Multiple myeloma is a plasma cell malignancy characterized by an abnormal increase in monoclonal immunoglobulins. Despite significant advances in treatment, some patients progress to more aggressive forms of multiple myeloma, including extramedullary disease or plasma cell leukemia. Although the exact molecular mechanisms are not known, several studies have confirmed the involvement of small extracellular vesicle-enriched microRNAs in multiple myeloma progression. Therefore, we performed expression profiling of these molecules in bone marrow plasma of multiple myeloma, extramedullary disease, and plasma cell leukemia patients using small RNA sequencing to identify novel molecules involved in disease pathogenesis. In total, 42 microRNAs were significantly dysregulated among analyzed subgroups. Independent validation by RT-qPCR confirmed elevated levels of miR-140-3p, miR-584-5p, miR-191-5p, and miR-143-3p in multiple myeloma patients compared to extramedullary disease and plasma cell leukemia patients. Subsequent statistical analysis revealed significant correlations between patient clinical characteristics or flow cytometry parameters and microRNA expression. These results indicate that dysregulation of microRNAs could contribute to multiple myeloma progression.
- Publikační typ
- časopisecké články MeSH
BACKGROUND: Multiple myeloma (MM) represents the second most common hematological malignancy characterized by the infiltration of the bone marrow by plasma cells that produce monoclonal immunoglobulin. While the quality and length of life of MM patients have significantly increased, MM remains a hard-to-treat disease; almost all patients relapse. As MM is highly heterogenous, patients relapse at different times. It is currently not possible to predict when relapse will occur; numerous studies investigating the dysregulation of non-coding RNA molecules in cancer suggest that microRNAs could be good markers of relapse. RESULTS: Using small RNA sequencing, we profiled microRNA expression in peripheral blood in three groups of MM patients who relapsed at different intervals. In total, 24 microRNAs were significantly dysregulated among analyzed subgroups. Independent validation by RT-qPCR confirmed changed levels of miR-598-3p in MM patients with different times to relapse. At the same time, differences in the mass spectra between groups were identified using matrix-assisted laser desorption/ionization time of flight mass spectrometry. All results were analyzed by machine learning. CONCLUSION: Mass spectrometry coupled with machine learning shows potential as a reliable, rapid, and cost-effective preliminary screening technique to supplement current diagnostics.
- Publikační typ
- časopisecké články MeSH
Multiple myeloma (MM) is the second most prevalent hematological malignancy, characterized by infiltration of the bone marrow by malignant plasma cells. Extramedullary disease (EMD) represents a more aggressive condition involving the migration of a subclone of plasma cells to paraskeletal or extraskeletal sites. Liquid biopsies could improve and speed diagnosis, as they can better capture the disease heterogeneity while lowering patients' discomfort due to minimal invasiveness. Recent studies have confirmed alterations in the proteome across various malignancies, suggesting specific changes in protein classes. In this study, we show that MALDI-TOF mass spectrometry fingerprinting of peripheral blood can differentiate between MM and primary EMD patients. We constructed a predictive model using a supervised learning method, partial least squares-discriminant analysis (PLS-DA) and evaluated its generalization performance on a test dataset. The outcome of this analysis is a method that predicts specifically primary EMD with high sensitivity (86.4%), accuracy (78.4%), and specificity (72.4%). Given the simplicity of this approach and its minimally invasive character, this method provides rapid identification of primary EMD and could prove helpful in clinical practice.
- MeSH
- lidé středního věku MeSH
- lidé MeSH
- mnohočetný myelom * krev diagnóza MeSH
- nádorové biomarkery krev MeSH
- senioři MeSH
- spektrometrie hmotnostní - ionizace laserem za účasti matrice metody MeSH
- tekutá biopsie metody MeSH
- Check Tag
- lidé středního věku MeSH
- lidé MeSH
- mužské pohlaví MeSH
- senioři MeSH
- ženské pohlaví MeSH
- Publikační typ
- časopisecké články MeSH
BACKGROUND/AIM: Brain metastases (BMs) are the most frequent intracranial tumors in adults and one of the greatest challenges for modern oncology. Most are derived from lung, breast, renal cell, and colorectal carcinomas and melanomas. Up to 14% of patients are diagnosed with BMs of unknown primary, which are commonly characterized by an early and aggressive metastatic spread. It is important to discover novel biomarkers for early identification of BM origin, allowing better management of patients with this disease. Our study focused on microRNAs (miRNAs), which are very stable in frozen native and FFPE tissues and have been shown to be sensitive and specific diagnostic biomarkers of cancer. We aimed to identify miRNAs with significantly different expression in the five most frequent groups of BMs and develop a diagnostic classifier capable of sensitive and specific classification of BMs. MATERIALS AND METHODS: Total RNA enriched for miRNAs was isolated using the mirVana miRNA Isolation Kit from 71 fresh-frozen histopathologically confirmed BM tissues originating in 5 cancer types. Sequencing libraries were prepared using the QIAseq miRNA Library Kit and sequenced on the NextSeq 500 platform. MiRNA expression was further validated by RT-qPCR. RESULTS: Differential analysis identified 373 miRNAs with significantly different expression between 5 BM groups (p<0.001). A classifier model was developed based on the expression of 6 miRNAs (hsa-miR-141-3p, hsa-miR-141-5p, hsa-miR-146a-5p, hsa-miR-194-5p, hsa-miR-200b-3p and hsa-miR-365b-5p) with the ability to correctly classify 91.5% of samples. Subsequent validation confirmed both significantly different expression of selected miRNAs in 5 BM groups as well as their diagnostic potential. CONCLUSION: To date, our study is the first to analyze miRNA expression in various types of BMs using small RNA sequencing to develop a diagnostic classifier and, thus, to help stratify BMs of unknown primary. The presented results confirm the importance of studying the dysregulated expression of miRNAs in BMs and the diagnostic potential of the validated 6-miRNA signature.
- MeSH
- biologické markery MeSH
- dospělí MeSH
- lidé MeSH
- melanom * MeSH
- mikro RNA * genetika metabolismus MeSH
- nádory mozku * genetika MeSH
- nádory neznámé primární lokalizace * MeSH
- Check Tag
- dospělí MeSH
- lidé MeSH
- Publikační typ
- časopisecké články MeSH
Glioblastoma (GBM) is regarded as an aggressive brain tumor that rarely develops extracranial metastases. Despite well-investigated molecular alterations in GBM, there is a limited understanding of these associated with the metastatic potential. We herein present a case report of a 43-year-old woman with frontal GBM with primitive neuronal component who underwent gross total resection followed by chemoradiation. Five months after surgery, the patient was diagnosed with an intraspinal GBM metastasis. Next-generation sequencing analysis of both the primary and metastatic GBM tissues was performed using the Illumina TruSight Tumor 170 assay. The number of single nucleotide variants observed in the metastatic sample was more than two times higher. Mutations in TP53, PTEN, and RB1 found in the primary and metastatic tissue samples indicated the mesenchymal molecular GBM subtype. Among others, there were two inactivating mutations (Arg1026Ile, Trp1831Ter) detected in the NF1 gene, two novel NOTCH3 variants of unknown significance predicted to be damaging (Pro1505Thr, Cys1099Tyr), one novel ARID1A variant of unknown significance (Arg1046Ser), and one gene fusion of unknown significance, EIF2B5-KIF5B, in the metastatic sample. Based on the literature evidence, the alterations of NF1, NOTCH3, and ARID1A could explain, at least in part, the acquired invasiveness and metastatic potential in this particular GBM case.
- Publikační typ
- kazuistiky MeSH
Východiska: Mozkové metastázy (BM) jsou nejčastějšími intrakraniálními nádory u dospělých onkologických pacientů. Zatímco dříve byly BM léčeny pouze symptomaticky, přístup k terapii se mění v důsledku zvyšující se incidence, vyplývající z efektivnější léčby primárních nádorů a časnějšího záchytu malých asymptomatických BM. Protože prognóza pacientů s BM je vysoce variabilní, bylo by užitečné zlepšit diagnostické a prognostické nástroje začleněním nových výkonných biomarkerů. MikroRNA (miRNA) jsou v tomto ohledu slibné a díky své vysoké stabilitě vhodné jak pro sekvenování (RNA-Seq), tak pro retrospektivní analýzy v tkáních fixovaných formalinem a zalitých v parafinu (formalin-fixed and paraffin embedded – FFPE). Materiál a metody: Celková RNA obohacená o miRNA byla izolována ze 71 čerstvě zmrazených histopatologicky potvrzených tkání BM s původem v pěti typech nádorů (karcinom plic, 37 %; melanom, 23 %; karcinom prsu, 18 %; renální karcinom, 15 %; kolorektální karcinom, 7 %). Před léčbou byl od každého pacienta získán informovaný souhlas schválený místní etickou komisí. Z RNA byly připraveny knihovny pro sekvenování na platformě NextSeq 500 (Illumina). Mapování čtení na referenci bylo provedeno pomocí nástroje miraligner a databáze miRBase a diferenciální analýza pro 2 437 maturovaných miRNA pomocí nástroje limma. Molekuly miRNA ze vzorků celkové RNA izolovaných z retrospektivního souboru 119 FFPE tkání byly reverzně transkribovány a exprese vybraných diferenciálně exprimovaných miRNA (miR-122-5p, miR-141-3p, miR-146a-5p, miR-194-5p, miR-200c-3p, miR-211-3p, miR-215-5p, miR-514b-3p, miR-934, miR-1270) byla validována pomocí qPCR. Výsledky: Diferenciální analýzou bylo identifikováno 373 miRNA s významně odlišnou expresí mezi pěti skupinami BM (p < 0,001). Následnou validací byla ověřena významně odlišná exprese vybraných miRNA v pěti skupinách BM. Závěr: Prezentované výsledky potvrzují důležitost studia dysregulované exprese miRNA v BM a diagnostický potenciál validovaných miRNA.
Background: Brain metastases (BM) are the most common intracranial tumors in adult cancer patients. While previously BMs were only treated symptomatically, the approach to therapy is changing due to the increasing incidence resulting from more effective treatment of primary tumors and earlier detection of small asymptomatic BMs. As the prognosis of patients with BM is highly variable, it would be useful to improve diagnostic and prognostic tools by incorporating new powerful biomarkers. MicroRNAs (miRNAs) are promising in this regard and, due to their high stability, suitable both for sequencing (RNA-Seq) and for retrospective analyses in formalin-fixed and paraffin-embedded (FFPE) tissues. Material and methods: Total RNA enriched for miRNAs was isolated from 71 fresh-frozen histopathologically confirmed BM tissues originating from 5 tumor types (lung cancer, 37%; melanoma, 23%; breast cancer, 18%; renal cell carcinoma, 15%; colorectal carcinoma, 7%). Informed consent approved by the local ethics committee was obtained from each patient before treatment. Libraries were prepared from RNA for sequencing on the NextSeq 500 platform (Illumina). Read-to-reference mapping was performed using the tool miraligner and the database miRBase, and differential analysis for 2 437 matured miRNAs was done using the tool limma. MiRNA molecules from total RNA samples isolated from a retrospective set of 119 FFPE tissues were reverse transcribed and the expression of selected differentially expressed miRNAs (miR-122-5p, miR-141-3p, miR-146a-5p, miR-194-5p, miR-200c -3p, miR-211-3p, miR-215-5p, miR-514b-3p, miR-934, miR-1270) was validated by qPCR. Results: Differential analysis identified 373 miRNAs with significantly different expression between the five BM groups (P < 0.001). Subsequent pilot validation verified significantly different expression of selected miRNAs in five BM groups. Conclusion: The presented results confirm the importance of studying dysregulated miRNA expression in BM and the diagnostic potential of validated miRNAs.
Brain metastases are the most frequent intracranial tumors in adults and the cause of death in almost one-fourth of cases. The incidence of brain metastases is steadily increasing. The main reason for this increase could be the introduction of new and more efficient therapeutic strategies that lead to longer survival but, at the same time, cause a higher risk of brain parenchyma infiltration. In addition, the advances in imaging methodology, which provide earlier identification of brain metastases, may also be a reason for the higher recorded number of patients with these tumors. Metastasis is a complex biological process that is still largely unexplored, influenced by many factors and involving many molecules. A deeper understanding of the process will allow the discovery of more effective diagnostic and therapeutic approaches that could improve the quality and length of patient survival. Recent studies have shown that microRNAs (miRNAs) are essential molecules that are involved in specific steps of the metastatic cascade. MiRNAs are endogenously expressed small non-coding RNAs that act as post-transcriptional regulators of gene expression and thus regulate most cellular processes. The dysregulation of these molecules has been implicated in many cancers, including brain metastases. Therefore, miRNAs represent promising diagnostic molecules and therapeutic targets in brain metastases. This review summarizes the current knowledge on the importance of miRNAs in brain metastasis, focusing on their involvement in the metastatic cascade and their potential clinical implications.
- Publikační typ
- časopisecké články MeSH
- přehledy MeSH
BACKGROUND/AIM: Glioblastoma (GBM) is one of the deadliest human cancers responding very poorly to therapy. Although the central nervous system has been traditionally considered an immunologically privileged site with an enhanced immune response, GBM appears to benefit from this immunosuppressive milieu. Immunomodulatory molecules play an important role in immune tumor-host interactions. Non-classical human leukocyte antigens (HLA) class Ib molecules HLA-E, HLA-F, and HLA-G have been previously described to be involved in protecting semi-allogeneic fetal allografts from the maternal immune response and in transplant tolerance as well as tumoral immune escape. Unfortunately, their role in GBM remains poorly understood. Our study, therefore, aimed to characterize the relationship between the expression of these molecules in GBM on the transcriptional level and clinicopathological and molecular features of GBM as well as the effect of ionizing radiation. MATERIALS AND METHODS: We performed the analysis of HLA-E, HLA-F, and HLA-G mRNA expression in 69 GBM tissue samples and 21 non-tumor brain tissue samples (controls) by reverse transcription polymerase chain reaction. Furthermore, two primary GBM cell cultures had been irradiated to identify the effect of ionizing radiation on the expression of non-classical HLA molecules. RESULTS: Analyses revealed that both HLA-E and HLA-F are significantly up-regulated in GBM samples. Subsequent survival analysis showed a significant association between low expression of HLA-E and shorter survival of GBM patients. The dysregulated expression of both molecules was also observed between patients with methylated and unmethylated O-6-methylguanine-DNA methyltransferase (MGMT) promoter. Finally, we showed that ionizing radiation increased HLA-E expression level in GBM cells in vitro. CONCLUSION: HLA-E and HLA-F play an important role in GBM biology and could be used as diagnostic biomarkers, and in the case of HLA-E also as a prognostic biomarker.
