	US 12,239,647 B2
	ALS treatment using induced regulatory T (iTREG) cells
Daniel Harding Fowler, Bethesda, MD (US)
Assigned to Rapa Therapeutics, LLC, Rockville, MD (US)
Filed by RAPA THERAPEUTICS, LLC, Rockville, MD (US)
Filed on May 14, 2021, as Appl. No. 17/320,891.
Application 17/320,891 is a continuation of application No. PCT/US2019/061818, filed on Nov. 15, 2019.
Claims priority of provisional application 62/927,075, filed on Oct. 28, 2019.
Claims priority of provisional application 62/768,176, filed on Nov. 16, 2018.
Prior Publication US 2021/0275591 A1, Sep. 9, 2021
Int. Cl. A61P 25/28 (2006.01); A61K 9/00 (2006.01); A61K 31/513 (2006.01); A61K 31/675 (2006.01); A61K 31/7056 (2006.01); A61K 38/17 (2006.01); A61K 39/00 (2006.01); C12N 5/0783 (2010.01); G01N 33/50 (2006.01)

CPC A61K 31/513 (2013.01) [A61K 9/0019 (2013.01); A61K 31/675 (2013.01); A61K 31/7056 (2013.01); A61K 38/1793 (2013.01); A61K 39/4611 (2023.05); A61K 39/4621 (2023.05); A61K 39/46432 (2023.05); A61P 25/28 (2018.01); C12N 5/0637 (2013.01); G01N 33/5088 (2013.01); A61K 2239/31 (2023.05); A61K 2239/38 (2023.05); A61K 2239/47 (2023.05); C12N 2501/06 (2013.01); C12N 2501/15 (2013.01); C12N 2501/2302 (2013.01); C12N 2501/2304 (2013.01); G01N 2333/46 (2013.01); G01N 2800/52 (2013.01)]

17 Claims

1. A method for manufacturing induced hybrid T_REG/Th2 cells, comprising:

culturing T cells in a first culture medium comprising vitamin D, temsirolimus, and an IL-2 signaling inhibitor for a first period of time sufficient to yield de-differentiated T cells;

separating said de-differentiated T cells from the first culture medium;

culturing de-differentiated T cells in a second culture medium comprising IL-2, IL-4, and TGF-β;

adding anti-CD3/anti-CD28 coated magnetic beads at a ratio of 3:1 (bead:T cell ratio) to the second culture medium;

incubating said de-differentiated T cells in the second culture medium for a second period of time to yield induced hybrid T_REG/Th2 cells.