3D-organoid culture supports differentiation of human CAR+ iPSCs into highly functional CAR T cells

Human organoids: New strategies and methods for analyzing human development and disease

“…A variety of tumor organoids derived from different patient cancers have been co-cultured with CAR T cells to test their specificity and efficacy in attacking cancer cells, including glioblastoma (Jacob et al, 2020c) and colorectal cancer organoids (Schnalzger et al, 2019). Besides modeling different types of cancers, organoid technology can facilitate the production of immune cells for cancer immunotherapy (Wang et al, 2022). Using a multi-step artificial thymic organoids (ATO) differentiation protocol allowed successful generation of CAR T cells with potent antitumor activity against leukemia in vivo (Wang et al, 2022), an approach that could facilitate CAR T cell production and enable "off-the-shelf" manufacturing strategies.…”

“…Besides modeling different types of cancers, organoid technology can facilitate the production of immune cells for cancer immunotherapy (Wang et al, 2022). Using a multi-step artificial thymic organoids (ATO) differentiation protocol allowed successful generation of CAR T cells with potent antitumor activity against leukemia in vivo (Wang et al, 2022), an approach that could facilitate CAR T cell production and enable "off-the-shelf" manufacturing strategies.…”

Applications of human organoids in the personalized treatment for digestive diseases

“… 431 Organoids have also been shown to be effective platforms for evaluating CAR-T-cell potency and tumor specificity. 417 , 432 , 433 In HER2-expressing colorectal cancer organoids obtained from patient biopsies, the addition of anti-HER2 CAR-T cells alone resulted in minimal killing, while the combination of CAR-T cells with the apoptosis antagonist birinapant induced powerful lethality of organoids. 434 Colonic organoids were also used to track CAR-mediated NK-cell cytotoxicity.…”

Stem cell-based multi-tissue platforms to model human autoimmune diabetes

“…Current protocols progress through intermediate CD34 + hematopoietic progenitors, and invariably transfer CD34 + progenitors to mouse stromal lines that overexpress signals critical for T cell specification, such as the NOTCH ligands Delta-like 1 (DLL1) or Delta-like 4 (DLL4) [ [85] , [86] , [87] , [88] , [89] ] ( Figure 3 ). In two-dimensional (2D) co-culture methods using these stromal cells, there is limited positive selection of thymocytes and the cells generated are usually immature CD4 + CD8 + T cell progenitors or mature CD8 + effector T cells with very few CD4 + T cells [ [94] , [95] , [96] ]. Furthermore, T cell development is biased towards unconventional T cells that express the CD8αα + homodimer, an inefficient T cell receptor (TCR) coreceptor, instead of the HLA class I-restricted CD8αβ + that is expressed on conventional T cells [ 86 , 92 , 97 ].…”

“…Furthermore, T cell development is biased towards unconventional T cells that express the CD8αα + homodimer, an inefficient T cell receptor (TCR) coreceptor, instead of the HLA class I-restricted CD8αβ + that is expressed on conventional T cells [ 86 , 92 , 97 ]. To circumvent this issue, a three-dimensional (3D) artificial thymic organoid (ATO) system has been developed, which promotes positive selection and can support the development of functional conventional CD8αβ + T cells [ [94] , [95] , [96] ]. High efficiency differentiation of CD4+ T cells as well as negative selection, the intra-thymic process of eliminating thymocytes that express T cell receptors with high affinity for self-antigens, is nonetheless still lacking in this xenogeneic culture system.…”