IP Library › Granted Patent US 12,485,187
Granted Patent B2
US 12,485,187 · App. 18/597,488 · Granted Dec 2, 2025

Method for improving retroviral transduction and gene editing in hematopoietic stem cells using clyclosporin H and UM171

Inventors: Anna Christina Kajaste-Rudnitski (Milan, IT); Carolina Petrillo (Milan, IT); Bernhard Rudolf Gentner (Milan, IT); Luigi Naldini (Milan, IT); Pietro Genovese (Milan, IT); Giulia Schiroli (Milan, IT)
Assignees: Ospedale San Raffaele S.R.L.; Fondazione Telethon
A61K48/0008C07K7/645C12N5/0647C12N15/69C12N15/86C12N2740/15043
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Quick Facts
Patent No.
US 12,485,187
App. No.
18/597,488
Granted
Dec 2, 2025
Kind
B2
Abstract

Use of cyclosporin H (CsH) or a derivative thereof for increasing the efficiency of transduction of an isolated population of cells by a viral vector and/or increasing the efficiency of gene editing of an isolated population of cells when transduced by a viral vector.

Claims (30)

1 . A method of transducing a population of CD34+ HSPCs comprising the steps of:

(a) culturing the population of HSPCs in the presence of UM171;

(b) contacting the population of HSPCs with cyclosporin H (CsH); and

(c) transducing the population of HSPCs with a viral vector, wherein the viral vector is a lentiviral or gammaretroviral vector, and wherein the viral vector is pseudotyped to enter cells via an endocytosis-dependent mechanism.

2 . The method of claim 1 , wherein the viral vector is a VSV-g pseudotyped vector.

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

4 . The method of claim 1 , wherein the viral vector is an integration-defective lentiviral vector (IDLV).

5 . The method of claim 1 , wherein the HSPCs are stimulated HSPCs.

6 . The method of claim 1 , wherein the population of HSPCs is cultured in step (a) in the further presence of StemRegenin 1 (SR-1).

7 . The method of claim 1 , wherein the population of HSPCs is cultured in step (a) in the further presence of prostaglandin E2.

8 . The method of claim 7 , wherein the prostaglandin E2 is 16-16 dimethyl prostaglandin E2.

9 . The method of claim 1 , wherein the viral vector comprises a nucleotide of interest.

10 . The method of claim 9 , wherein the nucleotide of interest is a donor template.

11 . The method of claim 1 , wherein the method further comprises the step:

(d) contacting the population of HSPCs with a CRISPR/Cas system.

12 . The method of claim 11 , wherein the CRISPR/Cas system comprises a ribonucleoprotein (RNP) comprising Cas protein and gRNA.

13 . The method of claim 11 , wherein step (d) comprises electroporating the population of HSPCs.

14 . The method of claim 1 , wherein efficiency of transduction of the population of HSPCs by the viral vector is increased and/or efficiency of gene editing of the population of HSPCs when transduced by the viral vector is increased.

15 . The method of claim 1 , wherein the percentage of cells transduced by the viral vector is increased and/or the vector copy number per cell is increased.

16 . The method of claim 1 , wherein the method is carried out in vitro or ex vivo.

17 . The method of claim 1 , wherein the CsH is at a concentration of about 1-50 μM.

18 . The method of claim 1 , comprising a further step of enriching the population for CD34+/CD38-cells.

19 . The method of claim 1 , wherein the method comprises the steps of:

(a) culturing the population of HSPCs in the presence of UM171;

(b) contacting the population of HSPCs with CsH;

(c) transducing the population of HSPCs with an IDLV comprising a donor template;

(d) contacting the population of HSPCs with a CRISPR/Cas system.

20 . The method of claim 19 , wherein the CRISPR/Cas system comprises a ribonucleoprotein (RNP) comprising Cas protein and gRNA.

21 . The method of claim 1 , wherein the HSPCs are from umbilical cord blood, mobilized peripheral blood, or bone marrow.

22 . The method of claim 1 , wherein the HSPCs are CD34+CD38-HSPCs.

Assignments (2)
CHANGE OF NAME Recorded Jul 13, 2024
From: FONDAZIONE TELETHON
To: FONDAZIONE TELETHON ETS
Reel/Frame 068298/0069 →
ASSIGNMENT OF ASSIGNOR'S INTEREST Recorded Apr 3, 2024
From: KAJASTE-RUDNITSKI, ANNA CHRISTINA; PETRILLO, CAROLINA; GENTNER, BERNHARD RUDOLPH; NALDINI, LUIGI; GENOVESE, PIETRO; SCHIROLI, GIULIA
To: OSPEDALE SAN RAFFAELE S.R.L.; FONDAZIONE TELETHON
Reel/Frame 066996/0528 →
Priority Claims (1)
GB 1706394 · Apr 21, 2017 · national
Continuity (2)
Division 16606633
Related Publication 20240350661A1 · Oct 24, 2024
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