<i>JAK2‐V617F</i> mutation analysis of granulocytes and platelets from patients with chronic myeloproliferative disorders: advantage of studying platelets

Toyama, Kohtaro; Karasawa, Masamitsu; Yamane, Arito; Irisawa, Hiroyuki; Yokohama, Akihiko; Saitoh, Takayuki; Handa, Hiroshi; Maekawa, Takafumi; Sawamura, Morio; Miyawaki, Shuichi; Murakami, Hirokazu; Nojima, Yoshihisa; Tsukamoto, Norifumi

doi:10.1111/j.1365-2141.2007.06755.x

Cited by 24 publications

(13 citation statements)

References 31 publications

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“…T-cells recovered from the remaining mononuclear cells were further positively selected using anti-CD3 immunoconjugated magnetic beads (Miltenyi Biotec, Bergisch Gladbach Germany). Granulocytes were isolated from the pellet after the removal of red blood cells by hypotonic lysis (5).…”

Section: Isolation Of Platelets Granulocytes and T-cells In Vitro Cmentioning

confidence: 99%

“…We have previously reported the advantage of studying the JAK2-V617F mutation in platelets in MPN patients, as we have found that approximately 10% of essential thrombocythemia (ET) patients only have the JAK2-V617F mutation in their platelets (5). Based on these results, we decided to investigate the JAK2-V617F mutation status in four cell lineages of granulocytes, platelets, T-cells, and erythroid colonies, and then determine whether there was a difference in the cell lineages that are involved with the JAK2-V617F mutation in each of the MPN subtypes.…”

Section: Introductionmentioning

confidence: 99%

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Differences in the JAK2 and MPL Mutation Status in the Cell Lineages of the bcr/abl-negative Chronic Myeloproliferative Neoplasm Subtypes

et al. 2011

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Objective While the somatic mutation of Janus Kinase 2 (JAK2) and the thrombopoietin receptor (c-MPL) gene are thought to affect the pathogenesis of bcr/abl negative chronic myeloproliferative neoplasm (MPN), the relationship between the mutation and the clinical features remain obscure. Methods The mutation status of these genes in granulocytes, platelets, T-cells, and erythroid colonies (BFU-E) was obtained from 115 MPN patients, and then the clinical features of the MPN subtypes were compared. Results The JAK2-V617F mutation was observed in three lineages of granulocytes, platelets, and BFU-E in almost all polycythemia vera (PV) and primary myelofibrosis (PMF) patients. In contrast, 68% of essential thrombocythemia (ET) patients have the JAK2-V617F mutation in at least one of the lineages, of which 70% of these patients have the JAK2-V617F mutation in three lineages; the remaining ET patients with the JAK2-V617F mutation only exhibited the mutation in one or two lineages. Further, the ET patients that exhibited the JAK2-V617F mutation in three lineages had higher WBC and granulocyte counts as compared to the ET patients that did not have the JAK2-V617F mutation or only had the mutation in one or two lineages. Concerning the MPL gene, two ET patients had the MPL-W515L gene mutation in their platelets, although the lineage of the JAK2-V617F mutation involved differed from case to case. Conclusion The progenitor cells that are involved with the JAK2-V617F mutation in MPNs are different in each subtype and this difference may also affect the clinical features of MPNs.

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Section: Isolation Of Platelets Granulocytes and T-cells In Vitro Cmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 99%

Differences in the JAK2 and MPL Mutation Status in the Cell Lineages of the bcr/abl-negative Chronic Myeloproliferative Neoplasm Subtypes

et al. 2011

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“…Prior work suggested that analysis of platelets may be a more sensitive approach for detecting JAK2 V617F in BCR-ABL1-negative MPN patients as compared with analysis of granulocytes alone. 25 We suggest that examination of RNA from buffy coats (which include platelets and granulocytes) may also offer increased sensitivity when compared with looking at a single cell population. Finally, unlike many of the more sensitive published assays, our assay does not require isolation of granulocytes, because this procedure is difficult to integrate into most laboratories' workflow.…”

Section: Discussionmentioning

confidence: 99%

“…The inclusion of platelet RNA may be particularly relevant for ET, because prior studies have indicated that the analysis of platelet RNA may improve the clinical sensitivity of JAK2 V617F assays. 25 Based on these considerations, we developed an assay that uses allele-specific PCR and real-time detection with hydrolysis probes for the quantification of JAK2 V617F and wild-type JAK2 transcript levels. Our initial evaluation indicates the method is sufficiently sensitive, specific, and reproducible to be used in clinical molecular pathology laboratories.…”

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confidence: 99%

Design and Evaluation of a Real-Time PCR Assay for Quantification of JAK2 V617F and Wild-Type JAK2 Transcript Levels in the Clinical Laboratory

Merker

Jones

et al. 2010

The Journal of Molecular Diagnostics

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The somatic mutation JAK2 V617F is associated with BCR-ABL1-negative myeloproliferative neoplasms. Detection of this mutation aids diagnosis of these neoplasms , and quantification of JAK2 V617F may provide a method to monitor response to therapy. For these reasons , we designed a clinical assay that uses allele-specific PCR and real-time detection with hydrolysis probes for the quantification of JAK2 V617F , wild-type JAK2 , and GAPDH transcripts. Mutant and wild-type JAK2 were quantified by using external plasmid standards that contain the relevant JAK2 V617F or JAK2 sequence , respectively. We tested 55 peripheral blood specimens from patients with suspected myeloproliferative neoplasms and 55 peripheral blood specimens from patients not known to have myeloproliferative neoplasms. Low-level , nonspecific amplification was detected in reactions containing a high copy number of plasmid standards and in specimens from patients not known to have myeloproliferative neoplasms , necessitating the use of a laboratory-established mutant to wild-type cutoff. The limit of detection established by using cell line dilutions is 0.1% , and this method identified three JAK2 V617F-positive patients who were not detected by a less sensitive method. The assay characteristics and our initial evaluation indicate this method can be used for the detection and quantification of JAK2 V617F, which should be useful for diagnosis of myeloproliferative neoplasms and potentially for monitoring minimal residual disease in future trials of therapies targeted to myeloproliferative neoplasms.

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“…Using a direct sequencing method we described before, the JAK2-V617F mutation status was investigated [23]. To confirm the existence of the JAK2-V617F mutation, an allele-specific polymerase chain reaction (AS-PCR) was performed as previously described [11].…”

Section: Dna Preparation and Jak2 Mutation Analysismentioning

confidence: 99%

Low burden of a JAK2-V617F mutated clone in monoclonal haematopoiesis in a Japanese woman with Budd-Chiari syndrome

et al. 2009

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Approximately one-half of the cases of Budd-Chiari syndrome (BCS) are caused by bcr/abl negative chronic myeloproliferative disorders (CMPDs). Furthermore, a mutation in the Janus kinase protein (JAK2-V617F) is detected in half of the patients with BCS. However, whether the JAK2 mutation is the primary event leading to CMPDs and BCS is controversial. We present a report concerning a young woman who suffered from BCS prior to the onset of CMPDs. Analysis of X-chromosome inactivation patterns in this patient, using the human androgen receptor gene demonstrated monoclonal haematopoiesis in her granulocytes. In contrast, she had a low burden of a JAK2-V617F mutation positive clone among granulocyte populations. These results suggest that the JAK2-V617F mutation occurs after the onset of monoclonal haematopoiesis; thus the V617F mutation of JAK2 may not be the primary event in the induction of BCS.

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JAK2‐V617F mutation analysis of granulocytes and platelets from patients with chronic myeloproliferative disorders: advantage of studying platelets

Cited by 24 publications

References 31 publications

Differences in the JAK2 and MPL Mutation Status in the Cell Lineages of the bcr/abl-negative Chronic Myeloproliferative Neoplasm Subtypes

Differences in the JAK2 and MPL Mutation Status in the Cell Lineages of the bcr/abl-negative Chronic Myeloproliferative Neoplasm Subtypes

Design and Evaluation of a Real-Time PCR Assay for Quantification of JAK2 V617F and Wild-Type JAK2 Transcript Levels in the Clinical Laboratory

Low burden of a JAK2-V617F mutated clone in monoclonal haematopoiesis in a Japanese woman with Budd-Chiari syndrome

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