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An Optimized Single Nucleotide Polymorphism-Based Detection Method Suggests That Allelic Variants in the 3’ Untranslated Region of RRAS2 Correlate with Treatment Response in Chronic Lymphocytic Leukemia Patients

SIMPLE SUMMARY: Overexpression of unmutated RRAS2 could be behind the development of chronic lymphocytic leukemia (CLL). This is, in part, supported by the presence of a single-nucleotide polymorphism (SNP) in the 3’ untranslated region of the RRAS2 mRNA that is linked to higher RRAS2 overexpression...

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Autores principales: Hortal, Alejandro, Lacuna, Marta, Cifuentes, Claudia, Alcoceba, Miguel, Bustelo, Xosé R., González, Marcos, Alarcón, Balbino
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2023
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9913312/
https://www.ncbi.nlm.nih.gov/pubmed/36765602
http://dx.doi.org/10.3390/cancers15030644
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author Hortal, Alejandro
Lacuna, Marta
Cifuentes, Claudia
Alcoceba, Miguel
Bustelo, Xosé R.
González, Marcos
Alarcón, Balbino
author_facet Hortal, Alejandro
Lacuna, Marta
Cifuentes, Claudia
Alcoceba, Miguel
Bustelo, Xosé R.
González, Marcos
Alarcón, Balbino
author_sort Hortal, Alejandro
collection PubMed
description SIMPLE SUMMARY: Overexpression of unmutated RRAS2 could be behind the development of chronic lymphocytic leukemia (CLL). This is, in part, supported by the presence of a single-nucleotide polymorphism (SNP) in the 3’ untranslated region of the RRAS2 mRNA that is linked to higher RRAS2 overexpression and worse prognosis. Determining the nucleotide composition (G or C) at the SNP position might be of prognostic value and helpful in orienting the choice for therapies. To make SNP analysis more feasible, we show here the implementation of a new PCR method that allows the characterization of the SNP allele composition with a little amount of genomic DNA. As an example, we show in a small subset of CLL patients that treatment with the drug Ibrutinib results in a stronger reduction in circulating leukemic cells if the patients bear at least one RRAS2 allele with a C at the SNP position. ABSTRACT: Unlike classical RAS genes, oncogenic mutations on RRAS2 are seldomly found in human cancer. By contrast, RRAS2 is frequently found overexpressed in a number of human tumors, including B and T cell lymphomas, breast, gastric, head and neck cancers. In this regard, we have recently shown that overexpression of wild-type RRAS2 drives the formation of both chronic lymphocytic leukemia (CLL) and breast cancer in mice. In support for the relevance of overexpression of wild type RRAS2 in human cancer, we have found that RRAS2 expression is influenced by the presence of a specific single nucleotide polymorphism (SNP) located in the 3’-untranslated region (UTR) of the RRAS2 mRNA. Perhaps more importantly, the presence of the alternate C, rather than the G allele, at the RRAS2 SNP designated as rs8570 is also associated with worse patient prognosis in CLL. This indicates that the detection of this SNP allelic variants can be informative to predict RRAS2 expression levels and disease long-term evolution in patients. Here, we describe a polymerase chain reaction (PCR)-based method that facilitates the rapid and easy determination of G and C allelic variants of the SNP. Using this approach, we confirm that the C allelic variant is associated with higher expression levels of RRAS2 transcripts and poor patient prognosis. However, we have also found that expression of the C allelic variants correlates with better response to ibrutinib, a Bruton kinase inhibitor commonly used in CLL treatments. This suggests that this method for detecting the RRAS2 rs8570 SNP might be a useful as a tool to predict both patient prognosis and response to targeted therapy in CLL.
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spelling pubmed-99133122023-02-11 An Optimized Single Nucleotide Polymorphism-Based Detection Method Suggests That Allelic Variants in the 3’ Untranslated Region of RRAS2 Correlate with Treatment Response in Chronic Lymphocytic Leukemia Patients Hortal, Alejandro Lacuna, Marta Cifuentes, Claudia Alcoceba, Miguel Bustelo, Xosé R. González, Marcos Alarcón, Balbino Cancers (Basel) Article SIMPLE SUMMARY: Overexpression of unmutated RRAS2 could be behind the development of chronic lymphocytic leukemia (CLL). This is, in part, supported by the presence of a single-nucleotide polymorphism (SNP) in the 3’ untranslated region of the RRAS2 mRNA that is linked to higher RRAS2 overexpression and worse prognosis. Determining the nucleotide composition (G or C) at the SNP position might be of prognostic value and helpful in orienting the choice for therapies. To make SNP analysis more feasible, we show here the implementation of a new PCR method that allows the characterization of the SNP allele composition with a little amount of genomic DNA. As an example, we show in a small subset of CLL patients that treatment with the drug Ibrutinib results in a stronger reduction in circulating leukemic cells if the patients bear at least one RRAS2 allele with a C at the SNP position. ABSTRACT: Unlike classical RAS genes, oncogenic mutations on RRAS2 are seldomly found in human cancer. By contrast, RRAS2 is frequently found overexpressed in a number of human tumors, including B and T cell lymphomas, breast, gastric, head and neck cancers. In this regard, we have recently shown that overexpression of wild-type RRAS2 drives the formation of both chronic lymphocytic leukemia (CLL) and breast cancer in mice. In support for the relevance of overexpression of wild type RRAS2 in human cancer, we have found that RRAS2 expression is influenced by the presence of a specific single nucleotide polymorphism (SNP) located in the 3’-untranslated region (UTR) of the RRAS2 mRNA. Perhaps more importantly, the presence of the alternate C, rather than the G allele, at the RRAS2 SNP designated as rs8570 is also associated with worse patient prognosis in CLL. This indicates that the detection of this SNP allelic variants can be informative to predict RRAS2 expression levels and disease long-term evolution in patients. Here, we describe a polymerase chain reaction (PCR)-based method that facilitates the rapid and easy determination of G and C allelic variants of the SNP. Using this approach, we confirm that the C allelic variant is associated with higher expression levels of RRAS2 transcripts and poor patient prognosis. However, we have also found that expression of the C allelic variants correlates with better response to ibrutinib, a Bruton kinase inhibitor commonly used in CLL treatments. This suggests that this method for detecting the RRAS2 rs8570 SNP might be a useful as a tool to predict both patient prognosis and response to targeted therapy in CLL. MDPI 2023-01-19 /pmc/articles/PMC9913312/ /pubmed/36765602 http://dx.doi.org/10.3390/cancers15030644 Text en © 2023 by the authors. https://creativecommons.org/licenses/by/4.0/Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (https://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Hortal, Alejandro
Lacuna, Marta
Cifuentes, Claudia
Alcoceba, Miguel
Bustelo, Xosé R.
González, Marcos
Alarcón, Balbino
An Optimized Single Nucleotide Polymorphism-Based Detection Method Suggests That Allelic Variants in the 3’ Untranslated Region of RRAS2 Correlate with Treatment Response in Chronic Lymphocytic Leukemia Patients
title An Optimized Single Nucleotide Polymorphism-Based Detection Method Suggests That Allelic Variants in the 3’ Untranslated Region of RRAS2 Correlate with Treatment Response in Chronic Lymphocytic Leukemia Patients
title_full An Optimized Single Nucleotide Polymorphism-Based Detection Method Suggests That Allelic Variants in the 3’ Untranslated Region of RRAS2 Correlate with Treatment Response in Chronic Lymphocytic Leukemia Patients
title_fullStr An Optimized Single Nucleotide Polymorphism-Based Detection Method Suggests That Allelic Variants in the 3’ Untranslated Region of RRAS2 Correlate with Treatment Response in Chronic Lymphocytic Leukemia Patients
title_full_unstemmed An Optimized Single Nucleotide Polymorphism-Based Detection Method Suggests That Allelic Variants in the 3’ Untranslated Region of RRAS2 Correlate with Treatment Response in Chronic Lymphocytic Leukemia Patients
title_short An Optimized Single Nucleotide Polymorphism-Based Detection Method Suggests That Allelic Variants in the 3’ Untranslated Region of RRAS2 Correlate with Treatment Response in Chronic Lymphocytic Leukemia Patients
title_sort optimized single nucleotide polymorphism-based detection method suggests that allelic variants in the 3’ untranslated region of rras2 correlate with treatment response in chronic lymphocytic leukemia patients
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC9913312/
https://www.ncbi.nlm.nih.gov/pubmed/36765602
http://dx.doi.org/10.3390/cancers15030644
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