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Involvement of miR-3117 in pediatric acute lymphoblastic leukemia susceptibility
EACR24 Poster Sessions / European Journal of Cancer 61, Suppl. 1 (2016) S9–S218
S29
172 14q32 miRNA cluster: A hot spot for osteosarcoma risk in a Spanish population
174 Involvement of miR-3117 in pediatric acute lymphoblastic leukemia susceptibility
I. Mart´ın-Guerrero1 , N. Bilbao-Aldaiturriaga1 , J. Uriz2 , A. Garc´ıa-Orad1,3 . University of the Basque Country, Genetics- Physical Anthropology and Animal Physiology, Bilbao, Spain, 2 University Hospital Donostia, Unit of Pediatric Oncohematology, San Sebastian, Spain, 3 BioCruces Health Research Institute, Pediatrics, Barakaldo, Spain
1 A. Gutierrez-Camino1 , S. Varona-Fernandez ´ , N. Garc´ıa de Andoin2 , 5 ˜ , V. Dolzan6 , I. Astigarraga3,7 , A. Navajas3 , A. Sastre4 , A. Carbone´ Baneres I. Mart´ın-Guerrero1 , A. Garc´ıa-Orad1,3 . 1 University of the Basque Country, Genetics- Physical Anthropology and Animal Physiology, Bilbao, Spain, 2 Hospital Donostia, Pediatrics, San Sebastian, Spain, 3 BioCruces Health Research Institute, Pediatrics, Barakaldo, Spain, 4 University Hospital La Paz, Oncohaematology, Madrid, Spain, 5 Hospital Miguel Servet, Pediatrics, Zaragoza, Spain, 6 Institute of Biochemistry, Faculty of Medicine, Ljubljana, Slovenia, 7 University Hospital Cruces, Pediatrics, Bilbao, Spain
1
Background: Recent genome wide association study (GWAS) in osteosarcoma risk showed that several significant results were located in intergenic positions. These results suggest that non-coding regions could play an important role in the risk of osteosarcoma. One of the most studied noncoding molecules are microRNAs (miRNAs), whose deregulation has been associated with osteosarcoma evolution. Therefore, genetic variants affecting miRNA function could contribute to the risk of the disease. To date, only three polymorphisms in miRNAs have been analyzed in relation to the risk of osteosarcoma, and two of them showed significant association. In this context, this study aimed to evaluate the involvement of all genetic variants in premiRNAs in the risk of osteosarcoma. Material and Methods: We selected all SNPs described in miRNAs with a MAF >0.1. A total of 213 SNPs in 206 pre-miRNAs were analyzed in a cohort of patients (n = 74) and their corresponding controls (n = 160) using Goldengate Veracode technology. c2 or Fisher and FDR were used. Results: The most remarkably finding was the association detected between 4 SNPs located at 14q32 miRNA cluster. Interestingly, miRNAs in this cluster have been associated with MYC deregulation. Therefore, genetic variations in these miRNAs could lead to their downregulation, which in turn would lead to the overexpression of MYC. This result supports the idea that this region is a hotspot for the development of the disease. Conclusion: In conclusion, different variants in 14q32 miRNA cluster could be implicated in osteosarcoma susceptibility. Acknowledgement: This project was supported by RETICS (RD12/0036/0060, RD12/0036/0036), UPV/EHU (UFI 11/35) and Basque Government (IT66113). No conflict of interest. 173 Genetic variant in miR-618 in the risk of chronic lymphocytic leukemia I. Mart´ın-Guerrero1 , A. D´ıaz-Navarro1 , A. Gutierrez-Camino1 , A. Garc´ıa-Orad1,2 . 1 University of the Basque Country, Genetics- Physical Anthropology and Animal Physiology, Bilbao, Spain, 2 BioCruces Health Research Institute, Pediatrics, Barakaldo, Spain Background: Recent Genome Wide Association Studies (GWAS) have found new genetic variants associated with chronic lymphocytic leukemia (CLL) susceptibility, several of which were located in non-coding regions. In fact, nowadays it is known that more than 40% of significant variants associated with cancer risk are in non-coding regions, where non-coding RNAs are located. MicroRNAs (miRNAs) are one of the most studied non-coding RNAs that have been involved in the risk of CLL. In a previous study of our group including 46 SNPs in 42 pre-miRNAs, we observed rs11614913 and rs2114358 associated with the risk of CLL. Considering these results, we aimed to determine the involvement in the risk of CLL of all the SNPs described in pre-miRNAs with a MAF >1% in a larger population. Material and Methods: A total of 213 SNPs in 206 miRNAs were genotyped in 164 CLL patients and 237 cancer-free controls using Veracode GoldenGate Technology (Illumina). Results: One polymorphism at miR-618 showed the most significant association under the dominant model (OR = 0.49; 95% CI: 0.29–0.81; p = 0.005). Remarkably, this polymorphism was previously found to be associated with follicular lymphoma, altering the expression of miR-618. In addition, this miRNA was observed, in silico, to regulate genes associated with CLL, such as BCL2, LEF1 or QPCT. Conclusion: These findings suggest that polymorphisms in pre-miRNAs contribute to the risk of CLL. Large-scale studies are needed to validate the current findings. Acknowledgement: This project was supported by RETICS (RD12/0036/0060, RD12/0036/0036), UPV/EHU (UFI11/35) and Basque Government (IT661-13, S-PE13UN079). No conflict of interest.
Background: Recently, several Genome wide associations studies (GWAS) have found genetic variants associated with pediatric acute lymphoblastic leukemia (ALL) risk. The interest of these studies was mainly focused in coding regions. However, nowadays it is known that more than 40% of significant variants associated with cancer risk are situated in non-coding regions, where non-coding RNAs are located. One of the non-coding RNAs more related with cancer are microRNAs (miRNAs), which have been shown to be dysregulated in ALL, suggesting that they may have a role in ALL risk. Therefore, variants in miRNAs that may alter its function could contribute to childhood B-ALL predisposition. The aim of this study was to determine if SNPs in miRNAs are involved in B-ALL susceptibility. Material and Methods: Blood samples of 296 B-cell ALL patients in complete remission and 426 healthy controls of Spanish and a Slovenian cohort were analyzed. We selected all the SNPs described in pre-miRNAs with a MAF >1% (213 SNPs in 206 miRNAs). VeraCode GoldenGate platform was used. Results: Among the most interesting results, a variant in the seed region of miR-3117 was associated with B-ALL in both populations (p = 0.006). Of note is that in silico analyses show that miR-3117 could regulate genes of the MAPK pathway, which is known to promote leukemogenesis. Conclusion: Our results suggest that a SNP in miR-3117 may be involved in B-ALL susceptibility and give new keys to understand its biology. Acknowledgement: This project was supported by RETICS (RD12/0036/0060, RD12/0036/0036), UPV/EHU (UFI 11/35) and Basque Government (IT661-13, S-PE12UN060). No conflict of interest.
175 The KDM5B demethylase in the normal and malignant mammary gland F. Kogera1 , C. Steven1 , B. Spencer-Dene2 , G. Picco1 , V. Tajadura-Ortega1 , Y.H. Chen3 , E. Bennett3 , J. Quist4 , J. Taylor-Papadimitriou1 , J. Burchell1 . 1 KCL, Research Oncology, London, United Kingdom, 2 Cancer Research UKCrick Institute, Experimental Histopathology Lab, London, United Kingdom, 3 University of Copenhagen, Copenhagen Centre for Glycomics, Copenhagen, Denmark, 4 KCL, Breast Cancer Now Unit, London, United Kingdom Background: It is becoming clear that changes occurring in cancer, including breast cancer reflect a dysregulated epigenome. Histone marks play a major role in determining the chromatin state of a cell, and the H3K4me3 mark is associated with active transcription. The KDM5 family of histone demethylases specifically demethylate the H3K4me3 mark and therefore act as transcriptional repressors. KDM5B, a KDM5 family member, was cloned in our lab as being down regulated when a cell over expressing HER2 was treated with Herceptin, and has been found to be widely expressed in breast cancer. Here, we investigate the role of KDM5B in the normal gland and in breast cancer, using breast cancer cell lines and a transgenic mouse model. Materials and Methods: Expression of KDM5B in breast cancers has been documented in samples from Cancer databases. Cell lines derived from different breast cancer subtypes have been analysed for KDM5B expression and the effect of a KO and KD of KDM5B examined. While constitutive KO of KDM5B in the mouse is embryonic lethal, a transgenic strain carrying a demethylase −ve mutant (D ARID domain deleted) is viable. This strain has been used to document an important function for KDM5B in the developing and pregnant mammary gland. Results: High expression of KDM5B was observed in the ER+ and HER2+ breast cancer subtypes. In the DARID mouse mammary gland, a transient delay in mammary tree expansion at mid pregnancy is observed. At this stage, expansion of the luminal progenitor cells is evident and involves the JAK/STAT5 signalling pathway. However, in the DARID mouse, STAT5 activation is dramatically reduced, implicating a role of KDM5B in this activation. We find that, decreased STAT5 activation in the DARID gland is due to increased levels of Caveolin1 (CAV1), an inhibitor of STAT5 activation. Thus in the normal gland KDM5B represses CAV1 expression during development of the luminal cell lineage. The effect of CAV1 on STAT5 activation must be mediated in a paracrine fashion, as CAV1 is not expressed in the luminal cells where STAT5 is functional but is limited to stromal and myoepithelial cells. KDM5B is expressed in these cells as well as in the luminal cells and thus could repress CAV1 expression.
S29
172 14q32 miRNA cluster: A hot spot for osteosarcoma risk in a Spanish population
174 Involvement of miR-3117 in pediatric acute lymphoblastic leukemia susceptibility
I. Mart´ın-Guerrero1 , N. Bilbao-Aldaiturriaga1 , J. Uriz2 , A. Garc´ıa-Orad1,3 . University of the Basque Country, Genetics- Physical Anthropology and Animal Physiology, Bilbao, Spain, 2 University Hospital Donostia, Unit of Pediatric Oncohematology, San Sebastian, Spain, 3 BioCruces Health Research Institute, Pediatrics, Barakaldo, Spain
1 A. Gutierrez-Camino1 , S. Varona-Fernandez ´ , N. Garc´ıa de Andoin2 , 5 ˜ , V. Dolzan6 , I. Astigarraga3,7 , A. Navajas3 , A. Sastre4 , A. Carbone´ Baneres I. Mart´ın-Guerrero1 , A. Garc´ıa-Orad1,3 . 1 University of the Basque Country, Genetics- Physical Anthropology and Animal Physiology, Bilbao, Spain, 2 Hospital Donostia, Pediatrics, San Sebastian, Spain, 3 BioCruces Health Research Institute, Pediatrics, Barakaldo, Spain, 4 University Hospital La Paz, Oncohaematology, Madrid, Spain, 5 Hospital Miguel Servet, Pediatrics, Zaragoza, Spain, 6 Institute of Biochemistry, Faculty of Medicine, Ljubljana, Slovenia, 7 University Hospital Cruces, Pediatrics, Bilbao, Spain
1
Background: Recent genome wide association study (GWAS) in osteosarcoma risk showed that several significant results were located in intergenic positions. These results suggest that non-coding regions could play an important role in the risk of osteosarcoma. One of the most studied noncoding molecules are microRNAs (miRNAs), whose deregulation has been associated with osteosarcoma evolution. Therefore, genetic variants affecting miRNA function could contribute to the risk of the disease. To date, only three polymorphisms in miRNAs have been analyzed in relation to the risk of osteosarcoma, and two of them showed significant association. In this context, this study aimed to evaluate the involvement of all genetic variants in premiRNAs in the risk of osteosarcoma. Material and Methods: We selected all SNPs described in miRNAs with a MAF >0.1. A total of 213 SNPs in 206 pre-miRNAs were analyzed in a cohort of patients (n = 74) and their corresponding controls (n = 160) using Goldengate Veracode technology. c2 or Fisher and FDR were used. Results: The most remarkably finding was the association detected between 4 SNPs located at 14q32 miRNA cluster. Interestingly, miRNAs in this cluster have been associated with MYC deregulation. Therefore, genetic variations in these miRNAs could lead to their downregulation, which in turn would lead to the overexpression of MYC. This result supports the idea that this region is a hotspot for the development of the disease. Conclusion: In conclusion, different variants in 14q32 miRNA cluster could be implicated in osteosarcoma susceptibility. Acknowledgement: This project was supported by RETICS (RD12/0036/0060, RD12/0036/0036), UPV/EHU (UFI 11/35) and Basque Government (IT66113). No conflict of interest. 173 Genetic variant in miR-618 in the risk of chronic lymphocytic leukemia I. Mart´ın-Guerrero1 , A. D´ıaz-Navarro1 , A. Gutierrez-Camino1 , A. Garc´ıa-Orad1,2 . 1 University of the Basque Country, Genetics- Physical Anthropology and Animal Physiology, Bilbao, Spain, 2 BioCruces Health Research Institute, Pediatrics, Barakaldo, Spain Background: Recent Genome Wide Association Studies (GWAS) have found new genetic variants associated with chronic lymphocytic leukemia (CLL) susceptibility, several of which were located in non-coding regions. In fact, nowadays it is known that more than 40% of significant variants associated with cancer risk are in non-coding regions, where non-coding RNAs are located. MicroRNAs (miRNAs) are one of the most studied non-coding RNAs that have been involved in the risk of CLL. In a previous study of our group including 46 SNPs in 42 pre-miRNAs, we observed rs11614913 and rs2114358 associated with the risk of CLL. Considering these results, we aimed to determine the involvement in the risk of CLL of all the SNPs described in pre-miRNAs with a MAF >1% in a larger population. Material and Methods: A total of 213 SNPs in 206 miRNAs were genotyped in 164 CLL patients and 237 cancer-free controls using Veracode GoldenGate Technology (Illumina). Results: One polymorphism at miR-618 showed the most significant association under the dominant model (OR = 0.49; 95% CI: 0.29–0.81; p = 0.005). Remarkably, this polymorphism was previously found to be associated with follicular lymphoma, altering the expression of miR-618. In addition, this miRNA was observed, in silico, to regulate genes associated with CLL, such as BCL2, LEF1 or QPCT. Conclusion: These findings suggest that polymorphisms in pre-miRNAs contribute to the risk of CLL. Large-scale studies are needed to validate the current findings. Acknowledgement: This project was supported by RETICS (RD12/0036/0060, RD12/0036/0036), UPV/EHU (UFI11/35) and Basque Government (IT661-13, S-PE13UN079). No conflict of interest.
Background: Recently, several Genome wide associations studies (GWAS) have found genetic variants associated with pediatric acute lymphoblastic leukemia (ALL) risk. The interest of these studies was mainly focused in coding regions. However, nowadays it is known that more than 40% of significant variants associated with cancer risk are situated in non-coding regions, where non-coding RNAs are located. One of the non-coding RNAs more related with cancer are microRNAs (miRNAs), which have been shown to be dysregulated in ALL, suggesting that they may have a role in ALL risk. Therefore, variants in miRNAs that may alter its function could contribute to childhood B-ALL predisposition. The aim of this study was to determine if SNPs in miRNAs are involved in B-ALL susceptibility. Material and Methods: Blood samples of 296 B-cell ALL patients in complete remission and 426 healthy controls of Spanish and a Slovenian cohort were analyzed. We selected all the SNPs described in pre-miRNAs with a MAF >1% (213 SNPs in 206 miRNAs). VeraCode GoldenGate platform was used. Results: Among the most interesting results, a variant in the seed region of miR-3117 was associated with B-ALL in both populations (p = 0.006). Of note is that in silico analyses show that miR-3117 could regulate genes of the MAPK pathway, which is known to promote leukemogenesis. Conclusion: Our results suggest that a SNP in miR-3117 may be involved in B-ALL susceptibility and give new keys to understand its biology. Acknowledgement: This project was supported by RETICS (RD12/0036/0060, RD12/0036/0036), UPV/EHU (UFI 11/35) and Basque Government (IT661-13, S-PE12UN060). No conflict of interest.
175 The KDM5B demethylase in the normal and malignant mammary gland F. Kogera1 , C. Steven1 , B. Spencer-Dene2 , G. Picco1 , V. Tajadura-Ortega1 , Y.H. Chen3 , E. Bennett3 , J. Quist4 , J. Taylor-Papadimitriou1 , J. Burchell1 . 1 KCL, Research Oncology, London, United Kingdom, 2 Cancer Research UKCrick Institute, Experimental Histopathology Lab, London, United Kingdom, 3 University of Copenhagen, Copenhagen Centre for Glycomics, Copenhagen, Denmark, 4 KCL, Breast Cancer Now Unit, London, United Kingdom Background: It is becoming clear that changes occurring in cancer, including breast cancer reflect a dysregulated epigenome. Histone marks play a major role in determining the chromatin state of a cell, and the H3K4me3 mark is associated with active transcription. The KDM5 family of histone demethylases specifically demethylate the H3K4me3 mark and therefore act as transcriptional repressors. KDM5B, a KDM5 family member, was cloned in our lab as being down regulated when a cell over expressing HER2 was treated with Herceptin, and has been found to be widely expressed in breast cancer. Here, we investigate the role of KDM5B in the normal gland and in breast cancer, using breast cancer cell lines and a transgenic mouse model. Materials and Methods: Expression of KDM5B in breast cancers has been documented in samples from Cancer databases. Cell lines derived from different breast cancer subtypes have been analysed for KDM5B expression and the effect of a KO and KD of KDM5B examined. While constitutive KO of KDM5B in the mouse is embryonic lethal, a transgenic strain carrying a demethylase −ve mutant (D ARID domain deleted) is viable. This strain has been used to document an important function for KDM5B in the developing and pregnant mammary gland. Results: High expression of KDM5B was observed in the ER+ and HER2+ breast cancer subtypes. In the DARID mouse mammary gland, a transient delay in mammary tree expansion at mid pregnancy is observed. At this stage, expansion of the luminal progenitor cells is evident and involves the JAK/STAT5 signalling pathway. However, in the DARID mouse, STAT5 activation is dramatically reduced, implicating a role of KDM5B in this activation. We find that, decreased STAT5 activation in the DARID gland is due to increased levels of Caveolin1 (CAV1), an inhibitor of STAT5 activation. Thus in the normal gland KDM5B represses CAV1 expression during development of the luminal cell lineage. The effect of CAV1 on STAT5 activation must be mediated in a paracrine fashion, as CAV1 is not expressed in the luminal cells where STAT5 is functional but is limited to stromal and myoepithelial cells. KDM5B is expressed in these cells as well as in the luminal cells and thus could repress CAV1 expression.