NOTCH1 INHIBITION REGULATES EVOLUTIONARY CONSERVED MIRNAS IN T-CELL LYMPHOBLASTIC LEUKEMIA
(Abstract release date: 05/19/16)
EHA Library. Saccomani V. 06/09/16; 134496; PB1596
Dr. Valentina Saccomani
Contributions
Contributions
Abstract
Abstract: PB1596
Type: Publication Only
Background
Activating mutations in the NOTCH1 ligand-activated transcription factor oncogene are found in over 60% of human T-cell lymphoblastic leukemia (T-ALL), where they result in high levels of NOTCH1 signaling. Among multiple functions of oncogenic NOTCH1, there are evidences of the role of non-coding RNAs in NOTCH-induced leukemia. MicroRNAs (miRNAs) are short, single-stranded RNA molecules of approximately 22 nucleotides in length that regulate gene expression by directing their target mRNAs for degradation or translational repression. Dual translocations, that simultaneously affect the 17-92 cluster and NOTCH1, highlight the oncogenic importance of this interaction in T-ALL. Moreover, miR-19 was found to play a crucial role in promoting leukemogenesis in NOTCH1-induced T-ALL. In addition, miR-451 and miR-223 have been described to play an important role downstream NOTCH1 in T-ALL cells both in mouse and human models. Beside these relevant discoveries, data regarding the miRNAs that are significantly regulated following NOTCH1 inhibition are still ill-defined.
Aims
In this study we analyzed NOTH1-regulated miRNAs following NOTCH1 inhibition in T-ALL cells in view of future therapies that may combine NOTCH1 inhibition with microRNA based therapy. Specifically we pursued the following aims: - To generate microarray profiles of NOTCH1-regulated microRNAs from T-ALL cells using a mouse model of NOTCH1-induced leukaemia; - To combine analysis of NOTCH1-regulated microRNAs and gene expression profiling following in vivo inhibition of NOTCH1 signalling pathway; -To identify and validate evolutionary conserved differentially regulated miRNAs.
Methods
We thus used a mouse model of NOTCH1-induced leukemia, that carries a NOTCH1 mutation recurrently found in human T-ALL patients (L1601P/DPEST). NOTCH1 inhibition was performed in vivo using a gamma secretase inhibitor (DBZ) and the treatment schedule was sufficient to determine a significant reduction of cleaved-NOTCH1. miRNAs profiling was performed using a mouse array (8X60K release 19.0; Agilent) that detected 1,247 mouse miRNAs. In parallel, gene expression analysis was performed using SurePrint G3 Mouse Gene Expression v2 array (Agilent) for the identification of about 27000 genes and 4578 lncRNAs.
Results
MYC and NOTCH signature resulted strongly down-regulated following NOTCH1 inhibition by Gene Set Enriched Analysis (GSEA) demonstrating the efficacy of our experimental model. From the miRNAs prolife, we obtained 27 up-regulated and 22 down-regulated miRNAs following NOTCH1 inhibition. Notably, among the NOTCH1 down-regulated miRNAs, we found the miR-17-92 cluster, previously reported highly expressed in T-ALL samples. Their regulation was also confirmed in another mouse model of NOTCH1-induced T-ALL and in human T-leukemia cells. Notably, we identified miR-34a-5p, miR-22a-3p, miR-199a-5p and miR-29a-3p significantly up-regulated following NOTCH1-inhibition suggesting a putative role as tumor suppressors in NOTCH1-driven leukemia. In particular, miR-22a-3p resulted significantly up-regulated following NOTCH1 inhibition both in mouse and human T-ALL cells.
Conclusion
In conclusion, the cluster 17-92 resulted to have a prominent role amongst the down-regulated miRNAs both in mouse and human T-ALL cells. Importantly, we identified novel up-regulated miRNAs downstream NOTCH1 inhibition. In particular, miRNAs 22a-3p resulted significantly up-regulated in human T-ALLs upon GSIs treatment. Further bioinformatics and functional analysis will be performed to elucidate the role of these novel miRNAs in NOTCH1-induced leukemia.
Session topic: E-poster
Keyword(s): Microarray analysis, Notch signaling, T-ALL
Type: Publication Only
Background
Activating mutations in the NOTCH1 ligand-activated transcription factor oncogene are found in over 60% of human T-cell lymphoblastic leukemia (T-ALL), where they result in high levels of NOTCH1 signaling. Among multiple functions of oncogenic NOTCH1, there are evidences of the role of non-coding RNAs in NOTCH-induced leukemia. MicroRNAs (miRNAs) are short, single-stranded RNA molecules of approximately 22 nucleotides in length that regulate gene expression by directing their target mRNAs for degradation or translational repression. Dual translocations, that simultaneously affect the 17-92 cluster and NOTCH1, highlight the oncogenic importance of this interaction in T-ALL. Moreover, miR-19 was found to play a crucial role in promoting leukemogenesis in NOTCH1-induced T-ALL. In addition, miR-451 and miR-223 have been described to play an important role downstream NOTCH1 in T-ALL cells both in mouse and human models. Beside these relevant discoveries, data regarding the miRNAs that are significantly regulated following NOTCH1 inhibition are still ill-defined.
Aims
In this study we analyzed NOTH1-regulated miRNAs following NOTCH1 inhibition in T-ALL cells in view of future therapies that may combine NOTCH1 inhibition with microRNA based therapy. Specifically we pursued the following aims: - To generate microarray profiles of NOTCH1-regulated microRNAs from T-ALL cells using a mouse model of NOTCH1-induced leukaemia; - To combine analysis of NOTCH1-regulated microRNAs and gene expression profiling following in vivo inhibition of NOTCH1 signalling pathway; -To identify and validate evolutionary conserved differentially regulated miRNAs.
Methods
We thus used a mouse model of NOTCH1-induced leukemia, that carries a NOTCH1 mutation recurrently found in human T-ALL patients (L1601P/DPEST). NOTCH1 inhibition was performed in vivo using a gamma secretase inhibitor (DBZ) and the treatment schedule was sufficient to determine a significant reduction of cleaved-NOTCH1. miRNAs profiling was performed using a mouse array (8X60K release 19.0; Agilent) that detected 1,247 mouse miRNAs. In parallel, gene expression analysis was performed using SurePrint G3 Mouse Gene Expression v2 array (Agilent) for the identification of about 27000 genes and 4578 lncRNAs.
Results
MYC and NOTCH signature resulted strongly down-regulated following NOTCH1 inhibition by Gene Set Enriched Analysis (GSEA) demonstrating the efficacy of our experimental model. From the miRNAs prolife, we obtained 27 up-regulated and 22 down-regulated miRNAs following NOTCH1 inhibition. Notably, among the NOTCH1 down-regulated miRNAs, we found the miR-17-92 cluster, previously reported highly expressed in T-ALL samples. Their regulation was also confirmed in another mouse model of NOTCH1-induced T-ALL and in human T-leukemia cells. Notably, we identified miR-34a-5p, miR-22a-3p, miR-199a-5p and miR-29a-3p significantly up-regulated following NOTCH1-inhibition suggesting a putative role as tumor suppressors in NOTCH1-driven leukemia. In particular, miR-22a-3p resulted significantly up-regulated following NOTCH1 inhibition both in mouse and human T-ALL cells.
Conclusion
In conclusion, the cluster 17-92 resulted to have a prominent role amongst the down-regulated miRNAs both in mouse and human T-ALL cells. Importantly, we identified novel up-regulated miRNAs downstream NOTCH1 inhibition. In particular, miRNAs 22a-3p resulted significantly up-regulated in human T-ALLs upon GSIs treatment. Further bioinformatics and functional analysis will be performed to elucidate the role of these novel miRNAs in NOTCH1-induced leukemia.
Session topic: E-poster
Keyword(s): Microarray analysis, Notch signaling, T-ALL
Abstract: PB1596
Type: Publication Only
Background
Activating mutations in the NOTCH1 ligand-activated transcription factor oncogene are found in over 60% of human T-cell lymphoblastic leukemia (T-ALL), where they result in high levels of NOTCH1 signaling. Among multiple functions of oncogenic NOTCH1, there are evidences of the role of non-coding RNAs in NOTCH-induced leukemia. MicroRNAs (miRNAs) are short, single-stranded RNA molecules of approximately 22 nucleotides in length that regulate gene expression by directing their target mRNAs for degradation or translational repression. Dual translocations, that simultaneously affect the 17-92 cluster and NOTCH1, highlight the oncogenic importance of this interaction in T-ALL. Moreover, miR-19 was found to play a crucial role in promoting leukemogenesis in NOTCH1-induced T-ALL. In addition, miR-451 and miR-223 have been described to play an important role downstream NOTCH1 in T-ALL cells both in mouse and human models. Beside these relevant discoveries, data regarding the miRNAs that are significantly regulated following NOTCH1 inhibition are still ill-defined.
Aims
In this study we analyzed NOTH1-regulated miRNAs following NOTCH1 inhibition in T-ALL cells in view of future therapies that may combine NOTCH1 inhibition with microRNA based therapy. Specifically we pursued the following aims: - To generate microarray profiles of NOTCH1-regulated microRNAs from T-ALL cells using a mouse model of NOTCH1-induced leukaemia; - To combine analysis of NOTCH1-regulated microRNAs and gene expression profiling following in vivo inhibition of NOTCH1 signalling pathway; -To identify and validate evolutionary conserved differentially regulated miRNAs.
Methods
We thus used a mouse model of NOTCH1-induced leukemia, that carries a NOTCH1 mutation recurrently found in human T-ALL patients (L1601P/DPEST). NOTCH1 inhibition was performed in vivo using a gamma secretase inhibitor (DBZ) and the treatment schedule was sufficient to determine a significant reduction of cleaved-NOTCH1. miRNAs profiling was performed using a mouse array (8X60K release 19.0; Agilent) that detected 1,247 mouse miRNAs. In parallel, gene expression analysis was performed using SurePrint G3 Mouse Gene Expression v2 array (Agilent) for the identification of about 27000 genes and 4578 lncRNAs.
Results
MYC and NOTCH signature resulted strongly down-regulated following NOTCH1 inhibition by Gene Set Enriched Analysis (GSEA) demonstrating the efficacy of our experimental model. From the miRNAs prolife, we obtained 27 up-regulated and 22 down-regulated miRNAs following NOTCH1 inhibition. Notably, among the NOTCH1 down-regulated miRNAs, we found the miR-17-92 cluster, previously reported highly expressed in T-ALL samples. Their regulation was also confirmed in another mouse model of NOTCH1-induced T-ALL and in human T-leukemia cells. Notably, we identified miR-34a-5p, miR-22a-3p, miR-199a-5p and miR-29a-3p significantly up-regulated following NOTCH1-inhibition suggesting a putative role as tumor suppressors in NOTCH1-driven leukemia. In particular, miR-22a-3p resulted significantly up-regulated following NOTCH1 inhibition both in mouse and human T-ALL cells.
Conclusion
In conclusion, the cluster 17-92 resulted to have a prominent role amongst the down-regulated miRNAs both in mouse and human T-ALL cells. Importantly, we identified novel up-regulated miRNAs downstream NOTCH1 inhibition. In particular, miRNAs 22a-3p resulted significantly up-regulated in human T-ALLs upon GSIs treatment. Further bioinformatics and functional analysis will be performed to elucidate the role of these novel miRNAs in NOTCH1-induced leukemia.
Session topic: E-poster
Keyword(s): Microarray analysis, Notch signaling, T-ALL
Type: Publication Only
Background
Activating mutations in the NOTCH1 ligand-activated transcription factor oncogene are found in over 60% of human T-cell lymphoblastic leukemia (T-ALL), where they result in high levels of NOTCH1 signaling. Among multiple functions of oncogenic NOTCH1, there are evidences of the role of non-coding RNAs in NOTCH-induced leukemia. MicroRNAs (miRNAs) are short, single-stranded RNA molecules of approximately 22 nucleotides in length that regulate gene expression by directing their target mRNAs for degradation or translational repression. Dual translocations, that simultaneously affect the 17-92 cluster and NOTCH1, highlight the oncogenic importance of this interaction in T-ALL. Moreover, miR-19 was found to play a crucial role in promoting leukemogenesis in NOTCH1-induced T-ALL. In addition, miR-451 and miR-223 have been described to play an important role downstream NOTCH1 in T-ALL cells both in mouse and human models. Beside these relevant discoveries, data regarding the miRNAs that are significantly regulated following NOTCH1 inhibition are still ill-defined.
Aims
In this study we analyzed NOTH1-regulated miRNAs following NOTCH1 inhibition in T-ALL cells in view of future therapies that may combine NOTCH1 inhibition with microRNA based therapy. Specifically we pursued the following aims: - To generate microarray profiles of NOTCH1-regulated microRNAs from T-ALL cells using a mouse model of NOTCH1-induced leukaemia; - To combine analysis of NOTCH1-regulated microRNAs and gene expression profiling following in vivo inhibition of NOTCH1 signalling pathway; -To identify and validate evolutionary conserved differentially regulated miRNAs.
Methods
We thus used a mouse model of NOTCH1-induced leukemia, that carries a NOTCH1 mutation recurrently found in human T-ALL patients (L1601P/DPEST). NOTCH1 inhibition was performed in vivo using a gamma secretase inhibitor (DBZ) and the treatment schedule was sufficient to determine a significant reduction of cleaved-NOTCH1. miRNAs profiling was performed using a mouse array (8X60K release 19.0; Agilent) that detected 1,247 mouse miRNAs. In parallel, gene expression analysis was performed using SurePrint G3 Mouse Gene Expression v2 array (Agilent) for the identification of about 27000 genes and 4578 lncRNAs.
Results
MYC and NOTCH signature resulted strongly down-regulated following NOTCH1 inhibition by Gene Set Enriched Analysis (GSEA) demonstrating the efficacy of our experimental model. From the miRNAs prolife, we obtained 27 up-regulated and 22 down-regulated miRNAs following NOTCH1 inhibition. Notably, among the NOTCH1 down-regulated miRNAs, we found the miR-17-92 cluster, previously reported highly expressed in T-ALL samples. Their regulation was also confirmed in another mouse model of NOTCH1-induced T-ALL and in human T-leukemia cells. Notably, we identified miR-34a-5p, miR-22a-3p, miR-199a-5p and miR-29a-3p significantly up-regulated following NOTCH1-inhibition suggesting a putative role as tumor suppressors in NOTCH1-driven leukemia. In particular, miR-22a-3p resulted significantly up-regulated following NOTCH1 inhibition both in mouse and human T-ALL cells.
Conclusion
In conclusion, the cluster 17-92 resulted to have a prominent role amongst the down-regulated miRNAs both in mouse and human T-ALL cells. Importantly, we identified novel up-regulated miRNAs downstream NOTCH1 inhibition. In particular, miRNAs 22a-3p resulted significantly up-regulated in human T-ALLs upon GSIs treatment. Further bioinformatics and functional analysis will be performed to elucidate the role of these novel miRNAs in NOTCH1-induced leukemia.
Session topic: E-poster
Keyword(s): Microarray analysis, Notch signaling, T-ALL
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