Shwachman–Bodian–Diamond syndrome (SBDS) protein is a direct inhibitor of protein phosphatase 2A (PP2A) activity and overexpressed in acute myeloid leukaemia

Dun, Matthew D.; Mannan, Abdul; Rigby, Callum J; Butler, Stephen; Toop, Hamish D.; Beck, Dominik; Connerty, Patrick; Sillar, Jonathan R; Kahl, Richard; Duchatel, Ryan J.; Germon, Zacary P.; Faulkner, Sam; Chi, Mengna; Skerrett‐Byrne, David A.; Murray, Heather C; Flanagan, Hayley; Almazi, Juhura G.; Hondermarck, Hubert; Nixon, Brett; Iuliis, Geoffry N. De; Chamberlain, Janis; Alvaro, Frank; Bock, Charles E. de; Morris, Jonathan C.; Enjeti, Anoop K; Verrills, Nicole M.

doi:10.1038/s41375-020-0814-0

Cited by 16 publications

(22 citation statements)

References 15 publications

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“…Samples underwent reduction and alkylation with 10 mM dithiothreitol (30 min at room temperature) and 20 mM iodoacetamide (30 min protected from light at room temperature) respectively. Peptide populations were digested with 1:30 Lys-C/Trypsin Mix (Promega) ( 26 , 27 , 28 , 29 , 30 ) at room temperature for 3 h. Using 50 mM triethylammonium bicarbonate, the concentration of urea was brought below 1 M and digested overnight at 37 °C. Lipids were precipitated from peptide suspensions using formic acid (2% v/v final concentration), and the resultant peptides were purified using Oasis 1 cc desalting columns (Waters Corp).…”

Section: Methodsmentioning

confidence: 99%

Proteomic Dissection of the Impact of Environmental Exposures on Mouse Seminal Vesicle Function

Skerrett‐Byrne

Trigg

Bromfield

et al. 2021

Molecular & Cellular Proteomics

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Seminal vesicles are an integral part of the male reproductive accessory gland system. They produce a complex array of secretions containing bioactive constituents that support gamete function and promote reproductive success, with emerging evidence suggesting these secretions are influenced by our environment. Despite their significance, the biology of seminal vesicles remains poorly defined. Here, we complete the first proteomic assessment of mouse seminal vesicles and assess the impact of the reproductive toxicant acrylamide. Mice were administered acrylamide (25 mg/kg bw/day) or control daily for five consecutive days prior to collecting seminal vesicle tissue. A total of 5013 proteins were identified in the seminal vesicle proteome with bioinformatic analyses identifying cell proliferation, protein synthesis, cellular death, and survival pathways as prominent biological processes. Secreted proteins were among the most abundant, and several proteins are linked with seminal vesicle phenotypes. Analysis of the effect of acrylamide on the seminal vesicle proteome revealed 311 differentially regulated (FC ± 1.5, p ≤ 0.05, 205 up-regulated, 106 downregulated) proteins, orthogonally validated via immunoblotting and immunohistochemistry. Pathways that initiate protein synthesis to promote cellular survival were prominent among the dysregulated pathways, and rapamycin-insensitive companion of mTOR (RICTOR, p = 6.69E-07) was a top-ranked upstream driver. Oxidative stress was implicated as contributing to protein changes, with acrylamide causing an increase in 8-OHdG in seminal vesicle epithelial cells (fivefold increase, p = 0.016) and the surrounding smooth muscle layer (twofold increase, p = 0.043). Additionally, acrylamide treatment caused a reduction in seminal vesicle secretion weight (36% reduction, p = 0.009) and total protein content (25% reduction, p = 0.017). Together these findings support the interpretation that toxicant exposure influences male accessory gland physiology and highlights the need to consider the response of all male reproductive tract tissues when interpreting the impact of environmental stressors on male reproductive function.

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Section: Methodsmentioning

confidence: 99%

Proteomic Dissection of the Impact of Environmental Exposures on Mouse Seminal Vesicle Function

Skerrett‐Byrne

Trigg

Bromfield

et al. 2021

Molecular & Cellular Proteomics

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“… 15 Cell growth and proliferation were determined as described, 16 using 2.5 × 10 4 cell/well, plated the day before the addition of ONC201 or GsONC201 to allow for neurosphere formation. Drug sensitivity was assessed over 96 h. Annexin V/PI assays were performed as described, 17 using 5 μM ONC201 or GsON201 over 72 h. Western blotting was performed as described. 18 …”

Section: Methodsmentioning

confidence: 99%

“…Assessment of ONC201 and GsONC201 into their putative targets was performed as described, 17 using GOLD Protein-Ligand Docking Software (version 2020.2.0). 19 , 20 …”

Section: Methodsmentioning

confidence: 99%

Preclinical and clinical evaluation of German-sourced ONC201 for the treatment of H3K27M-mutant diffuse intrinsic pontine glioma

Duchatel

Mannan

Woldu

et al. 2021

Neuro-Oncology Advances

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Background Diffuse intrinsic pontine glioma (DIPG) is a fatal childhood brainstem tumor for which radiation is the only treatment. Case studies report a clinical response to ONC201 for patients with H3K27M-mutant gliomas. Oncoceutics (ONC201) is only available in the United States and Japan; however, in Germany, DIPG patients can be prescribed and dispensed a locally produced compound—ONC201 German-sourced ONC201 (GsONC201). Pediatric oncologists face the dilemma of supporting the administration of GsONC201 as conjecture surrounds its authenticity. Therefore, we compared GsONC201 to original ONC201 manufactured by Oncoceutics Inc. Methods Authenticity of GsONC201 was determined by high-resolution mass spectrometry and nuclear magnetic resonance spectroscopy. Biological activity was shown via assessment of on-target effects, in vitro growth, proliferation, and apoptosis analysis. Patient-derived xenograft mouse models were used to assess plasma and brain tissue pharmacokinetics, pharmacodynamics, and overall survival (OS). The clinical experience of 28 H3K27M+ mutant DIPG patients who received GsONC201 (2017–2020) was analyzed. Results GsONC201 harbored the authentic structure, however, was formulated as a free base rather than the dihydrochloride salt used in clinical trials. GsONC201 in vitro and in vivo efficacy and drug bioavailability studies showed no difference compared to Oncoceutics ONC201. Patients treated with GsONC201 (n = 28) showed a median OS of 18 months (P = .0007). GsONC201 patients who underwent reirradiation showed a median OS of 22 months compared to 12 months for GsONC201 patients who did not (P = .012). Conclusions This study confirms the biological activity of GsONC201 and documents the OS of patients who received the drug; however, GsONC201 was never used as a monotherapy.

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“…One such phosphatase, a serine/threonine phosphatase PP2A, is commonly inactivated in myeloid leukemias including in about ~78% of AML [11][12][13][14] . Cancer associated PP2A inactivation is often due to either subunit deregulation [15][16][17][18][19] , or overexpression of PP2A inhibitors including SETBP1, CIP2A, SBDS and SET which are frequently associated with poor overall survival 12,[20][21][22][23][24] . PP2A was first characterized as a tumor suppressor in SV40 mediated oncogenic transformation 25,26 .…”

Section: Figures 7 Introductionmentioning

confidence: 99%

“…G) Increase in percent CD11b + cells with 10µM PPZ in HL-60 (48hrs) (n=5, p=0.0002) and U937 cells (72hrs) (n=7, p=0.0012) compared to vehicle control (Veh). H) Increase in percent CD11b + cells in primary AML cells (n=5,AML 15,18,24,38,and 44) with FTY720 (2.5µM, 48hrs, p=0.0375, 7.054% increase) and PPZ treatment (10µM, 48hrs, p=0.0181, 5.668% increase) compared to vehicle control (Veh). I) Representative images of mass cytometric analysis of primary AML cells treated with Veh or OSU-2S (3μM, 48hrs) reveals increased surface differentiation markers CD14 and CD15.…”

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confidence: 99%

PP2A is a therapeutically targetable driver of cell fate decisions via a c-Myc/p21 axis in human and murine acute myeloid leukemia

Goswami

Mani

Nunes

et al. 2022

Blood

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Dysregulated cellular differentiation is a hallmark of acute leukemogenesis. Phosphatases are widely suppressed in cancers but have not been traditionally associated with differentiation. Herein, we identified that the silencing of Protein Phosphatase 2A (PP2A) directly contributes to differentiation block in acute myeloid leukemia (AML). Gene expression and mass cytometric profiling reveal that PP2A activation modulates cell cycle and transcriptional regulators that program terminal myeloid differentiation. Using a novel pharmacological agent OSU-2S in parallel with genetic approaches, we discovered that PP2A enforces c-Myc and p21 dependent terminal differentiation, proliferation arrest and apoptosis in AML. Finally, we demonstrate that PP2A activation decreases leukemia initiating stem cells, increases leukemic blast maturation, and improves overall survival in murine Tet2-/-Flt3ITD/WT and human AML models in-vivo. Our findings identify the PP2A/c-Myc/p21 axis as a critical regulator of the differentiation/proliferation switch in AML that can be therapeutically targeted in malignancies with dysregulated maturation fate.

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Shwachman–Bodian–Diamond syndrome (SBDS) protein is a direct inhibitor of protein phosphatase 2A (PP2A) activity and overexpressed in acute myeloid leukaemia

Cited by 16 publications

References 15 publications

Proteomic Dissection of the Impact of Environmental Exposures on Mouse Seminal Vesicle Function

Proteomic Dissection of the Impact of Environmental Exposures on Mouse Seminal Vesicle Function

Preclinical and clinical evaluation of German-sourced ONC201 for the treatment of H3K27M-mutant diffuse intrinsic pontine glioma

PP2A is a therapeutically targetable driver of cell fate decisions via a c-Myc/p21 axis in human and murine acute myeloid leukemia

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