unsubstituted heteroarylene, or a combination of two or more thereof.

14 . The neural stem cell of claim 1 , wherein the linking group comprises a substituted 6 to 60 membered heteroalkylene.

15 . The neural stem cell of claim 1 , wherein the linking group comprises a substituted heteroalkylene of the formula:

wherein each X is independently —OH or —O—, and n is an integer from 1 to 10.

16 . The neural stem cell of claim 1 , wherein the exogenous nucleic acid comprises a nucleic acid sequence having at least 90% sequence identity to SEQ ID NO:1, SEQ ID NO:2, SEQ ID NO:3, SEQ ID NO:4, SEQ ID NO:5, SEQ ID NO:6, SEQ ID NO:7, SEQ ID NO: 8, SEQ ID NO:9, SEQ ID NO: 10, SEQ ID NO:11, SEQ ID NO: 12, SEQ ID NO: 13, SEQ ID NO: 14, SEQ ID NO: 15, SEQ ID NO:16, SEQ ID NO:33, or SEQ ID NO:34.

17 . The neural stem cell of claim 1 , wherein the exogenous nucleic acid further comprises a detectable moiety selected from the group consisting of an enzyme, biotin, digoxigenin, a paramagnetic molecule, a contrast agent, gadolinium, a radioisotope, radionuclide, fluorodeoxyglucose, barium sulfate, thorium dioxide, gold, a fluorophore, a hapten, a protein, a fluorescent moiety, and a combination of two or more thereof.

18 . A pharmaceutical composition comprising the neural stem cell of claim 1 and a pharmaceutically acceptable excipient.

19 . A method of treating cancer, a neurodegenerative disease, an inflammatory disease, or a viral infection in a patient in need thereof, the method comprising administering to the patient an effective amount of the neural stem cell of claim 1 .

20 . A process for producing the neural stem cell of claim 1 , the process comprising contacting the neural stem cell with the exogenous nucleic acid for a time sufficient for the neural stem cell to uptake the exogenous nucleic acid.

21 . The neural stem cell of claim 1 , wherein the neural stem cell-loading sequence is CpG ODN 1585, CpG ODN 2216, CpG ODN 2336, CpG ODN 1668, CpG ODN 1826, CpG ODN 2006, CpG ODN 2007, CpG ODN BW006, CpG ODN D-SL01, CpG ODN 2395, CpG ODN M362, CpG ODN D-SL03, or CpG ODN D19; and wherein the immunostimulatory sequence is a nucleic acid sequence set forth by SEQ ID NO:17, SEQ ID NO: 18, SEQ ID NO: 19, SEQ ID NO:20, SEQ ID NO:21, SEQ ID NO:22, SEQ ID NO:23, SEQ ID NO: 24, SEQ ID NO:25, SEQ ID NO:26, SEQ ID NO:27, SEQ ID NO:28, SEQ ID NO:29, SEQ ID NO: 30, SEQ ID NO:31, SEQ ID NO:32, or SEQ ID NO:35.

22 . The neural stem cell of claim 1 , wherein the neural stem cell comprises a vesicle encapsulated therein and wherein the exogenous nucleic acid is encapsulated within the vesicle.

23 . The neural stem of claim 1 , wherein the neural stem cell-loading sequence is a CpG oligodeoxynucleotide and the immunostimulatory sequence is a STAT3 antisense oligonucleotide.

Assignments (1)

Continuity (2)

References Cited (33)