Depleting interferon regulatory factor‐1(IRF‐1) with CRISPR/Cas9 attenuates inducible oxidative metabolism without affecting RA‐induced differentiation in HL‐60 human AML cells

Zhu, Kaiyuan; Yue, Jianbo; Yen, Andrew

doi:10.1096/fba.2020-00004

Cited by 2 publications

(4 citation statements)

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“…The CD11b and NUMB knockout cell lines were constructed as previously described with CRISPR/Cas9 [35]. The sequence for CD11b sgRNA is Forward: 5-CAC CGA CCT TCC AAG AGA ACG CAA G-3; Reverse: 5-AAA CCT TGC GTT CTC TTG GAA GGT C-3.…”

Section: Construction Of Crispr-mediated Stable Cell Linesmentioning

confidence: 99%

“…The sequence for Numb sgRNA is Forward : 5-CAC CGG ATG AAG AAG GCG TTC GCA C-3; Reverse: 5-AAA CGT GCG AAC GCC TTC TTC ATC C-3. The sg-NC control cell was the one used in our previous report [35].…”

Section: Construction Of Crispr-mediated Stable Cell Linesmentioning

confidence: 99%

“…The western blots were performed as previously described [35]. The antibodies used for western blots: anti-CD38 was from BD Biosciences (San Jose, CA), GAPDH, c-Raf, Phospho-c-Raf (S259), Mek, p-Mek, Erk, p-Erk, FAK, Lyn, Fgr, p47phox, Slp-76, Vav1, pY416-c-Src, horseradish peroxidase anti-mouse, and anti-rabbit antibodies, were from Cell Signaling (Danvers, MA), p-Tyr and c-Cbl antibodies were from Santa Cruz Biotechnology (Santa Cruz, CA), Phospho-c-Raf (S621) was from Thermo Fisher Scientific (Waltham, MA).…”

Section: Western Blot Analysis and Antibodiesmentioning

confidence: 99%

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Vacuolin-1 enhances RA-induced differentiation of human myeloblastic leukemia cells: evidence for involvement of a CD11b/FAK/LYN/SLP-76 axis subject to endosomal regulation that drives late differentiation steps

et al. 2022

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Background Retinoic acid(RA), an embryonic morphogen, regulates cell differentiation. Endocytosis regulates receptor signaling that governs such RA-directed cellular processes. Vacuolin-1 is a small molecule that disrupts endocytosis, motivating interest in its effect on RA-induced differentiation/arrest. In HL-60 myeloblastic-leukemia cells, RA causes differentiation evidenced by a progression of cell-surface and functional markers, CD38, CD11b, and finally reactive oxygen species(ROS) production and G1/0 cell cycle arrest in mature cells. Results We found that Vacuolin-1 enhanced RA-induced CD11b, ROS and G1/0 arrest, albeit not CD38. Enhanced CD11b expression was associated with enhanced activation of Focal Adhesion Kinase(FAK). Adding vacuolin-1 enhanced RA-induced tyrosine phosphorylation of FAK, Src Family Kinases(SFKs), and the adaptor protein, SLP-76, expression of which is known to drive RA-induced differentiation. Depleting CD11b cripples late stages of progressive myeloid differentiation, namely G1/0 arrest and inducible ROS production, but not expression of CD38. Loss of NUMB, a protein that supports early endosome maturation, affected RA-induced ROS and G1/0 arrest, but not CD38 expression. Conclusion Hence there appears to be a novel CD11b/FAK/LYN/SLP-76 axis subject to endosome regulation which contributes to later stages of RA-induced differentiation. The effects of vacuolin-1 thus suggest a model where RA-induced differentiation consists of progressive stages driven by expression of sequentially-induced receptors.

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Section: Construction Of Crispr-mediated Stable Cell Linesmentioning

confidence: 99%

Section: Construction Of Crispr-mediated Stable Cell Linesmentioning

confidence: 99%

Section: Western Blot Analysis and Antibodiesmentioning

confidence: 99%

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Vacuolin-1 enhances RA-induced differentiation of human myeloblastic leukemia cells: evidence for involvement of a CD11b/FAK/LYN/SLP-76 axis subject to endosomal regulation that drives late differentiation steps

et al. 2022

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“…CRISPR/Cas9 technology allowed the excision of the interferon regulatory factor-1 (IRF-1) gene from HL-60 cells. Thus, it was found that IRF-1 is not essential for differentiation induced by all-transretinoic acid or 1,25-Dihydroxyvitamin D3, but is involved in the upregulation of p47phox expression required for mature-cell ROS production in granulocyte metabolism 18 .…”

Section: Fbxo1allows the Progression Of Mds To Aml 8 2 Bend3mentioning

confidence: 99%

The high-performance technology CRISPR/Cas9 improves knowledge and management of acute myeloid leukemia

Mihăilă¹,

Topircean²

2021

Biomed Pap Med Fac Univ Palacky Olomouc Czech Repub

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Knowledge on acute myeloid leukemia pathogenesis and treatment has progressed recently, but not enough to provide ideal management. Improving the prognosis of acute myeloid leukemia patients depends on advances in molecular biology for the detection of new therapeutic targets and the production of effective drugs. The CRISPR/Cas9 technology allows gene insertions and deletions and it is the first step in investigating the function of their encoded proteins. Thus, new experimental models have been developed and progress has been made in understanding protein metabolism, antitumor activity, leukemic cell maintenance, differentiation, growth, apoptosis, and self-renewal, the combined pathogenetic mechanisms involved in leukemogenesis. The CRISPR/Cas9 system is used to understand drug resistance and find solutions to overcome it. The therapeutic progress achieved using the CRISPR/Cas9 system is remarkable. FST gene removal inhibited acute myeloid leukemia cell growth. Lysine acetyltransferase gene deletion contributed to decreased proliferation rate, increased apoptosis, and favored differentiation of acute myelid leukemia cells carrying MLL-X gene fusions. The removal of CD38 gene from NK cells decreased NK fratricidal cells contributing to increased efficacy of new CD38 CAR-NK cells to target leukemic blasts. BCL2 knockout has synergistic effects with FLT3 inhibitors. Exportin 1 knockout is synergistic with midostaurin treatment in acute myeloid leukemia with FLT3-ITD mutation. Using the results of CRISPR/Cas9 libraries and technology application will allow us to get closer to achieving the goal of curing acute myeloid leukemia in the coming decades.

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Depleting interferon regulatory factor‐1(IRF‐1) with CRISPR/Cas9 attenuates inducible oxidative metabolism without affecting RA‐induced differentiation in HL‐60 human AML cells

Abstract: This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

Cited by 2 publications

References 46 publications

Vacuolin-1 enhances RA-induced differentiation of human myeloblastic leukemia cells: evidence for involvement of a CD11b/FAK/LYN/SLP-76 axis subject to endosomal regulation that drives late differentiation steps

Vacuolin-1 enhances RA-induced differentiation of human myeloblastic leukemia cells: evidence for involvement of a CD11b/FAK/LYN/SLP-76 axis subject to endosomal regulation that drives late differentiation steps

The high-performance technology CRISPR/Cas9 improves knowledge and management of acute myeloid leukemia

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