Page 148 - Haematologica-5

Haematologica 2018 Volume 103(5):880-889
1Cancer Research Center-IBMCC (USAL-CSIC), Salamanca, Spain, 2Institute of Biomedical Research of Salamanca (IBSAL), Spain; 3National Medicines Institute, Warsaw, Poland; 4Hematology Department, Hospital 12 de Octubre, CNIO, Complutense University, CIBERONC, Madrid, Spain; 5Hospital Universitario de Salamanca, CIBERONC, Spain; 6Hospital 12 de Octubre, Madrid, Spain; 7Hospital Germans Trias i Pujol, Barcelona, Spain; 8Hospital Clinic, Barcelona, Spain and 9Clínica Universidad de Navarra, CIMA, IDISNA, CIBERONC, Pamplona, Spain
Ferrata Storti Foundation
Plasma Cell Disorders
A novel nano-immunoassay method for quantification of proteins from CD138-purified myeloma cells: biological and clinical utility
Irena Misiewicz-Krzeminska,1,2,3 Luis Antonio Corchete,1,2 Elizabeta A. Rojas,1,2 Joaquín Martínez-López,4 Ramón García-Sanz,1,2,5 Albert Oriol,7 Joan Bladé,8 Juan-José Lahuerta,6 Jesús San Miguel,9 María-Victoria Mateos1,2,5
and Norma C. Gutiérrez1,2,5
ABSTRACT
Protein analysis in bone marrow samples from patients with multiple myeloma has been limited by the low concentration of proteins
+
obtained after CD138 cell selection. A novel approach based on
capillary nano-immunoassay could make it possible to quantify dozens of proteins from each myeloma sample in an automated manner. Here we present a method for the accurate and robust quantification of the expression of multiple proteins extracted from CD138-purified multiple myeloma samples frozen in RLT Plus buffer, which is commonly used for nucleic acid preservation and isolation. Additionally, the biological and clinical value of this analysis for a panel of 12 proteins essential to the pathogenesis of multiple myeloma was evaluated in 63 patients with newly diagnosed multiple myeloma. The analysis of the prognostic impact of CRBN/Cereblon and IKZF1/Ikaros mRNA/protein showed that only the protein levels were able to predict progression-free survival of patients; mRNA levels were not associated with prognosis. Interestingly, high levels of Cereblon and Ikaros proteins were associated with longer progression-free survival only in patients who received immunomodulatory drugs and not in those treated with other drugs. In conclusion, the capillary nano-immunoassay platform provides a novel opportunity for automated quantification of the expression of more than 20 proteins in CD138+ primary multiple myeloma samples.
Introduction
Genomics has come to dominate biomedical research in recent years. For exam- ple, high-throughput genomic technologies have been used for the comprehensive analysis of multiple myeloma (MM). In particular, gene expression profiling has enabled the molecular classification of MM, which is widely used in biological research.1 However, a knowledge of protein expression is essential for identifying therapeutic targets, since proteins are the molecules through which most new drugs achieve their efficacy. The limited amount of sample remaining after plasma cell purification means that messenger RNA (mRNA) quantification is still used as an indirect measure of protein expression in most cases. However, several studies have shown that protein levels cannot be predicted from mRNA measurements.2
Immunohistochemistry and flow cytometry have been used to analyze expres- sion at the protein level, although to a limited extent. These methods are of great value and are of proven clinical utility, but they have some limitations that make them less useful for studying intracellular protein levels. They mostly use directly
Correspondence:
normagu@usal.es
Received: September 27, 2017. Accepted: January 31, 2018. Pre-published: March 15, 2018.
doi:10.3324/haematol.2017.181628
Check the online version for the most updated information on this article, online supplements, and information on authorship & disclosures: www.haematologica.org/content/103/5/880
©2018 Ferrata Storti Foundation
Material published in Haematologica is covered by copyright. All rights are reserved to the Ferrata Storti Foundation. Use of published material is allowed under the following terms and conditions: https://creativecommons.org/licenses/by-nc/4.0/legalcode. Copies of published material are allowed for personal or inter- nal use. Sharing published material for non-commercial pur- poses is subject to the following conditions: https://creativecommons.org/licenses/by-nc/4.0/legalcode, sect. 3. Reproducing and sharing published material for com- mercial purposes is not allowed without permission in writing from the publisher.
880
haematologica | 2018; 103(5)
ARTICLE