The helix-loop-helix transcription factor TWIST is dysregulated in myelodysplastic syndromes

Li, Xiang; Marcondes, A. Mario; Gooley, Theodore A.; Deeg, H. Joachim

doi:10.1182/blood-2009-09-242313

Cited by 45 publications

(67 citation statements)

References 54 publications

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“…The TWIST-1-p53 axis is an important regulator for the control of apoptosis in many disorders. 11,16,17 However, the association between p53 and TWIST-1-dependent miRs, an important inhibitory system of transcription/protein translation in the development of MDS, had not been investigated so far. Our data show that miRs-10a/b were expressed in MDS at levels significantly higher than in healthy controls ( Figure 1A and B) Figure S1A and B).…”

Section: Resultsmentioning

confidence: 99%

“…We had previously 11 shown that expression of TWIST-1 is dysregulated in CD34 + MDS cells and showed increased levels in KG1a, PL-21 and MDS-L cell lines. The present data in KG1a (mutant p53), PL-21 (wild-type p53), MDS-L (mutant p53) and ML1 cells (wild-type p53) show that expressions of miR10a/b correlated with TWIST-1 expression and was independent from the p53 mutational status.…”

Section: Resultsmentioning

confidence: 99%

“…11 We also showed that marrow stroma-dependent signals resulted in downregulation of TWIST-1 and sensitivity to tumor-necrosis-factor alpha (TNFα)-induced apoptosis in clonal myeloid cell lines and in CD34 + marrow cells from patients with MDS. 11 In the present study, we investigated whether TWIST-1-regulated miRs might be involved. Results show that interactions between TWIST-1-dependent miR-10 family members and p53 play a central role in regulating TNFα-mediated apoptosis in MDS clonal cells.…”

Section: Introductionmentioning

confidence: 89%

“…At 48 h apoptosis was assessed by flow cytometry using Annexin V-FITC labeling. 11 Proliferation and cell cycle phase were determined using the APC-BrdU Flow Kit (BD Biosciences, Mountainview, CA, USA).…”

Section: Apoptosis Proliferation and Cell Cycle Analysismentioning

confidence: 99%

“…21,22 TWIST-1/NF-kB/p53 interactions are central to the control of apoptosis and proliferation, as shown in other disorders. 11,16,17 A possible role for miRs in modulating this axis has not yet been investigated. To assess p53 function in our TWIST-1-dependent miR10a/b working model we used a luciferase reporter assay in unmodified and KD modified KG1a and PL-21 cells.…”

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confidence: 99%

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Transcriptional regulation of miR-10a/b by TWIST-1 in myelodysplastic syndromes

Chang

et al. 2012

Haematologica

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Correspondence: mmarcond@fhcrc.orgThe transcription factor TWIST-1 is up-regulated in CD34 + cells in myelodysplastic syndrome and is involved in resistance to apoptosis. There is evidence that TWIST-1 affects apoptosis via microRNAs (miRs). Expression of miRs was determined in myeloid cell lines and primary CD34 + marrow cells from patients with myelodysplastic syndrome and healthy donors using NanoString/array and validated by real-time-polymerase chain reaction. Expression levels of miR10a and miR10b were significantly higher in CD34 + marrow cells from 28 patients with myelodysplastic syndrome than in CD34 + cells from healthy donors (P=0.05 and P=0.012, respectively). Levels of miR10a/b correlated with TWIST-1 miR levels in CD34 + myelodysplastic marrow cells (miR10a, R=+0.69, P<0.0001; miR10b, R=+0.56, P=0.0008). Inhibition of miR10a/10b in clonal cells interfered with proliferation and enhanced sensitivity to apoptosis, which involved NF-kB-dependent p53 activation. These data support a role for miR10a/10b in the regulation of apoptosis in myelodysplastic syndrome and suggest the TWIST-1/miR10a/b-axis as a therapeutic target in myelodysplastic syndrome. ABSTRACTration reactions. All sample preparations and hybridization reactions were carried out according to the manufacturer's instructions, using 5 mL of the 5-fold diluted sample. All hybridization reactions were incubated at 65°C for a minimum of 18 h. Hybridized probes were purified and counted on the nCounter Prep Station and Digital Analyzer (NanoString, Seattle, WA, USA). For each assay, a high-density scan (600 fields of view) was performed. Real-time polymerase chain reactionRNA was extracted using the RNeasy Mini kit (QIAGEN, Valencia, CA, USA). Primers for miR-10a, miR-10b and U6 were synthesized by Applied Biosystems (Carlsbad, CA, USA). Total RNA (100 ng) was reverse transcribed using the Taqman MiR Reverse Transcriptase kit (Applied Biosystems, Carlsbad, CA, USA). Real-time quantitative (q)RT-PCR was performed and assessed as described. 11Lentivirus-mediated knockdown of miR-10a/b and conditional knockdown of TWIST-1 Lentiviral vectors mZ-CTRL (miRZip-control) and mZ-10a and mZ-10b (miRZip-10a/b) were from System Biosciences (Mountain View, CA, USA). The miRZip delivers short anti-sense RNAs that are stably expressed and competitively bind their endogenous miR targets, thereby inhibiting their function. For conditional inhibition of TWIST-1 a doxycyclin inducible lentiviral construct pTRIPZ (Openbiosystems, Huntsville, AL, USA; RHS4696-99682454) was used. Apoptosis, proliferation and cell cycle analysisLentivirally transfected KG1a or PL-21 cells were plated in triplicates in 24-well plates (2×10 5 cells/well) containing TNFα at 0.5, 15, or 25 ng/mL. At 48 h apoptosis was assessed by flow cytometry using Annexin V-FITC labeling.11 Proliferation and cell cycle phase were determined using the APC-BrdU Flow Kit (BD Biosciences, Mountainview, CA, USA). pGL3-p53 luciferase reporter assayFor a dual-luciferase reporter assay, cells were plated in 1...

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Section: Resultsmentioning

confidence: 99%

Section: Resultsmentioning

confidence: 99%

Section: Introductionmentioning

confidence: 89%

Section: Apoptosis Proliferation and Cell Cycle Analysismentioning

confidence: 99%

mentioning

confidence: 99%

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Transcriptional regulation of miR-10a/b by TWIST-1 in myelodysplastic syndromes

Chang

et al. 2012

Haematologica

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Targeting the Spleen as an Alternative Site for Hematopoiesis

et al. 2019

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Bone marrow is the main site for hematopoiesis in adults. It acts as a niche for hematopoietic stem cells (HSCs) and contains non‐hematopoietic cells that contribute to stem cell dormancy, quiescence, self‐renewal, and differentiation. HSC also exist in resting spleen of several species, although their contribution to hematopoiesis under steady‐state conditions is unknown. The spleen can however undergo extramedullary hematopoiesis (EMH) triggered by physiological stress or disease. With the loss of bone marrow niches in aging and disease, the spleen as an alternative tissue site for hematopoiesis is an important consideration for future therapy, particularly during HSC transplantation. In terms of harnessing the spleen as a site for hematopoiesis, here the remarkable regenerative capacity of the spleen is considered with a view to forming additional or ectopic spleen tissue through cell engraftment. Studies in mice indicate the potential for such grafts to support the influx of hematopoietic cells leading to the development of normal spleen architecture. An important goal will be the formation of functional ectopic spleen tissue as an aid to hematopoietic recovery following clinical treatments that impact bone marrow. For example, expansion or replacement of niches could be considered where myeloablation ahead of HSC transplantation compromises treatment outcomes.

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Twist-1, A Novel Regulator of Hematopoietic Stem Cell Self-Renewal and Myeloid Lineage Development

et al. 2014

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Transcription factor Twist-1 plays essential roles in specification and differentiation of mesoderm-derived tissues. Growing evidences now link Twist-1 to the acquisition of stem-celllike properties. However, the role of Twist-1 in hematopoietic stem cell (HSC) remains largely uncharacterized. We report that Twist-1 is more highly expressed in murine HSC and its expression declines with differentiation. To investigate Twist-1 gene function, retroviral-mediated overexpression or removal experiments are performed. Competitive repopulation studies demonstrate that enforced expression of Twist-1 in HSC-enriched Lin 2 c-Kit 1 Sca-1 1 (LKS) cells results in an increase in the size of the G 0 population, and in their reconstitution ability after the first and a second transplantation. Conversely, removal of Twist-1 in LKS cells impairs their ability to repopulate. In addition, increased Twist-1 expression causes a shift toward production of myeloid cells. Twist-1 transduction in LKS cells activates myeloid lineage-determining factors PU.1 and GATA-1 and downregulates lymphoid factor GATA-3 in vitro, suggesting that Twist-1-mediated myeloid skewing occurs in hematopoietic stem and progenitor cells (HSPCs). These findings indicate that Twist-1 is not only involved in the maintenance of HSC dormancy and self-renewal capacity but also implicated in the myeloid lineage fate choice of HSPCs. Exploration of the underlying mechanisms reveals that Runx1/c-Mpl/Tie2 regulatory pathway could possibly account for the observed effects caused by Twist-1 overexpression. Our study provides the first evidence supporting a role for Twist-1 in hematopoiesis. STEM CELLS

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The helix-loop-helix transcription factor TWIST is dysregulated in myelodysplastic syndromes

Cited by 45 publications

References 54 publications

Transcriptional regulation of miR-10a/b by TWIST-1 in myelodysplastic syndromes

Transcriptional regulation of miR-10a/b by TWIST-1 in myelodysplastic syndromes

Targeting the Spleen as an Alternative Site for Hematopoiesis

Twist-1, A Novel Regulator of Hematopoietic Stem Cell Self-Renewal and Myeloid Lineage Development

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