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In Vitro Screening for Drug-Induced Depression and/or Suicidal Adverse Effects: A New Toxicogenomic Assay Based on CE-SSCP Analysis of HTR2C mRNA Editing in SH-SY5Y Cells

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Abstract

Many drugs in clinical trials, or already on the market, can have psychiatric side effects, independently of their therapeutic indication (e.g., Acomplia, Taranabant, and Roaccutane). There is currently no in vitro or in vivo approved test for the detection/prediction of such adverse effects, and the Food and Drugs Administration (FDA) can only issue general alerts on specific therapeutic classes. The development of a screening assay is therefore of real interest. The anti-viral and anti-tumor action of human interferon-alpha (hIFNα) is associated with a variety of neuropsychiatric side effects, including major depression, suicidal ideation and suicide. RNA editing of the serotonin 2C receptor (HTR2C) by adenosine deaminases acting on RNA (ADARs) is a post-transcriptional modification, the regulation of which is altered in depressed suicide victims. In this study, we show that in the SH-SY5Y neuroblastoma cell line, hIFNα specifically activates the ADAR1a isoform and thereby modifies the HTR2C mRNA editing profile. As this hIFNα-induced altered profile partly overlaps with that observed in the brain of depressed suicide victims, we investigated whether it could be used as a signature to identify drugs with depression and/or suicidal side effects. By means of the Biocortech proprietary screening assay, which allows the relative quantification of all the edited HTR2C isoforms in a sample, we blind-tested the effect of 50 marketed molecules on HTR2C mRNA editing in SH-SY5Y cells and identified 17 compounds with an IFN-like editing profile. This new toxicogenomic assay can identify compounds with potential psychiatric adverse events with a positive predictive value of 90 %.

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Acknowledgments

We gratefully acknowledge the helpful discussions and suggestions, and the constant support throughout this study from Dr. Ernie Bush from the Drug Safety Executive Council.

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Correspondence to Dinah Weissmann.

Additional information

Laurent Cavarec and Laurent Vincent contributed equally to this work.

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12640_2012_9324_MOESM1_ESM.ppt

Figure S1: Best decision tree for FDA alert prediction by using the CART algorithm. By convention, at each decision node, drugs that satisfy the decision rule are listed by alphabetical order on the left (black box) and drugs which do not satisfy the rule are listed on the right (dotted box). For Ketamine-HCl and Lithium carbonate results were those obtained with 1μM and 100 μM respectively. TP = true positive, FP=false positive, TN=true negative, FN=false negative. (PPT 43 kb)

12640_2012_9324_MOESM2_ESM.xls

Table S1: List of the 50 tested molecules. Drugs are ranked by alphabetical order. The CAS number and the therapeutic class are indicated for each drug. Human IFNα was included in the Table. The seventeen drugs with an IFN-like profile are boxed in gray, and the eleven drugs presenting a non-IFN-like profile are boxed in green with a thick line. For each drug, FDA warning labels or safety reports on suicidal behavior and/or depression are specified (YES or NO). EHNA-HCl was not considered (NC). (XLS 98 kb)

12640_2012_9324_MOESM3_ESM.xls

Table S2: Compilation of the results of the three unsupervised classification methods for the IFN-like editing profile. The drugs concentration was 10 μM except for Ketamine-HCl (1 μM) and Lithium carbonate (100 μM). PMS: drugs were clustered based on our “PMS” method; PTM and K-means: drugs were clustered by using the PTM or the K-means approach. For a given test, a positive result is indicated by x and boxed in gray. The 17 drugs in bold were classified as IFN-like by at least two of the three clustering methods. (XLS 35 kb)

12640_2012_9324_MOESM4_ESM.xls

Table S3: Compilation of the results obtained with three unsupervised classification methods for drugs without an IFN-like editing profile. The drugs concentration was 10 μM except for Lithium carbonate (100 μM). PMS: drugs were clustered based on our “profile-match score” method; PTM and K-means: drugs were clustered by using the PTM or the K-means approach. For a given test and drug, a non-IFN-like profile is indicated by x and boxed in gray. The 11 drugs in bold were classified as non-IFN-like by at least two of the three clustering method. (XLS 34 kb)

12640_2012_9324_MOESM5_ESM.xls

Table S4: Ranking of the molecules by using the mROC program for FDA warning labels (FDA). The decision rules established with the selected combination of HTR2C isoforms are presented above the Table. The threshold value Z=0.154 is shown. Z values that are greater or equal to the threshold value are boxed in orange and correspond to drugs sorted as having an alert. Z values below the threshold are boxed in blue and correspond to drugs sorted as without alert. In the Table the molecules are subdivided in drugs with (yes, salmon-colored boxes) or without (no, green-colored boxes) alerts. The NPV and PPV are calculated for the two classes. (XLS 27 kb)

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Cavarec, L., Vincent, L., Le Borgne, C. et al. In Vitro Screening for Drug-Induced Depression and/or Suicidal Adverse Effects: A New Toxicogenomic Assay Based on CE-SSCP Analysis of HTR2C mRNA Editing in SH-SY5Y Cells. Neurotox Res 23, 49–62 (2013). https://doi.org/10.1007/s12640-012-9324-9

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  • DOI: https://doi.org/10.1007/s12640-012-9324-9

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