Bhoj PS et al. (2018) Parasitol Res "Apoptotic impact on Brugia malayi by sulphonamido-quinoxaline: search for a novel therapeutic rationale."

Ingle RG, Goswami K, Jena L, Wadher S, Bhoj PS

[Parasitol Res, 2018]

Human lymphatic filariasis although not fatal but poses serious socioeconomic burden due to associated disability. This is reflected by the huge magnitude of the estimated disability-adjusted life years of about 5.09 million. Therefore, following WHO mandate, our earlier studies on antifilarial drug development revealed the significance of apoptosis. Apoptotic impact has been implicated in anticancer rationale of several drugs. In this study, we explored the antifilarial potential of sulphonamido-quinoxaline compounds, shown to be specific inhibitor for c-Met kinase in human cancer cells. Out of studied compounds, Q4, showing favorable drug-likeness and medicinal chemistry properties on bioinformatics platform along with subsequently recorded lowest IC₁₀₀value, was considered as a suitable antifilarial candidate. Significant apoptosis due to mitochondrial involvement was recorded in drug-treated parasite unlike untreated control. In spite of homology between human c-Met kinase and Brugia malayi counterpart, comparative docking result of this compound showed more favorable binding parameters with the parasitic target. The wide gap between IC₁₀₀and LD₅₀values further confirmed the therapeutic safety. We propose sulphonamido-quinoxaline derivative as a lead candidate for antifilarial drug development. Further study is warranted to authenticate parasitic c-Met kinase as a novel therapeutic target reminiscent of anticancer rationale implicating inhibition of proliferation.

Bhoj PS et al. (2020) Acta Parasitol "Role of Glutathione in Chalcone Derivative Induced Apoptosis of Brugia malayi and its Possible Therapeutic Implication."

Bahekar S, Singh N, Dash D, Togre NS, Khatri V, Bhoj PS, Chandak HS, Goswami K

[Acta Parasitol, 2020]

PURPOSE: Oxidative stress is an essential component of innate response against microbes. The oxidative impact has a very subtle connection with apoptosis. Our previous work indicated presumptive evidence of apoptosis by the chalcone derivatives against the human lymphatic filarial parasite. Evidence suggests the involvement of glutathione-S-transferase (GST) in the mechanism of action of chalcone drugs. In the present study, we explored the implications of redox status in apoptosis of the parasite by this drug. RESULTS: Treatment with the representative drug, 4t, significantly decreased GSH level and increased GST activity in the Brugia malayi microfilariae (Mf) in comparison to Mf without 4t treatment. Drug-induced loss of motility of the parasites was reversed by the treatment with GSH (41%) and NAC (19%). A significant fall in rGST activity was observed due to drug addition, which could be reversed by the addition of GSH co-substrate, but not with the re-addition of rGST, indicating a vital role of GSH. In silico study demonstrated a favorable drug-GST enzyme interaction. Oxidative stress was reflected by increased protein carbonylation and intracellular reactive oxygen species level, in the drug-treated parasite. Mitochondrial oxygen consumption was reduced by the drug, which was reversed on the addition of GSH. Mitochondrial dysfunction was confirmed by MTT and cytochrome c assay. Apoptosis was confirmed by the inhibition in PARP activity. CONCLUSION: We conclude that the depletion of GSH by chalcone with concomitant mitochondrial dysfunction revealed a novel rationale of apoptosis in the parasite. Such a mechanism might have wide therapeutic implications.

Bhoj PS et al. (2019) Parasitol Res "In vitro apoptotic effect on human lymphatic filarial parasite by piperidine derivatives and thymidine reversal study."

Goswami K, Togre NS, Rao S, Patil MB, Sharma R, Agrawal NR, Chandak HS, Bahekar SP, Bhoj PS

[Parasitol Res, 2019]

A novel library of synthetic piperidine derivatives was used to screen against human lymphatic filarial parasite Brugia malayi. Piperidine has earlier been reported to have effect against parasites including rodent filarial nematodes. Compounds with hydroxyl substitutions (4Q and 4H) showed marked antifilarial effect. Molecular docking of 4H derivative showed more favorable thermodynamic parameters against thymidylate synthase of B. malayi than human counterpart. A wide difference between IC₅₀ and LD₅₀ ensured the therapeutic safety of the candidates against the filarial parasites. Addition of thymidine to the treatment regimen led to a significant reversal of antifilarial effect of 4H that confirmed inhibition of thymidylate synthase as pharmacological rationale. Apoptosis induced in the parasite as a consequence of probable inhibition of thymidylate synthase was studied by acridine orange/ethidium bromide fluorescent staining and poly (ADP-ribose) polymerase activity inhibition. Involvement of mitochondria was confirmed by decreased 3-[4,5-dimethylthiazol-2-yl]-2,5 diphenyl tetrazolium bromide (MTT) conversion and increased cytosolic cytochrome c level in 4H treated microfilariae, compared with the untreated microfilariae. Moreover, Michael adduct of chalcone targeting dihydrofolate reductase and piperidine targeting thymidylate synthase demonstrated synergistic effect on the parasite, indicating the importance of inhibition of DNA synthesis by combined effect. In conclusion, piperidine derivatives with hydroxyl substitution have a great therapeutic potential with an apoptotic rationale involving mitochondrial pathway, due to possible inhibition of parasitic thymidylate synthase.

Brockhaus M et al. (1998) Neuroreport "Caspase-mediated cleavage is not required for the activity of presenilins in amyloidogenesis and NOTCH signaling."

Loetscher H, Wittenburg N, Baumeister R, Rohrig S, Haass C, Brockhaus M, Grunberg J, Jacobsen H

[Neuroreport, 1998]

The Alzheimer's disease (AD) associated presenilin (PS) proteins are proteolytically processed. One of the processing pathways involves cleavage by caspases. Pharmacological inhibition of caspases is currently being discussed as a treatment for a variety of neurodegenerative diseases, including AD. We therefore inhibited caspase mediated processing of PS-1 and PS-2 in cells transfected with wt and mutant PS by mutagenizing the substrate recognition site or by using specific peptide aldehydes known to block caspases. We found that the inhibition of caspase mediated processing of PS proteins does not decrease its amyloidogenic activity. PS cDNA constructs with mutations in the caspase cleavage site are biologically active in Caenorhabditis elegans such as the wt human PS proteins, demonstrating that caspase-mediated cleavage is not required for the physiological PS function in NOTCH signaling.

Kim HM et al. (2020) J Hazard Mater "Comprehensive phenotyping and multi-omic profiling in the toxicity assessment of nanopolystyrene with different surface properties."

Min JE, Kwon SW, Long NP, Yoon SJ, Anh NH, Kim HM, Kim SJ

[J Hazard Mater, 2020]

There is a growing concern regarding the toxic effects of terrestrial nanoplastic contaminants. However, an all-encompassing phenotyping- and omics-based strategy for the toxicity assessment of nanoplastics with different surface properties on soil living organisms remains to be established. Herein, we devised a comprehensive phenotyping and multi-omic profiling method to examine the molecular disturbance of nanopolystyrene (PS)-exposed Caenorhabditis elegans. The exposure time was 24 h with either 1 g/mL or 10 g/mL of PS. We found that PS considerably affected the reproduction and locomotion, as well as increased the oxidative stress of worms regardless of their surface properties. Nevertheless, each type of PS affected the metabolome and lipidome of the nematodes differently. Uncharged PS (PS-N) triggered significant metabolic disturbances, whereas the metabolic influences from PS-NH₂ and PS-COOH were subtle. The dysregulated transcriptome profiles of PS-N were strongly associated with the metabolic pathways. Besides, the altered expression of several genes associated with autophagy and longevity was observed. Collectively, we demonstrated that comprehensive phenotyping and omics-based profiling establish a practical framework that allows us to gain deeper insights into the maladaptive consequences of PS in nematodes. It can be utilized for the evaluation of other environmental contaminants in the terrestrial ecosystem.

Zhou T et al. (2023) Int J Mol Sci "Seawater Accelerated the Aging of Polystyrene and Enhanced Its Toxic Effects on Caenorhabditis elegans."

Wu J, Liu Y, Xu A, Zhou T

[Int J Mol Sci, 2023]

Microplastics (MPs) are emerging pollutants and pose a significant threat to marine ecosystems. Although previous studies have documented the mechanisms and toxic effects of aging MPs in various environments, the impact of the marine environment on MPs remains unclear. In the present study, the aging process of polystyrene (PS) in seawater was simulated and the changes in its physicochemical properties were investigated. Our results showed that the surface of the PS eroded in the seawater, which was accompanied by the release of aged MPs with a smaller size. In situ optical photothermal infrared microspectroscopy revealed that the mechanism of PS aging was related to the opening of the carbonyl group and breaking of the bond between carbon and benzene removal. To verify the toxic effects of aged PS, <i>Caenorhabditis elegans</i> was exposed to PS. Aged PS resulted in a greater reduction in locomotion, vitality, and reproduction than virgin PS. Mechanistically, aged PS led to oxidative stress, high glutathione s-transferase activity, and high total glutathione in worms. Together, our findings provided novel information regarding the accelerated aging of PS in seawater and the increased toxicity of aged PS, which could improve our understanding of MPs' ecotoxicity in the marine environment.

Yu Y et al. (2020) Sci Total Environ "Polystyrene microplastics (PS-MPs) toxicity induced oxidative stress and intestinal injury in nematode Caenorhabditis elegans."

Chen X, Ding P, Li H, Yu Y, Han Y, Hua X, Dang Y, Yu Z, Chen H

[Sci Total Environ, 2020]

To understand the toxicity and mechanism of polystyrene microplastics (PS-MPs) exposure, Caenorhabditis elegans (C. elegans) was exposed to various concentrations (0, 0.1, 1, 10, and 100g/L) of PS-MPs, and the levels physiological, biochemical, and molecular parameters were measured as endpoints. Subacute exposure to 1-100g/L of PS-MPs resulted in adverse physiological effects in C. elegans, and PS-MPs were ingested and accumulated in the intestine of C. elegans. Exposure to 100g/L of PS-MPs significantly increased reactive oxygen species (ROS) production, lipofuscin accumulation, and the expression oxidative stress-related genes, which suggests that PS-MPs exposure induced oxidative stress by ROS. In addition, exposure to 100g/L of PS-MPs caused a hyperpermeable state of the intestinal barrier and altered the expression of genes related to intestinal development, which indicates intestinal damage in C. elegans. According to Pearson correlation analyses, oxidative stress and intestinal damage were significantly correlated with adverse effects of PS-MPs in C. elegans. Therefore, it was speculated that the toxicity induced by PS-MPs resulted from the combination of oxidative stress and intestinal injury.

Zhao Y et al. (2021) Ecotoxicol Environ Saf "Induction of protective response to polystyrene nanoparticles associated with dysregulation of intestinal long non-coding RNAs in Caenorhabditis elegans."

Wang J, Zhao Y, Rui Q, Wang D, Xu R, Chen X

[Ecotoxicol Environ Saf, 2021]

Intestinal barrier plays a crucial function during the response to polystyrene nanoparticles (PS-NPs) in nematode Caenorhabditis elegans. Long non-coding RNAs (lncRNAs) are involved in the control of various biological processes, including stress response. We here used C. elegans to determine intestinal lncRNAs dysregulated by PS-NPs (1-100g/L). In intestine of PS-NPs exposed worms, we found four lncRNAs (linc-61, linc-50, linc-9, and linc-2) in response to PS-NPs and with the function in controlling PS-NPs toxicity. The alteration in expressions of these four intestinal lncRNAs reflected a protective response to PS-NPs exposure. During the response to PS-NPs, limited number of transcriptional factors functioned as the downstream targets of these four lncRNAs. linc-2 acted upstream of DAF-16, linc-9 acted upstream of NHR-77, linc-50 functioned upstream of DAF-16, and linc-61 regulated the functions of DAF-16, DVE-1, and FKH-2 to control PS-NPs toxicity. The obtained data demonstrated the important role of lncRNAs in intestinal barrier to mediate a protective response to PS-NPs exposure at low concentrations.

Liu H et al. (2021) J Hazard Mater "Acetylation regulation associated with the induction of protective response to polystyrene nanoparticles in Caenorhabditis elegans."

Liu H, Tian L, Wang D, Qu M

[J Hazard Mater, 2021]

Caenorhabditis elegans is a useful animal model to assess nanoplastic toxicity. Using polystyrene nanoparticles (PS-NPs) as the example of nanoplastics, we found that exposure to PS-NPs (1-100g/L) from L1-larvae for 6.5 days increased expression of cbp-1 encoding an acetyltransferase. The susceptibility to PS-NPs toxicity was observed in cbp-1(RNAi) worms, suggesting that CBP-1-mediated histone acetylation regulation reflects a protective response to PS-NPs. The functions of CBP-1 in intestine, neurons, and germline were required for formation of this protective response. In intestinal cells, CBP-1 controlled PS-NPs toxicity by modulating functions of insulin and p38 MAPK signaling pathways. In neuronal cells, CBP-1 controlled PS-NPs toxicity by affecting functions of DAF-7/TGF- and JNK MAPK signaling pathways. In germline cells, CBP-1 controlled PS-NPs toxicity by suppressing NHL-2 activity, and NHL-2 further regulated PS-NPs toxicity by modulating insulin communication between germline and intestine. Therefore, our data suggested that the CBP-1-mediated histone acetylation regulation in certain tissues is associated with the induction of protective response to PS-NPs in C. elegans.

Liu H et al. (2021) Sci Total Environ "Size-dependent transgenerational toxicity induced by nanoplastics in nematode Caenorhabditis elegans."

Wang S, Wang D, Tian L, Liu H

[Sci Total Environ, 2021]

Nanoplastic exposure can potentially cause the severe transgenerational toxicity in organisms. However, the transgenerational nanoplastic toxicity and the underlying mechanisms are still largely unclear. Using Caenorhabditis elegans as an animal model, we here compared the transgenerational toxicity of two sizes of polystyrene nanoparticles (PS-NPs, 20 and 100 nm). The nematodes were exposed to PS-NPs at the P0 generation, and from the F1 generation the nematodes were grown under the normal condition. Exposure to 20 nm PS-NPs resulted in more severe transgenerational toxicity than exposure to 100 nm PS-NPs. At the concentration of 100 g/L, the toxicity of 20 nm PS-NPs on locomotion and reproduction was detected at the F1-F6 generations, whereas the toxicity of 100 nm PS-NPs could only be observed at the F1-F3 generations. The difference in transgeneration toxicity between PS-NPs (20 nm) and PS-NPs (100 nm) was associated with the difference in transgenerational activation of oxidative stress. Based on observations on SOD-3::GFP, HSP-6::GFP, and HSP-4::GFP expressions, PS-NPs (20 nm) and PS-NPs (100 nm) further induced different transgenerational responses of anti-oxidation, mt UPR, and ER UPR. Our data suggested that the induction of transgenerational toxicity of PS-NPs was size dependent in nematodes. The results are helpful for our understanding the cellular mechanisms for the induction of transgenerational nanoplastic toxicity in organisms.