Depletion of the ER chaperone ENPL-1 sensitizes<i><i>C. elegans</i></i>to the anticancer drug cisplatin

Natarajan, Balasubramanian; Gaur, Rahul; Hemmingsson, Oskar; Kao, Gautam; Naredi, Peter

doi:10.4161/worm.24059

Cited by 15 publications

(19 citation statements)

References 37 publications

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“…asna-1 mutants have multiple phenotypes including elevated autophagy levels ( Fig. S2D ), high ER stress (Natarajan et al, 2013), and low insulin signaling levels (Hemmingsson et al, 2010; Kao et al, 2007). We asked whether these phenotypes contribute to the TAP targeting defect in asna-1(ok938) mutants.…”

Section: Resultsmentioning

confidence: 99%

“…This analysis of ASNA-1 demonstrates that cisplatin perturbs ER function, which might also explain other effects of cisplatin on signaling pathways, Ca 2+ pump function, and membrane properties. We previously showed that while cisplatin does not induce ER stress, combinatorial use with an ER stress inducer sensitizes resistant worms to cisplatin (Natarajan et al, 2013). This is consistent with the idea that cisplatin treatment sensitizes the ER in a metastable state due to TAP targeting defects and if ER function is further compromised then the cells will become hypersensitive to cisplatin cytotoxicity.…”

Section: Discussionmentioning

confidence: 99%

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ASNA-1 oxidation induced by cisplatin exposure enhances its cytotoxicity by selectively perturbing tail anchored protein targeting

Raj

Billing

Podraza

et al. 2019

Preprint

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protein SUMMARY STATEMENTSensitizing tumors to cisplatin would be of considerable therapeutic benefit. Here we show a novel mechanism of cisplatin sensitization via oxidation of ASNA-1 in a Caenorhabditis elegans model. ABSTRACTCisplatin is a frontline cancer treatment, but intrinsic or acquired resistance is common. We previously showed that ASNA-1/TRC40 inactivation increases cisplatin sensitivity in mammalian cells and a Caenorhabditis elegans asna-1 knockdown model. ASNA-1 has conserved tail-anchored protein (TAP) targeting and insulin secretion functions. Here we examined the mechanism of ASNA-1 action. We show that ASNA-1 exists in two physiologically-responsive redox states with separable TAP-targeting and insulin secretion functions. Cisplatin-generated ROS targeted ASNA-1 oxidation, resulting in a selective targeting defect of an ASNA-1-dependent TAP. Increased ASNA-1 oxidation sensitized worms to cisplatin cytotoxicity. Mutants with a redox balance favoring oxidized ASNA-1 were cisplatin sensitive as null mutants by diverting ASNA-1 away from its TAP-targeting role and 2 instead perturbing endoplasmic reticulum (ER) function. Mutations in the ASNA-1 receptor required for TAP insertion induced equivalent cisplatin sensitivity. We reveal a previously undescribed cellular dysfunction induced by cisplatin, identify a cisplatin target, and show that drug exposure causes TAP targeting-induced ER dysfunction. Therapeutic oxidation of ASNA-1 could be a clinically useful means to increase cisplatin sensitivity, reduce cytotoxic drug doses, and counteract cisplatin resistance. AUTHOR SUMMARYCisplatin is a very effective anti-cancer drug and is widely used as a frontline treatment.However, tumor resistance limits its use. Tumor re-sensitization would improve cancer treatment. ASNA-1/TRC40 knockdown in Caenorhabditis elegans and mammals results in cisplatin hypersensitivity, but the underlying mechanistic details are largely unknown. We show that in C. elegans ASNA-1 mutants, increased cisplatin killing is coupled with delocalization of a tail-anchored protein, SEC-61b, a membrane protein that should reach the ER and is instead mistargeted. Like its homologs, the reduced form of worm ASNA-1 is needed for targeting activity. Targeting is blocked upon ASNA-1 oxidation after cisplatin treatment, likely via reactive oxygen species (ROS) generated by cisplatin treatment. Nevertheless, the oxidized form of the protein can execute other functions like insulin secretion. We show also that mutants with high oxidized ASNA-1 levels are cisplatin sensitive. Additionally, cisplatin induced mistargeting strictly acts through ASNA-1 inactivation. Thus, we define a pathway from cisplatin exposure that targets protein (ASNA-1) inactivation, consequently leading to mis-targeting of proteins that need ASNA-1 for their maturation. This multi-step process provides vital information about likely proteins that can be targeted by drugs to enhance cisplatin mediated killing and improve chemotherapy.

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Section: Resultsmentioning

confidence: 99%

Section: Discussionmentioning

confidence: 99%

ASNA-1 oxidation induced by cisplatin exposure enhances its cytotoxicity by selectively perturbing tail anchored protein targeting

Raj

Billing

Podraza

et al. 2019

Preprint

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“…We did not find enrichment of particular pathways among the species-specific response genes, but this may not be surprising as stress response pathways are often regulated by a small number of key master regulator genes (Haynes et al 2010;Li et al 2011;Natarajan et al 2013;Mahat et al 2016;Quiros et al 2017). However, several of the genes with significant species by condition interactions have functions related to G-protein signaling, TGF-β signaling, and metabolism ( Figure S12).…”

Section: Species-specific Transcriptional Changes In Response To Hypoxiamentioning

confidence: 92%

Inter-species differences in response to hypoxia in iPSC-derived cardiomyocytes from humans and chimpanzees

Ward

Gilad

2018

Preprint

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Despite anatomical similarities, there appear to be differences in susceptibility to cardiovascular disease between primates. For example, humans are prone to ischemia-induced myocardial infarction unlike chimpanzees, which tend to suffer from fibrotic disease. However, it is challenging to determine the relative contributions of genetic and environmental effects to complex disease phenotypes within and between primates. The ability to differentiate cardiomyocytes from induced pluripotent stem cells (iPSCs), now allows for direct inter-species comparisons of the gene regulatory response to disease-relevant perturbations. A consequence of ischemia is oxygen deprivation. Therefore, in order to understand human-specific regulatory adaptations in the heart, and to potentially gain insight into the evolution of disease susceptibility and resistance, we developed a model of hypoxia in human and chimpanzee cardiomyocytes. We differentiated eight human and seven chimpanzee iPSC lines into cardiomyocytes under normoxic conditions, and subjected these cells to 6 hours of hypoxia, followed by 6 or 24 hours of re-oxygenation. We collected genome-wide gene expression data as well as measurements of cellular stress at each time-point. The overall cellular and transcriptional response to hypoxic stress is generally conserved across species. Supporting the functional importance of precise regulatory response to hypoxia, we found that genes that respond to hypoxic stress in both species are depleted for association with expression quantitative trait loci (eQTLs) in the heart, and cardiovascular-related genes. We also identified hundereds of inter-species regulatory differences in our study. In particular, RASD1, which is associated with coronary artery disease, is up-regulated specifically in humans following hypoxia.

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“…C. elegans is less complex than the mammalian system, while still sharing high genetic homology (60–80%) 28 . The model organism C. elegans has been previously used in a limited number of studies to identify interactions between Cisplatin and signaling pathways in vivo 29–31 . Hypersensitivity to Cisplatin was noted in worms with mutations in asna-1 , a metalloregulated ATPase gene promoting insulin secretion and membrane insertion of tail-anchored proteins, leading the authors to suggest that drugs targeting ASNA-1 may sensitize cancer cells to Cisplatin 30 .…”

Section: Introductionmentioning

confidence: 99%

“…Hypersensitivity to Cisplatin was noted in worms with mutations in asna-1 , a metalloregulated ATPase gene promoting insulin secretion and membrane insertion of tail-anchored proteins, leading the authors to suggest that drugs targeting ASNA-1 may sensitize cancer cells to Cisplatin 30 . In a follow up study, the same authors identified that depletion of the endoplasmic reticulum (ER) resident chaperone ENPL-1/ GRP94 renders worms sensitive to Cisplatin 31 . Since DNA repair mechanisms are known to occur in the genetically amenable nematode 32, 33 , deletion mutants have been used to identify various genes of the DNA repair pathways that are protective against Cisplatin toxicity.…”

Section: Introductionmentioning

confidence: 99%

Elemental bioimaging of Cisplatin in Caenorhabditis elegans by LA-ICP-MS

et al. 2015

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Cis-diamminedichloroplatinum(II) (Cisplatin) is one of the most important and frequently used cytostatic drugs for the treatment of various solid tumors. Herein, a laser ablation-inductively coupled plasma-mass spectrometry (LA-ICP-MS) method incorporating a fast and simple sample preparation protocol was developed for the elemental mapping of Cisplatin in the model organism Caenorhabditis elegans (C. elegans). The method allows imaging of the spatially-resolved elemental distribution of platinum in the whole organism with respect to the anatomic structure in L4 stage worms at a lateral resolution of 5 µm. In addition, a dose- and time-dependent Cisplatin uptake was corroborated quantitatively by a total reflection X-ray fluorescence spectroscopy (TXRF) method, and the elemental mapping indicated that Cisplatin is located in the intestine and in the head of the worms. Better understanding of the distribution of Cisplatin in this well-established model organism will be instrumental in deciphering Cisplatin toxicity and pharmacokinetics. Since the cytostatic effect of Cisplatin is based on binding the DNA by forming intra- and interstrand crosslinks, the response of poly(ADP-ribose)metabolism enzyme 1 (pme-1) deletion mutants to Cisplatin was also examined. Loss of pme-1, which is the C. elegans ortholog of human poly(ADP-ribose) polymerase 1 (PARP-1) led to disturbed DNA damage response. With respect to survival and brood size, pme-1 deletion mutants were more sensitive to Cisplatin as compared to wildtype worms, while Cisplatin uptake was indistinguishable.

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Depletion of the ER chaperone ENPL-1 sensitizesC. elegansto the anticancer drug cisplatin

Cited by 15 publications

References 37 publications

ASNA-1 oxidation induced by cisplatin exposure enhances its cytotoxicity by selectively perturbing tail anchored protein targeting

ASNA-1 oxidation induced by cisplatin exposure enhances its cytotoxicity by selectively perturbing tail anchored protein targeting

Inter-species differences in response to hypoxia in iPSC-derived cardiomyocytes from humans and chimpanzees

Elemental bioimaging of Cisplatin in Caenorhabditis elegans by LA-ICP-MS

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