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Event: 1009

Key Event Title

The KE title should describe a discrete biological change that can be measured. It should generally define the biological object or process being measured and whether it is increased, decreased, or otherwise definably altered relative to a control state. For example “enzyme activity, decreased”, “hormone concentration, increased”, or “growth rate, decreased”, where the specific enzyme or hormone being measured is defined. More help

Inhibition, Deiodinase 1

Short name
The KE short name should be a reasonable abbreviation of the KE title and is used in labelling this object throughout the AOP-Wiki. The short name should be less than 80 characters in length. More help
Inhibition, Deiodinase 1

Biological Context

Structured terms, selected from a drop-down menu, are used to identify the level of biological organization for each KE. Note, KEs should be defined within a particular level of biological organization. Only KERs should be used to transition from one level of organization to another. Selection of the level of biological organization defines which structured terms will be available to select when defining the Event Components (below). More help

Cell term

Further information on Event Components and Biological Context may be viewed on the attached pdf.The biological context describes the location/biological environment in which the event takes place.  For molecular/cellular events this would include the cellular context (if known), organ context, and species/life stage/sex for which the event is relevant. For tissue/organ events cellular context is not applicable.  For individual/population events, the organ context is not applicable. More help

Organ term

Further information on Event Components and Biological Context may be viewed on the attached pdf.The biological context describes the location/biological environment in which the event takes place.  For molecular/cellular events this would include the cellular context (if known), organ context, and species/life stage/sex for which the event is relevant. For tissue/organ events cellular context is not applicable.  For individual/population events, the organ context is not applicable. More help

Key Event Components

Further information on Event Components and Biological Context may be viewed on the attached pdf.Because one of the aims of the AOP-KB is to facilitate de facto construction of AOP networks through the use of shared KE and KER elements, authors are also asked to define their KEs using a set of structured ontology terms (Event Components). In the absence of structured terms, the same KE can readily be defined using a number of synonymous titles (read by a computer as character strings). In order to make these synonymous KEs more machine-readable, KEs should also be defined by one or more “event components” consisting of a biological process, object, and action with each term originating from one of 22 biological ontologies (Ives, et al., 2017; See List). Biological process describes dynamics of the underlying biological system (e.g., receptor signalling). The biological object is the subject of the perturbation (e.g., a specific biological receptor that is activated or inhibited). Action represents the direction of perturbation of this system (generally increased or decreased; e.g., ‘decreased’ in the case of a receptor that is inhibited to indicate a decrease in the signalling by that receptor).Note that when editing Event Components, clicking an existing Event Component from the Suggestions menu will autopopulate these fields, along with their source ID and description. To clear any fields before submitting the event component, use the 'Clear process,' 'Clear object,' or 'Clear action' buttons. If a desired term does not exist, a new term request may be made via Term Requests. Event components may not be edited; to edit an event component, remove the existing event component and create a new one using the terms that you wish to add. More help
Process Object Action
catalytic activity type I iodothyronine deiodinase decreased

Key Event Overview

AOPs Including This Key Event

All of the AOPs that are linked to this KE will automatically be listed in this subsection. This table can be particularly useful for derivation of AOP networks including the KE. Clicking on the name of the AOP will bring you to the individual page for that AOP. More help
AOP Name Role of event in AOP Point of Contact Author Status OECD Status
DIO1i posterior swim bladder MolecularInitiatingEvent Dries Knapen (send email) Open for adoption EAGMST Under Review
DIO1i anterior swim bladder MolecularInitiatingEvent Dries Knapen (send email) Open for adoption EAGMST Under Review
DIO1 inhib alters metamorphosis MolecularInitiatingEvent Jonathan Haselman (send email) Under Development: Contributions and Comments Welcome

Stressors

This is a structured field used to identify specific agents (generally chemicals) that can trigger the KE. Stressors identified in this field will be linked to the KE in a machine-readable manner, such that, for example, a stressor search would identify this as an event the stressor can trigger. NOTE: intermediate or downstream KEs in one AOP may function as MIEs in other AOPs, meaning that stressor information may be added to the KE description, even if it is a downstream KE in the pathway currently under development.Information concerning the stressors that may trigger an MIE can be defined using a combination of structured and unstructured (free-text) fields. For example, structured fields may be used to indicate specific chemicals for which there is evidence of an interaction relevant to this MIE. By linking the KE description to a structured chemical name, it will be increasingly possible to link the MIE to other sources of chemical data and information, enhancing searchability and inter-operability among different data-sources and knowledgebases. The free-text section “Evidence for perturbation of this MIE by stressor” can be used both to identify the supporting evidence for specific stressors triggering the MIE as well as to define broad chemical categories or other properties that classify the stressors able to trigger the MIE for which specific structured terms may not exist. More help

Taxonomic Applicability

Latin or common names of a species or broader taxonomic grouping (e.g., class, order, family) can be selected from an ontology. In many cases, individual species identified in these structured fields will be those for which the strongest evidence used in constructing the AOP was available in relation to this KE. More help
Term Scientific Term Evidence Link
rat Rattus norvegicus High NCBI
mouse Mus musculus Moderate NCBI
pigs Sus scrofa Moderate NCBI
Ovis orientalis aries Ovis aries Moderate NCBI
fathead minnow Pimephales promelas Moderate NCBI
killifish Fundulus heteroclitus Moderate NCBI
gilthead bream Sparus aurata Moderate NCBI
African clawed frog Xenopus laevis Moderate NCBI
human Homo sapiens High NCBI
Oreochromis niloticus Oreochromis niloticus Moderate NCBI
zebrafish Danio rerio Moderate NCBI

Life Stages

The structured ontology terms for life-stage are more comprehensive than those for taxa, but may still require further description/development and explanation in the free text section. More help
Life stage Evidence
All life stages Moderate

Sex Applicability

The authors must select from one of the following: Male, female, mixed, asexual, third gender, hermaphrodite, or unspecific. More help
Term Evidence
Unspecific Moderate

Key Event Description

A description of the biological state being observed or measured, the biological compartment in which it is measured, and its general role in the biology should be provided. For example, the biological state being measured could be the activity of an enzyme, the expression of a gene or abundance of an mRNA transcript, the concentration of a hormone or protein, neuronal activity, heart rate, etc. The biological compartment may be a particular cell type, tissue, organ, fluid (e.g., plasma, cerebrospinal fluid), etc. The role in the biology could describe the reaction that an enzyme catalyses and the role of that reaction within a given metabolic pathway; the protein that a gene or mRNA transcript codes for and the function of that protein; the function of a hormone in a given target tissue, physiological function of an organ, etc. Careful attention should be taken to avoid reference to other KEs, KERs or AOPs. Only describe this KE as a single isolated measurable event/state. This will ensure that the KE is modular and can be used by other AOPs, thereby facilitating construction of AOP networks. More help

Disruption of the thyroid hormone system is increasingly being recognized as an important toxicity pathway, as it can cause many adverse outcomes. Thyroid hormones do not only play an important role in the adult individual, but they are also critical during embryonic development. Thyroid hormones (THs) play an important role in a wide range of biological processes in vertebrates including growth, development, reproduction, cardiac function, thermoregulation, response to injury, tissue repair and homeostasis. Numerous chemicals are known to disturb thyroid function, for example by inhibiting thyroperoxidase (TPO) or deiodinase (DIO), upregulating excretion pathways or modifying gene expression. The two major thyroid hormones are triiodothyronine (T3) and thyroxine (T4), both iodinated derivatives of tyrosine. Most TH actions depend on the binding of T3 to its nuclear receptors. Active and inactive THs are tightly regulated by enzymes called iodothyronine deiodinases (DIO). The activation occurs via outer ring deiodination (ORD), i.e. removing iodine from the outer, phenolic ring of T4 to form T3, while inactivation occurs via inner ring deiodination (IRD), i.e. removing iodine from the inner tyrosol ring of T4 or T3.

Three types of iodothyronine deiodinases (DIO1-3) have been described in vertebrates that activate or inactivate THs and are therefore important mediators of TH action. All deiodinases are integral membrane proteins of the thioredoxin superfamily that contain selenocysteine in their catalytic centre. Type I deiodinase is capable of both ORD and IRD, including the conversion of T4 into T3, as well as the conversion of rT3 to the inactive thyroid hormone 3,3’ T2. rT3, rather than T4, is the preferred substrate for DIO1. furthermore, DIO1 has a very high Km (µM range, compared to nM range for DIO2) (Darras and Van Herck, 2012). Type II deiodinase (DIO2) is only capable of ORD activity with T4 as a preferred substrate (i.e., activation of T4 to T3). DIO3 can inner ring deiodinate T4 and T3 to the inactive forms of THs, reverse T3, (rT3) and 3,3’-T2 respectively. DIO1 is a plasma membrane protein with its catalytic domain facing the cytosol.

How It Is Measured or Detected

One of the primary considerations in evaluating AOPs is the relevance and reliability of the methods with which the KEs can be measured. The aim of this section of the KE description is not to provide detailed protocols, but rather to capture, in a sentence or two, per method, the type(s) of measurements that can be employed to evaluate the KE and the relative level of scientific confidence in those measurements. Methods that can be used to detect or measure the biological state represented in the KE should be briefly described and/or cited. These can range from citation of specific validated test guidelines, citation of specific methods published in the peer reviewed literature, or outlines of a general protocol or approach (e.g., a protein may be measured by ELISA).Key considerations regarding scientific confidence in the measurement approach include whether the assay is fit for purpose, whether it provides a direct or indirect measure of the biological state in question, whether it is repeatable and reproducible, and the extent to which it is accepted in the scientific and/or regulatory community. Information can be obtained from the OECD Test Guidelines website and the EURL ECVAM Database Service on Alternative Methods to Animal Experimentation (DB-ALM). ?

At this time, there are no approved OECD or EPA guideline protocols for measurement of DIO inhibition. Deiodination is the major pathway regulating T3 bioavailability in mammalian tissues. In vitro assays can be used to examine inhibition of deiodinase 1 (DIO1) activity upon exposure to thyroid disrupting compounds.

Several methods for deiodinase activity measurements are available. A first in vitro assay measures deiodinase activities by quantifying the radioactive iodine release from iodine-labelled substrates, depending on the preferred substrates of the isoforms of deiodinases (Ferreira et al., 2002; Forhead et al., 2006; Freyberger and Ahr, 2006; Pavelka, 2010; Stinckens et al., 2018). Another assay uses a chromatography-based method coupled to mass spectroscopy to measure products of thyroxin resulting from deiodinase type-1 activity (Butt et al., 2011). A colorimetric method (Renko et al., 2012), the Sandell-Kolthoff method, that measures the release of iodine from T4 is also available. Each of these assays requires a source of deiodinase which can be obtained for example using unexposed pig liver tissue (available from slaughterhouses) or rat liver tissue. Hornung et al. (2018) and Olker et al. (2019) on the other hand used an adenovirus expression system to produce the DIO1 enzyme and developed an assay for nonradioactive measurement of iodide released using the Sandell-Kolthoff method in a 96-well plate format. This assay was then used to screen the ToxCast Phase 1 chemical library. The specific synthesis of DIO1 through the adenovirus expression system provides an important advantage over other methods where activity of the different deiodinase isoforms needs to be distinguished in other ways, such as based on differences in enzyme kinetics.

Measurements of in vivo deiodinase activity in tissues collected from animal experiments are scarce. Noyes et al. (2011) showed decreased rate of outer ring deiodination (mediated by DIO1 and DIO2) in whole fish microsomes after exposure to BDE-209. After incubation with the substrate, thyroid hormone levels were measured using LC-MS/MS. Houbrechts et al. (2016) confirmed decreased DIO1 activity in a DIO1-DIO2 knockdown zebrafish at the ages of 3 and 7 days post fertilization. Decreased T3 levels are often used as evidence of DIO inhibition, for example after exposure to iopanoic acid, in fish species such as zebrafish (Stinckens et al., 2020) and fathead minnow (Cavallin et al., 2017). It should be noted that it is difficult to make the distinction between decreased T3 levels caused by outer ring deiodination mediated by DIO2 inhibition or DIO1 inhibition.

Domain of Applicability

This free text section should be used to elaborate on the scientific basis for the indicated domains of applicability and the WoE calls (if provided). While structured terms may be selected to define the taxonomic, life stage and sex applicability (see structured applicability terms, above) of the KE, the structured terms may not adequately reflect or capture the overall biological applicability domain (particularly with regard to taxa). Likewise, the structured terms do not provide an explanation or rationale for the selection. The free-text section on evidence for taxonomic, life stage, and sex applicability can be used to elaborate on why the specific structured terms were selected, and provide supporting references and background information.  More help

Taxonomic: Deiodination by DIO enzymes is known to exist in a wide range of vertebrates and invertebrates. Therefore, this KE is plausibly applicable across vertebrates. Studies reporting DIO1 inhibition have used human liver (Kuiper et al., 2006), human recombinant DIO1 enzyme (Olker et al., 2019), rat (Rattus norvegicus) liver (Klaren et al., 2005; Freyberger and Ahr, 2006; Kuiper et al., 2006; Pavelka, 2010) and thyroid gland (Ferreira et al., 2002), mouse (Mus musculus) brain (hernandez et al., 2006), hog (Sus scrofa domesticus) liver (Stinckens et al., 2018), sheep (Ovis orientalis aries) fetal hepatic, renal and perirenal adipose tissue (Forhead et al., 2006), tadpole (Xenopus laevis) liver (Kuiper et al., 2006), fathead minnow (Pimephales promelas) whole fish (Noyes et al., 2011), Nile tilapia (Oreochromis niloticus) liver (Walpita et al., 2007), Gilthead Seabream (Sparus aurata) kidney (Klaren et al., 2005), and killifish (Fundulus heteroclitus) liver (Orozco et al., 2003) among others. The latter teleostean DIO1 enzymes as well as amphibian enzymes differ from other vertebrate DIO1 enzymes in their lower sensitivity to propylthiouracil (PTU), a typical DIO1 inhibitor in mammals.

Deiodinase 1 in liver is the main supplier of T3 to circulation in mammals (Marsili et al., 2011), and the same appears to be true for birds. By contrast, DIO1 function in teleostean and amphibian T3 plasma regulation is less clear (Finnson et al. 1999, Kuiper et al. 2006). The presence of DIO1 in the liver of teleosts has been a controversial issue, and both the high level of DIO2 activity and its expression in the liver of teleosts are unique among vertebrates (Orozco and Valverde, 2005). This could explain why DIO2 inhibition seems to be more important than DIO1 inhibition in determining the adverse outcome in zebrafish (Stinckens et al., 2018).

Life stage: Deiodinase activity is important for all vertebrate life stages. Already during early embryonic development, deiodinase activity is needed to regulate thyroid hormone concentrations and coordinate developmental processes. However, the role of DIO1 and DIO2 seems to be distinct. The fact that DIO1 knockdown during zebrafish development only causes developmental defects when combined with DIO2 knockdown (Walpita et al., 2010), suggests that DIO1 is only important in cases of increased TH need during specific stages of development, as supported by increased expression during such stages (Vergauwen et al., 2018), and in cases of thyroid hormone depletion in fish.

Sex: This KE is plausibly applicable to both sexes. Deiodinases are important for TH homeostasis and identical in both sexes. Therefore inhibition of deiodinases is not expected to be sex-specific.

Evidence for Perturbation by Stressor

Overview for Molecular Initiating Event

When a specific MIE can be defined (i.e., the molecular target and nature of interaction is known), in addition to describing the biological state associated with the MIE, how it can be measured, and its taxonomic, life stage, and sex applicability, it is useful to list stressors known to trigger the MIE and provide evidence supporting that initiation. This will often be a list of prototypical compounds demonstrated to interact with the target molecule in the manner detailed in the MIE description to initiate a given pathway (e.g., 2,3,7,8-TCDD as a prototypical AhR agonist; 17α-ethynyl estradiol as a prototypical ER agonist). Depending on the information available, this could also refer to chemical categories (i.e., groups of chemicals with defined structural features known to trigger the MIE). Known stressors should be included in the MIE description, but it is not expected to include a comprehensive list. Rather initially, stressors identified will be exemplary and the stressor list will be expanded over time. For more information on MIE, please see pages 32-33 in the User Handbook.

Propylthiouracil (PTU) is the prototypical DIO1 inhibitor in mammals, although teleostean and amphibian DIO1 enzymes are less sensitive to inhibition by PTU (Orozco et al., 2003; Kuiper et al., 2006). Olker et al. (2019) identified 22 DIO1-specific inhibitors using a human recombinant DIO1 enzyme (e.g., genistein, 6-methyl-2-thiouracil, sulfasalazine). Another well-known inhibitor of DIO1 (and DIO2 and 3) is iopanoic acid (IOP). Renko et al. (2003, 2015) pointed out that IOP is actually a substrate of DIO1 (and DIO2 and 3) which is in line with its action as a competitive inhibotor. In fact, many compounds inhibit all three DIO isoforms. Olker et al. (2019) identified 93 compounds that inhibit DIOs 1, 2 and 3.

References

List of the literature that was cited for this KE description. Ideally, the list of references, should conform, to the extent possible, with the OECD Style Guide (https://www.oecd.org/about/publishing/OECD-Style-Guide-Third-Edition.pdf) (OECD, 2015). More help

Butt, C.M., Wang, D.L., Stapleton, H.M., 2011. Halogenated Phenolic Contaminants Inhibit the In Vitro Activity of the Thyroid-Regulating Deiodinases in Human Liver. Toxicological Sciences 124, 339-347.

Cavallin JE, Ankley GT, Blackwell BR, Blanksma CA, Fay KA, Jensen KM, Kahl MD, Knapen D, Kosian PA, Poole ST et al. 2017. Impaired swim bladder inflation in early life stage fathead minnows exposed to a deiodinase inhibitor, iopanoic acid. Environmental Toxicology and Chemistry. 36(11):2942-2952.

Darras, V.M., Van Herck, S.L.J., 2012. Iodothyronine deiodinase structure and function: from ascidians to humans. Journal of Endocrinology 215, 189-206.

Ferreira, A.C.F., Lisboa, P.C., Oliveira, K.J., Lima, L.P., Barros, I.A., Carvalho, D.P., 2002. Inhibition of thyroid type 1 deiodinase activity by flavonoids. Food and Chemical Toxicology 40, 913-917.

Finnson, K.W., McLeese, J.M., Eales, J.G., 1999. Deiodination and deconjugation of thyroid hormone conjugates and type I deiodination in liver of rainbow trout, Oncorhynchus mykiss. General and Comparative Endocrinology 115, 387-397.

Forhead, A.J., Curtis, K., Kaptein, E., Visser, T.J., Fowden, A.L., 2006. Developmental control of iodothyronine deiodinases by cortisol in the ovine fetus and placenta near term. Endocrinology 147, 5988-5994.

Freyberger, A., Ahr, H.J., 2006. Studies on the goitrogenic mechanism of action of N,N,N',N'-tetramethylthiourea. Toxicology 217, 169-175.

Hernandez, A., Martinez, M.E., Fiering, S., Galton, V.A., St Germain, D., 2006. Type 3 deiodinase is critical for the maturation and function of the thyroid axis. J Clin Invest 116, 476-484.

Hornung, M.W., Korte, J.J., Olker, J.H., Denny, J.S., Knutsen, C., Hartig, P.C., Cardon, M.C., Degitz, S.J., 2018. Screening the ToxCast Phase 1 Chemical Library for Inhibition of Deiodinase Type 1 Activity. Toxicological Sciences 162, 570-581.

Houbrechts AM, Vergauwen L, Bagci E, Van Houcke J, Heijlen M, Kulemeka B, Hyde DR, Knapen D, Darras VM. 2016. Deiodinase knockdown affects zebrafish eye development at the level of gene expression, morphology and function. Molecular and Cellular Endocrinology. 424(C):81-93.

Klaren, P.H.M., Haasdijk, R., Metz, J.R., Nitsch, L.M.C., Darras, V.M., Van der Geyten, S., Flik, G., 2005. Characterization of an iodothyronine 5 '-deiodinase in gilthead seabream (Sparus auratus) that is inhibited by dithiothreitol. Endocrinology 146, 5621-5630.

Kuiper, G., Klootwijk, W., Dubois, G.M., Destree, O., Darras, V.M., Van der Geyten, S., Demeneix, B., Visser, T.J., 2006. Characterization of recombinant Xenopus laevis type I iodothyronine deiodinase: substitution of a proline residue in the catalytic center by serine (Pro132Ser) restores sensitivity to 6-propyl-2-thiouracil. Endocrinology 147, 3519-3529.

Marsili, A., Zavacki, A.M., Harney, J.W., Larsen, P.R., 2011. Physiological role and regulation of iodothyronine deiodinases: A 2011 update. Journal of Endocrinological Investigation 34, 395-407.

Noyes, P.D., Hinton, D.E., Stapleton, H.M., 2011. Accumulation and Debromination of Decabromodiphenyl Ether (BDE-209) in Juvenile Fathead Minnows (Pimephales promelas) Induces Thyroid Disruption and Liver Alterations. Toxicological Sciences 122, 265-274.

Olker, J.H., Korte, J.J., Denny, J.S., Hartig, P.C., Cardon, M.C., Knutsen, C.N., Kent, P.M., Christensen, J.P., Degitz, S.J., Hornung, M.W., 2019. Screening the ToxCast Phase 1, Phase 2, and e1k Chemical Libraries for Inhibitors of Iodothyronine Deiodinases. Toxicological Sciences 168, 430-442.

Orozco, A., Valverde, R.C., 2005. Thyroid hormone deiodination in fish. Thyroid 15, 799-813.

Orozco, A., Villalobos, P., Jeziorski, M.C., Valverde, C., 2003. The liver of Fundulus heteroclitus expresses deiodinase type 1 mRNA. General and Comparative Endocrinology 130, 84-91.

Pavelka, S., 2010. Radiometric enzyme assays: development of methods for extremely sensitive determination of types 1, 2 and 3 iodothyronine deiodinase enzyme activities. Journal of Radioanalytical and Nuclear Chemistry 286, 861-865.

Renko, K., Hoefig, C.S., Hiller, F., Schomburg, L., Kohrle, J., 2012. Identification of Iopanoic Acid as Substrate of Type 1 Deiodinase by a Novel Nonradioactive Iodide-Release Assay. Endocrinology 153, 2506-2513.

Stinckens, E., Vergauwen, L., Ankley, G.T., Blust, R., Darras, V.M., Villeneuve, D.L., Witters, H., Volz, D.C., Knapen, D., 2018. An AOP-based alternative testing strategy to predict the impact of thyroid hormone disruption on swim bladder inflation in zebrafish. Aquatic Toxicology 200, 1-12.

Stinckens E, Vergauwen L, Blackwell BR, Anldey GT, Villeneuve DL, Knapen D. 2020. Effect of thyroperoxidase and deiodinase inhibition on anterior swim bladder inflation in the zebrafish. Environmental Science & Technology. 54(10):6213-6223.

Vergauwen, L., Cavallin, J.E., Ankley, G.T., Bars, C., Gabriels, I.J., Michiels, E.D.G., Fitzpatrick, K.R., Periz-Stanacev, J., Randolph, E.C., Robinson, S.L., Saari, T.W., Schroeder, A.L., Stinckens, E., Swintek, J., Van Cruchten, S.J., Verbueken, E., Villeneuve, D.L., Knapen, D., 2018. Gene transcription ontogeny of hypothalamic-pituitary-thyroid axis development in early-life stage fathead minnow and zebrafish. General and Comparative Endocrinology 266, 87-100.

Visser, T.J., Van Overmeeren, E., Fekkes, D., Docter, R., Hennemann, G. 1979. Inhibition of iodothyronine 5'-deiodinase by thioureylenes: structure-activity relationship. FEBS Letters, 103, 2.

Walpita, C.N., Crawford, A.D., Darras, V.M., 2010. Combined antisense knockdown of type 1 and type 2 iodothyronine deiodinases disrupts embryonic development in zebrafish (Danio rerio). Gen Comp Endocrinol 166, 134-141.

Walpita, C.N., Grommen, S.V., Darras, V.M., Van der Geyten, S., 2007. The influence of stress on thyroid hormone production and peripheral deiodination in the Nile tilapia (Oreochromis niloticus). Gen Comp Endocrinol 150, 18-25.