Unravelling the relevance of CLEC12A as a cancer stem cell marker in myelodysplastic syndrome.

Evidence of distinct disease propagating stem cells in myelodysplastic syndrome (MDS) has emerged in recent years. However, immunophenotypic characterization of these cancer stem cells remains sparse. In acute myeloid leukaemia (AML), we have previously described aberrant expression of the C-type lectin domain family 12, member A (CLEC12A) as a stable and reliable marker of leukaemia blasts and as a tool for assessing minimal residual disease. Furthermore, CLEC12A has been proposed as a promising marker of leukaemic stem cells in AML. The role of CLEC12A in MDS, however, remains to be elucidated. In this study, we found CLEC12A aberrantly expressed on the CD34+ CD38- cell compartment in 71% (22/31) of MDS patients, distributed across all Revised International Prognostic Scoring System risk groups. We showed that the CD34+ CD38- CLEC12A+ cells were indeed malignant and possessed functional stem cell properties in the long-term colony-initiating cell assay. As opposed to reported findings in AML, we showed that cancer stem cells from MDS samples derived from both CLEC12A positive and negative CD34+ CD38- subpopulations. Due to the absence of CLEC12A on normal haematopoietic stem cells, CLEC12A stem cell immunophenotyping may contribute to diagnosing and monitoring MDS patients and could furthermore add knowledge about disease propagating cells in MDS.

Unraveling stem cell and progenitor subsets in autologous grafts according to methods of mobilization: implications for prediction of hematopoietic recovery.

BACKGROUND AIMS: In the autologous setting, granulocyte colony-stimulating factor (G-CSF) (G), or, when failing, G plus plerixafor (G+P), are common regimens for mobilization of stem cells into peripheral blood. To delineate mobilization effects on graft composition and hematopoietic recovery, we compared contents of stem cells and progenitor cells in products of G+P- and G patients. Paired samples of G+P patients and prior insufficient G mobilization were available for analyses. METHODS: Subset analyses of grafts were performed by flow cytometry and myeloid colony-forming assay. In search of new markers to ascertain graft quality, we determined the fractions of aldehyde dehydrogenase bright (ALDH(br)) cells. RESULTS: G grafts contained higher percentages of CD34+ cells, CD34+CD38- cells, and committed progenitors (CD34+CD38+) compared with G+P grafts. A detailed characterization of the mobilized CD34+ cell subset showed higher percentages of CD38- among the CD34+ cells of the G+P group (P = 0.032). In contrast, the CD34+ cell subset in G grafts was characterized by a higher percentage of ALDH(br) cells (P < 0.0001). Studying engraftment and day +100 graft function the G and G+P transplanted patients were comparable with respect to neutrophils, whereas in platelets they differed. In the prediction of engraftment and hematopoietic recovery, the dose of infused ALDH(br) cells correlated best to both platelet (r = 0.565, P = 0.002) and neutrophil reconstitution (r = 0.366, P = 0.06). CONCLUSIONS: Besides showing dissimilar distributions of CD34+CD38- cells and progenitors in G and G+P grafts, this study further designated ALDH(br) as a promising marker in determination and prediction of graft quality and hematopoietic recovery.

The CLEC12A receptor marks human basophils: Potential implications for minimal residual disease detection in acute myeloid leukemia.

BACKGROUND: The transmembrane receptor C-type lectin domain family 12, member A (CLEC12A) is known to be highly expressed on monocytes and neutrophils and is a reliable leukemia associated marker in acute myeloid leukemia. Consequently, detailed knowledge of the various normal cell types expressing this receptor is essential. We have observed CLEC12A to be expressed on CD45lowSSClowCD14-CD123+ basophils in peripheral blood (PB) and in this study, we aimed at verifying this observation and further delineate the CD45lowSSClowCD14-CD123 + CLEC12A+ subpopulation. METHODS: We analyzed PB from 20 diagnostic chronic myeloid leukemia (CML) samples and eight healthy donors in a six-color multicolor flowcytometry (FCM) based assay. Furthermore, we performed fluorescence activated cell sorting on one CML sample to morphologically confirm the CD45lowSSClowCD14-CD123 + CLEC12A+ subset to be highly enriched for basophils. Finally, to further delineate the CD45lowSSClowCD14-CD123 + CLEC12A+ subpopulation in normal PB, we examined three healthy donors in a 10-color FCM assay enabling further separation of the cell subset into basophils and dendritic cells. RESULTS: The CLEC12A receptor is expressed on basophils. CONCLUSIONS: Identification and enumeration of basophils is of high relevance in diagnostic hematology and immunology. We here show that CLEC12A in a simple FCM assay consistently marks basophils. Importantly, as basophils are characterized by a CD45lowSSClow profile similar to the "blast-gate" used for the evaluation of hematological disorders, awareness of minor normal CLEC12A+ subpopulations is crucial when using CLEC12A as a minimal residual disease marker in myeloid malignancies. © 2017 International Clinical Cytometry Society.

A novel del(8)(q23.2q24.11) contributing to disease progression in a case of JAK2/TET2 double mutated chronic myelomonocytic leukemia.

We have identified a novel 7.7 Mb del(8)(q23.2q24.11) in a patient progressing to acute myeloid leukemia (AML) following a 12-year stable phase of chronic myelomonocytic leukemia (CMML). A surprisingly high JAK2+ allelic burden of 92% at the time of AML led us to delineate the molecular aberrations relevant for leukemogenesis. While a frameshift mutation in the TET2 gene was stably present throughout the course of disease the JAK2 mutation was acquired after initial diagnosis of CMML. At progression aCGH revealed del(8q)(q23.2q24.11) encompassing various cancer relevant genes of which RAD21 and CSMD3 are of particular interest.

[Melkersson-Rosenthal syndrome treated with clofazimine]. / Melkersson-Rosenthals syndrom behandlet med clofazimin.

Melkersson-Rosenthal syndrome (MRS) is a triad of facial palsy, lingua plicata and chronic orofacial swelling. Mono- and oligosymptomatic forms are common and cheilitis granulomatosa is often considered a monosymptomatic form of MRS. In advanced stages the histological findings are noncaseating granulomas. The etiology remains unclear and treatment is difficult. A few uncontrolled studies have shown corticosteroids and clofazimine to be effective. We report on a 31 year-old man with complete MRS who responded well to treatment with clofazimine.