Application of the FICTION technique for the simultaneous detection of immunophenotype and chromosomal abnormalities in routinely fixed, paraffin wax embedded bone marrow trephines

Epidemiological findings suggesting that formaldehyde exposure is associated with a higher risk of acute myelogenous leukemia (AML) and other hematological cancers have led to consideration of the potential mechanism of action by which inhalation of this rapidly reactive agent can cause bone marrow cancer. Two major mechanism-based arguments against formaldehyde as a leukemogen have been the difficulty in envisioning how inhaled formaldehyde might penetrate to the bone marrow; and the lack of similarity of non-cancer effects to other known human myeloleukemogens, particularly the absence of pancytopenia in humans or laboratory animals exposed to high levels. However, both of these arguments have been addressed by the recent finding of a pancytopenic effect and chromosomal abnormalities in heavily exposed Chinese workers which, if replicated, are indicative of a genotoxic effect of formaldehyde on hematopoietic stem cells that is in keeping with other known human leukemogens. Review of the body of evidence suggests an apparent discrepancy between studies in laboratory animals, which generally fail to show evidence of penetration of formaldehyde into the blood or evidence of blood or bone marrow genotoxicity, and studies of exposed humans in which there tends to be evidence of genotoxicity in circulating blood cells. One possible explanation for this discrepancy is species difference. Another possible explanation is that myeloid precursors within the nasal mucosa may be the site for leukemogenesis. However, chloromas, which are local collections of myeloid tumor cells, are rarely if ever found in the nose. Other proposed mechanisms for formaldehyde leukemogenesis are reviewed, and dose issues at the interface between the epidemiological and hematotoxicological findings are explored.

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Hemophagocytic Lymphohistiocytosis Complicating Erythroleukemia in a Child with Monosomy 7: A Case Report and Review of the Literature

Case Reports in Hematology ◽

10.1155/2013/581073 ◽

2013 ◽

Vol 2013 ◽

pp. 1-3

Author(s):

Samin Alavi ◽

Maryam Ebadi ◽

Alireza Jenabzadeh ◽

M. T. Arzanian ◽

Sh. Shamsian

Keyword(s):

Bone Marrow ◽

Case Report ◽

Hemophagocytic Lymphohistiocytosis ◽

Chromosomal Abnormalities ◽

Bone Marrow Aspiration ◽

Review Of The Literature ◽

Monosomy 7 ◽

Persistent Fever ◽

First Case ◽

Erythroid Precursors

Herein, the first case of childhood erythrophagocytosis following chemotherapy for erythroleukemia in a child with monosomy 7 is reported. A 5-year-old boy presented with anemia, thrombocytopenia, and hepatosplenomegaly in whom erythroleukemia was diagnosed. Prolonged pancytopenia accompanied by persistent fever and huge splenomegaly and hepatomegaly became evident after 2 courses of chemotherapy. On bone marrow aspiration, macrophages phagocytosing erythroid precursors were observed and the diagnosis of HLH was established; additionally, monosomy 7 was detected on bone marrow cytogenetic examination. In conclusion, monosomy 7 can lead to erythrophagocytosis associated with erythroid leukemia and should be considered among the chromosomal abnormalities contributing to the association.

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A New Approach to Phenotyping Disseminated Tumor Cells: Methodological advances and Clinical Implications

The International Journal of Biological Markers ◽

10.1177/172460080001500119 ◽

2000 ◽

Vol 15 (1) ◽

pp. 100-104 ◽

Cited By ~ 15

Author(s):

F. Noack ◽

M. Schmitt ◽

J. Bauer ◽

D. Helmecke ◽

W. Krüger ◽

...

Keyword(s):

Bone Marrow ◽

Tumor Cells ◽

Laser Scanning ◽

Simultaneous Detection ◽

Disseminated Tumor Cells ◽

Laser Scanning Microscopy ◽

Confocal Laser ◽

Prognostic Impact ◽

Primary Therapy ◽

Double Immunofluorescence

At the time of primary therapy (surgery, systemic chemotherapy and/or radiation), disseminated tumor cells in the bone marrow can be found in almost one-third of patients with cancer of the breast, ovary, esophagus, stomach, colon, and other solid tumors. Whereas the prognostic impact of the mere presence of these cells is still a matter of debate, it has been shown that expression of tumor-associated antigens in disseminated tumor cells is linked to more aggressive disease. Therefore, further characterization of disseminated tumor cells at the protein and gene level has become increasingly important. To date, the most common detection method for disseminated tumor cells in the bone marrow is an immunocytochemical approach using cytokeratin-directed antibodies for detection of epithelial cells and the APAAP system for their visualization. We have established a new double immunofluorescence technique enabling simultaneous detection, phenotyping, and antigen quantification of disseminated tumor cells. Mononuclear cells from bone marrow are enriched by Ficoll gradient centrifugation and cytospins are prepared. Double immunofluorescence is performed using antibodies against cytokeratins 8/18/19 (mAb A45B/B3) and the uPA receptor CD87 (pAb HU277). CD87 expression is recorded by confocal laser scanning microscopy (CLSM) using fluorescence labeled latex beads as the reference; staining intensities of all the scans are then summed and quantified (extended focus). This protocol, originally designed for disseminated tumor cells in bone marrow, can also be applied to disseminated tumor cells in blood, to leukapheresis cells or to cells present in malignant ascites or other malignant effusions. The tumor cells detected may be used for gene and mRNA analyses. Furthermore, disseminated tumor cells also represent interesting targets for clinical studies on patient prognosis or prediction of therapy response as well as for specific tumor-biological therapies.

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Chromosome 10 Abnormality Predicts Prognosis of Neuroblastoma Patients With Bone Marrow Metastasischromosome 10 Abnormality Predicts Prognosis of Neuroblastoma Patients With Bone Marrow Metastasis

10.21203/rs.3.rs-146698/v1 ◽

2021 ◽

Author(s):

Chiyi Jiang ◽

Xiao Xu ◽

Binglin Jian ◽

Xue Zhang ◽

Zhixia Yue ◽

...

Keyword(s):

Bone Marrow ◽

Clinical Characteristics ◽

Chromosomal Abnormalities ◽

Abnormal Chromosome ◽

Mycn Amplification ◽

Chromosome 10 ◽

Orbital Metastases ◽

One Year ◽

The Relationship

Abstract Background Neuroblastoma (NB) is the most common extracranial solid tumor in children with high heterogeneity and concealed onset. The mechanism for its occurrence and development has not been revealed. The purpose of this study was to summarize the clinical characteristics of children with NB and abnormal chromosome 10. To investigate the relationship between the number and structure of chromosome 10 abnormality and NB prognosis.MethodsWe used chromosome G-banding in the first diagnosis to evaluate the genetics of chromosomes in patients with NB, and follow up their clinical characteristics and prognosis. All participants were diagnosed with NB in Hematology Oncology Center, Beijing Children’s Hospital from May 2015 to December 2018, and were followed up for at least one year. ResultsOf all 150 patients with bone marrow metastases, 42 were clearly diagnosed with chromosomal abnormalities. There were 13 patients with chromosome 10 abnormalities definitely, and the loss of chromosome 10 was the most common decrease in the number of chromosomes. These 13 patient had higher LDH, lower OS and EFS than that of children in abnormal group without chromosome 10 abnormality. Eight patients both had MYCN amplification and 1p36 deletion. Two of them had optic nerve damage and no vision, and 1 had left supraorbital metastases five months after treatment. Among the 16 children with suspected chromosome 10 abnormalities, 3 also had orbital metastases. ConclusionsThe above results showed that chromosome 10 might be a new prognostic marker. MYCN amplification and 1p36 deletion may be related with chromosome 10 abnormalities in NB. And NB patients with abnormal chromosome 10 were prone to have orbital metastases.

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Myelofibrosis osteoclasts are clonal and functionally impaired

Blood ◽

10.1182/blood-2018-10-878926 ◽

2019 ◽

Vol 133 (21) ◽

pp. 2320-2324 ◽

Cited By ~ 1

Author(s):

Ivo Veletic ◽

Taghi Manshouri ◽

Asha S. Multani ◽

C. Cameron Yin ◽

Lei Chen ◽

...

Keyword(s):

Bone Marrow ◽

Bone Remodeling ◽

Chromosomal Abnormalities ◽

Cathepsin K ◽

Janus Kinase ◽

Tartrate Resistant Acid Phosphatase ◽

Healthy Individuals ◽

Functionally Impaired ◽

Calr Mutations ◽

Mineral Resorption

Abstract Bone marrow (BM) sclerosis is commonly found in patients with late-stage myelofibrosis (MF). Because osteoclasts (OCs) and osteoblasts play a key role in bone remodeling, and MF monocytes, the OC precursors, are derived from the neoplastic clone, we wondered whether decreased OC numbers or impairment in their osteolytic function affects the development of osteosclerosis. Analysis of BM biopsies from 50 MF patients showed increased numbers of multinucleated tartrate-resistant acid phosphatase (TRAP)/cathepsin K+ OCs expressing phosphorylated Janus kinase 2 (JAK2). Randomly microdissected TRAP+ OCs from 16 MF patients harbored JAK2 or calreticulin (CALR) mutations, confirming MF OCs are clonal. To study OC function, CD14+ monocytes from MF patients and healthy individuals were cultured and differentiated into OCs. Unlike normal OCs, MF OCs appeared small and round, with few protrusions, and carried the mutations and chromosomal abnormalities of neoplastic clones. In addition, MF OCs lacked F-actin–rich ring-like structures and had fewer nuclei and reduced colocalization signals, compatible with decreased fusion events, and their mineral resorption capacity was significantly reduced, indicating impaired osteolytic function. Taken together, our data suggest that, although the numbers of MF OCs are increased, their impaired osteolytic activity distorts bone remodeling and contributes to the induction of osteosclerosis.

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Tissue methylation and demethylation influence translesion synthesis DNA polymerases (TLS) contributing to the genesis of chromosomal abnormalities in myelodysplastic syndrome

Journal of Clinical Pathology ◽

10.1136/jclinpath-2020-207131 ◽

2020 ◽

pp. jclinpath-2020-207131

Author(s):

Gabrielle Melo Cavalcante ◽

Daniela Paula Borges ◽

Roberta Taiane Germano de Oliveira ◽

Cristiana Libardi Miranda Furtado ◽

Ana Paula Negreiros Nunes Alves ◽

...

Keyword(s):

Dna Methylation ◽

Bone Marrow ◽

Myelodysplastic Syndrome ◽

Chromosomal Abnormalities ◽

Dna Polymerases ◽

Translesion Synthesis ◽

Normal Karyotype ◽

Dna Demethylation ◽

Repair System ◽

Low Expression

AimsDNA methylation has its distribution influenced by DNA demethylation processes with the catalytic conversion of 5-methylcytosine (5mC) into 5-hydroxymethylcytosine (5hmC). Myelodysplastic syndrome (MDS) has been associated with epigenetic dysregulation of genes related to DNA repair system, chronic immune response and cell cycle.MethodsWe evaluated the tissue DNA methylation/hydroxymethylation in bone marrow trephine biopsies of 73 patients with MDS, trying to correlate with the mRNA expression of 21 genes (POLH, POLL, REV3L, POLN, POLQ, POLI, POLK, IRF-1, IRF-2, IRF-3, IRF-4, IRF-5, IRF6, IRF-7, IRF-8,IRF-9, MAD2, CDC20, AURKA, AURKB and TPX2).ResultsThe M-score (5mC) was significantly higher in patients with chromosomal abnormalities than patients with normal karyotype (95% CI –27.127779 to –2.368020; p=0.022). We observed a higher 5mC/5hmC ratio in patients classified as high-risk subtypes compared with low-risk subtypes (95% CI –72.922115 to –1.855662; p=0.040) as well as patients with hypercellular bone marrow compared with patients with normocellular/hypocellular bone marrow (95% CI –69.189259 to –0.511828; p=0.047) and with the presence of dyserythropoiesis (95% CI 17.077703 to 51.331388; p=0.001). DNA pols with translesion activity are significantly influenced by methylation. As 5mC immunoexpression increases, the expressions of POLH (r=−0.816; r2 =0.665; p=0.000), POLQ (r=−0.790; r2=0.624; p=0.001), PCNA (r=−0.635; r2=0.403; p=0.020), POLK (r=−0.633; r2=0.400; p=0.036 and REV1 (r=−0.578; r2=0.334; p=0.049) decrease.ConclusionsOur results confirm that there is an imbalance in the DNA methylation in MDS, influencing the development of chromosomal abnormalities which may be associated with the low expression of DNA polymerases with translesion synthesis polymerases activity.

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Chromosomal Abnormalities in Multiple Myeloma

Blood ◽

10.1182/blood.v30.6.738.738 ◽

1967 ◽

Vol 30 (6) ◽

pp. 738-748 ◽

Cited By ~ 18

Author(s):

K. C. DAS ◽

B. K. AIKAT

Keyword(s):

Multiple Myeloma ◽

Bone Marrow ◽

Chromosomal Aberrations ◽

Chromosomal Abnormality ◽

Chromosomal Abnormalities ◽

Acute Leukemias ◽

Leukocyte Culture ◽

Peripheral Leukocyte ◽

Direct Preparation ◽

Chromosomal Changes

Abstract Chromosome studies were carried out in five cases of multiple myeloma by peripheral leukocyte culture and direct preparation from bonemarrow cells. No chromosomal abnormality was detected by leukocyte culture in any of these cases. Analysis of bone marrow metaphase plates revealed abnormalities in four out of five cases. Chromosomal changes were characterized by their variability from case to case, as well as among different groups of cells in individual cases. Both numerical and structural anomalies were encountered. The chromosome series more frequently involved were C, A, G and B in decreasing order. The diverse chromosomal aberrations encountered in these four cases of multiple myeloma are reminiscent of those described in acute leukemias.

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Biosystem Analysis of the Hypoxia Inducible Domain Family Member 2A: Implications in Cancer Biology

Genes ◽

10.3390/genes11020206 ◽

2020 ◽

Vol 11 (2) ◽

pp. 206 ◽

Cited By ~ 2

Author(s):

Celia Salazar ◽

Osvaldo Yañez ◽

Alvaro A. Elorza ◽

Natalie Cortes ◽

Olimpo García-Beltrán ◽

...

Keyword(s):

Gene Expression ◽

Dna Methylation ◽

Bone Marrow ◽

Cancer Biology ◽

Chromosomal Abnormalities ◽

Human Cancer ◽

Expression Profiles ◽

Homology Modelling ◽

B Cell Lymphoma ◽

Qrt Pcr

The expression of HIGD2A is dependent on oxygen levels, glucose concentration, and cell cycle progression. This gene encodes for protein HIG2A, found in mitochondria and the nucleus, promoting cell survival in hypoxic conditions. The genomic location of HIGD2A is in chromosome 5q35.2, where several chromosomal abnormalities are related to numerous cancers. The analysis of high definition expression profiles of HIGD2A suggests a role for HIG2A in cancer biology. Accordingly, the research objective was to perform a molecular biosystem analysis of HIGD2A aiming to discover HIG2A implications in cancer biology. For this purpose, public databases such as SWISS-MODEL protein structure homology-modelling server, Catalogue of Somatic Mutations in Cancer (COSMIC), Gene Expression Omnibus (GEO), MethHC: a database of DNA methylation and gene expression in human cancer, and microRNA-target interactions database (miRTarBase) were accessed. We also evaluated, by using Real-Time Quantitative Reverse Transcription Polymerase Chain Reaction (qRT-PCR), the expression of Higd2a gene in healthy bone marrow-liver-spleen tissues of mice after quercetin (50 mg/kg) treatment. Thus, among the structural features of HIG2A protein that may participate in HIG2A translocation to the nucleus are an importin α-dependent nuclear localization signal (NLS), a motif of DNA binding residues and a probable SUMOylating residue. HIGD2A gene is not implicated in cancer via mutation. In addition, DNA methylation and mRNA expression of HIGD2A gene present significant alterations in several cancers; HIGD2A gene showed significant higher expression in Diffuse Large B-cell Lymphoma (DLBCL). Hypoxic tissues characterize the “bone marrow-liver-spleen” DLBCL type. The relative quantification, by using qRT-PCR, showed that Higd2a expression is higher in bone marrow than in the liver or spleen. In addition, it was observed that quercetin modulated the expression of Higd2a gene in mice. As an assembly factor of mitochondrial respirasomes, HIG2A might be unexpectedly involved in the change of cellular energetics happening in cancer. As a result, it is worth continuing to explore the role of HIGD2A in cancer biology.

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