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Cited by 7Acute Myeloid Leukemia ResearchRetinoids in leukemia and cellular processesProtein Kinase Regulation and GTPase Signaling

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Bimodal expression of RHOH during myelomonocytic differentiation: Implications for the expansion of AML differentiation therapy

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Galiègue‐Zouitina, Sylvie, Fu, Qiangwei, Carton‐Latreche, Céline, Poret, Nicolas, Cheok, Meyling, Leprêtre, Frédéric, Figeac, Martin, Quesnel, Bruno, El Bouazzati, Hassiba, Shelley, Carl S·2021·eJHaem
DOI 10.1002/jha2.128PMC PMC9175762
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Abstract

RhoH is an unusual member of the Rho family of small GTP‐binding proteins in that it lacks GTPase activity. Since the RhoH protein is constantly bound by GTP, it is constitutively active and controlled predominantly by changes in quantitative expression. Abnormal levels of RHOH gene transcripts have been linked to a range of malignancies including acute myeloid leukemia (AML). One of the hallmarks of AML is a block in the normal program of myeloid differentiation. Here we investigate how myeloid differentiation is controlled by the quantitative expression of RHOH. Our analysis demonstrates that increasingly mature myeloid cells express progressively lower levels of RHOH. However, as monocytic myeloid cells terminally differentiate into macrophages, RHOH expression is up‐regulated. This up‐regulation is not apparent in AML where myeloid differentiation is blocked at stages of low RHOH expression. Nevertheless, when the up‐regulation of RHOH is forced, then terminal macrophage differentiation is induced and the Cdc42 and Wnt intracellular signalling pathways are repressed. These results indicate that RHOH induction is a driver of terminal differentiation and might represent a means of effecting AML differentiation therapy. The potential of this therapeutic strategy is supported by forced up‐regulation of RHOH reducing the ability of AML cells to produce tumours in vivo.

Overview

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TL;DR

RHOH expression is bimodal during myelomonocytic differentiation: it declines through early/intermediate stages but is induced during terminal monocyte-to-macrophage differentiation. Forced RHOH expression in AML cells drives terminal differentiation, represses Cdc42/Wnt signalling, and reduces tumour growth in a xenograft model.

Key findings

  • RHOH mRNA and protein progressively decline with myeloid maturation: M0/M1 vs M5 mRNA was 0.802 ± SD 0.603 vs 0.037 ± SD 0.033 (21.7-fold, P = 0.0067); patient protein was 2.12 ± SD 0.42 vs 1.29 ± SD 0.18 (P = 0.0012); cell-line protein was 2.06 ± SD 0.46 vs 0.94 ± SD 0.40 (P = 0.0008).
  • Terminal differentiation induces RHOH: PMA or LPS treatment of HL60/OCI-AML3, and PMA treatment of primary monocytes and AML FAB M4 myeloblasts, increased RhoH protein while JunD was down-regulated.
  • Forced RHOH in OCI-AML3 (OCI-RhoH vs OCI-Empty) reduced CD93 and CD11b and altered 72 transcripts; 42 were linked to mature myeloid functions, with 9 down-regulated Cdc42/Wnt-positive effectors and 4 up-regulated negative effectors plus RMRP.
  • In xenografts, tumours developed in 12/13 mice injected with OCI-Empty vs 5/12 with OCI-RhoH; OCI-RhoH tumours had average volumes 3.6-fold smaller (P = 0.004).

Methods

  • RHOH expression was analysed in FAB-subtyped de novo AML transcriptomes from five microarray datasets and one RNA-seq dataset (ALFA-0701, GEO GDS1064/GDS3057/GDS3312/GDS3329, TCGA), plus AML cell lines, primary monocytes, and primary AML blasts using Q-RT-PCR and Western blotting.
  • OCI-AML3 daughter lines stably expressing empty vector (OCI-Empty) or RHOH (OCI-RhoH) were compared by Western blot and Agilent whole-human-genome microarray; 72 genes differed significantly (P ≤ 0.05; GEO GSE138479).
  • OCI-Empty or OCI-RhoH cells (5 or 10 × 10^6) were injected subcutaneously into SHrNTM Hairless NOD.SCID mice; tumour volumes were measured 3 weeks later and compared by Mann-Whitney test.

Limitations

  • The paper has no dedicated limitations section. It notes that cell lines may not mimic in vivo biology, so key findings were verified in primary monocytes and AML myeloblasts.
  • Evident from the text: the in vivo therapeutic evidence comes from a single AML cell line (OCI-AML3) in immunocompromised mice, with small groups (13 OCI-Empty and 12 OCI-RhoH injected; tumour volume analysis n = 12 vs n = 11 after outlier removal) and a short 3-week endpoint.
  • The conflict-of-interest statement notes that Carl S. Shelley is President and CEO of Leukemia Therapeutics, LLC, which is developing AML treatments based on targeting consequences of aberrant RHOH repression.

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Integrity signals

Registry

Crossref · 10.1002/jha2.128

Cites

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Figures

FIGURE 1 Repression of RHOH mRNA expression during the intermediate phases of myelomonocytic differentiation as assessed by the FAB subtype of AML patient samples and cell lines. (A– D) Regression curves of the means of relative RHOH mRNA levels as a function of FAB subtype established from the NCBI GEO Datasets GDS1064 [46] and GDS3057 [47] composed of AML patients with all karyotypes and GDS3312 and GDS3329 composed only of AML patients with a normal karyotype [48]. (E) Regression curves of the means of relative RHOH mRNA levels as a function of FAB subtype established from microarray analyses of AML patients recruited by the ALFA‐0701 study (ALFA) [43, 44, 45]. (F) Regression curve of the means of relative RHOH mRNA levels as a function of FAB subtype established from normalized RNA‐seq data generated by The Cancer Genome Atlas (TCGA) [45, 49]. The GDS DataSets as well as the ALFA‐0701 and TCGA data all reveal a linear decrease in RHOH mRNA expression as differentiation stage progresses from AML FAB subtype M0 to M5 (R2 values are presented at the bottom left of each panel in black text). In addition, all sets of data demonstrate a significant difference in RHOH mRNA expression between the early M0 and M1 stages of AML differentiation compared to the later M5 stage (P values presented at the bottom left of each panel in blue text). Mean RHOH mRNA levels for individual FAB subtypes are depicted as open blue lozenges with their standard deviations (SD) presented as vertical bars above and below. In addition, the number (n) of patients analysed in each FAB subtype is presented. (G) Total RNA was extracted from the cell lines Kasumi‐3 (K3), KG‐1 (KG) and KG‐1a (Ka) established from patients diagnosed with AML FAB subtype M0; from the cell lines GF‐D8 (GF) and YNH‐1 (YN) representing AML FAB subtype M1; and from THP‐1 (TH), Nomo‐1 (N1), Mono Mac‐1 (M1), MV4‐11 (MV), Molm‐14 (MO), SHI‐1 (SH) and SKM‐1 (SK) representing FAB subtype M5. The relative level of RHOH mRNA expression was evaluated for each cell line by Q‐RT‐PCR as previously described [37]. The mean relative expression of RHOH mRNA for three independent cultures of each cell line, each analysed twice in duplicate by Q‐RT‐PCR, are depicted as histograms with SD presented as vertical lines above. The mean relative RHOH mRNA expression in FAB subtypes M0 and M1 combined is 0.802 ± SD 0.603 while that of M5 is 0.037 ± SD 0.033. These two means are depicted as blue horizontal bars and their difference of 21.7 fold was determined to be significant (P = 0.0067). The P values presented in all panels were calculated using an unpaired Student's t‐test
FIGURE 1 Repression of RHOH mRNA expression during the intermediate phases of myelomonocytic differentiation as assessed by the FAB subtype of AML patient samples and cell lines. (A– D) Regression curves of the means of relative RHOH mRNA levels as a function of FAB subtype established from the NCBI GEO Datasets GDS1064 [46] and GDS3057 [47] composed of AML patients with all karyotypes and GDS3312 and GDS3329 composed only of AML patients with a normal karyotype [48]. (E) Regression curves of the means of relative RHOH mRNA levels as a function of FAB subtype established from microarray analyses of AML patients recruited by the ALFA‐0701 study (ALFA) [43, 44, 45]. (F) Regression curve of the means of relative RHOH mRNA levels as a function of FAB subtype established from normalized RNA‐seq data generated by The Cancer Genome Atlas (TCGA) [45, 49]. The GDS DataSets as well as the ALFA‐0701 and TCGA data all reveal a linear decrease in RHOH mRNA expression as differentiation stage progresses from AML FAB subtype M0 to M5 (R2 values are presented at the bottom left of each panel in black text). In addition, all sets of data demonstrate a significant difference in RHOH mRNA expression between the early M0 and M1 stages of AML differentiation compared to the later M5 stage (P values presented at the bottom left of each panel in blue text). Mean RHOH mRNA levels for individual FAB subtypes are depicted as open blue lozenges with their standard deviations (SD) presented as vertical bars above and below. In addition, the number (n) of patients analysed in each FAB subtype is presented. (G) Total RNA was extracted from the cell lines Kasumi‐3 (K3), KG‐1 (KG) and KG‐1a (Ka) established from patients diagnosed with AML FAB subtype M0; from the cell lines GF‐D8 (GF) and YNH‐1 (YN) representing AML FAB subtype M1; and from THP‐1 (TH), Nomo‐1 (N1), Mono Mac‐1 (M1), MV4‐11 (MV), Molm‐14 (MO), SHI‐1 (SH) and SKM‐1 (SK) representing FAB subtype M5. The relative level of RHOH mRNA expression was evaluated for each cell line by Q‐RT‐PCR as previously described [37]. The mean relative expression of RHOH mRNA for three independent cultures of each cell line, each analysed twice in duplicate by Q‐RT‐PCR, are depicted as histograms with SD presented as vertical lines above. The mean relative RHOH mRNA expression in FAB subtypes M0 and M1 combined is 0.802 ± SD 0.603 while that of M5 is 0.037 ± SD 0.033. These two means are depicted as blue horizontal bars and their difference of 21.7 fold was determined to be significant (P = 0.0067). The P values presented in all panels were calculated using an unpaired Student's t‐testCC BY · PubMed Central
FIGURE 2 Repression of RhoH protein expression during the intermediate phases of myelomonocytic differentiation as assessed by the FAB subtype of AML patient samples and cell lines. (A) Western blot evaluation of relative RhoH protein expression in lysates prepared from peripheral blood mononuclear cells of patients diagnosed with AML FAB subtype M0 (P1 and P2), M1 (P3 ‐ 6) or M5 (P7 ‐ 12). RhoH protein expression is normalized to the expression of GAPDH protein and presented as histograms representing the mean ± SD of three (P1, P2, P5, P6, and P8 ‐ 12) or four (P3, P4, and P7) independent Western blot analyses. Blue horizontal bars represent the means of relative RhoH protein expression in FAB subtypes M0 and M1 (2.12 ± SD 0.42) compared to FAB subtype M5 (1.29 ± SD 0.18). The difference between these two means was calculated by an unpaired Student's t‐test as being significant (P = 0.0012). (B) Representative Western blots evaluating RhoH and GAPDH protein expression in patients with AML FAB subtype M0 (P1 and P2), M1 (P4 and P5), and M5 (P8 and P9). (C) Western blot evaluation of relative RhoH protein expression in lysates prepared from cell lines Kasumi‐3 (K3), KG‐1 (KG), and KG‐1a (Ka) representing AML FAB subtype M0, cell lines GF‐D8 (GF) and YNH‐1 (YN) representing FAB subtype M1 and THP‐1 (TH), Nomo‐1 (N1), Mono Mac‐1 (M1), MV4‐11 (MV), Molm‐14 (MO), SHI‐1 (SH), SKM‐1 (SK), and Mono Mac‐6 (M6) representing FAB subtype M5. RhoH protein expression is normalized to the expression of GAPDH protein and presented as histograms representing the mean ± SD of the results obtained from two or three independent cultures of each cell line and each of these subjected to between two and eight Western blot analyses. Blue horizontal bars represent the means of relative RhoH protein expression in FAB subtypes M0 and M1 (2.06 ± SD 0.46) compared to FAB subtype M5 (0.94 ± SD 0.40). The difference between these two means was calculated by an unpaired Student's t‐test as being significant (P = 0.0008). (D) Representative Western blots evaluating RhoH and GAPDH protein expression in cell lines established from patients with AML FAB subtype M0 (K3 and KG), M1 (GF and YN), and M5 (MO and SH)
FIGURE 2 Repression of RhoH protein expression during the intermediate phases of myelomonocytic differentiation as assessed by the FAB subtype of AML patient samples and cell lines. (A) Western blot evaluation of relative RhoH protein expression in lysates prepared from peripheral blood mononuclear cells of patients diagnosed with AML FAB subtype M0 (P1 and P2), M1 (P3 ‐ 6) or M5 (P7 ‐ 12). RhoH protein expression is normalized to the expression of GAPDH protein and presented as histograms representing the mean ± SD of three (P1, P2, P5, P6, and P8 ‐ 12) or four (P3, P4, and P7) independent Western blot analyses. Blue horizontal bars represent the means of relative RhoH protein expression in FAB subtypes M0 and M1 (2.12 ± SD 0.42) compared to FAB subtype M5 (1.29 ± SD 0.18). The difference between these two means was calculated by an unpaired Student's t‐test as being significant (P = 0.0012). (B) Representative Western blots evaluating RhoH and GAPDH protein expression in patients with AML FAB subtype M0 (P1 and P2), M1 (P4 and P5), and M5 (P8 and P9). (C) Western blot evaluation of relative RhoH protein expression in lysates prepared from cell lines Kasumi‐3 (K3), KG‐1 (KG), and KG‐1a (Ka) representing AML FAB subtype M0, cell lines GF‐D8 (GF) and YNH‐1 (YN) representing FAB subtype M1 and THP‐1 (TH), Nomo‐1 (N1), Mono Mac‐1 (M1), MV4‐11 (MV), Molm‐14 (MO), SHI‐1 (SH), SKM‐1 (SK), and Mono Mac‐6 (M6) representing FAB subtype M5. RhoH protein expression is normalized to the expression of GAPDH protein and presented as histograms representing the mean ± SD of the results obtained from two or three independent cultures of each cell line and each of these subjected to between two and eight Western blot analyses. Blue horizontal bars represent the means of relative RhoH protein expression in FAB subtypes M0 and M1 (2.06 ± SD 0.46) compared to FAB subtype M5 (0.94 ± SD 0.40). The difference between these two means was calculated by an unpaired Student's t‐test as being significant (P = 0.0008). (D) Representative Western blots evaluating RhoH and GAPDH protein expression in cell lines established from patients with AML FAB subtype M0 (K3 and KG), M1 (GF and YN), and M5 (MO and SH)CC BY · PubMed Central
FIGURE 3 Induction of RhoH protein expression during terminal differentiation of AML cell lines. (A) The AML cell line HL60 or (B) the AML cell line OCI‐AML3 were cultured in the presence of 20 ng/ml PMA or its vehicle DMSO (Ctrl) for 1, 9 or 48 h or 100 ng/ml LPS or its vehicle PBS (Ctrl) for 7 or 14 days. Total protein lysates were prepared and Western blot analysis of RhoH and GAPDH protein expression performed. Histograms represent the means ± SD of RhoH protein expression normalized to GAPDH protein in cells treated with PMA or LPS calculated relative to values of 100% assigned to normalized levels of RhoH protein expressed in cells treated with the appropriate vehicle. Protein lysates were prepared from three independent cell cultures for each treatment condition. Each lysate was analyzed for RhoH and GAPDH expression one or two times by Western blotting. One of these analyses is presented below each histogram. P values indicating significant induction of RhoH protein expression in cells treated with PMA for 48 hours or LPS for 14 days compared to cells treated with the appropriate vehicle were calculated using an unpaired Student's t‐test
FIGURE 3 Induction of RhoH protein expression during terminal differentiation of AML cell lines. (A) The AML cell line HL60 or (B) the AML cell line OCI‐AML3 were cultured in the presence of 20 ng/ml PMA or its vehicle DMSO (Ctrl) for 1, 9 or 48 h or 100 ng/ml LPS or its vehicle PBS (Ctrl) for 7 or 14 days. Total protein lysates were prepared and Western blot analysis of RhoH and GAPDH protein expression performed. Histograms represent the means ± SD of RhoH protein expression normalized to GAPDH protein in cells treated with PMA or LPS calculated relative to values of 100% assigned to normalized levels of RhoH protein expressed in cells treated with the appropriate vehicle. Protein lysates were prepared from three independent cell cultures for each treatment condition. Each lysate was analyzed for RhoH and GAPDH expression one or two times by Western blotting. One of these analyses is presented below each histogram. P values indicating significant induction of RhoH protein expression in cells treated with PMA for 48 hours or LPS for 14 days compared to cells treated with the appropriate vehicle were calculated using an unpaired Student's t‐testCC BY · PubMed Central
FIGURE 4 Induction of RhoH protein expression during terminal differentiation of freshly isolated monocytes and myeloblasts; RhoH induction drives terminal differentiation (A) Normal monocytes were isolated from the peripheral blood of healthy donors. Total protein lysates were prepared from these cells (Mono). In addition, total protein lysates were prepared after these monocytes were induced to differentiate into macrophages with 5 ng/ml of PMA for 48 hours (Mac). Next, Western blot analysis was performed to determine the expression of RhoH, CD93, CD11b, and JunD proteins relative to the control protein HSC70. Depicted are representative examples of analyses performed on five independent preparations of monocytes and macrophages. Three preparations were analysed for RhoH and CD93 expression by four and two Western blots, respectively. Two preparations were analysed for CD11b and JunD expression by one and two Western blots, respectively. (B) Myeloblasts were isolated at diagnosis from patients with AML FAB M4. Total protein lysates were prepared from these cells (Myelo). In addition, total protein lysates were prepared after these myeloblasts were induced to differentiate into macrophages with 5 ng/ml of PMA for 48 hours (Mac). Western blot analysis was performed to determine the expression of RhoH, CD93, CD11b and JunD proteins relative to the control protein HSC70. Depicted are representative examples of analyses performed on two independent preparations of myeloblasts and macrophages. Each preparation was analysed by two Western blots for RhoH and CD93 expression and by one Western blot for CD11b and JunD expression. (C) Total protein lysates were prepared from the AML cell line OCI‐AML3 stably transfected either with the empty vector pMEP4 (Empty) or pMEP4‐RhoH that constitutively expresses RHOH (RhoH). Next, Western blot analysis was performed to determine the expression of RhoH, CD93, CD11b and JunD proteins relative to the control proteins GAPDH or HSC70. Depicted are representative examples of analyses performed on three independent cultures of each stable transfectant. Each independent culture was analysed by three Western blots for RhoH expression and two Western blots for CD93, CD11b and JunD expression
FIGURE 4 Induction of RhoH protein expression during terminal differentiation of freshly isolated monocytes and myeloblasts; RhoH induction drives terminal differentiation (A) Normal monocytes were isolated from the peripheral blood of healthy donors. Total protein lysates were prepared from these cells (Mono). In addition, total protein lysates were prepared after these monocytes were induced to differentiate into macrophages with 5 ng/ml of PMA for 48 hours (Mac). Next, Western blot analysis was performed to determine the expression of RhoH, CD93, CD11b, and JunD proteins relative to the control protein HSC70. Depicted are representative examples of analyses performed on five independent preparations of monocytes and macrophages. Three preparations were analysed for RhoH and CD93 expression by four and two Western blots, respectively. Two preparations were analysed for CD11b and JunD expression by one and two Western blots, respectively. (B) Myeloblasts were isolated at diagnosis from patients with AML FAB M4. Total protein lysates were prepared from these cells (Myelo). In addition, total protein lysates were prepared after these myeloblasts were induced to differentiate into macrophages with 5 ng/ml of PMA for 48 hours (Mac). Western blot analysis was performed to determine the expression of RhoH, CD93, CD11b and JunD proteins relative to the control protein HSC70. Depicted are representative examples of analyses performed on two independent preparations of myeloblasts and macrophages. Each preparation was analysed by two Western blots for RhoH and CD93 expression and by one Western blot for CD11b and JunD expression. (C) Total protein lysates were prepared from the AML cell line OCI‐AML3 stably transfected either with the empty vector pMEP4 (Empty) or pMEP4‐RhoH that constitutively expresses RHOH (RhoH). Next, Western blot analysis was performed to determine the expression of RhoH, CD93, CD11b and JunD proteins relative to the control proteins GAPDH or HSC70. Depicted are representative examples of analyses performed on three independent cultures of each stable transfectant. Each independent culture was analysed by three Western blots for RhoH expression and two Western blots for CD93, CD11b and JunD expressionCC BY · PubMed Central
FIGURE 5 Regulators of Cdc42 and Wnt signalling controlled by RHOH. Differential microarray analysis identified 72 transcripts that are significantly changed in their expression when RHOH is reconstituted in the M4 AML cell line OCI‐AML3 (GEO Accession number GSE138479). Of these transcripts, 13 encode proteins and 1 the long non‐coding RNA RMRP known to regulate Cdc42 and/or Wnt signalling (Supplemental References 2). Core components of these signalling pathways are depicted as blue ovals. Agonists of these signalling pathways that are down‐regulated by RHOH induction are depicted as green ovals and antagonists up‐regulated by RHOH are depicted as red ovals. Overlapping ovals signify the direct binding of proteins
FIGURE 5 Regulators of Cdc42 and Wnt signalling controlled by RHOH. Differential microarray analysis identified 72 transcripts that are significantly changed in their expression when RHOH is reconstituted in the M4 AML cell line OCI‐AML3 (GEO Accession number GSE138479). Of these transcripts, 13 encode proteins and 1 the long non‐coding RNA RMRP known to regulate Cdc42 and/or Wnt signalling (Supplemental References 2). Core components of these signalling pathways are depicted as blue ovals. Agonists of these signalling pathways that are down‐regulated by RHOH induction are depicted as green ovals and antagonists up‐regulated by RHOH are depicted as red ovals. Overlapping ovals signify the direct binding of proteinsCC BY · PubMed Central
FIGURE 6 RHOH reconstitution inhibits the growth of AML tumours in vivo. The AML cell line OCI‐RhoH that stably expresses the RHOH vector pMEP‐RhoH and its sister line OCI‐Empty that stably expresses the same vector empty of RHOH were injected subcutaneously into SHrN™ Hairless NOD.SCID mice (Harlan Laboratories, Inc., Indianapolis, Indiana, USA). In one experiment 5 million OCI‐Empty cells were injected into 8 mice and in parallel 5 million OCI‐RhoH cells were injected into 7 mice. In a second experiment 10 million OCI‐Empty or OCI‐RhoH cells were each injected into 5 mice. The volumes of palpable tumours were measured 3 weeks after injection. Tumour volumes beyond 1.5 times the interquartile ranges of the ordered OCI‐Empty and OCI‐RhoH datasets were removed (Supplemental Table 1). Histograms represent the means ± SEM of the remaining volumes (OCI‐Empty n = 12; OCI‐RhoH n = 11). A Mann‐Whitney test determined that the mean volume of tumours produced by OCI‐RhoH was significantly smaller than that produced by OCI‐Empty (P = 0.004)
FIGURE 6 RHOH reconstitution inhibits the growth of AML tumours in vivo. The AML cell line OCI‐RhoH that stably expresses the RHOH vector pMEP‐RhoH and its sister line OCI‐Empty that stably expresses the same vector empty of RHOH were injected subcutaneously into SHrN™ Hairless NOD.SCID mice (Harlan Laboratories, Inc., Indianapolis, Indiana, USA). In one experiment 5 million OCI‐Empty cells were injected into 8 mice and in parallel 5 million OCI‐RhoH cells were injected into 7 mice. In a second experiment 10 million OCI‐Empty or OCI‐RhoH cells were each injected into 5 mice. The volumes of palpable tumours were measured 3 weeks after injection. Tumour volumes beyond 1.5 times the interquartile ranges of the ordered OCI‐Empty and OCI‐RhoH datasets were removed (Supplemental Table 1). Histograms represent the means ± SEM of the remaining volumes (OCI‐Empty n = 12; OCI‐RhoH n = 11). A Mann‐Whitney test determined that the mean volume of tumours produced by OCI‐RhoH was significantly smaller than that produced by OCI‐Empty (P = 0.004)CC BY · PubMed Central
FIGURE 7 Nucleotide sequence of the P3 promoter of the human RHOH gene. The site of transcription initiated by the promoter is indicated as +1 with a black horizontal arrow depicting the 5′ end of exon 4 of the RHOH gene [14]. The location of potential binding sites for the transcription factor proteins c‐Myc, E2F1, HIF1, LEF1 and SRY that could activate the promoter are indicated with green horizontal bars and text [71, 72]. Each of these transcription factors is regulated by Wnt and/or Cdc42 signalling (Supplemental References 3). The location of binding sites for the JunD protein that represses the promoter are indicated with red horizontal bars and text [14]. The 7 CpG dinucleotides that could be methylated in AML are depicted in bold red text [70]
FIGURE 7 Nucleotide sequence of the P3 promoter of the human RHOH gene. The site of transcription initiated by the promoter is indicated as +1 with a black horizontal arrow depicting the 5′ end of exon 4 of the RHOH gene [14]. The location of potential binding sites for the transcription factor proteins c‐Myc, E2F1, HIF1, LEF1 and SRY that could activate the promoter are indicated with green horizontal bars and text [71, 72]. Each of these transcription factors is regulated by Wnt and/or Cdc42 signalling (Supplemental References 3). The location of binding sites for the JunD protein that represses the promoter are indicated with red horizontal bars and text [14]. The 7 CpG dinucleotides that could be methylated in AML are depicted in bold red text [70]CC BY · PubMed Central

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Authors
Galiègue‐Zouitina, S., Fu, Q., Carton‐Latreche, C., Poret, N., Cheok, M., Leprêtre, F., Figeac, M., Quesnel, B., El Bouazzati, H., & Shelley, C. S.
Affiliations
  • Inserm (FR)
  • California Institute for Biomedical Research (US)
  • JPARC UMRS 1172 Inserm Lille University Lille France
  • Centre National de la Recherche Scientifique (FR)
  • Université de Lille (FR)
  • LLC Leukaemia Therapeutics Hull Massachusetts USA
Corresponding author
  • Galiègue‐Zouitina, Sylvie
  • Shelley, Carl S
ORCID
Venue
eJHaem
Publisher
Wiley
Published
2021-01-20
Volume
2
Issue
2
Pages
196-210
ISSN
2688-6146 (ISSN-L)
DOI
10.1002/jha2.128
PMC
PMC9175762
OpenAlex
W3122045750
Semantic Scholar
15fe3a033339383a91c14d85f9077b30e17446da
Cited by
7
References
73
Citations by year
2026: 3 · 2025: 1 · 2024: 2 · 2021: 1
FWCI
0.49
Type
journal article
Language
en
Open access
Licence
Locations
  • eJHaem (publishedVersion, cc-by)
  • PubMed (publishedVersion)
  • PubMed Central (publishedVersion, cc-by)
  • HAL (submittedVersion)
  • DOAJ (DOAJ: Directory of Open Access Journals) (submittedVersion)
Funders
Institut National de la Santé et de la Recherche Médicale
Topics
  • Health Sciences › Medicine › Hematology › Acute Myeloid Leukemia Research
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Retinoids in leukemia and cellular processes
  • Life Sciences › Biochemistry, Genetics and Molecular Biology › Molecular Biology › Protein Kinase Regulation and GTPase Signaling
Keywords
RhoH, acute myeloid leukemia, myelomonocytic differentiation, macrophage differentiation, terminal differentiation, Cdc42 signaling, Wnt signaling, differentiation therapy

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Galiègue‐Zouitina, S., Fu, Q., Carton‐Latreche, C., Poret, N., Cheok, M., Leprêtre, F., Figeac, M., Quesnel, B., El Bouazzati, H., & Shelley, C. S. (2021). Bimodal expression of RHOH during myelomonocytic differentiation: Implications for the expansion of AML differentiation therapy. eJHaem. https://doi.org/10.1002/jha2.128