Digital.CSIC. Repositorio Institucional del CSIC

oai:digital.csic.es:10261/351982

Supplementary table 1. Analysis of CD10, CD184, CD19, CD133, CD166 and CD44 positive subpopulations by FACS in HNSCC cell lines.-- Supplementary Table 2. Differential genes common to CD10, CD184, CD19 and NAMPT subpopulations obtained from transcriptomic analysis. All the genes in color are related to tumorigenic process, acting as oncogenes (in red), tumor suppressor genes (in blue), ambiguous genes depending on the type of the tumor (in green).-- Supplementary Table 3: IC50s for NAMPT inhibitors in parental and NAMPT CRISPR clones of both cell lines.-- Supplementary figure 1: Verification of NAMPT overexpression in RPMI and Detroit HNSCC cell lines. A: Western blot showing increased NAMPT ectopic overexpression. B: increased NAD total and NAD+ pools in cells overexpressing NAMPT (in orange).-- Supplementary Figure 2. Overall survival of HNSCC patients from the TCGA database. Kaplan-Meier curves show the overall survival of HNSCC patients with high and low expression levels of CD10, CD184, CD19, CD133, CD166 and NAMPT genes from the TCGA (The Cancer Genome Atlas) database, N=520.-- Supplementary Figure 3. GO term analysis. Analysis of the genes by the terms GO biological process, molecular function and cellular component (p<0.05).-- Supplementary Figure 4. Transcriptomic analysis of differential genes common to CD10, CD184, CD19 and NAMPT subpopulations in HNSCC cell lines. Venn diagram represents the differential genes common to positive and negative CD10, CD184 and CD19 populations and NAMPT overexpression and CRISPRs in RPMI-2650 and Detroit-562 cell lines obtained by NGS sequencing.-- Supplementary Figure 5: Densitometric quantification of NAMPT expression in the WB of figure 2C.-- Supplementary Figure 6: Densitometric quantification of NAMPT and NAPRT expression in the WB of Figure 5C.-- Supplementary materials: Original images of western blots., Head and neck squamous cell carcinoma (HNSCC) is a heterogeneous group of tumors that affect different anatomical locations. Despite this heterogeneity, HNSCC treatment depends on the anatomical location, TNM stage and resectability of the tumor. Classical chemotherapy is based on platinum-derived drugs (cisplatin, carboplatin and oxaliplatin), taxanes (docetaxel, paclitaxel) and 5-fluorouracil1. Despite advances in HNSCC treatment, the rate of tumor recurrence and patient mortality remain high. Therefore, the search for new prognostic identifiers and treatments targeting therapy-resistant tumor cells is vital. Our work demonstrates that there are different subgroups with high phenotypic plasticity within the CSC population in HNSCC. CD10, CD184, and CD166 may identify some of these CSC subpopulations with NAMPT as a common metabolic gene for the resilient cells of these subpopulations. We observed that NAMPT reduction causes a decrease in tumorigenic and stemness properties, migration capacity and CSC phenotype through NAD pool depletion. However, NAMPT-inhibited cells can acquire resistance by activating the NAPRT enzyme of the Preiss-Handler pathway. We observed that coadministration of the NAMPT inhibitor with the NAPRT inhibitor cooperated inhibiting tumor growth. The use of an NAPRT inhibitor as an adjuvant improved NAMPT inhibitor efficacy and reduced the dose and toxicity of these inhibitors. Therefore, it seems that the reduction in the NAD pool could have efficacy in tumor therapy. This was confirmed by in vitro assays supplying the cells with products of inhibited enzymes (NA, NMN or NAD) and restoring their tumorigenic and stemness properties. In conclusion, the coinhibition of NAMPT and NAPRT improved the efficacy of antitumor treatment, indicating that the reduction in the NAD pool is important to prevent tumor growth., This research was funded by Grants RTI2018-097455-B-I00 and PID2021-122629OB-I00 funded by MCIN/AEI/10.13039/501100011033 and by “ERDF A way of making Europe”, by the “European Union”. Additional grants from CIBER de Cáncer (CB16/12/00275), from Consejeria de Salud (PI-0397–2017) and Project P18-RT-2501 from 2018 competitive research projects call within the scope of PAIDI 2020—80% co-financed by the European Regional Development Fund (ERDF) from the Regional Ministry of Economic Transformation, Industry, Knowledge and Universities. Junta de Andalucía. Special thanks to the AECC (Spanish Association of Cancer Research) Founding Ref. GC16173720CARR for supporting this work. AES was funded by a grant from the Fundación AECC. EMVS was funded by a postdoctoral fellowship from Junta de Andalucía (CTEICU/PAIDI 2020). LEN, ES-M and LS-D were funded by Spanish ministry of education (FPU16/0290; FPU17/02173; FPU18/01009)., Peer reviewed