US 12,258,580 B2
Process for generating therapeutic compositions of engineered cells
Sarah Y. Lee, Mercer Island, WA (US); Pascal Beauchesne, Seattle, WA (US); Mark L. Bonyhadi, Sammamish, WA (US); Ryan L. Crisman, Seattle, WA (US); Ryan P. Larson, Seattle, WA (US); Mary Mallaney, Seattle, WA (US); Christopher Glen Ramsborg, Seattle, WA (US); Clinton Weber, Seattle, WA (US); John Matthew Wesner, Seattle, WA (US); and Nathan Yee, Seattle, CA (US)
Assigned to Juno Therapeutics, Inc., Seattle, WA (US)
Appl. No. 16/760,240
Filed by Juno Therapeutics, Inc., Seattle, WA (US)
PCT Filed Oct. 31, 2018, PCT No. PCT/US2018/058590
§ 371(c)(1), (2) Date Apr. 29, 2020,
PCT Pub. No. WO2019/089855, PCT Pub. Date May 9, 2019.
Claims priority of provisional application 62/721,603, filed on Aug. 22, 2018.
Claims priority of provisional application 62/596,771, filed on Dec. 8, 2017.
Claims priority of provisional application 62/580,409, filed on Nov. 1, 2017.
Prior Publication US 2020/0354677 A1, Nov. 12, 2020
Int. Cl. C12N 5/0783 (2010.01); A61K 39/00 (2006.01)
CPC C12N 5/0636 (2013.01) [A61K 39/4611 (2023.05); A61K 39/4631 (2023.05); A61K 39/464412 (2023.05); A61K 2239/48 (2023.05); C12N 2501/2302 (2013.01); C12N 2501/2307 (2013.01); C12N 2501/2315 (2013.01); C12N 2501/999 (2013.01); C12N 2510/00 (2013.01)] 41 Claims
 
1. A method for producing a composition of engineered cells, the method comprising:
(a) incubating an input composition comprising T cells enriched for one or both of CD4+ and CD8+ primary human T cells, thereby generating a stimulated composition, wherein:
the primary human T cells are from a human subject having a disease or condition; and
the incubating is carried out under one or more stimulating conditions, the stimulating conditions comprising the presence of (i) an anti-CD3 antibody or antibody fragment thereof and an anti-CD28 antibody or antibody fragment thereof, and (ii) recombinant IL-2 and recombinant IL-15, wherein the concentration of recombinant IL-2 is from about 10 IU/mL to about 200 IU/mL and the concentration of recombinant IL-15 is from about 1 IU/mL to about 25 IU/mL;
(b) introducing a recombinant receptor into cells of the stimulated composition, thereby generating an engineered composition comprising engineered T cells, wherein the recombinant receptor is capable of binding to a target antigen that is expressed on a cell of the disease or condition; and
(c) cultivating the engineered composition under conditions to promote expansion of the engineered T cells, thereby producing an output composition comprising the engineered T cells that is suitable for an autologous cell therapy to treat the disease or condition of the subject, wherein:
(i) the cultivating is carried out in the presence of:
(1) recombinant IL-2 and recombinant IL-15, wherein the concentration of recombinant IL-2 is from about 50 IU/mL to about 500 IU/mL and the concentration of recombinant IL-15 is from about 5 IU/mL to about 50 IU/mL; and
(2) a surfactant, wherein the surfactant is a poloxamer added during the cultivating step;
(ii) at least a portion of the cultivating is performed with perfusion; and
(iii) the cultivating is performed for 2 days to 8 days, inclusive, wherein the cultivating is performed at least until the output composition comprises 4-fold or greater viable T cells compared to the composition prior to the cultivating; and
(d) collecting cells of the output composition after at most 8 days of the cultivating.