Reinvent Aliphatic Arsenicals as Reversible Covalent Warheads toward Targeted Kinase Inhibition and Non-acute Promyelocytic Leukemia Cancer Treatment.

The success of arsenic in acute promyelocytic leukemia (APL) treatment is hardly transferred to non-APL cancers, mainly due to the low selectivity and weak binding affinity of traditional arsenicals to oncoproteins critical for cancer survival. We present herein the reinvention of aliphatic trivalent arsenicals (As) as reversible covalent warheads of As-based targeting inhibitors toward Bruton's tyrosine kinase (BTK). The effects of As warheads' valency, thiol protection, methylation, spacer length, and size on inhibitors' activity were studied. We found that, in contrast to the bulky and rigid aromatic As warhead, the flexible aliphatic As warheads were well compatible with the well-optimized guiding group to achieve nanomolar inhibition against BTK. The optimized As inhibitors effectively blocked the BTK-mediated oncogenic signaling pathway, leading to elevated antiproliferative activities toward lymphoma cells and xenograft tumor. Our study provides a promising strategy enabling rational design of new aliphatic arsenic-based reversible covalent inhibitors toward non-APL cancer treatment.

Hesperidin and chlorogenic acid mitigate arsenic-induced oxidative stress via redox regulation, photosystems-related gene expression, and antioxidant efficiency in the chloroplasts of Zea mays.

The ubiquitous metalloid arsenic (As), which is not essential, can be found extensively in the soil and subterranean water of numerous nations, raising substantial apprehensions due to its impact on both agricultural productivity and sustainability. Plants exposed to As often display morphological, physiological, and growth-related abnormalities, collectively leading to reduced productivity. Polyphenols, operating as secondary messengers within the intricate signaling networks of plants, assume integral functions in the acquisition of resistance to diverse environmental stressors, including but not limited to drought, salinity, and exposure to heavy metals. The pivotal roles played by polyphenols in these adaptive processes underscore their profound significance in plant biology. This study aims to elucidate the impact of hesperidin (HP) and chlorogenic acid (CA), recognized as potent bioactive compounds, on maize plants exposed to As. To achieve this objective, the study examined the physiological and biochemical impacts, including growth parameters, photosynthesis, and chloroplastic antioxidants, of HP (100 µM) and CA (50 µM) on Zea mays plants exposed to arsenate stress (AsV, 100 µM - Na2HAsO4⋅7H2O). As toxicity led to reductions in fresh weight (FW) and dry weight (DW) by 33% and 26%, respectively. However, the application of As+HP and As + CA increased FW by 22% and 40% and DW by 14% and 17%, respectively, alleviating the effects of As stress. As toxicity resulted in the up-regulation of PSII genes (psbA and psbD) and PSI genes (psaA and psaB), indicating a potential response to the re-formation of degraded regions, likely driven by the heightened demand for photosynthesis. Exogenous HP or/and CA treatments effectively counteracted the adverse effects of As toxicity on the photochemical quantum efficiency of PSII (Fv/Fm). H2O2 content showed a 23% increase under As stress, and this increase was evident in guard cells when examining confocal microscopy images. In the presence of As toxicity, the chloroplastic antioxidant capacity can exhibit varying trends, with either a decrease or increase observed. After the application of CA and/or HP, a significant increase was observed in the activity of GR, APX, GST, and GPX enzymes, resulting in decreased levels of H2O2 and MDA. Additionally, the enhanced functions of MDHAR and DHAR have modulated the redox status of ascorbic acid (AsA) and glutathione (GSH). The HP or CA-mediated elevated levels of AsA and GSH content further contributed to the preservation of redox homeostasis in chloroplasts facing stress induced by As. In summary, the inclusion of HP and CA in the growth medium sustained plant performance in the presence of As toxicity by regulating physiological and biochemical characteristics, chloroplastic antioxidant enzymes, the AsA-GSH cycle and photosynthesis processes, thereby demonstrating their significant potential to confer resistance to maize through the mitigation of As-induced oxidative damage and the safeguarding of photosynthetic mechanisms.

Iron oxide nanoparticles improving multimetal phytoextraction in Helianthus annuus.

This study assessed the phytotoxicity of a mixture of five different trace elements (TEs) frequently found as pollutants in soils: arsenic, cadmium, copper, lead and zinc. On the other hand, the plant response to a magnetite (Fe3O4) nanoparticle amendment on this mixture as well as nanomagnetite remediation potential has been tested. Sunflower (Helianthus annuus) plants were grown for 90 days in soil contaminated with the five mentioned TEs at the limit levels of TEs in soils likely to receive sludge established by French legislation. Depending on the conditions, experimental set-ups were amended or not with 1% dry weight nanomagnetite (NPsMagn), citric acid-coated nanomagnetite (NPsMagn@CA) or micro-sized magnetite (µPs) in order to assess the behavior of nanomagnetites in a TEs-contaminated water-soil-plant system under repeated water-deficiency stress. The mixture of TEs did not induce phytotoxicity as estimated by plant growth, pigment content, maximum quantum yield of photosynthesis, oxidative impact and antioxidant response. Furthermore, both nanomagnetites treatments in a TEs-contaminated soil significantly increased biomass production by 64 % compared to control and antioxidant enzyme activities compared to control and TEs-treated plants. NPsMagn and NPsMagn@CA particularly enhance phytoextraction of Cd and Cu, increasing the amounts of TEs in aerial parts from 1.5 to 4.5 times compared to set-ups without nanomagnetites. Based on Cd, Cu, Pb and Zn contents in soil solutions, both nanomagnetites treatments improved TEs phytoextraction without increasing groundwater contamination. On the contrary, nanomagnetites significantly reduce arsenic uptake by plants and solubilization in dissolved phase. Our results show that modifying surface physicochemical properties of NPsMagn with citric acid coating does not improve their effects compared to bare NPsMagn. NPsMagn and NPsMagn@CA also appear to mitigate the effects of drought stress. This work highlights several positive environmental aspects related to the use of nanomagnetites in phytoremediation.

Arsenic album 30C exhibits crystalline nano structure of arsenic trioxide and modulates innate immune markers in murine macrophage cell lines.

Macrophages are associated with innate immune response and M1-polarized macrophages exhibit pro-inflammatory functions. Nanoparticles of natural or synthetic compounds are potential triggers of innate immunity. As2O3 is the major component of the homeopathic drug, Arsenic album 30C.This has been claimed to have immune-boosting activities, however, has not been validated experimentally. Here we elucidated the underlying mechanism of Ars. alb 30C-mediated immune priming in murine macrophage cell line. Transmission Electron Microscopy (TEM) and X-ray diffraction (XRD) used for the structural analysis of the drug reveals the presence of crystalline As2O3 nanoparticles of cubic structure. Similarly, signatures of M1-macrophage polarization were observed by surface enhanced Raman scattering (SERS) in RAW 264.7 cells with concomitant over expression of M1 cell surface marker, CD80 and transcription factor, NF-κB, respectively. We also observed a significant increase in pro-inflammatory cytokines like iNOS, TNF-α, IL-6, and COX-2 expression with unaltered ROS and apoptosis in drug-treated cells. Enhanced expression of Toll-like receptors 3 and 7 were observed both in transcriptional and translational levels after the drug treatment. In sum, our findings for the first time indicated the presence of crystalline As2O3 cubic nanostructure in Ars. alb 30C which facilitates modulation of innate immunity by activating macrophage polarization.

Nephroprotective effect of diosmin against sodium arsenite-induced renal toxicity is mediated via attenuation of oxidative stress and inflammation in mice.

Arsenic compounds, which are used in different industries like pesticide manufacturing, cause severe toxic effects in almost all organs, including the kidneys. Since the primary route of exposure to arsenic is through drinking water, and millions of people worldwide are exposed to unsafe levels of arsenic that can pose a threat to their health, this research was performed to investigate the nephroprotective effects of Diosmin (Dios), a flavonoid found in citrus fruits, against nephrotoxicity induced by sodium arsenite (SA). To induce nephrotoxicity, SA (10 mg/kg, oral gavage) was administered to mice for 30 days. Dios (25, 50, and 100 mg/kg, oral gavage) was given to mice for 30 days prior to SA administration. After the study was completed, animals were euthanized and blood and kidney samples were taken for biochemical and histopathological assessments. Results showed that SA-treated mice significantly increased the blood urea nitrogen and creatinine levels in the serum. This increase was associated with significant kidney tissue damage in SA-treated mice, which was confirmed by histopathological studies. Furthermore, SA enhanced the amounts of renal thiobarbituric acid reactive substances and decreased total thiol reserves, as well as the activity of antioxidant enzymes such as catalase, superoxide dismutase, and glutathione peroxidase. Also, in the SA-exposed group, an increase in the levels of kidney inflammatory biomarkers, including nitric oxide and tumor necrosis factor-alpha was observed. The western blot analysis indicated an elevation in the protein expression of kidney injury molecule-1 and nuclear factor-kappa B in SA-treated mice. However, pretreatment with Dios ameliorated the SA-related renal damage in mice. Our findings suggest that Dios can protect the kidneys against the nephrotoxic effects of SA by its antioxidant and anti-inflammatory characteristics.

Macrophage-hitchhiked arsenic/AB bionic preparations for liver cancer.

Macrophage-hitchhiked arsenic/AB bionic preparations were developed to improve the therapeutic effect on liver cancer by means of the tumor-targeting ability of macrophages in vivo. In vitro and in vivo cellular uptake assays demonstrated that arsenic/AB, with negatively charged particles of around 100-200 nm size, could hitchhike to macrophages. Dissolution experiments of arsenic/AB showed that arsenic/AB could delay the release of arsenic and ensure the safety of macrophages during its transport. Histological examination confirmed the safety of the preparations for major organs. In vivo distribution experiment showed that the arsenic/AB bionic preparations could rapidly accumulate in tumors, and in vivo treatment experiment showed a significant tumor inhibition of arsenic/AB. The therapeutic mechanism of liver cancer might be that the arsenic/AB bionic preparations could inhibit tumor growth by reducing inflammatory response and inhibiting CSF1 secretion to block CSF1R activation to induce more differentiation of tumor-associated macrophages (TAMs) towards the anti-tumor M1 phenotype. Therefore, we concluded that the arsenic/AB bionic preparations could improve the distribution of arsenic in vivo by hitchhiking on macrophages as well as make it have tumor targeting and deep penetration abilities, thus increasing the therapeutic effect of arsenic on liver cancer with reduced side effects.

Association of urinary arsenic with the oxidative DNA damage marker 8-hydroxy-2 deoxyguanosine: A meta-analysis.

BACKGROUND: The International Agency for Research on Cancer has classified arsenic as a class I carcinogen. Oxidative DNA damage is a typical early precursor to recognized malignancies. The most sensitive early independent marker of oxidative DNA damage is believed to be 8-hydroxy-2 deoxyguanosine (8-OHdG). To date, research on the link between urinary arsenic and 8-OHdG has not been consistent. OBJECTIVE: This study was aimed at exploring the effects of urinary arsenic on 8-OHdG in human urine. METHODS: A literature search until January 2023 was performed on the PubMed, Cochrane Library, Web of Science, Embase, and Scopus databases through a combination of computer and manual retrieval. Stata 12.0 was used to examine the degree of heterogeneity among included studies. The percentage change and 95 % confidence interval (95 % CI) of 8-OHdG were calculated between populations exposed to different doses. We used a random effect model because the degree of heterogeneity exceeded 50 %. Sensitivity analysis and testing for publication bias were performed. RESULTS: This meta-analysis included nine studies, most of which were performed in China. After exposure to arsenic, urinary arsenic (per 10 µg/g creatinine increase) was associated with the increased 8-OHdG (% change = 41.49 %, 95 % CI: 19.73 %, 63.25 %). Subgroup analysis indicated that the percentage change in 8-OHdG in urine was more pronounced in people exposed to arsenic <50 µg/L (% change = 24.60 %, 95 % CI: 17.35 %, 37.85 %). In studies using total urinary arsenic content as an indicator, the percentage change in 8-OHdG in urine was more significant (% change = 60.38 %, 95 % CI: 15.08 %, 105.68 %). CONCLUSION: The 8-OHdG levels in human urine significantly increased after exposure to environmental arsenic, thus suggesting that arsenic exposure is correlated with oxidative DNA damage.

Protective effects of macromolecular polyphenols, metals (zinc, selenium, and copper) - Polyphenol complexes, and different organs with an emphasis on arsenic poisoning: A review.

For the potential health benefits and nutritional value, polyphenols are one of the secondary metabolites of plants that have received extensive research. It has anti-inflammatory and cytotoxicity-reducing properties in addition to a high antioxidant content. Macromolecular polyphenols and polysaccharides are biologically active natural polymers with antioxidant and anti-inflammatory potential. Arsenic is an ecologically toxic metalloid. Arsenic in drinking water is the most common way people come into contact with this metalloid. While arsenic is known to cause cancer, it is also used to treat acute promyelocytic leukemia (APL). The treatment's effectiveness is hampered by the adverse effects it can cause on the body. Oxidative stress, inflammation, and the inability to regulate cell death cause the most adverse effects. Polyphenols and other macromolecules like polysaccharides act as neuroprotectants by mitigating free radical damage, inhibiting nitric oxide (NO) production, lowering A42 fibril formation, boosting antioxidant levels, and controlling apoptosis and inflammation. To prevent the harmful effects of toxins, polyphenols and pectin lower oxidative stress, boost antioxidant levels, improve mitochondrial function, control apoptosis, and suppress inflammation. Therefore, it prevents damage to the heart, liver, kidneys, and reproductive system. This review aims to identify the effects of the polyphenols in conjugation with polysaccharides as an ameliorative strategy for arsenic-induced toxicity in various organs.

Structural Basis of PML-RARA Oncoprotein Targeting by Arsenic Unravels a Cysteine Rheostat Controlling PML Body Assembly and Function.

PML nuclear bodies (NB) are disrupted in PML-RARA-driven acute promyelocytic leukemia (APL). Arsenic trioxide (ATO) cures 70% of patients with APL, driving PML-RARA degradation and NB reformation. In non-APL cells, arsenic binding onto PML also amplifies NB formation. Yet, the actual molecular mechanism(s) involved remain(s) elusive. Here, we establish that PML NBs display some features of liquid-liquid phase separation and that ATO induces a gel-like transition. PML B-box-2 structure reveals an alpha helix driving B2 trimerization and positioning a cysteine trio to form an ideal arsenic-binding pocket. Altering either of the latter impedes ATO-driven NB assembly, PML sumoylation, and PML-RARA degradation, mechanistically explaining clinical ATO resistance. This B2 trimer and the C213 trio create an oxidation-sensitive rheostat that controls PML NB assembly dynamics and downstream signaling in both basal state and during stress response. These findings identify the structural basis for arsenic targeting of PML that could pave the way to novel cancer drugs. SIGNIFICANCE: Arsenic curative effects in APL rely on PML targeting. We report a PML B-box-2 structure that drives trimer assembly, positioning a cysteine trio to form an arsenic-binding pocket, which is disrupted in resistant patients. Identification of this ROS-sensitive triad controlling PML dynamics and functions could yield novel drugs. See related commentary by Salomoni, p. 2505. This article is featured in Selected Articles from This Issue, p. 2489.

Polyamine, 1,3-diaminopropane, regulates defence responses on growth, gas exchange, PSII photochemistry and antioxidant system in wheat under arsenic toxicity.

The metalloid arsenic (As) is extremely hazardous to all living organisms, including plants. Pollution with As is very detrimental to the photosynthetic machinery, cell division, energy generation, and redox status. In order to cope with stress, the use of growth regulators such as polyamines (PA), which strengthen the antioxidant system of plants, has become widespread in recent years. PAs can modulate the plant growth through basic mechanisms common to all living organisms, such as membrane stabilization, free radical scavenging, deoxyribonucleic acid (DNA), ribonucleic acid (RNA) and protein synthesis, enzyme activities and second messengers. However, the effect of 1,3- diaminopropane (Dap), which is a product of PA catabolism, is not clear enough in plants exposed to As toxicity. In the current study, the different concentrations of 1,3-diaminopropane (0.1, 0.5 and 1 mM Dap) were hydroponically treated to wheat (Triticum aestivum) under arsenic stress (100 µM As) and then relative growth rate (RGR), relative water content (RWC), proline content (Pro), gas exchange parameters, PSII photochemistry, chlorophyll fluorescence kinetics, antioxidant activity and lipid peroxidation were assessed. RGR, RWC, osmotic potential and Pro content decreased in As-applied plants. The inhibition of these parameters could be reversed by Dap treatments. Besides, Dap applications mitigated the As toxicity-induced suppression on chlorophyll fluorescence (Fv/Fm, Fv/Fo and Fo/Fm) and the performance of PSII photochemistry. As impaired the balance on antioxidant capacity by decreased activities of catalase (CAT), peroxidase (POX), glutathione peroxidase (GPX), ascorbate peroxidase (APX), monodehydroascorbate reductase (MDHAR), dehydroascorbate reductase (DHAR), and the contents of ascorbate (AsA) and glutathione (GSH) and then lipid peroxidation (TBARS content) increased. In the presence of Dap under As stress, the plants exhibited an increase in superoxide dismutase (SOD), POX, and GPX. Dap treatments contributed to the maintenance of cellular redox state (AsA/DHA and GSH/GSSG) by regulating the activities/contents of enzyme/non-enzyme involved in the AsA-GSH cycle. After Dap applications against stress, ROS accumulation (H2O2 content) and lipid peroxidation (TBARS) were effectively reduced. The findings showed that by eliminating As-induced oxidative damage and protecting the biochemical processes of photosynthesis, Dap treatments have a substantial potential to give resistance to wheat.

Alleviation of arsenic toxicity in pepper plants by aminolevulinic acid and heme through modulating its sequestration and distribution within cell organelles.

Aminolevulinic acid (ALA) is essential for chlorophyll and heme synthesis. However, whether heme interacts with ALA to elicit antioxidants in arsenic (As)-exposed plants is still unknown. ALA was applied daily to pepper plants for 3 days prior to beginning As stress (As-S). Then, As-S was initiated for 14 days by employing sodium hydrogen arsenate heptahydrate (0.1 mM AsV). Arsenic treatment decreased photosynthetic pigments (chl a by 38% and chl b by 28%), biomass by 24%, and heme by 47% content, but it elevated contents of malondialdehyde (MDA) by 3.3-fold, hydrogen peroxide (H2O2) by 2.3-fold, glutathione (GSH), methylglyoxal (MG), and phytochelatins (PCs) and electrolyte leakage (EL) by 2.3-fold along with enhanced subcellular As concentration in the pepper plant's roots and leaves. The supplementation of ALA to the As-S-pepper seedlings enhanced the amount of chlorophyll, heme content, and antioxidant enzyme activity as well as plant growth, while it reduced the levels of H2O2, MDA, and EL. ALA boosted GSH and phytochelates (PCs) in the As-S-seedlings by controlling As sequestration and rendering it harmless. The addition of ALA enhanced the amount of As that accumulated in the root vacuoles and reduced the poisonousness of the soluble As in the vacuoles. The ALA treatment facilitated the deposition and fixation of As in the vacuoles and cell walls, thereby reducing the transport of As to other cell organelles. This mechanism may have contributed to the observed decrease in As accumulation in the leaves. The administration of 0.5 mM hemin (H) (a source of heme) significantly enhanced ALA-induced arsenic stress tolerance. Hemopexin (Hx, 0.4 µg L-1), a heme scavenger, was treated with the As-S plants along with ALA and ALA + H to observe if heme was a factor in ALA's increased As-S tolerance. Heme synthesis/accumulation in the pepper plants was reduced by Hx, which counteracted the positive effects of ALA. Supplementation of H along with ALA + Hx reversed the negative effects of Hx, demonstrating that heme is required for ALA-induced seedling As-S tolerance.

New insights in to the ameliorative effects of zinc and iron oxide nanoparticles to arsenic stressed spinach (Spinacia oleracea L.).

Nanotechnology is capturing great interest worldwide due to their stirring applications in various fields and also individual application of iron oxide nanoparticle (FeO-NPs) and zinc oxide nanoparticle (ZnO-NPs) have been studied in many literatures. However, the combined application of FeO and ZnO-NPs is a novel approach and studied in only few studies. For this purpose, a pot experiment was conducted to examine the plant growth and biomass, photosynthetic pigments, gas exchange attributes, oxidative stress and response of antioxidant compounds (enzymatic and nonenzymatic), sugars, nutritional status of the plant, organic acid exudation pattern As accumulation from the different parts of the plants in spinach (Spinacia oleracea L.) under the different As concentrations i.e., 0 (no As), 60 and 120 µM] which were primed with combined application of two levels of FeO-NPs (10 and 20 mg L-1) and ZnO-NPs (20 and 40 mg L-1). Results from the present study showed that the increasing levels of As in the soil significantly (P < 0.05) decreased plant growth and biomass, photosynthetic pigments, gas exchange attributes, sugars, and nutritional contents from the roots and shoots of the plants. In contrast, increasing levels of As in the soil significantly (P < 0.05) increased oxidative stress indicators in term of malondialdehyde, hydrogen peroxide, and electrolyte leakage, and also increased organic acid exudation patter in the roots of S. oleracea. The negative impact of As toxicity can overcome the combined application of ZnO-NPs and FeO-NPs, which ultimately increased plant growth and biomass by capturing the reactive oxygen species, and decreased oxidative stress in S. oleracea by decreasing the As contents in the roots and shoots of the plants. Research findings, therefore, suggest that the combined application of ZnO-NPs and FeO-NPs can ameliorate As toxicity in S. oleracea, resulting in improved plant growth and composition under As stress, as depicted by balanced exudation of organic acids.

Targeting HDAC3 to overcome the resistance to ATRA or arsenic in acute promyelocytic leukemia through ubiquitination and degradation of PML-RARα.

Acute promyelocytic leukemia (APL) is driven by the oncoprotein PML-RARα, which recruits corepressor complexes, including histone deacetylases (HDACs), to suppress cell differentiation and promote APL initiation. All-trans retinoic acid (ATRA) combined with arsenic trioxide (ATO) or chemotherapy highly improves the prognosis of APL patients. However, refractoriness to ATRA and ATO may occur, which leads to relapsed disease in a group of patients. Here, we report that HDAC3 was highly expressed in the APL subtype of AML, and the protein level of HDAC3 was positively associated with PML-RARα. Mechanistically, we found that HDAC3 deacetylated PML-RARα at lysine 394, which reduced PIAS1-mediated PML-RARα SUMOylation and subsequent RNF4-induced ubiquitylation. HDAC3 inhibition promoted PML-RARα ubiquitylation and degradation and reduced the expression of PML-RARα in both wild-type and ATRA- or ATO-resistant APL cells. Furthermore, genetic or pharmacological inhibition of HDAC3 induced differentiation, apoptosis, and decreased cellular self-renewal of APL cells, including primary leukemia cells from patients with resistant APL. Using both cell line- and patient-derived xenograft models, we demonstrated that treatment with an HDAC3 inhibitor or combination of ATRA/ATO reduced APL progression. In conclusion, our study identifies the role of HDAC3 as a positive regulator of the PML-RARα oncoprotein by deacetylating PML-RARα and suggests that targeting HDAC3 could be a promising strategy to treat relapsed/refractory APL.

Bacterial consortium (Priestia endophytica NDAS01F, Bacillus licheniformis NDSA24R, and Priestia flexa NDAS28R) and thiourea mediated amelioration of arsenic stress and growth improvement of Oryza sativa L.

The present study analyzed the effects of individual microbes and their consortium (Priestia endophytica NDAS01F, Bacillus licheniformis NDSA24R, and P. flexa NDAS28R) either alone or in interaction with thiourea (TU) on growth and responses of rice plants subjected to As stress (50 mg kg-1 in soil) in a pot experiment. The bacteria used in the experiment were isolated from As contaminated fields of Nadia, West Bengal and showed significant As removal potential in in vitro experiment. The results revealed significant growth improvement, biomass accumulation, and decline in malondialdehyde levels in rice plants in bacterial and TU treatments as compared to control As treatment. The best results were observed in a bacterial consortium (B1-2-3), which induced a profound increase of 65%, 43%, 127% and 83% in root length, shoot length, leaf width and fresh weight, respectively. Sulfur metabolism and cell wall synthesis were stimulated upon bacterial and TU amendment in plants. The maximum reduction in As concentration was observed in B2 in roots (-55%) and in B1-2-3 in shoot (-83%). The combined treatment of B1-2-3 + TU proved to be less effective as compared to that of B1-2-3 in terms of As reduction and growth improvement. Hence, the usage of bacterial consortium obtained in the present work is a sustainable approach, which might find relevance in field conditions to achieve As reduction in rice grains and to attain higher growth of plants without the need for additional TU supplementation.

Effect of phosphate on arsenic species uptake in plants under hydroponic conditions.

Monothioarsenate (MTA) is a newly discovered arsenic (As) compound that can be formed under reduced sulfur conditions, mainly in paddy soil pore waters. It is structurally similar to arsenate As(V) and inorganic phosphate (Pi), which is taken up through phosphate transporters. Due to the similarity between As(V) and Pi, As(V) enters into plants instead of Pi. The important role played by phytochelatin (PC), glutathione (GSH), and the PC-vacuolar transporters ABCC1 and ABCC2 under As stress in plants is well known. However, the plant uptake and mechanisms surrounding MTA still have not been completely addressed. This investigation was divided in two stages: first, several hydroponic assays were set up to establish the sensibility-tolerance of wild-type Arabidopsis thaliana (accession Columbia-0, Col-0). Then Col-0 was used as a control plant to evaluate the effects of As(V) or MTA in (PC)-deficient mutant (cad1-3), glutathione biosynthesis mutant (cad2), and PC transport (abcc1-2). The inhibitory concentration (IC50) root length was calculated for both As species. According to the results, both arsenic species (As(V) and MTA) exhibited high toxicity for the genotypes evaluated. This could mean that these mechanisms play a constitutive role in MTA detoxification. Second, for the Pi-MTA and As(V)-Pi competition assays, a series of experiments on hydroponic seedlings of A. thaliana were carried out using Col-0 and a pht1;1. The plants were grown under increasing Pi concentrations (10 µM, 0.1 mM, or 1 mM) at 10 µM As(V) or 50 µM MTA. The total As concentration in the roots was significantly lower in plants exposed to MTA, there being less As content in the pht1;1 mutant at the lowest Pi concentrations tested compared with the As(V)/Pi treatments. In addition, a higher rate of As translocation from the roots to the shoots under MTA was observed in comparison to the As(V)-treatments.

A novel organic arsenic derivative MZ2 remodels metabolism and triggers mtROS-mediated apoptosis in acute myeloid leukemia.

PURPOSE: Acute myeloid leukemia (AML) is one of the most common neoplasms in adults, and it is difficult to achieve satisfactory results with conventional drugs. Here, we synthesized a novel organic arsenic derivative MZ2 and evaluated its ability to remodel energy metabolism to achieve anti-leukemia. METHODS: MZ2 was characterized by the average 1-min full mass spectra analysis. Biological methods such as Western blot, qPCR, flow cytometry and confocal microscopy were used to assess the mode and mechanism of MZ2-induced death. The in vivo efficacy of MZ2 was assessed by constructing a patient-derived xenograft (PDX) AML model. RESULTS: Unlike the precursor organic arsenical Z2, MZ2 can effectively reduce the level of aerobic glycolysis. Our in-depth found that MZ2 inhibited the expression of PDK2 in a dose-dependent manner and did not affect the expression of LDHA, another key enzyme of the glycolytic pathway. MZ2 reconstituted energy metabolism to induce the generation of mitochondrial ROS (mtROS) and then triggerd intrinsic apoptosis pathway. We also assessed whether MZ2 generates autophagy and results showed that MZ2 can induce autophagy of AML cells, which may be associated with the precursor organic arsenic drug. In vivo, MZ2 effectively attenuated leukemia progression in mice, and immunohistochemical results suggested its PDK2 inhibitory effect. CONCLUSION: In summary, the novel organic arsine derivative MZ2 exhibited excellent anti-tumor effects in acute myeloid leukemia, which may provide a potential strategy for the treatment of acute myeloid leukemia.

Subcellular Partitioning of Arsenic Trioxide Revealed by Label-Free Imaging.

Subcellular partitioning of therapeutic agents is highly relevant to their interactions with target molecules and drug efficacy, but studying subcellular partitioning is an enormous challenge. Here, we describe the application of nanoscale secondary ion mass spectrometry (NanoSIMS) analysis to define the subcellular pharmacokinetics of a cytotoxic chemotherapy drug, arsenic trioxide (ATO). We reasoned that defining the partitioning of ATO would yield valuable insights into the mechanisms underlying ATO efficacy. NanoSIMS imaging made it possible to define the intracellular fate of ATO in a label-free manner─and with high resolution and high sensitivity. Our studies of ATO-treated cells revealed that arsenic accumulates in the nucleolus. After prolonged ATO exposure, ∼40 nm arsenic- and sulfur-rich protein aggregates appeared in the cell nucleolus, nucleus, and membrane-free compartments in the cytoplasm, and our studies suggested that the partitioning of nanoscale aggregates could be relevant to cell survival. All-trans retinoic acid increased intracellular ATO levels and accelerated the nanoscale aggregate formation in the nucleolus. This study yielded fresh insights into the subcellular pharmacokinetics of an important cancer therapeutic agent and the potential impact of drug partitioning and pharmacokinetics on drug activity.

Self-activated arsenic manganite nanohybrids for visible and synergistic thermo/immuno-arsenotherapy.

Arsenotherapy has been clinically exploited to treat a few types of solid tumors despite of acute promyelocytic leukemia using arsenic trioxide (ATO), however, its efficacy is hampered by inadequate delivery of ATO into solid tumors owing to the absence of efficient and biodegradable vehicles. Precise spatiotemporal control of subcellular ATO delivery for potent arsenotherapy thus remains challengeable. Herein, we report the self-activated arsenic manganite nanohybrids for high-contrast magnetic resonance imaging (MRI) and arsenotherapeutic synergy on triple-negative breast cancer (TNBC). The nanohybrids, composed of arsenic­manganese-co-biomineralized nanoparticles inside albumin nanocages (As/Mn-NHs), switch signal-silent background to high proton relaxivity, and simultaneously afford remarkable subcellular ATO level in acidic and glutathione environments, together with reduced ATO resistance against tumor cells. Then, the nanohybrids enable in vivo high-contrast T1-weighted MRI signals in various tumor models for delineating tumor boundary, and simultaneously yield efficient arsenotherapeutic efficacy through multiple apoptotic pathways for potently suppressing subcutaneous and orthotopic breast models. As/Mn-NHs exhibited the maximum tumor-to-normal tissue (T/N) contrast ratio of 205% and tumor growth inhibition rate of 88% at subcutaneous 4T1 tumors. These nanohybrids further yield preferable synergistic antitumor efficacy against both primary and metastatic breast tumors upon combination with concurrent thermotherapy. More importantly, As/Mn-NHs considerably induce immunogenic cell death (ICD) effect to activate the immunogenically "cold" tumor microenvironment into "hot" one, thus synergizing with immune checkpoint blockade to yield the strongest tumor inhibition and negligible metastatic foci in the lung. Our study offers the insight into clinically potential arsenotherapeutic nanomedicine for potent therapy against solid tumors.

Realgar (As4S4), a traditional Chinese medicine, induces acute promyelocytic leukemia cell death via the Bcl-2/Bax/Cyt-C/AIF signaling pathway in vitro.

Acute promyelocytic leukemia (APL) is a specific subtype of acute myelogenous leukemia (AML) characterized by the proliferation of abnormal promyelocytes. Realgar, a Chinese medicine containing arsenic, can be taken orally. Traditional Chinese medicine physicians have employed realgar to treat APL for over a thousand years. Therefore, realgar may be a promising candidate for the treatment of APL. Nevertheless, the underlying mechanism behind realgar therapy is largely unclear. The present study aimed to investigate the effect of realgar on cell death in the APL cell line (NB4) in vitro and to elucidate the underlying mechanism. In this study, after APL cells were treated with different concentrations of realgar, the cell survival rate, apoptotic assay, morphological changes, ATP levels and cell cycle arrest were assessed. The expression of Bcl-2, Bax, Cytochrome C (Cyt-C) and apoptosis-inducing factor (AIF) at the mRNA and protein levels were also measured by immunofluorescence, quantitative PCR (qPCR) and Western blotting. We found that realgar could significantly inhibit APL cell proliferation and cell death in a time- and dose-dependent manner. Realgar effectively decreased the ATP levels in APL cells. Realgar also induced APL cell cycle arrest at the S and G2/M phases. Following realgar treatment, the mRNA and protein levels of Bcl-2 were significantly downregulated, whereas the levels of Bax, Cyt-C, and AIF were significantly upregulated. In summary, realgar can induce APL cell death via the Bcl-2/Bax/Cyt-C/AIF signaling pathway, suggesting that realgar may be an effective therapeutic for APL.

Selenite and selenate showed contrasting impacts on the fate of arsenic in rice (Oryza sativa L.) regardless of the formation of iron plaque.

The different effects of selenite and selenate on the fate of As and the function of iron plaque in the interaction between Se and As are poorly understood. Rice seedlings (Oryza sativa L.) were selected as experimental plants in this study, the hydroponic experiments were conducted to investigate the possible regulatory roles of selenite and selenate on the uptake, translocation, and transformation of arsenite or arsenate accompanied by iron plaque. In arsenite- and arsenate-treated rice, the Fe30 treatments stimulated root uptake by 12.4-39.8% and 18.6-37.0%, respectively, but inhibited the movement of As from iron plaque to the roots, resulting in the absorption of a considerable amount of As on iron plaque. Regardless of the iron plaque formation, selenite (selenate) significantly increased (decreased) the root uptake of arsenite and arsenate by 28.1-53.0% and 40.0%-61.7%, respectively (45.6-56.3% and 42.5-47.7%, respectively). Interestingly, the supply of selenite significantly reduced root-to-shoot As translocation by 71.9-77.3% and 66.2-67.7%, respectively, in arsenite- and arsenate-treated rice seedlings; however, a significant increase (90.5-122.9%) was induced by selenate was found only in the arsenate-treated plants. Furthermore, the translocation of As from iron plaque to the roots was significantly increased (decreased) by selenite (selenate). As and Fe in iron plaque were significantly positively correlated in all As-treated rice plants, and this correlation was more profound than that in the shoots and roots. However, neither Fe treatments nor inorganic Se addition affected the interconversion between As(III) and As(V) obviously; and As(III) was the dominant species in both shoots (68.3-84.9%) and roots (90.7-98.2%). Our results indicate selenite and selenate are effective in reducing the As accumulation in an opposite way, and the presence of iron plaque had no obvious impact on the interaction between Se and As in rice plants.