A comparison of TBI and non-TBI conditioning regimens in patients undergoing matched-unrelated bone marrow transplantation

published_or_final_versio

Functional characterisation of a novel nucleoporin gene NUP98 in zebrafish embryo.

Oral PresentationINTRODUCTION: The nucleoporin gene nup98 is important for the regulation of cytoplasmic-nuclear trafficking. Frequent disruptions of NUP98 during chromosomal translocation in acute myeloid leukaemia suggest that it may play a role in normal haematopoiesis. nup98-knockout mice has resulted in early embryonic lethality. Therefore, its role in embryonic haematopoiesis remains unclear. In this study, we have cloned a zebrafish nup98 gene and examined its role in embryonic development, with particular reference to haematopoiesis. METHODS: Two expressed sequence tags with translated sequence homologous to human NUP98 were identified. The gene was cloned by PCR from cDNA of zebrafish embryos. Expression of nup98 in zebrafish embryos was investigated spatially by whole-mount in-situ hybridisation and temporally by RT-PCR. The functions of nup98 were examined by morpholino knockdown and the effects on embryonic development evaluated by gene expression studies and confocal microscopy. Cellular functions of zebrafish nup98 were investigated in HeLa cells. RESULTS: Zebrafish nup98 gene shared 65% identity to human NUP98 homolog in protein sequence. The gene was expressed during early embryonic development since 1-cell stage and diffusely in eyes and the developing brain since 18 hpf. About 30% nup98-knockdown embryos developed intracranial haemorrhage at 48 hpf, resulting from disrupted blood vessels. nup98-knockdown upregulated pu.1 and scl as evaluated by quantitative RT-PCR. Moreover, ectopic expression of zebrafish nup98 rescued the defective mRNA export due to NUP98 knockdown in HeLa cells. CONCLUSION: A novel zebrafish nup98 gene was shown to exhibit conserved function in mRNA trafficking. Its role in embryonic development should be further evaluated.published_or_final_versio

INDELseek: Detection Of Complex Insertions And Deletions From Next-generation Sequencing Data

BACKGROUND: Complex insertions and deletions (indels) from next-generation sequencing (NGS) data were prone to escape detection by currently available variant callers as shown by large-scale human genomics studies. Somatic and germline complex indels in key disease driver genes could be missed in NGS-based genomics studies. RESULTS: INDELseek is an open-source complex indel caller designed for NGS data of random fragments and PCR amplicons. The key differentiating factor of INDELseek is that each NGS read alignment was examined as a whole instead of 'pileup' of each reference position across multiple alignments. In benchmarking against the reference material NA12878 genome (n = 160 derived from high-confidence variant calls), GATK, SAMtools and INDELseek showed complex indel detection sensitivities of 0%, 0% and 100%, respectively. INDELseek also detected all known germline (BRCA1 and BRCA2) and somatic (CALR and JAK2) complex indels in human clinical samples (n = 8). Further experiments validated all 10 detected KIT complex indels in a discovery cohort of clinical samples. In silico semi-simulation showed sensitivities of 93.7-96.2% based on 8671 unique complex indels in >5000 genes from dbSNP and COSMIC. We also demonstrated the importance of complex indel detection in accurately annotating BRCA1, BRCA2 and TP53 mutations with gained or rescued protein-truncating effects. CONCLUSIONS: INDELseek is an accurate and versatile tool for complex indel detection in NGS data. It complements other variant callers in NGS-based genomics studies targeting a wide spectrum of genetic variations.published_or_final_versio

Differential NOD/SCID mouse engraftment of peripheral blood CD34 + cells and JAK2V617F clones from patients with myeloproliferative neoplasms

We evaluated the NOD/SCID engraftment of CD34 + cells from polycythemia vera (PV) and secondary polycythemia patients (SP) and the JAK2V617F clone before and after transplantation. Peripheral blood CD34 + cells were transplanted intra-femorally. In the injected BM, successful engraftment (>0.1%) occurred in 8/26 mice transplanted with CD34+ cells from 5/13 PV patients (median: 4.26%, range: 0.3-5.56%), in contrast to 0/14 mice from 9 SP patients (P=0.017). The engrafting PV cells were of multi-lineage. JAK2V617F/total JAK2 ratios decreased after transplantation (initial: 65.9% versus 6-week: 13.0%, P=0.001). Essential thrombocythemia (ET) BM cells also exhibited a similar decrease in JAK2V617F clone. The results suggested that events in addition to JAK2V617F are involved in the pathogenesis of PV and ET. © 2010 Elsevier Ltd.postprin

ACER eNews Issue 04 April 2013

Cell mechanics, in particular mechanical properties, has been suggested as a new biomarker indicative of cell state and phenotype. Acute myeloid leukemia (AML) is characterized by the abnormal increase of myeloblasts in blood and bone marrow. While AML has been extensively studied from the perspectives of biochemical and genetic aspects, little is known about its cellular biophysical properties. In this study, optical tweezer technology was used to examine the micromechanical properties of myeloblasts from bone marrow of AML patients at single cell level. The myeloblasts were separately analyzed according to their expression of CD34 +, a marker of primitive hematopoietic cells. To extract the intrinsic properties from the relationship between the stretching force and the induced deformation, a theoretical approach was developed to model the mechanical responses of cells and further characterize their mechanical properties. The preliminary results show that the area compressibility modulus of CD34 + myeloblasts was significantly less than that of CD34 - cells, which indicate that micromechanical properties are unique features of myeloblasts and provide us with an insight into the cell mechanics of primitive AML cells. © 2010 IEEE.published_or_final_versio

Helper T-lymphocyte precursor frequency (HTLPf) predicts the occurrence of graft-versus-host disease (GVHD) and disease relapse after allogeneic bone marrow transplantation

published_or_final_versio

Unmanipulated bone marrow transplantation from one-Hla antigen mismatched siblings carries high transplant-related mortality compared with the Hla-identical counterparts

published_or_final_versio

Clinical characteristics and risk factors of herpes zoster after bone marrow transplantation

published_or_final_versio

Distinctive functions of methionine aminopeptidase II in embryonic haematopoiesis in zebrafish embryos

Oral presentationINTRODUCTION: Methionine aminopeptidase 2 (MetAP-2) is known to be the target of an anti-angiogenesis compound fumagillin and has been investigated for its robust expression in human cancers and its antiproliferative effects on endothelial cells. Together with a related member MetAP-1, they are proteases which remove the initiator NH2-terminal methionine from the nascent peptides during protein translation. Despite its pathogenetic significance in cancers, the physiological role of MetAP-2, particularly during embryonic …published_or_final_versionThe 15th Annual Research Conference of the Department of Medicine, The University of Hong Kong, Hong Kong, 16 January 2010. In Hong Kong Medical Journal, 2010, v. 16, suppl. 1, p. 44, abstract no. 7