Tox21: Transforming Environmental Health Protection Raymond Tice, Ph.D. Chief, Biomolecular Screening Branch (tice@niehs.nih.gov) LINCS Consortium Kick-Off Meeting Rockville, MD October 23-24, 2011 - PowerPoint PPT Presentation

Loading...

PPT – Tox21: Transforming Environmental Health Protection Raymond Tice, Ph.D. Chief, Biomolecular Screening Branch (tice@niehs.nih.gov) LINCS Consortium Kick-Off Meeting Rockville, MD October 23-24, 2011 PowerPoint presentation | free to download - id: 607f68-Yzc3Y



Loading


The Adobe Flash plugin is needed to view this content

Get the plugin now

View by Category
About This Presentation
Title:

Tox21: Transforming Environmental Health Protection Raymond Tice, Ph.D. Chief, Biomolecular Screening Branch (tice@niehs.nih.gov) LINCS Consortium Kick-Off Meeting Rockville, MD October 23-24, 2011

Description:

Title: Presentation Title Author: buhrman Last modified by: Raymond Tice Created Date: 6/21/2011 7:24:34 PM Document presentation format: On-screen Show (4:3) – PowerPoint PPT presentation

Number of Views:83
Avg rating:3.0/5.0

less

Write a Comment
User Comments (0)
Transcript and Presenter's Notes

Title: Tox21: Transforming Environmental Health Protection Raymond Tice, Ph.D. Chief, Biomolecular Screening Branch (tice@niehs.nih.gov) LINCS Consortium Kick-Off Meeting Rockville, MD October 23-24, 2011


1
Tox21 Transforming Environmental Health
Protection Raymond Tice, Ph.D.Chief,
Biomolecular Screening Branch(tice_at_niehs.nih.gov)
LINCS Consortium Kick-Off MeetingRockville,
MD October 23-24, 2011
2
The Tox21 Screening Timeline
Tox21 MOU
NTP Vision Roadmap
FDA joins Tox21
EPAs Strategic Plan
NAS Report Tox in the 21st Century
NCGC
EPA NCCT
Tox21 - a Community Resource Project
2
3
Tox21 Partners
Area of Expertise NIEHS NCGC EPA FDA
Lab Animal Toxicology ? ? ?
Human Toxicology/Exposure Assessment ? ? ?
Ultra High Throughput Screening ?
Low to Mid Throughput Assays ? ? ? ?
Stem Cell Assay Development ? ? ? ?
Epigenetic Assays ? ?
Engineered Tissue Models ? ? ?
Omic Based Systems ? ? ? ?
Lower Organism Systems ? ? ?
Genetic Variability in Response ? ?
Databases Informatic Tools ? ? ? ?
Validation Experience ? ? ? ?
3
4
Tox21 Goals
  • Identify patterns of compound-induced biological
    response in order to
  • characterize toxicity/disease pathways
  • facilitate cross-species extrapolation
  • model low-dose extrapolation
  • Prioritize compounds for more extensive
    toxicological evaluation
  • Develop predictive models for biological response
    in humans

4
5
5
6
Tox21 Phase I Proof of Principle
  • NCGC screened 1408 compounds (1353 unique) from
    NTP and 1462 compounds (1384 unique) from EPA in
    gt100 qHTS at 14 conc (5 nM to 92 ?M typical).
  • EPA via ToxCast screened 320 compounds (309
    unique, primarily pesticide actives and some
    endocrine active compounds) in 550 assays.
  • Data released to the scientific community via
  • EPA ACToR (Aggregated Computational Toxicology
    Resource http//epa.gov/actor)
  • NLM PubChem (http//pubchem.ncbi.nlm.nih.gov/)
  • NTP CEBS (Chemical Effects in Biological Systems
    http//www.niehs.nih.gov/research/resources/databa
    ses/cebs/index.cfm)

6
7
ToxCastTM Phase I Testing
  • ToxCast 1.2 (June, 2008)
  • NR Activation and translocation (CellzDirect)
  • HTS Genotoxicity (Gentronix)
  • Organ toxicity dosimetry (Hamner Institutes)
  • Toxicity and signaling pathways (Invitrogen)
  • C. elegans WormTox (NIEHS)
  • Gene markers from microscale cultured hepatocytes
    (MIT)
  • 3D Cellular microarray with metabolism (Solidus)
  • Zebrafish vascular/cardiotoxicity (Zygogen)
  • HTS stress response (NHEERLNCGC)
  • ToxCast 1.0 (April, 2007)
  • Enzyme inhibition/receptor binding HTS
    (Novascreen)
  • NR/transcription factors (Attagene, NCGC)
  • Cellular impedance (ACEA)
  • Complex cell interactions (BioSeek)
  • Hepatocelluar HCS (Cellumen)
  • Hepatic, renal and airway cytotoxicity (IVAL)
  • In vitro hepatogenomics (IVAL, Expression
    Analysis)
  • Zebrafish developmental toxicity (Phylonix)
  • ToxCast 1.1 (January, 2008)
  • Neurite outgrowth HCS (NHEERL)
  • Cell proliferation (NHEERL)
  • Zebrafish developmental toxicity (NHEERL)

8
Phase I NCGC qHTS Assays
  • Phenotypic readouts
  • Cytotoxicity
  • Apoptosis caspase 3/7, 8, 9
  • Membrane integrity LDH, protease release
  • Mitochondrial toxicity (membrane potential)
  • Genetox p53, ELG1, DNA damage gene deficient
    lines (DT40 lines and mouse)
  • Cell Signaling
  • Stress response ARE, ESRE, HSP, Hypoxia, AP-1
  • Immune response IL-8, TNF?, TTP
  • Other AP-1, CRE, ERK, HRE, JNK3, NFkB, LDR
  • Epigenetics
  • Locus DeRepression (LDR)
  • Drug metabolism
  • CYP1A2, CYP2C19, CYP2C9, CYP2D6, CYP3A4
  • Target specific assays
  • Nuclear receptors AR, AhR, ER?, FXR, GR, LXR,
    PPAR?, PPARd, PPAR?, PXR, RXR, TRß, VDR, ROR?,
    ROR?
  • hERG channel
  • Isolated molecular targets 12hLO, 15hLO1,
    15hLO2, ALDH1A1, HADH560, HPGD, HSD17b4, APE1,
    TDP1, DNA polymerase III, RECQ1 helicase, RGS4,
    BRCA, IMPase, O-Glc NAc Transferase, Caspase-1/7,
    CBFß-RUNX1, PK, Tau, Cruzain, ß-Lactamase, PRX,
    YjeE , NPS, Proteasome, SF1, SMN2, beta-globin
    splicing, Anthrax Lethal Factor, TSHR
  • Genetic variation 87 HapMap CEPH Panel

8
9
Differential Compound Toxicity Among 13 Cell Types
IC50 Values
H
H
M
H
H
H
R
M
H
H
H
H
R
Xia et al., EHP 116284, 2008
9
10
Tox21 Phase II
  • EPAs ToxCast Phase II 1000 compounds in 550
    assays.
  • NCGC qHTS Phase II gt10K compounds 3x at 14 conc
    for
  • nuclear receptor activation or inhibition (AR,
    AhR, ER, FXR, GR, LXR, PPAR, PXR, RXR, TR, VDR,
    ROR)
  • induction of stress response pathways (e.g., DNA
    damage, heat shock, hypoxia, inflammation,
    oxidative)
  • Assay selection based on
  • Information from in vivo toxicological
    investigations
  • Phase I experience, advice of basic researchers,
    and nominated assays
  • Maps of disease-associated cellular pathways
  • Future focus on disease-associated pathways
    (e.g., obesity/diabetes, autism) using stem
    cells/differentiated cells and high throughput
    gene array assays

10
11
Tox21 Phase II qHTS 10K Library
  • NCGC
  • Drugs
  • Drug-like compounds
  • Active pharmaceutical ingredients
  • EPA
  • ToxCast I and II compounds
  • Antimicrobial Registration Program
  • Endocrine Disruptor Screening Program
  • OECD Molecular Screening Working Group List
  • FDA Drug Induced Liver Injury Project
  • Failed Drugs
  • NTP
  • NTP-studied compounds
  • NTP nominations and related compounds
  • NICEATM/ICCVAM validation reference compounds for
    regulatory tests
  • External collaborators (e.g., Silent Spring
    Institute, U.S. Army Public Health Command)
  • Formulated mixtures

12
  • The Tox21 Genomes Project (with I. Rusyn, UNC)
  • Assessment of variation within and between
    populations
  • Mapping of genomic regions associated with
    variation of responses to individual chemicals or
    classes
  • In a cell-based system, with carefully controlled
    growth and environmental conditions, the assay
    may serve as an endo-phenotype, with a greater
    proportion of variation explained by genomic
    variation than for a typical complex trait
  • Status
  • Phase I 87 CEPH panel x 240 cmpds x 12 conc x 2
    assays (cytotox caspase 3/7)
  • Phase II 1090 lines (9 racial groups) x 180
    cmpds x 8 conc x 1 assay (cytotox)

12
13
The NTP DrugMatrix Rat Toxicogenomics Database
  • Integrated Collection of Data
  • 637 unique chemicals (mostly drugs)
  • 5600 drug-treatment transcript profiles in rat
    organs
  • 127,000 histopathology measurements
  • 100,000 blood chemistry measurements
  • 60,000 literature facts
  • Over 500 validated signatures
  • Mode of action and pathology
  • Comprehensive data mining
  • Formulate 100,000s questions (phenotypes)
  • Test for ability to classify using transcript
    data only
  • 122,000 frozen tissues
  • Automated genomics analysis
  • Drugmatrix website https//ntp.niehs.nih.gov/drug
    matrix
  • ToxFx website https//ntp.niehs.nih.gov/toxfx/

13
14
Detailed view of a pathway
The NCGC Universe of Human Pathways
Pathways
Gene information
1100 human pathways mapped to the pathway globe
14
15
Development of an Integrated Prediction System
  • Collaboration between NIEHS, Leadscope Inc.,
    Lhasa Limited, and MultiCASE Inc.
  • System to support the prioritization of chemicals
    through human-relevant toxicity predictions
  • Designed for use by scientists with different
    backgrounds
  • Brings together toxicity data and predictions
    from multiple geographically distributed
    locations
  • qHTS data from the Tox21 project
  • Historical in vitro and in vivo data
  • (Q)SAR models for human adverse event endpoints
    as well as in vitro and in vivo endpoints

16
Identifying Disease Pathways
http//cerhr.niehs.nih.gov/evals/diabetesobesity/
16
17
Success depends on
  • Well-characterized chemical libraries (identity,
    purity, concentration, stability)
  • Well-characterized assays in terms of reliability
    and relevance
  • Ability to incorporate xenobiotic metabolism
  • Informatic tools to integrate and mine robust
    data from multiple sources
  • Understanding the relationships between pathways
    and disease in animal models and humans
  • Making the data freely accessible as quickly as
    possible
  • Scientific outreach and training the next
    generation

17
About PowerShow.com