US 12,343,397 B2
Drug-resistant immune cells and methods of use thereof
Mark J. Osborn, St. Louis Park, MN (US); Keli Hippen, Minneapolis, MN (US); and Bruce R. Blazer, Minneapolis, MN (US)
Assigned to Regents of the University of Minnesota, Minneapolis, MN (US)
Appl. No. 17/056,233
Filed by Regents of the University of Minnesota, Minneapolis, MN (US)
PCT Filed May 16, 2019, PCT No. PCT/US2019/032686
§ 371(c)(1), (2) Date Nov. 17, 2020,
PCT Pub. No. WO2019/222513, PCT Pub. Date Nov. 21, 2019.
Claims priority of provisional application 62/672,868, filed on May 17, 2018.
Prior Publication US 2021/0213062 A1, Jul. 15, 2021
Int. Cl. A61P 37/06 (2006.01); A61K 31/436 (2006.01); A61K 31/573 (2006.01); A61K 35/28 (2015.01); A61K 38/13 (2006.01); A61K 40/10 (2025.01); A61K 40/11 (2025.01); A61K 40/22 (2025.01); A61K 40/41 (2025.01); A61K 45/06 (2006.01); C07K 14/705 (2006.01); C12N 5/0783 (2010.01); C12N 9/16 (2006.01); C12N 9/22 (2006.01); C12N 15/11 (2006.01); C12N 15/90 (2006.01)
CPC A61K 45/06 (2013.01) [A61K 31/436 (2013.01); A61K 31/573 (2013.01); A61K 35/28 (2013.01); A61K 38/13 (2013.01); A61K 40/10 (2025.01); A61K 40/11 (2025.01); A61K 40/22 (2025.01); A61K 40/418 (2025.01); A61P 37/06 (2018.01); C07K 14/705 (2013.01); C12N 5/0637 (2013.01); C12N 9/16 (2013.01); C12N 9/22 (2013.01); C12N 15/11 (2013.01); C12N 15/907 (2013.01); C12Y 301/03016 (2013.01); A61K 2239/31 (2023.05); A61K 2239/38 (2023.05); C12N 2310/20 (2017.05); C12N 2800/80 (2013.01)] 13 Claims
 
1. A method for generating a modified cell, comprising:
introducing a gene editing system into a regulatory T (Treg) cell that produces a modification in an exon, a splice donor, or a splice acceptor of a gene locus encoding for NR3C1, wherein the modification downregulates gene expression of NR3C1 and renders the modified cell steroid-resistant; and
inserting an exogenous calcineurin inhibitor resistance gene into the genome of the cell to render the modified cell calcineurin inhibitor (CNI)-resistant,
thereby generating a modified Treg cell.