Inhibition of <scp>MEK</scp>/<scp>ERK</scp> signalling pathway promotes erythroid differentiation and reduces <scp>HSC</scp>s engraftment in <i>ex vivo</i> expanded haematopoietic stem cells

Zarrabi, Morteza; Afzal, Elaheh; Asghari, Mohammad Hossein; Mohammad, Monireh; Es, Hamidreza Aboulkheyr; Ebrahimi, Marzieh

doi:10.1111/jcmm.13379

Cited by 20 publications

(14 citation statements)

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“…12,13 Extracellular signal-regulated kinase 1/2 (ERK1/ 2), one of the members of the MAPK family, is able to mediate many physiological and pathological processes, such as cell proliferation, protein synthesis, cell survival, differentiation, migration and apoptosis. [14][15][16][17] Previous studies have shown that activated ERK1/2 is present in both the cytoplasm and the nucleus, 18 whereas inhibiting (MEK1/2) activation significantly reduces inner hair cells (IHCs) death caused by neomycin treatment, suggesting that ERK activation in supporting cells might promote HC death. 15,18 Moreover, ERK1/2 inhibition in cochlear explants causes a loss of outer hair cells (OHCs).…”

Section: Introductionmentioning

confidence: 99%

STK33 alleviates gentamicin‐induced ototoxicity in cochlear hair cells and House Ear Institute‐Organ of Corti 1 cells

Zhou

Sun

Zhang

et al. 2018

J Cellular Molecular Medi

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Serine/threonine kinase 33 (STK33), a member of the calcium/calmodulin‐dependent kinase (CAMK), plays vital roles in a wide spectrum of cell processes. The present study was designed to investigate whether STK33 expressed in the mammalian cochlea and, if so, what effect STK33 exerted on aminoglycoside‐induced ototoxicity in House Ear Institute‐Organ of Corti 1 (HEI‐OC1) cells. Immunofluorescence staining and western blotting were performed to investigate STK33 expression in cochlear hair cells (HCs) and HEI‐OC1 cells with or without gentamicin treatment. CCK8, flow cytometry, immunofluorescence staining and western blotting were employed to detect the effects of STK33 knockdown, and/or U0126, and/or N‐acetyl‐L‐cysteine (NAC) on the sensitivity to gentamicin‐induced ototoxicity in HEI‐OC1 cells. We found that STK33 was expressed in both mice cochlear HCs and HEI‐OC1 cells, and the expression of STK33 was significantly decreased in cochlear HCs and HEI‐OC1 cells after gentamicin exposure. STK33 knockdown resulted in an increase in the cleaved caspase‐3 and Bax expressions as well as cell apoptosis after gentamicin damage in HEI‐OC1 cells. Mechanistic studies revealed that knockdown of STK33 led to activated mitochondrial apoptosis pathway as well as augmented reactive oxygen species (ROS) accumulation after gentamicin damage. Moreover, STK33 was involved in extracellular signal‐regulated kinase 1/2 pathway in primary culture of HCs and HEI‐OC1 cells in response to gentamicin insult. The findings from this work indicate that STK33 decreases the sensitivity to the apoptosis dependent on mitochondrial apoptotic pathway by regulating ROS generation after gentamicin treatment, which provides a new potential target for protection from the aminoglycoside‐induced ototoxicity.

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Section: Introductionmentioning

confidence: 99%

STK33 alleviates gentamicin‐induced ototoxicity in cochlear hair cells and House Ear Institute‐Organ of Corti 1 cells

Zhou

Sun

Zhang

et al. 2018

J Cellular Molecular Medi

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“…2 a). The precise effect of PD on ex vivo expansion of CD34 + cells has been discussed before [ 16 ]. An additional round of small molecules removal showed that the deletion of NAM and Pur from the cocktail increased the fold expansion of TNCs and CD34 + cells.…”

Section: Resultsmentioning

confidence: 99%

Combination of SB431542, Chir9901, and Bpv as a novel supplement in the culture of umbilical cord blood hematopoietic stem cells

et al. 2020

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Background Small molecule compounds have been well recognized for their promising power in the generation, expansion, and maintenance of embryonic or adult stem cells. The aim of this study was to identify a novel combination of small molecules in order to optimize the ex vivo expansion of umbilical cord blood-derived CD34+ cells. Methods Considering the most important signaling pathways involved in the self-renewal of hematopoietic stem cells, CB-CD34+ cells were expanded with cytokines in the presence of seven small molecules including SB, PD, Chir, Bpv, Pur, Pμ, and NAM. The eliminativism approach was used to find the best combination of selected small molecules for effective ex vivo expansion of CD34+ cell. In each step, proliferation, self-renewal, and clonogenic potential of the expanded cells as well as expression of some hematopoietic stem cell-related genes were studied. Finally, the engraftment potential of expanded cells was also examined by the mouse intra-uterine transplantation model. Results Our data shows that the simultaneous use of SB431542 (TGF-β inhibitor), Chir9901 (GSK3 inhibitor), and Bpv (PTEN inhibitor) resulted in a 50-fold increase in the number of CD34+CD38− cells. This was further reflected in approximately 3 times the increase in the clonogenic potential of the small molecule cocktail-expanded cells. These cells, also, showed a 1.5-fold higher engraftment potential in the peripheral blood of the NMRI model of in utero transplantation. These results are in total conformity with the upregulation of HOXB4, GATA2, and CD34 marker gene as well as the CXCR4 homing gene. Conclusion Taken together, our findings introduce a novel combination of small molecules to improve the yield of existing protocols used in the expansion of hematopoietic stem cells.

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“…G protein expression levels increased and ERK1/2 activated during hemin-induced differentiation of K562 cells [65]. Inhibition of the MEK/ERK signaling pathway promoted erythroid differentiation and reduced HSCs engraftment in ex-vivo expanded haematopoietic stem cells [66]. The ERK/MAPK pathway is involved in megakaryocytic differentiation of K562 cells induced by 3-hydrogenkwadaphnin [67].…”

Section: Discussionmentioning

confidence: 99%

CRKL but not CRKII Inhibits Erythropoiesis and Megakaryopoiesis of CML via Inactivating Raf/MEK/ERK/Elk-1 Pathway

Guo

Zhang

Yan

et al. 2020

Preprint

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Background: As members of the CT10 regulation of kinase (CRK) adaptor protein family, CRK-like (CRKL) and CRKII are involved in cell proliferation, survival, adhesion, migration and differentiation. However, the exact role and underlying mechanism of CRKL and CRKII in leukemic cell differentiation are still unknown. Methods: Quantitative real-time qPCR (qRT-PCR) was used to detect the expression levels of CRKL and CRKII in chronic myeloid leukemia (CML) patients and complete remission (CR) patients; Western blotting (WB) was used to measure the expression levels of CRKL and CRKII during hemin-induced erythroid differentiation of K562 cells; Benzidine staining, isobaric tags for relative and absolute quantitation (iTRAQ) proteomic analysis, cDNA microarray assay, qRT-PCR and WB were used to examine the effects of CRKL and CRKII deregulation on erythroid and megakaryocyte differentiation of K562 cells; PD98059 was used to investigate the underlying mechanism of CRKL in erythropoiesis and megakaryopoiesis. Results: CRKL was found to be overexpressed in chronic myeloid leukemia (CML) patients compared with normal samples, while its expression level was lower in CR patients than in corresponding CML patients. The CRKL expression level was significantly decreased during the erythroid differentiation of K562 cells following hemin treatment. Moreover, CRKL downregulation promoted erythroid and megakaryocyte differentiation of K562 cells accompanied by increased expression level of the erythroid differentiation markers γ-globin, glycophorin (GPA) and the megakaryocyte differentiation markers CD41, CD61. Furthermore, gene microarray and iTRAQ quantitative proteomic analysis showed that CRKL downregulation increased hemoglobin (HB) molecules HBD, HBA1, HBA2, HBZ, HBE1, HBG1 and globin transcription factor 1 (GATA1), high-mobility group protein (HMGB2) expression levels. Mechanistically, CRKL inhibited erythroid and megakaryocyte differentiation of K562 cell via inactivating Raf/MEK/ERK/Elk-1 pathway. Conversely, CRKII was only slightly overexpressed in CML patients and had no effect on erythroid differentiation of K562 cells. Conclusions: Taken together, our results demonstrate that CRKL but not CRKII contributes to development, progression, erythropoiesis and megakaryopoiesis of CML, providing novel insights into effective diagnosis and therapy for CML patients.

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Inhibition of MEK/ERK signalling pathway promotes erythroid differentiation and reduces HSCs engraftment in ex vivo expanded haematopoietic stem cells

Cited by 20 publications

References 58 publications

STK33 alleviates gentamicin‐induced ototoxicity in cochlear hair cells and House Ear Institute‐Organ of Corti 1 cells

STK33 alleviates gentamicin‐induced ototoxicity in cochlear hair cells and House Ear Institute‐Organ of Corti 1 cells

Combination of SB431542, Chir9901, and Bpv as a novel supplement in the culture of umbilical cord blood hematopoietic stem cells

CRKL but not CRKII Inhibits Erythropoiesis and Megakaryopoiesis of CML via Inactivating Raf/MEK/ERK/Elk-1 Pathway

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