Escape from cell-cell and cell-matrix adhesion dependence underscores disease progression in gastric cancer organoid models

Abstract

ObjectiveCell-cell (CC) and cell-matrix (CM) adhesions are essential for epithelial cell survival, yet dissociation-induced apoptosis is frequently circumvented in malignant cells.DesignWe explored CC and CM dependence in 58 gastric cancer (GC) organoids by withdrawing either ROCK inhibitor, matrix or both to evaluate their tumorigenic potential in terms of apoptosis resistance, correlation with oncogenic driver mutations and clinical behaviour. We performed mechanistic studies to determine the role of diffuse-type GC drivers: ARHGAP fusions, RHOA and CDH1, in modulating CC (CCi) or CM (CMi) adhesion independence.Results97% of the tumour organoids were CMi, 66% were CCi and 52% were resistant to double withdrawal (CCi/CMi), while normal organoids were neither CMi nor CCi. Clinically, the CCi/CMi phenotype was associated with an infiltrative tumour edge and advanced tumour stage. Moreover, the CCi/CMi transcriptome signature was associated with poor patient survival when applied to three public GC datasets. CCi/CMi and CCi phenotypes were enriched in diffuse-type GC organoids, especially in those with oncogenic driver perturbation of RHO signalling via RHOA mutation or ARHGAP fusions. Inducible knockout of ARHGAP fusions in CCi/CMi tumour organoids led to resensitisation to CC/CM dissociation-induced apoptosis, upregulation of focal adhesion and tight junction genes, partial reversion to a more normal cystic phenotype and inhibited xenograft formation. Normal gastric organoids engineered with CDH1 or RHOA mutations became CMi or CCi, respectively.ConclusionsThe CCi/CMi phenotype has a critical role in malignant transformation and tumour progression, offering new mechanistic information on RHO-ROCK pathway inhibition that contributes to GC pathogenicity.

Divergent lineage trajectories and genetic landscapes in human gastric intestinal metaplasia organoids associated with early neoplastic progression

“…All 3 CMi IMOs remained sensitive to CC dissociation-induced apoptosis and died upon Y-27632 withdrawal (online supplemental figure 10B). As cell-cell adhesion independence (CCi) was previous observed mostly in advanced stage or diffuse type gastric cancer, 47 their absence in IMOs is consistent with their relatively early stage of neoplastic progression. We did not find any commonly mutated cancer driver genes among the 3 IM spheroids that correlated with the CMi phenotype (figure 6D, online supplemental table 11).…”

“…We recently showed that almost all GC organoids were cell-matrix adhesion independent (CMi), while normal gastric organoids were not, suggesting that gaining CMi represents the earliest critical step in malignant transformation. 47 Therefore, we sought to test whether IMOs were CMi by randomly selecting 10 IMOs and 10 AOs/BOs for spheroid formation assay (figure 1A). After 2 weeks in culture, there was a significant growth inhibition, irrespective of the organoid type (figure 7D).…”

Integrated multi‐omics profiling landscape of organising pneumonia

“… 22 While these adhesive dynamics are vital for normal cell survival, cancer cells often exploit this mechanism to evade apoptosis, highlighting potential therapeutic targets. 23 Our hypothesis was that the downregulation of cell matrix adhesion and developmental growth involved in the morphogenesis of macrophages led to epithelial cell death and morphological changes of OP. Apoptosis of lung alveolar epithelial cells (AECs) is an early pathogenic event in the development of OP, although its causative mechanism is still unclear.…”

Success rate of current human-derived gastric cancer organoids establishment and influencing factors: A systematic review and meta-analysis

“…Five studies that provided detailed clinical information on all lines of successful and failed construction of GCOs were further analyzed by different subgroups[ 21 - 27 , 37 ] (Table 5 ).…”