Reprogramming anchorage dependency by adherent-to-suspension transition promotes metastatic dissemination

Abstract

Background Although metastasis is the foremost cause of cancer-related death, a specialized mechanism that reprograms anchorage dependency of solid tumor cells into circulating tumor cells (CTCs) during metastatic dissemination remains a critical area of challenge. Methods We analyzed blood cell-specific transcripts and selected key Adherent-to-Suspension Transition (AST) factors that are competent to reprogram anchorage dependency of adherent cells into suspension cells in an inducible and reversible manner. The mechanisms of AST were evaluated by a series of in vitro and in vivo assays. Paired samples of primary tumors, CTCs, and metastatic tumors were collected from breast cancer and melanoma mouse xenograft models and patients with de novo metastasis. Analyses of single-cell RNA sequencing (scRNA-seq) and tissue staining were performed to validate the role of AST factors in CTCs. Loss-of-function experiments were performed by shRNA knockdown, gene editing, and pharmacological inhibition to block metastasis and prolong survival. Results We discovered a biological phenomenon referred to as AST that reprograms adherent cells into suspension cells via defined hematopoietic transcriptional regulators, which are hijacked by solid tumor cells to disseminate into CTCs. Induction of AST in adherent cells 1) suppress global integrin/ECM gene expression via Hippo-YAP/TEAD inhibition to evoke spontaneous cell–matrix dissociation and 2) upregulate globin genes that prevent oxidative stress to acquire anoikis resistance, in the absence of lineage differentiation. During dissemination, we uncover the critical roles of AST factors in CTCs derived from patients with de novo metastasis and mouse models. Pharmacological blockade of AST factors via thalidomide derivatives in breast cancer and melanoma cells abrogated CTC formation and suppressed lung metastases without affecting the primary tumor growth. Conclusion We demonstrate that suspension cells can directly arise from adherent cells by the addition of defined hematopoietic factors that confer metastatic traits. Furthermore, our findings expand the prevailing cancer treatment paradigm toward direct intervention within the metastatic spread of cancer.

Anchorage Dependence and Cancer Metastasis

“…2 ). 20 The gene IKZF1 is a key regulator of lymphocyte differentiation and is closely linked to leukemogenesis, particularly in acute lymphoblastic leukemia. 46 NFE2 is traditionally recognized for regulating erythropoiesis.…”

“…Moreover, the omission of any AST factor significantly diminishes the efficiency of AST, suggesting a cooperative role in facilitating this transition. 20 …”

“…Further investigation revealed that AST reprograms anchorage dependence via affecting cell-ECM interactions rather than cell-cell interactions. 20 The YAP/TEAD pathway is the key to this reprogramming, and cell detachment depends on its inactivation by AST factors ( Fig. 2 ).…”

“…The concept of an “adherent-to-suspension transition (AST)” has recently emerged as a significant factor in promoting survival in an anchorage-independent condition. 20 This transition entails a profound reconfiguration of cellular mechanisms, encompassing modifications in gene expression, signaling pathways, and metabolic activity that facilitate detachment and cell survival.…”

RXR Agonists Enhance Lenalidomide Anti-Myeloma Activity and T Cell Functions while Retaining Glucose-Lowering Effect

“…Immunodeficient (SCID) mice (male, 6-8 weeks old; OCI) were injected subcutaneously with 1 × 10 7 MM1.R in the lower right flank. When tumors were palpable (approximately 21 days after injection), the mice were randomly assigned into 4 groups (n = 7 in each group), receiving an intraperitoneal injection with 10 mg/kg/day [37,38] lenalidomide or in combination with LG100754 5 mg/kg three times a week. The tumors were measured with external calipers every 3 days, and the tumor volume (in mm 3 ) was calculated using the following standard formula: V = 0.5a × b 2 , where "a" and "b" are the long and short diameters of the tumor, respectively.…”