Loss of p53 enhances the tumor-initiating potential and drug resistance of clonogenic multiple myeloma cells
Ontology highlight
ABSTRACT:
ORGANISM(S): Homo sapiens
PROVIDER: GSE217769 | GEO | 2022/11/15
REPOSITORIES: GEO
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Project description:Tumor relapse and drug resistance are major factors that limit the curability of multiple myeloma (MM). New regimens have improved overall MM survival rates, but patients with high-risk features continue to have inferior outcomes. Chromosome 17p13 deletion (del17p) that includes the loss of the TP53 gene is a high-risk cytogenetic abnormality and is associated with poor clinical outcomes owing to relatively short remissions and the development of pan-drug resistant disease. Increased relapse rates suggest that del17p enhances clonogenic growth, and we found that the loss of p53 increased both the frequency and drug resistance of tumor-initiating MM cells (TICs). Subsequent RNA sequencing (RNA-seq) studies demonstrated significant activation of the Notch signaling pathway and upregulation of inhibitor of DNA binding (ID1/ID2) genes in p53-knock out (p53-KO) cells. We found that the loss of ID1 or HES-1 expression or treatment with a gamma-secretase inhibitor (GSI) significantly decreased the clonogenic growth of p53-KO but not p53 wild-type cells. GSI treatment in a small set of MM specimens also reduced the clonogenic growth in del17p samples but not in non-del17p samples. This effect was specific as overexpression of the Notch intracellular domain (NICD) rescued the effects of GSI treatment. Our study demonstrates that the Notch signaling and ID1 expression are required for TIC expansion in p53-KO MM cells. These findings also suggest that GSI may be specifically active in patients with p53 mutant MM.
| S-EPMC10368840 | biostudies-literature
Project description:Loss of p53 enhances the tumor-initiating potential and drug resistance of clonogenic multiple myeloma cells
| PRJNA900292 | ENA
Project description:In order to establish a strict p53-dependent gene expression profile, p53 negative clones were derived from two TP53+/+ and two TP53-/mut t(4;14) human myeloma cell lines (HMCLs) using CRISPR/Cas9 technology. From the 17 dysregulated genes shared between the 6 clones from the two TP53+/+ myeloma cell lines, we established a 13-gene functional p53 score, involving genes downregulated on p53-silencing. This functional score allowed us to segregate clones, myeloma cell lines and patients’ samples according to the presence or absence of a single or double hit in TP53 gene i.e., chromosomal deletion and/or mutation. The score also distinguished 1,162 cell lines from hematological and solid cancers according to hits in TP53. Moreover, the 13-gene score was predictive of overall survival in two independent cohorts of patients. Among the 13 genes, we showed that p53-regulated BAX expression correlated to, and directly impacted, the MCL1 BH3 mimetic S63845 sensitivity of myeloma cells by modulating the amount of MCL1-BAX complexes. Resistance to cell death induced by p53-silencing was overcome by combining S63845 with venetoclax, although the synergy was significantly lower in p53-deficient versus -proficient clones and patients’ samples. Finally, by using scRNAseq analysis, we further showed in patients’ bone marrow samples that myeloma cells surviving to the BH3 mimetic combination had an about 3-fold lower p53 score. This data established a functional p53 score and showed that p53-mediating BAX expression greatly impacts mitochondria-mediated cell death in myeloma cells.
2024-04-04 | GSE245049 | GEO
Project description:In order to establish a strict p53-dependent gene expression profile, p53 negative clones were derived from two TP53+/+ and two TP53-/mut t(4;14) human myeloma cell lines (HMCLs) using CRISPR/Cas9 technology. From the 17 dysregulated genes shared between the 6 clones from the two TP53+/+ myeloma cell lines, we established a 13-gene functional p53 score, involving genes downregulated on p53-silencing. This functional score allowed us to segregate clones, myeloma cell lines and patients’ samples according to the presence or absence of a single or double hit in TP53 gene i.e., chromosomal deletion and/or mutation. The score also distinguished 1,162 cell lines from hematological and solid cancers according to hits in TP53. Moreover, the 13-gene score was predictive of overall survival in two independent cohorts of patients. Among the 13 genes, we showed that p53-regulated BAX expression correlated to, and directly impacted, the MCL1 BH3 mimetic S63845 sensitivity of myeloma cells by modulating the amount of MCL1-BAX complexes. Resistance to cell death induced by p53-silencing was overcome by combining S63845 with venetoclax, although the synergy was significantly lower in p53-deficient versus -proficient clones and patients’ samples. Finally, by using scRNAseq analysis, we further showed in patients’ bone marrow samples that myeloma cells surviving to the BH3 mimetic combination had an about 3-fold lower p53 score. This data established a functional p53 score and showed that p53-mediating BAX expression greatly impacts mitochondria-mediated cell death in myeloma cells.
2024-04-04 | GSE245162 | GEO
Project description:The introduction of novel therapeutic agents has considerably improved the median survival of patients with multiple myeloma (MM). However, the natural history of the disease is characterized by continuous relapses over time. As a consequence, the development of new drugs is still required to treat MM recurrence. Here, we report for the first time the potent anti-myeloma activity of amiloride, an old potassium-sparing diuretic approved for the treatment of hypertension and edema due to heart failure. Amilorideinduced apoptosis was observed in a broad panel of MM cell lines and in xenograft mouse models. Moreover, amiloride also had a synergistic effect when combined with dexamethasone and melphalan. RNA-seq experiments showed that amiloride not only significantly altered the level of transcript isoforms and alternative splicing events, but also deregulated the spliceosomal machinery. Additionally, disruption of the splicing machinery in immunofluorescence studies was associated with the inhibition of myeloma cell viability after amiloride exposure. Although amiloride was able to induce apoptosis in myeloma cells lacking p53 expression, activation of p53 signaling was observed in wild-type and mutated TP53 cells after amiloride exposure. On the other hand, the manageable toxicity profile of amiloride is well known and we did not find a significant systemic toxicity in mice treated with amiloride. Overall, our results provide a mechanistic rationale for the use of amiloride as an alternative treatment option for relapsed MM patients.
2018-09-10 | GSE95077 | GEO
Project description:Deletion (Del) of 17p involving the TP53 tumor suppressor remains an adverse prognostic factor in multiple myeloma, and more effective targeted therapies are needed for these patients. Genomic studies revealed that del17p was associated with reduced copy number and mRNA expression of RNA polymerase II subunit alpha (POLR2A), which is located near TP53. We therefore studied HDP-101, an anti-B-cell maturation antigen (BCMA) antibody drug conjugate (ADC) with the POLR2A poison α-amanitin, which showed potent anti-proliferative and pro-apoptotic activity against cell lines and primary samples. POLR2A knockdown (KD) in both TP53 wild-type (WT) and knockout (KO) cells, with the latter being a model of del17p, enhanced sensitivity to HDP-101 compared to POLR2A WT cells. Gene expression profiling and proteomic studies showed evidence for activation of endoplasmic reticulum stress and the unfolded protein response, as well as a reduction of anti-apoptotic proteins such as Myeloid cell leukemia (MCL)-1. Bortezomib enhanced the activity of HDP-101, as did a gamma-secretase inhibitor, and this ADC overcame resistance both due to microenvironmental factors and in acquired drug resistance models. Finally, HDP-101 was well tolerated in vivo, reduced disease burden with single, 2-4 mg/kg doses, and no evidence of relapse was seen out to 100 days, even in xenografts that modeled del17p through dual TP53 KO/POLR2A KD, which grew more aggressively without treatment. Together, the data validate the efficacy of a new BCMA ADC that is ready for clinical translation, and suggest that it could be equipotent, or perhaps even more effective against myeloma with del17p. We used microarray to investigate the global changes caused by HDP-101 across p53 Wt and p53 KO multiple myeloma cell lines
2020-04-08 | GSE148242 | GEO
Project description:Inactivating TP53 mutations lead to a loss of function of p53, but can also often result in oncogenic gain-of-function (GOF) of mutant p53 (mutp53) proteins which promote tumor development and progression. The GOF activities of TP53 mutations are well documented, but the mechanisms involved remain poorly understood. Here, we study the mutp53 interactome with IP-MS.
2023-11-27 | PXD047094 | Pride
Project description:The tumor suppressor protein p53, encoded by TP53, inhibits tumorigenesis by inducing cell cycle arrest, senescence and apoptosis. Several genetic polymorphisms exist in TP53, including a proline to arginine variant at amino acid 72 (P72 and R72, respectively); this polymorphism alters p53 function. In general, the P72 variant shows increased ability to induce cell cycle arrest, while the R72 variant possesses increased ability to induce apoptosis, relative to P72. At present, the underlying mechanisms for these functional differences are not fully understood. Toward elucidating the molecular basis for these differences a gene expression microarray analysis was conducted on normal human fibroblast cells that are homozygous for P72 and R72 variants, along with subclones of these lines that express a p53 short hairpin (shp53). Approximately three dozen genes were identified whose transactivation is affected by the codon 72 polymorphism. One of these is the tripartite motif family-like 2 (TRIML2) gene, which is preferentially induced by the R72 variant. Importantly, the accumulated data indicate that TRIML2 interacts with p53, and facilitates the modification of p53 with SUMO2. TRIML2 also enhances the ability of p53 to transactivate a subset of pro-apoptotic target genes associated with prolonged oxidative stress, including PIDD, PIG3 (TP53I3) and PIG6 (PRODH). These data indicate that TRIML2 is part of a feed-forward loop that activates p53 in cells expressing the R72 variant, particularly after prolonged stress.
2014-12-01 | GSE61124 | GEO
Project description:MicroRNAs have been demonstrated to be deregulated in multiple myeloma (MM). We have previously reported the downregulation of miR-214 in MM compared to normal plasma cells. In the present study, we have explored the functional role of miR-214 in myeloma pathogenesis. Ectopic expression of miR-214 reduced cell growth and induced apoptosis of myeloma cells. In order to identify the potential direct target genes of miR-214 which could be involved in the biological pathways regulated by this miRNA, gene expression profiling of H929 myeloma cell line transfected with precursor miR-214 was carried out. Functional analysis revealed significant enrichment for DNA replication, cell cycle phase and DNA binding. We show that miR-214 directly down-regulates the expression of PSMD10, which encodes the oncoprotein gankyrin, and ASF1B, a histone chaperone required for DNA replication, by binding to their 3'-UTR. In addition, gankyrin inhibition induced an increase of P53 mRNA levels and subsequent up-regulation in CDKN1A (p21Waf1/Cip1) and BAX transcripts, which are direct transcriptional targets of p53. In conclusion, we demonstrate that miR-214 function as a tumor suppressor in myeloma by a positive regulation of p53 and inhibition of DNA replication.
2012-12-28 | GSE35948 | GEO
Project description:MicroRNAs have been demonstrated to be deregulated in multiple myeloma (MM). We have previously reported the downregulation of miR-214 in MM compared to normal plasma cells. In the present study, we have explored the functional role of miR-214 in myeloma pathogenesis. Ectopic expression of miR-214 reduced cell growth and induced apoptosis of myeloma cells. In order to identify the potential direct target genes of miR-214 which could be involved in the biological pathways regulated by this miRNA, gene expression profiling of H929 myeloma cell line transfected with precursor miR-214 was carried out. Functional analysis revealed significant enrichment for DNA replication, cell cycle phase and DNA binding. We show that miR-214 directly down-regulates the expression of PSMD10, which encodes the oncoprotein gankyrin, and ASF1B, a histone chaperone required for DNA replication, by binding to their 3'-UTR. In addition, gankyrin inhibition induced an increase of P53 mRNA levels and subsequent up-regulation in CDKN1A (p21Waf1/Cip1) and BAX transcripts, which are direct transcriptional targets of p53. In conclusion, we demonstrate that miR-214 function as a tumor suppressor in myeloma by a positive regulation of p53 and inhibition of DNA replication. H929 cell line was transfected with Pre-miR™ miRNA precursors pre-miR-214 or pre-miR™ miRNA negative, non-targeting control#1 (Ambion) at 50 nM concentration, using the nucleofector II system with C-16 program (Amaxa). The experiments were performed in triplicates.
2012-12-28 | E-GEOD-35948 | biostudies-arrayexpress