NOTCH pathway inactivation reprograms stem-like oral cancer cells to JAK-STAT dependent state and provides the opportunity of synthetic lethality

• Published in: Translational oncology Vol. 32; p. 101669
• Main Authors: Ghosh, Subhashis, Mitra, Paromita, Saha, Uday, Nandi, Rimpa, Jena, Subhashree, Ghosh, Arnab, Roy, Shantanu Saha, Acharya, Moulinath, Biswas, Nidhan Kumar, Singh, Sandeep
• Format: Journal Article
• Language: English
• Published: United States Elsevier Inc 01.06.2023; Neoplasia Press; Elsevier
• Subjects: Cellular plasticity; JAK-STAT; NOTCH; Oral cancer; Stemness; Synthetic lethal pair; NOTCH; Stemness; Oral cancer; Cellular plasticity; JAK-STAT; Synthetic lethal pair
• ISSN: 1936-5233; 1936-5233
• DOI: 10.1016/j.tranon.2023.101669

•Cellular plasticity drives non-genetic heterogeneity in stem-like cancer cells in oral cancer.•5-genes signature obtained from NOTCH-inactive population correlated with poor-prognosis.•NOTCH-HES and JAK-STAT pathway displayed synthetic lethal interaction against stemness.•JAK-inhibitors targeted the tumor-initiating ability of NOTCH-inactive oral cancer cells. We have recently provided the evidence of interconvertible cellular states, driving non-genetic heterogeneity among stem-like oral cancer cells (oral-SLCCs). Here, NOTCH pathway-activity status is explored as one of the possible mechanisms behind this stochastic plasticity. Oral-SLCCs were enriched in 3D-spheroids. Constitutively-active and inactive status of NOTCH pathway was achieved by genetic or pharmacological approaches. RNA sequencing and real-time PCR was performed for gene expression studies. in vitro cytotoxicity assessments were performed by AlamarBlue assay and in vivo effects were studied by xenograft growth in zebrafish embryo. We have observed stochastic plasticity in oral-SLCCs, spontaneously maintaining both NOTCH-active and inactive states. While cisplatin refraction was associated with post-treatment adaptation to the active-state of NOTCH pathway, oral-SLCCs with inactive NOTCH pathway status showed aggressive tumor growth and poor prognosis. RNAseq analysis clearly suggested the upregulation of JAK-STAT pathway in NOTCH pathway-inactive subset. The 3D-spheroids with lower NOTCH-activity status displayed significantly higher sensitivity to JAK-selective drugs, Ruxolitinib or Tofacitinib or siRNA mediated downregulation of tested partners STAT3/4. Oral-SLCCs were programmed to adapt the inactive status of NOTCH pathway by exposing to γ-secretase inhibitors, LY411575 or RO4929097, followed by targeting with JAK-inhibitors, Ruxolitinib or Tofacitinib. This approach resulted in a very significant inhibition in viability of 3D-spheroids as well as xenograft initiation in Zebrafish embryos. Study revealed for the first time that NOTCH pathway-inactive state exhibit activation of JAK-STAT pathways, as synthetic lethal pair. Therefore, co-inhibition of these pathway may serve as novel therapeutic strategy against aggressive oral cancer. [Display omitted]