Intraosseous intraneural perineurioma derived from the inferior alveolar nerve with an abnormality of chromosome 22 and expression of the BCR-ABL fusion gene: report of a case and review of recent literature.

BACKGROUND: Perineurioma (PN) is a peripheral nerve disease that primarily develops in the limbs and trunk and very rarely occurs in the oral cavity. PN is classified into two types: intraneural perineurioma (INPN) and soft tissue perineurioma (extraneural perineurioma, ENPN). In this article, we report a patient with mandibular body INPN derived from the perineurium of the inferior alveolar nerve. CASE PRESENTATION: The patient was a 43-year-old male. He consulted our department for a detailed examination of the right mandibular body. A biopsy was performed at another hospital and he was diagnosed with a schwannoma. At his first visit, hypesthesia extending from the right lower lip to the mental region was recognized and enlargement of the right mandibular canal was confirmed with X-ray CT and MRI. Considering the possibility of future tumor growth, we extirpated the tumor under general anesthesia. Cystic tumor was seen continuously in the inferior alveolar nerve. Immunohistologically, the tumor cells were positive for Glut-1, weakly positive for EMA, and weakly positive for Claudin-1, and the histopathological diagnosis was INPN. In addition, absence of the BCR region of chromosome 22 and expression of the BCR-ABL fusion gene were observed by fluorescent in situ hybridization (FISH), and a chromosome 22 abnormality was confirmed. These findings indicated that the disease was a neoplastic lesion. CONCLUSION: Expression of the BCR-ABL fusion gene in INPN that develops in the oral cavity is thought to be very rare, and to the best of our knowledge, ours is the first case to be reported in the literature. About three postoperative years have passed, but findings suggestive of recurrence have not been observed.

[Inhibitory effect and mechanism of platycodin D combined with imatinib on K562/R].

Platycodin D(PD) has a significantly inhibitory effect on multiple malignant tumors, and can inhibit the proliferation of leukemia cells K562 and induce apoptosis. However, its effect in improving the sensitivity of drug-resistant cells to imatinib and their molecular mechanism remained unclear. To investigate the effect and mechanism of PD alone or combined with imatinib (IM) in inhibiting CML imatinib resistant cell line K562/R, the cell proliferation was examined by CCK8 assay to reveal the effect of PD on the inhibitory function of imatinib. Cell apoptosis was detected by Annexin V-FITC/PI double staining. Protein expressions of cleaved caspase-3, cleaved caspase-9, PARP, cleaved PARP, Bcr/abl, p-AKT and p-mTOR were detected by Western blot. The results showed that the inhibitory effect of PD combined with imatinib on the proliferation and apoptosis of K562/R cells was significantly higher than that of the control group and the single drug group. Protein expressions of cleaved caspase-3, cleaved caspase-9 and cleaved PARP were significantly up-regulated in the combination group, and protein expressions of PARP, Bcr/abl, p-AKT and p-mTOR were down-regulated. The results indicated that PD increased the sensitivity of drug-resistant cells to imatinib, and the inhibitory effect of PD combined with imatinib was significantly better than the single drug on cell proliferation, induction of apoptosis, inhibition of Bcr/abl protein and PI3K/AKT/mTOR signaling pathway.

Magnetic Nanoparticles PCR Enzyme-Linked Gene Assay for Quantitative Detection of BCR/ABL Fusion Gene in Chronic Myelogenous Leukemia.

BACKGROUND: Magnetic nanoparticles (MNPs) have been widely used in medical diagnostic research. In this work, two technologies, MNPs and polymerase chain reaction (PCR), were combined to increase detection sensitivity and specificity. A novel technique based on the MNPs-PCR enzyme-linked gene assay (MELGA) was developed for detection of the BCR/ABL abnormal gene in chronic myelogenous leukemia (CML) patients. METHODS: An MNPs-labeled BCR forward primer and a biotin-labeled ABL reverse primer were used to specifically amplify the target gene. After magnetic separation, the PCR product bound to MNPs labeled with streptavidin-conjugated horseradish peroxidase was incubated with the peroxidase substrate and hydrogen peroxide to generate the colorimetric signal. RESULTS: When compared with real-time quantitative-PCR (RQ-PCR), the MELGA technique exhibited an increased sensitivity of <1 fg with high specificity for the BCR/ABL fusion gene in CML patients. In addition, MELGA colorimetric results correlated well with the number of copies obtained from RQ-PCR. CONCLUSION: This simple and cost-effective technique is suitable for monitoring CML patients during targeted therapy (tyrosine kinase inhibitors) especially in rural hospitals.

Acute Basophilic Leukemia: Recent Molecular and Diagnostic Update.

Acute basophilic leukemia (ABL) is an uncommon subtype of acute leukemia characterized by clinical signs and symptoms related to hyper-histaminemia. Patients usually present with bone marrow (BM) failure due to the infiltration of BM by the blasts and may or may not have circulating blasts. Myeloid markers such as CD13 and CD33 are expressed by leukemic blasts, which are also positive for CD123, CD203c, and CD11b, but KIT (CD117) and other monocytic markers are usually negative. t(X;6) (p11; q23) translocation resulting in the MYB-GATA1 fusion gene has been seen in sporadic cases of ABL. Early phases of hematopoiesis are characterized by high levels of MYB and low levels of GATA1; as differentiation develops, an inverse regulation occurs, resulting in high levels of GATA1 and low levels of MYB.The translocation t(X;6) produces the MYB-GATA1 fusion gene (p11; q23). In mouse lineage-negative cells, MYB-GATA1 expression commits them to the granulocyte lineage and inhibited differentiation at an early stage. Cells expressing MYB-GATA1 show enhanced expression of markers of immaturity (CD34), granulocytic lineage (CD33 and CD117), and basophilic differentiation (CD203c and FcRI). NTRK1 and IL1RL1 transcription is directly triggered by MYB and MYB-GATA1, resulting in basophilic skewing of the blasts.

Electrochemical DNA biosensor for chronic myelocytic leukemia based on hybrid nanostructure.

The present research refers to elaborating a new label-free electrochemical biosensor used to detect the BCR/ABL fusion gene. We used a hybrid nanocomposite composed of chitosan and zinc oxide nanoparticles (Chit-ZnONP) immobilized on a polypyrrole (PPy) film. DNA segments were covalently immobilized, allowing biomolecular recognition. Atomic force microscopy (AFM), cyclic voltammetry (CV), and electrochemical impedance spectroscopy (EIS) were used to evaluate the assembly stages of the biosensor. The biosensor's analytical performance was investigated using recombinant plasmids containing the target oncogene and clinical samples from patients with chronic myeloid leukemia (CML). A limit of detection (LOD) of 1.34 fM, limit of quantification (LOQ) of 4.08 fM, and sensitivity of 34.03 µA fM-1 cm2 were calculated for the BCR/ABL fusion oncogene. The sensing system exhibited high specificity, selectivity, and reproducibility with a standard deviation (SD) of 4.21%. Additionally, a linear response range was observed between 138.80 aM to 13.88 pM with a regression coefficient of 0.96. Also, the biosensor shows easy operationalization and fast analytical response, contributing to the early cancer diagnosis. The proposed nanostructured device is an alternative for the genetic identification BCR/ABL fusion gene.

CeO2/MXene heterojunction-based ultrasensitive electrochemiluminescence biosensing for BCR-ABL fusion gene detection combined with dual-toehold strand displacement reaction for signal amplification.

An "on-off" nonenzymatic and ultrasensitive electrochemiluminescence (ECL) biosensing platform has been constructed to detect BCR-ABL fusion gene based on CeO2/MXene heterojunction and configuration-entropy driven dual-toehold strand displacement reaction (DT-SDR) for signal amplification. The CeO2/MXene heterojunction were prepared via one-step hydrothermal method through in situ synthesis of CeO2 nanocubes on the surface of Ti3C2-MXene nanosheets. Surprisingly, the prepared CeO2/MXene heterojunction with good dispersion and excellent conductivity not only significantly enhanced ECL emission of S2O82-/O2 system, but also acted as good electrode modification materials to provide massive active sites for three-stranded ST/AS/BK complex immobilization. In the presence of target BCR-ABL fusion gene and Bio-FS, target BCR-ABL fusion gene bound to dual-toehold exposed at the ends of ST, replacing AS and BK and obtaining ST/target with a loop. Subsequently, Bio-FS bound to the loop (as toehold) in ST strand of ST/target to form ST/Bio-FS, replacing the target to further trigger a new SDA cycle. This configuration-entropy driven DT-SDR made three-stranded ST/AS/BK complex transform into dual-stranded ST/Bio-FS in the electrode interface. Ultimately, the quenching labels of streptavidin modified Pt nanoparticles functionalized polydopamine composites (SA-Pt@PDA) were introduced via biotin and streptavidin recognition, realizing ECL emission quenching of S2O82-/O2 system for "on-off" detection of BCR-ABL fusion gene. The developed ECL biosensor for BCR-ABL fusion gene detection achieves the wide concentration variation from 1 fM to 100 pM with low limit of detection down to 0.27 fM, which provides new enlightenment and basis for molecular diagnosis of chronic myelogenous leukemia in clinical practice.

Sudden extramedullary and extranodal Philadelphia-positive anaplastic large-cell lymphoma transformation during imatinib treatment for CML: A case report.

BACKGROUND: Chronic myeloid leukemia (CML) is a malignant hematologic malignancy that can progress to blast phase with a myeloid or lymphoid phenotype. Some patients with CML can also progress to blast crisis phase; however, the transformation of CML into Philadelphia-positive lymphoma is extremely rare. CASE SUMMARY: We present a patient with CML who experienced a sudden transformation to anaplastic large-cell lymphoma (ALCL) after 7 mo of treatment with imatinib, during which she had achieved partial cytogenetic response as well as early molecular response. The patient noticed a mass in her left shoulder, the biopsy data of which were consistent with ALCL; moreover, her lymphoma cells exhibited BCR-ABL gene fusion. The patient was diagnosed with Philadelphia-positive ALCL that progressed from CML, and was thus treated with the second generation tyrosine kinase inhibitor nilotinib. Six months later, the mass had totally disappeared and the BCR-ABL fusion gene was undetectable in the peripheral blood. To our knowledge, this is the first patient known to have developed Philadelphia-positive ALCL transformed from CML. CONCLUSION: Unexplained lymphadenopathy or an extramedullary mass in a patient with CML may warrant a biopsy and testing for BCR-ABL fusion.

Self-enhanced luminol-based electrochemiluminescent hydrogels: An ultrasensitive biosensing platform for fusion gene analysis coupled with target-initiated DNAzyme motor.

BCR/ABL fusion gene has been discovered as an important and reliable biomarker for early diagnosis of chronic myeloid leukemia (CML). Herein, a novel and switching electrochemiluminescence (ECL) biosensor was developed for ultrasensitive determination of the fusion gene based on the self-enhanced polyethyleneimine-luminol (PEI-Lum) hydrogels coupled with target-initiated DNAzyme motor. The facilely prepared PEI-Lum hydrogels could not only immobilize enormous luminol but shorten the distance of binary system, thus facilitating the mass and electron transfer efficiency of the sensing interface, so that the enhanced ECL signal was achieved. Moreover, the engineering DNA motor was powered by Mg2+-dependent DNAzyme for isothermal DNA signal amplification. As a result, the fabricated ECL biosensor enabled highly sensitive detection of BCR/ABL fusion gene with a broad linear range from 10.0 fM to 10.0 nM and a low detection limit of 3.75 fM (S/N = 3). Significantly, the developed biosensing method provides a potential tool for nucleic acid analysis in clinical diagnosis and a new avenue to design high-efficient ECL nanomaterials.

Association of BCR/ABL transcript variants with different blood parameters and demographic features in Iraqi chronic myeloid leukemia patients.

BACKGROUND: Chronic myeloid leukemia (CML) is a myeloproliferative neoplasm characterized by the presence of BCR-ABL fusion gene (GenBank accession NC_000022.11). In the vast majority of CML patients, the typical subtype of BCR-ABL transcript are b3a2, b2a2 or both. The aim of this study was to determine the different subtypes of BCR-ABL transcript and their impact on the demographic and hematological parameters in Iraqi patients with CML. METHODS: One hundred patients with chronic phase CML (11 newly diagnosed and 89 imatinib-resistant) were enrolled in this study. Ribonucleic acid (RNA) was extracted from leukocytes, and complementary DNA was created using reverse transcriptase polymerase chain reaction technique. A multiplex polymerase chain reaction with four specific primers was used to determine the BCR-ABL fusion subtypes in each patient. RESULTS: Male to female ratio was 1.38:1. Fifty-nine patients expressed b3a2 transcript, whereas 39 of the remaining cases were positive for b2a2 variant. One case expressed b2a3 transcript, while the last case coexpressed the two subtypes of mRNA b3a2/b2a2. Male and female were significantly associated with b3a2 and b2a2 subtypes, respectively. The b3a2 subtype showed higher total leukocyte count than b2a2 subgroup, while b2a2 variant demonstrated significantly elevated platelet counts compared to those with b3a2 transcript. A significantly higher plateletcrit percentage (PCT%) was found in patients with b2a2 transcript whereas. CONCLUSIONS: The testified Iraqi group expressed M-BCR-ABL type with preponderance of b3a2 over b2a2 subtype. There was a gender-skewed distribution in BCR-ABL transcript types with b3a2 transcript more prevalent in males. The type of BCR-ABL transcript is reflected by different leukocyte and platelet counts at diagnosis, which might represent a distinct phenotype and disease biology.

Attomolar electrochemical detection of the BCR/ABL fusion gene based on an amplifying self-signal metal nanoparticle-conducting polymer hybrid composite.

In the last ten years, conjugated polymers started to be used in the immobilization of nucleic acids via non-covalent interactions. In the present study, we describe the construction and use of an electrochemical DNA biosensor based on a nanostructured polyaniline-gold composite, specifically developed for the detection of the BCR/ABL chimeric oncogene. This chromosome translocation is used as a biomarker to confirm the clinical diagnosis of both chronic myelogenous leukemia (CML) and acute lymphocytic leukemia (ALL). The working principle of the biosensor rests on measuring the conductivity resulting from the non-covalent interactions between the hybrid nanocomposite and the DNA probe. The nanostructured platform exhibits a large surface area that enhances the conductivity. Positive cases, which result from the hybridization between DNA probe and targeted gene, induce changes in the amperometric current and in the charge transfer resistance (RCT) responses. Atomic force microscopy (AFM) images showed changes in the genosensor surface after exposure to cDNA sample of patient with leukemia, evidencing the hybridization process. This new hybrid sensing-platform displayed high specificity and selectivity, and its detection limit is estimated to be as low as 69.4 aM. The biosensor showed excellent analytical performance for the detection of the BCR/ABL oncogene in clinical samples of patients with leukemia. Hence, this electrochemical sensor appears as a simple and attractive tool for the molecular diagnosis of the BCR/ABL oncogene even in early-stage cases of leukemia and for the monitoring of minimum levels of residual disease.

Coupling a universal DNA circuit with graphene sheets/polyaniline/AuNPs nanocomposites for the detection of BCR/ABL fusion gene.

This article described a novel method by coupling a universal DNA circuit with graphene sheets/polyaniline/AuNPs nanocomposites (GS/PANI/AuNPs) for highly sensitive and specific detection of BCR/ABL fusion gene (bcr/abl) in chronic myeloid leukemia (CML). DNA circuit known as catalyzed hairpin assembly (CHA) is enzyme-free and can be simply operated to achieve exponential amplification, which has been widely employed in biosensing. However, application of CHA has been hindered by the need of specially redesigned sequences for each single-stranded DNA input. Herein, a transducer hairpin (HP) was designed to obtain a universal DNA circuit with favorable signal-to-background ratio. To further improve signal amplification, GS/PANI/AuNPs with excellent conductivity and enlarged effective area were introduced into this DNA circuit. Consequently, by combining the advantages of CHA and GS/PANI/AuNPs, bcr/abl could be detected in a linear range from 10 pM to 20 nM with a detection limit of 1.05 pM. Moreover, this protocol showed excellent specificity, good stability and was successfully applied for the detection of real sample, which demonstrated its great potential in clinical application.

Casos atípicos e14a3 (b3a3) en el gen de fusión BCR-ABL de la leucemia mieloide crónica en Cuba / Rare cases of e14a3 (b3a3) BCR-ABL fusion in chronic myeloid leukemia in Cuba

Introducción: La leucemia mieloide crónica es un desorden clonal maligno de células madres hematopoyéticas pluripotentes que se caracteriza por la presencia del cromosoma Filadelfia, consecuencia de la traslocación cromosómica recíproca entre los brazos largos de los cromosomas 9 y 22. El resultado de esta alteración cromosómica es un gen de fusión que contiene las uniones b2a2 (e13a2) o b3a2 (e14a2). En la mayor parte de los casos, las células de la leucemia mieloide crónica expresan uno de los dos transcritos (b2a2 o b3a2); sin embargo, el 5 por ciento de los pacientes tienen ambos tipos de ARNm como resultado de empalmes alternativos. Se han encontrado otros transcriptos como e19a2, e2a2, e1a3, e6a2, e13a3(b2a3), y e14a3(b3a3), que ocurren con menos frecuencia. Objetivo: Describir el comportamiento de dos pacientes con leucemia mieloide crónica que presentan un trascripto BCR/ABL atípico. Casos clínicos: En el estudio molecular por reacción en cadena de la polimerasa cualitativo realizado a los dos pacientes, se observó un punto de ruptura del gen de fusión BCR/ABL poco frecuente, el cual se correspondía al transcripto e14a3 (b3a3). Estos pacientes iniciaron tratamiento con mesilato de imatinib a dosis de 400 mg diarios. Al primer paciente a los dos meses de tratamiento se le detectó crisis blástica, por lo que se le cambió el tratamiento a nilotinib 400 mg diarios que mantiene hasta la actualidad. La segunda paciente mantuvo igual tratamiento, aunque en ocasiones ha sido necesario incorporar tratamiento citorreductor con hidroxiurea por presentar leucocitosis. Conclusiones: Los pacientes con BCR/ABL a3 presentan un curso más benigno de la enfermedad. Aunque en los pacientes estudiados no se observó una respuesta satisfactoria al tratamiento pues presentaron diversas complicaciones(AU)

Introduction: Chronic myeloid leukemia is a malignant clonal disorder of pluripotent hematopoietic stem cells and characterized by the presence of the Philadelphia chromosome, which is the product of a reciprocal translocation between the long arms of chromosomes 9 and 22. The result of this chromosomal alteration is a fusion gene that contains the e13a2 (b2a2) and e14a2 (b3a2) junctions. In most cases, chronic myeloid leukemia cells express one of the two transcripts (b2a2 or b3a2); however, 5 percent of patients have both types of mRNA, as a result of alternative junctions. Other transcripts have been identified, such as e19a2, e2a2, e1a3, e6a2, e13a3 (b2a3), and e14a3 (b3a3), which occur less frequently. Objective: To describe the behavior of two patients with chronic myeloid leukemia who have an atypical BCR-ABL transcript. Clinical cases: In a qualitative molecular study of polymerase chain reaction carried out with two patients, a BCR-ABL fusion gene breakpoint was observed, which corresponded to the e14a3 (b3a3) transcript. These patients started treatment with imatinib mesylate at a dose of 400mg/d. At two months, the first patient had a diagnose of blast crisis, so the treatment was changed to nilotinib at a dose of 400mg/d, which the patient maintained to date. The second patient maintained the same treatment, although it was sometimes necessary to incorporate cytoreductive treatment with hydroxyurea due to leukocytosis. Conclusions: Patients with BCR-ABL a3 present a more benign evolution of the disease. However, a satisfactory response to treatment was not observed in the patients studied, as long as they presented various complications(AU)

Graphene sheets, polyaniline and AuNPs based DNA sensor for electrochemical determination of BCR/ABL fusion gene with functional hairpin probe.

A sensitive and selective electrochemical DNA sensor was developed for the detection of BCR/ABL fusion gene in chronic myelogenous leukemia (CML). Firstly, graphene sheets (GS) suspension was prepared with the aid of chitosan (CS) solution and then fabricated onto the glassy carbon electrode (GCE), followed by the electro-polymerization of aniline to form the PANI layer, then, Au nanoparticles (AuNPs) were electro-deposited onto the modified GCE to immobilize the capture probes. The capture probe employed a hairpin structure and dually labeled with a 5'-SH and a 3'-biotin. After hybridization with the target DNA, hairpin structure was compelled to open and 3'-biotin was forced to stay away from the electrode surface. As a result, streptavidin-alkaline phosphatase (SA-AP) was covalently binded to the capture probe via biotin-avidin system. Reduction currents were then generated after catalyzing the hydrolysis of the electroinactive 1-naphthyl phosphate (1-NP) to 1-naphthol and monitored by differential pulse voltammetry (DPV). Under optimum conditions, the amperometric signals increased linearly with the target DNA concentrations (10 pM to 1000 pM), and the DNA sensor exhibited a detection limit as low as 2.11 pM (S/N=3) with an excellent differentiation ability, and the proposed method showed acceptable stability and reproducibility. It has been applied for assay of BCR/ABL fusion gene from real samples with satisfactory results.

Electrochemical determination of BCR/ABL fusion gene based on in situ synthesized gold nanoparticles and cerium dioxide nanoparticles.

An efficient DNA electrochemical biosensor, based on the gold nanoparticles (GNPs) in situ synthesized at the surface of multiwalled carbon nanotubes (MWCNTs), cerium dioxide (CeO2) and chitosan (Chits) composite membrane, was developed for the detection of BCR/ABL fusion gene in chronic myelogenous leukemia (CML). The capture probe was attached onto the nanocomposite membrane modified glassy carbon electrode (GCE) through the conjugated structure. Owing to the synergistic effects of CeO2 nanoparticles with a strong adsorption ability and MWCNTs with a large surface area and excellent electron transfer ability, the prepared composite membrane was demonstrated an efficient electron transfer ability. The biosensor was electrochemically characterized by cyclic voltammogram (CV) and differential pulse voltammetry (DPV), and the decrease of the peak currents upon hybridization was observed using methylene blue (MB) as the electroactive indicator. Under the optimized conditions, peak currents were linear over the range from 1 × 10(-9) M to 1 × 10(-)(12) M, with a detection limit of 5 × 10(-)(13) M (based on the 3σ). And the proposed method was successfully applied for the detection of PCR real samples with satisfactory results. Furthermore, the developed DNA biosensor was demonstrated a good selectivity, a reasonable stability and a favorable reproducibility, which could be regenerated easily.