US 12,403,110 B2
Methods and compositions for treating cancers
Frank Griscelli, Villejuif (FR); Ali Turhan, Villejuif (FR); Annelise Bennaceur Griscelli, Villejuif (FR); and Christophe Desterke, Villejuif (FR)
Assigned to INSTITUT NATIONAL DE LA SANTÉ ET DE LA RECHERCHE MÉDICALE (INSERM), Paris (FR); ASSISTANCE PUBLIQUE-HÔPITAUX DE PARIS, Paris (FR); UNIVERSITÉ PARIS-SACLAY, Paris (FR); and UNIVERSITÉ PARIS CITÉ, Paris (FR)
Filed by INSTITUT NATIONAL DE LA SANTÉ ET DE LA RECHERCHE MÉDICALE (INSERM), Paris (FR); ASSISTANCE PUBLIQUE—HÔPITAUX DE PARIS, Paris (FR); UNIVERSITÉ PARIS-SACLAY, Saint-Aubin (FR); and UNIVERSITÉ PARIS CITÉ, Paris (FR)
Filed on Jun. 9, 2023, as Appl. No. 18/207,752.
Application 18/207,752 is a division of application No. 16/766,438, granted, now 11,679,148, previously published as PCT/EP2018/082429, filed on Nov. 23, 2018.
Claims priority of application No. 17306635 (EP), filed on Nov. 24, 2017.
Prior Publication US 2023/0338494 A1, Oct. 26, 2023
Int. Cl. A61K 35/28 (2015.01); A61K 31/19 (2006.01); A61K 35/30 (2015.01); A61K 35/34 (2015.01); A61K 35/35 (2015.01); A61K 35/545 (2015.01); A61K 39/39 (2006.01); A61K 40/00 (2025.01); A61K 40/10 (2025.01); A61K 40/42 (2025.01); A61P 35/00 (2006.01); A61K 39/00 (2006.01)
CPC A61K 31/19 (2013.01) [A61K 39/39 (2013.01); A61K 40/10 (2025.01); A61K 40/4242 (2025.01); A61P 35/00 (2018.01); A61K 2039/515 (2013.01); A61K 2239/31 (2023.05); A61K 2239/38 (2023.05); A61K 2239/54 (2023.05)] 15 Claims
 
1. A method for producing a vaccine composition comprising a population of inactivated cells, comprising:
(a) expanding pluripotent stem cells, optionally in the presence of a mutagenic agent;
(b) differentiating the expanded pluripotent stem cells along an ectodermal, endodermal, or mesodermal lineage pathway;
(c) culturing the differentiated cells in a two-dimensional (2D) culture system or a three-dimensional (3D) organoid culture system while optionally exposing the differentiated cells to a mutagenic agent to further induce genetic changes;
(d) verifying that at least 70% of the differentiated cells express fetal cell markers, wherein:
the fetal cell markers expressed by the cells differentiated along the endoderm pathway are selected from SOX17, CXCR4, FOXA1, FOXA2, FOXA3, HHEX, GATA4, GATA6, HNF1B, HNF4A, TF, ALB, TBX3, AFP, TTR, CER1, MIXL1, LHX1, GSC, PAX9, NEPN, SHH, PYY, MNX1, KITL, CLDN4, CLDN8, GFPT2, KRT19, SORCS2, EPPK1, NEDD9, PLAT, VTN, PDX1, TMPRSS4, CLIC6, RIPK4, CLDN8, and ST1A;
the fetal cell markers expressed by the cells differentiated along the ectoderm pathway are selected from PCGF4, PAX6, PAX7, CXCR4, SOX1, SOX2, SOX10, ITGB1, FABP7, NES, FUT4, PROM1, MELK, MS11, MAP2, DCX, NCAM1, TUBB3, SLC1A3, CD44, S100B, VIM, GFAP, CNP, OLIG2, CA2, CSPG4, TAZ, MSX1, SPARC, ID2, NES, NKX2.2, NKX6-1, FOXP2, FOXD3, and ZIC1; and
the fetal cell markers expressed by the cells differentiated along the mesoderm pathway are selected from Brachury (T), MIXL1, SNA11, SNA12, HLX, EOMES, MESP1, MESP2, TBX6, MEST, NKX2-5, and KDR;
(e) optionally, verifying that the differentiated cells express at least one tumor associated antigen (TAA) or neo-antigen that is present in cancer cells; and
(f) inactivating the differentiated cells by irradiation so they lose their ability to divide, thereby obtaining a population of inactivated cells, wherein no more than 10% of the population of inactivated cells express the following makers characteristic of pluripotency: NANOG, POU5F1 (Oct4), SSEA4, Tra-1-81, and Tra-1-60; and
(g) formulating the inactivated cells into an effective vaccine composition.