ABSTRACT
As a promising alternative to bone marrow aspiration (BMA), mutational profiling on blood-derived circulating cell-free tumor DNA (cfDNA) is a harmless and simple technique to monitor molecular response and treatment resistance of patients with refractory/relapsed multiple myeloma (R/R MM). We evaluated the sensitivity and specificity of cfDNA compared to BMA CD138 positive myeloma plasma cells (PCs) in a series of 45 R/R MM patients using the 29-gene targeted panel (AmpliSeq) NGS. KRAS, NRAS, FAM46C, DIS3, and TP53 were the most frequently mutated genes. The average sensitivity and specificity of cfDNA detection were 65% and 97%, respectively. The concordance per gene between the two samples was good to excellent according to Cohen's κ coefficients interpretation. An increased number of mutations detected in cfDNA were associated with a decreased overall survival. In conclusion, we demonstrated cfDNA NGS analysis feasibility and accuracy in R/R MM patients who may benefit from early phase clinical trial.
Subject(s)
Biomarkers, Tumor , Circulating Tumor DNA , High-Throughput Nucleotide Sequencing , Multiple Myeloma , Mutation , Humans , Multiple Myeloma/genetics , Multiple Myeloma/diagnosis , Multiple Myeloma/mortality , Multiple Myeloma/blood , Male , Female , Middle Aged , Aged , Biomarkers, Tumor/genetics , Circulating Tumor DNA/genetics , Circulating Tumor DNA/blood , High-Throughput Nucleotide Sequencing/methods , Cell-Free Nucleic Acids/genetics , Cell-Free Nucleic Acids/blood , Sensitivity and Specificity , Aged, 80 and over , Adult , Neoplasm Recurrence, Local/genetics , Neoplasm Recurrence, Local/blood , Neoplasm Recurrence, Local/pathology , Drug Resistance, Neoplasm/genetics , DNA Mutational Analysis/methods , Prognosis , Reproducibility of ResultsABSTRACT
Isocitrate dehydrogenase 1 (IDH1) and 2 (IDH2) mutations in Myeloid Neoplams (MNs) exhibit DNA hypermethylation via 2-hydroxyglutarate (2HG) over-production. Clinical impact of azacitidine (AZA) remains inconsistent in IDH1/2-mutated MNs and the potential of serum 2HG as a suitable marker of response to AZA is unknown. To address these questions, we retrospectively analyzed 93 MNs patients (78 AML, 11 MDS, 4 CMML) with IDH1/2 mutations treated with AZA. After a median of 5 cycles of AZA, overall response rate was 28% (including 15% complete remission) and median OS was 12.3 months (significantly shorter in AML compared to MDS/CMML patients). In multivariate analysis of AML patients, DNMT3A mutation was associated with shorter OS while IDH1/2 mutation subtypes had no independent impact. No difference was observed in serum 2HG levels upon AZA treatment between responding and refractory patients suggesting that serum 2HG cannot be used as a surrogate marker of AZA response.
Subject(s)
Leukemia, Myeloid, Acute , Myelodysplastic Syndromes , Azacitidine/therapeutic use , Humans , Isocitrate Dehydrogenase/genetics , Leukemia, Myeloid, Acute/drug therapy , Leukemia, Myeloid, Acute/genetics , Mutation , Retrospective StudiesSubject(s)
Hematologic Neoplasms/pathology , Hematopoietic Stem Cells/pathology , Isocitrate Dehydrogenase/genetics , Lymphocytes/pathology , Mutation/genetics , Myeloid Cells/pathology , Animals , Cells, Cultured , Hematologic Neoplasms/metabolism , Hematopoietic Stem Cells/metabolism , Humans , Lymphocytes/metabolism , Mice , Myeloid Cells/metabolism , Retroviridae/geneticsABSTRACT
A chiral polymer easily obtained by treatment of a linear polyacrylamide with L-proline was adsorbed on a silica support treatment of a linear polyacrylamide with L-proline. After complexation with copper(II), and in spite of a rather low efficiency due to slow ligand exchange, such packing are very convenient for the resolution of amino acid enantiomers eluted in water (relative retention, k'L/k'D, higher than 2 may be observed). Other eluents may also be used. The chiral polymer was studied in solution by potentiometry and nuclear magnetic resonance spectroscopy (13C relaxation measurement). A model of the complex involved in the stereoselection is proposed which accounts for the chromatographic results obtained.