Cancer‐related mRNA expression analysis using a novel flow cytometry‐based assay

Depreter, Barbara; Philippé, Jan; Meul, Magali; Denys, Barbara; Vandepoele, Karl; Moerloose, Barbara De; Lammens, Tim

doi:10.1002/cyto.b.21593

Cited by 5 publications

(5 citation statements)

References 31 publications

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“…To date, more than 250 different molecular aberrancies have been found to be associated with leukemogenesis and a comprehensive knowledge about their molecular epidemiology is still missing. Incorporation of “old-fashion” techniques as conventional karyotyping or aneuploidy which lack in our study and the avant-garde NGS, 58 next generation flow cytometry (NGF) 59 and why not to start to think in combined technologies as PrimeFlow 60 to detect RNA targets in a particular population of cells by using flow cytometry, will help to enhance survival and will allow to find a more precise patient stratification and appropriate follow-up.…”

Section: Resultsmentioning

confidence: 98%

Poor Prognosis Biomolecular Factors Are Highly Frequent in Childhood Acute Leukemias From Oaxaca, Mexico

Juárez-Avendaño

Luna-Silva²,

Chargoy-Vivaldo

et al. 2020

Technol Cancer Res Treat

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Objective: To investigate the cellular and molecular epidemiology of acute leukemias in vulnerable populations of children and adolescents in Oaxaca de Juarez, Mexico. Material and Methods: Descriptive, cross-sectional and retrospective study, conducted from 2014 to 2018 in which profiles of molecular and immunophenotypic aberrations were investigated in children and adolescents diagnosed with acute leukemia, by evaluating 28 molecular abnormalities by HemaVision-Q28 multiplex RT-PCR kit and standardized EuroFlow Immunophenotyping of bone marrow cells. Results: We included 218 patients, with 82.5% younger than 14 years and 17.5% adolescents. The median age was 9 years and a main peak of incidence was recorded at age of 4 to 5 years. B-cell acute lymphoblastic leukemia was diagnosed in 70.64% of all cases, acute myeloid leukemia was in 22.48%, T-cell acute lymphoblastic leukemia in 6.42%, and mixed lineage acute leukemia in 0.46% of cases. Overall, chromosomal translocations were positive in 29.82% of cases. While 65.31% of patients with acute myeloid leukemia reported aberrancies, only in 18.83% of B-cell acute lymphoblastic leukemia cases genetic abnormalities were obvious. Surprisingly, most prevalent translocations in B-cell acute lymphoblastic leukemia were t(9;22) in 20.7%, followed by t(4;11) in 17.2% and t(6;11) in 13.8%, whereas patients with acute myeloid leukemia showed t(15;17) in 40.6% and t(8;21) in 21.9%. In contrast, an homogeneous expression of t(3;21) and t(6;11) was recorded for T-cell acute lymphoblastic leukemia and mixed lineage acute leukemia cases, respectively. Except for t(1;19), expressed only by pre-B cells, there was no association of any of the studied translocations with differentiation stages of the B-leukemic developmental pathway. Conclusion: Our findings identify near 50% of patients with acute lymphoblastic leukemia at debut with high-risk translocations and poor prognosis in B-cell acute lymphoblastic leukemia as well as an unexpected increase of acute myeloid leukemia cases in young children, suggesting a molecular shift that support a higher incidence of poor prognosis cases in Oaxaca.

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Section: Resultsmentioning

confidence: 98%

Poor Prognosis Biomolecular Factors Are Highly Frequent in Childhood Acute Leukemias From Oaxaca, Mexico

Juárez-Avendaño

Luna-Silva²,

Chargoy-Vivaldo

et al. 2020

Technol Cancer Res Treat

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“…Previous work by our group has consistently shown PIM1 gene expression in CD4+ T cells freshly isolated from peripheral blood to be significantly elevated during the development of RA compared with other diseases in an early arthritis clinic, being a component of a STAT3-regulated transcriptional program in these cells (15)(16)(17). Validation of a flow cytometry assay for PIM1 transcript measurement at a cellular level was undertaken in the absence of specific antibodies suitable for conventional flow cytometry (40). The assay circumvents the need to isolate leukocyte subsets, and validation of the technique against gold standard qRT-PCR readouts introduces a method that holds promise as a tractable test, using frozen PBMC samples.…”

Section: Discussionmentioning

confidence: 99%

Pim Kinases as Therapeutic Targets in Early Rheumatoid Arthritis

Maney

Lemos

Barron-Millar

et al. 2021

Arthritis & Rheumatology

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Objective As well as being an established oncoprotein and therapeutic target in cancer, proviral integration site for Moloney murine leukemia virus 1 (Pim‐1) is implicated in human autoimmunity. This study was undertaken to investigate Pim‐1 and its family members as potential therapeutic targets in early rheumatoid arthritis (RA). Methods A flow cytometry assay for PIM1 transcript measurement in peripheral blood mononuclear cells from patients with early arthritis was validated and applied as a biomarker of Pim‐1 activity at the cellular level. Synovial protein expression was similarly determined by multiplex immunofluorescence in tissue samples from untreated RA patients and non‐RA disease controls. Functional consequences of Pim kinase family manipulation in freshly isolated CD4+ T cells from these individuals were ascertained, along with the impact of Pim inhibition on mice with collagen‐induced arthritis (CIA). Results The percentage of circulating CD4+ T cells positive for PIM1 transcript by flow cytometry proved a faithful surrogate for gene expression and was significantly higher in patients with early RA than in those with other diseases. Pim‐1 protein levels were similarly up‐regulated in synovial CD4+ T cells from patients with early RA. Ex vivo, exposure of T cell receptor–stimulated early RA CD4+ T cells to Pim kinase inhibitors restrained their activation and proliferative capacity. Diminished production of proinflammatory cytokines (interferon‐γ and interleukin‐17) and an expanded CD25highFoxP3+ Treg cell fraction were also observed in exposed versus unexposed cells. Finally, administration of Pim inhibitors robustly limited arthritis progression and cartilage destruction in CIA. Conclusion Our findings indicate that Pim kinases are plausible therapeutic targets in a readily identifiable subgroup of patients with early RA. Repurposing of Pim inhibitors for this disease should be considered.

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“…In addition, quantification of multiple proteins in single cells using dyes or fluorescently-labeled antibodies can be achieved using flow cytometry. Similarly, flow cytometry has also been used to measure mRNA or microRNA expression levels using labeled antisense nucleotides [65, 66]. In combination with Fluorescence Activated Cell Sorting (FACS), target cells can be isolated and collected in a high-throughput manner, enabling further comprehensive gene expression and proteomic analysis.…”

Section: Analytical Approaches To Study Cell–cell Interactionsmentioning

confidence: 99%

Emerging approaches to study cell-cell interactions in tumor microenvironment

Nishida-Aoki

Gujral

2019

Oncotarget

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Cell–cell interactions are of crucial importance for tissue formation, homeostasis, regeneration processes, and immune response. Recent studies underlined contribution of cell–cell interaction in tumor microenvironment (TME) for tumor progression and metastasis. Cancer cells modify the host cells to tumor-supportive traits, and the modified host cells contribute to tumor progression by interacting with cancer cells and further modifying other normal cells. However, the complex interaction networks of cancer cells and host cells remained largely unknown. Recent advances in high throughput microscopy and single cells-based molecular analyses have unlocked a new era for studying cell–cell interactions in the complex tissue microenvironment at the resolution of a single cell. Here, we review various model systems and emerging experimental approaches that are used to study cell–cell interactions focusing on the studies of TME. We discuss strengths and weaknesses of each model system and each experimental approach, and how upcoming approaches can solve current fundamental questions of cell–cell interactions in TME.

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Cancer‐related mRNA expression analysis using a novel flow cytometry‐based assay

Abstract: PrimeFlow™ is a sensitive technique to investigate mRNA expressions, with high concordance to RT-qPCR. High (MYCN) and subtle (WT1) overexpressed mRNA targets can be quantified in heterogeneous and rare subpopulations e.g. LSCs. © 2017 International Clinical Cytometry Society.

Cited by 5 publications

References 31 publications

Poor Prognosis Biomolecular Factors Are Highly Frequent in Childhood Acute Leukemias From Oaxaca, Mexico

Poor Prognosis Biomolecular Factors Are Highly Frequent in Childhood Acute Leukemias From Oaxaca, Mexico

Pim Kinases as Therapeutic Targets in Early Rheumatoid Arthritis

Emerging approaches to study cell-cell interactions in tumor microenvironment

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