The Thalidomide Analogue, CC-4047, Induces Apoptosis Signaling and Growth Arrest in Childhood Acute Lymphoblastic Leukemia Cells <i>In vitro</i> and <i>In vivo</i>

Shalapour, Shabnam; Zelmer, Andrea; Pfau, Madlen; Moderegger, E; Costa-Blechschmidt, Cristiane; Landeghem, Frank K.H. van; Taube, Tillmann; Fichtner, Iduna; Bührer, Christoph; Henze, Günter; Seeger, Karl; Wellmann, Sven

doi:10.1158/1078-0432.ccr-06-0719

Cited by 25 publications

(24 citation statements)

References 28 publications

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“…Total cell protein extraction, quantification and immunoblotting were performed as described previously by our group (Shalapour et al, 2006;Wellmann et al, 2004). The following antibodies were used: goat polyclonal anti-RBM3 (#EB09201, Everestbiotech, Oxfordshire, UK), rabbit polyclonal anti-cleaved PARP (Asp214, Cell Signaling, Danvers, MA, USA), and rabbit polyclonal anti-DCX (#4604 Cell Signaling).…”

Section: Protein Extraction and Immunoblot Analysesmentioning

confidence: 99%

The RNA-binding protein RBM3 is involved in hypothermia induced neuroprotection

Chip

Zelmer

Ogunshola

et al. 2011

Neurobiology of Disease

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114

110

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Induced hypothermia is the only therapy with proven efficacy to reduce brain damage after perinatal asphyxia. While hypothermia down-regulates global protein synthesis and cell metabolism, low temperature induces a small subset of proteins that includes the RNA-binding protein RBM3 (RNA-binding motif protein 3), which has recently been implicated in cell survival. Here, immunohistochemistry of the developing postnatal murine brain revealed a spatio-temporal neuronal RBM3 expression pattern very similar to that of doublecortin, a marker of neuronal precursor cells. Mild hypothermia (32°C) profoundly promoted RBM3 expression and rescued neuronal cells from forced apoptosis as studied in primary neurons, PC12 cells, and cortical organotypic slice cultures. Blocking RBM3 expression in neuronal cells by specific siRNAs significantly diminished the neuroprotective effect of hypothermia while vector-driven RBM3 over-expression reduced cleavage of PARP, prevented internucleosomal DNA fragmentation, and LDH release also in the absence of hypothermia. Together, neuronal RBM3 up-regulation in response to hypothermia apparently accounts for a substantial proportion of hypothermia-induced neuroprotection. Together, neuronal RBM3 up-regulation in response to hypothermia apparently accounts for a substantial proportion of hypothermia-induced neuroprotection.

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Section: Protein Extraction and Immunoblot Analysesmentioning

confidence: 99%

The RNA-binding protein RBM3 is involved in hypothermia induced neuroprotection

Chip

Zelmer

Ogunshola

et al. 2011

Neurobiology of Disease

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114

110

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“…In addition, the stromal cellinduced proliferation of REH cells was less pronounced after treatment with VLA-4 blocking antibodies ( Figure 3B). To assess whether blocking of VLA-4 signaling overcomes the protection of leukemia cells from cytarabineinduced cytotoxicity by stromal cells, 21 we treated REH cells in mono-culture and in co-culture with cytarabine (Ara-C) in the presence or absence of anti-VLA-4 antibodies. Blocking of VLA-4 in leukemia cells significantly abolished the cytoprotective effect of stromal cells ( Figure 3B).…”

Section: Blocking Of Vla-4 Signaling Overcomes the Supportive Effect mentioning

confidence: 99%

High VLA-4 expression is associated with adverse outcome and distinct gene expression changes in childhood B-cell precursor acute lymphoblastic leukemia at first relapse

Shalapour¹,

Hof²,

Kirschner-Schwabe³

et al. 2011

Haematologica

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The online version of this article has a Supplementary Appendix. BackgroundResistance to therapy and subsequent relapse remain major challenges in the clinical management of relapsed childhood acute lymphoblastic leukemia. As the bone marrow environment plays an important role in survival and chemotherapy resistance of leukemia cells by activating different signaling pathways, such as the VLA-4 and PI3K/Akt pathways, we studied the prognostic and biological impact of VLA-4 expression in leukemia cells from children with relapsed B-cell precursor acute lymphoblastic leukemia and its influence on the sensitivity of the leukemia cells to drugs. Design and MethodsVLA-4 expression was quantified by real-time polymerase chain reaction in leukemia cells from 56 patients with relapsed acute lymphoblastic leukemia enrolled in the ALL-REZ BFM 2002 trial of the Berlin-Frankfurt-Münster study group. Gene expression changes related to VLA-4 expression were investigated by microarray-based mRNA profiling. The effect of VLA-4 signaling on proliferation and drug resistance was studied in co-cultures of leukemia and stromal cells. ResultsHigh expression of VLA-4 at first relapse was associated with adverse prognostic factors, poor molecular response to therapy and significantly worse probabilities of event-free and overall survival. VLA-4 expression was an independent prognostic parameter. Comparing gene expression profiles of leukemia cells with high versus low VLA-4 expression, we identified 27 differentially expressed genes primarily involved in the PI3K/Akt, ephrin and Rho GTPase pathways. Blocking of VLA-4 signaling in combination with cytarabine treatment abolished the growth supportive effect of stromal cells. ConclusionsOur results show that high VLA-4 expression is a marker of poor prognosis and a potential therapeutic target in children with relapsed acute lymphoblastic leukemia and confirm that cellular interactions and biological effects related to VLA-4 play a decisive role in the survival of leukemia cells and response to therapy. (ClinicalTrials.gov identifier: NCT00114348)

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“…Differential nuclear und cytosolic extracts from HEK293 and K562 cells were prepared by use of the TRansFactor Extraction Kit according to the manufactures recommendations (BD Biosciences, Allschwil, Switzerland). Total cell protein extraction, quantification, and immunoblotting were performed as described previously (2,12) using rabbit polyclonal antibodies against human RBM3 [kindly provided by Dr. Jun Fujita Department of Clinical Molecular Biology, Kyoto University, Kyoto 606-8507, Japan (2)], Caspase-3, Cleaved Caspase-3 (Asp175), or mouse MAb against CyclinD1 (DCS6, all from Cell Signaling, Danvers, MA,) or against COX-2 (Cayman Chemical, Ann Arbor, MI). As an internal control, blots were stripped and reprobed with mouse anti-␤-actin antibody (Sigma Chemical Co.-Aldrich).…”

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confidence: 99%

The RNA-Binding Protein RBM3 Is Required for Cell Proliferation and Protects Against Serum Deprivation-Induced Cell Death

et al. 2010

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Hypoxia and other adverse conditions are commonly encountered by rapidly growing cells. The RNA-binding protein RBM3 (RNA-binding motif protein 3), which is transcriptionally induced by low temperature and hypoxia, has recently been implicated in survival of colon cancer cells by mechanisms involving cyclooxygenase-2 (COX-2) signaling. Immunohistochemically, we found strong RBM3 expression in a variety of malignant and proliferating tissues but low expression in resting and terminally differentiated cells. RBM3 expression in fibroblasts and human embryonal kidney (HEK293) cells subjected to serum deprivation or contact inhibition closely paralleled proliferation rates, assessed by real-time RT-PCR and immunoblotting. siRNA-mediated RBM3 knockdown reduced cell viability and finally led to cell death, which did not involve caspase-3-mediated apoptosis, cell cycle arrest, or COX-2 regulation. In contrast, RBM3 over-expression rescued cells from death under serum starvation. This was associated with increased translation rates, as measured by 14 C serine and 3 H phenylalanine incorporation. Together, RBM3 is a critical factor providing cellular survival advantages in an adverse microenvironment presumably by restoring translation efficacy. We have previously demonstrated that both RBM3 mRNA and protein levels increase in response to hypoxia in a hypoxia inducible factor (HIF)-1-independent fashion (2). RBM3 is also one of the first proteins synthesized in response to cold shock (3,4) and consistently elevated during winter in hibernating animals, such as the ground squirrel (5).An involvement of RBM3 in the regulation of translation has been deduced from increased rates of protein synthesis associated with its over-expression in transfected cells or RBM3 up-regulation after hypothermia (6). Over-expression of RBM3 has also been shown to suppress cell death in cells harboring polyglutamine-huntingtin (7) and to stabilize cyclooxygenase-2 (COX-2) mRNA transcripts (8). Moreover, RBM3 was found to prevent cells from caspase-mediated apoptosis and mitotic catastrophe (9). On the other hand, down-regulation of RBM3 in human colon cancer cells has been reported to cause loss of COX-2 translation and decrease cell growth in culture, which was partially overcome with the COX-2 product, prostaglandin E2 (9).Here, we set out to further characterize the link between RBM3, cell proliferation, and apoptosis. We found that RBM3 expression paralleled proliferation rates in vitro and in vivo. Over-expression of RBM3 rescued human embryonal kidney (HEK293) cells from cell death triggered by serum starvation, and siRNA-mediated RBM3 knockdown-induced cell death occurred without caspase-3 activation. Both artificial up-and down-regulation of RBM3 in HEK293 cells had no effect on COX-2 expression, indicating that other mechanisms are involved that improve cell survival especially under adverse conditions. MATERIALS AND METHODSCell lines, lymphocyte preparation, culture conditions, and treatment.

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The Thalidomide Analogue, CC-4047, Induces Apoptosis Signaling and Growth Arrest in Childhood Acute Lymphoblastic Leukemia Cells In vitro and In vivo

Cited by 25 publications

References 28 publications

The RNA-binding protein RBM3 is involved in hypothermia induced neuroprotection

The RNA-binding protein RBM3 is involved in hypothermia induced neuroprotection

High VLA-4 expression is associated with adverse outcome and distinct gene expression changes in childhood B-cell precursor acute lymphoblastic leukemia at first relapse

The RNA-Binding Protein RBM3 Is Required for Cell Proliferation and Protects Against Serum Deprivation-Induced Cell Death

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