The EPA ToxCast program, with BioSeek as a contractor since 2007, focuses on a cost-effective approach for toxicity testing of thousands of chemicals through three phases: proof-of-concept, verification, and practical application. The program has developed a variety of data-rich signatures and has publicly posted its findings. The document further details biomap analysis outcomes, nanomaterial bioactivity, and related publications and presentations.

1. BioSeek – EPA ToxCastTM Program
Bridging the Gap From In Vitro to In Vivo
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2. EPA ToxCastTM Program
 EPA ToxCastTM Program
 Program to develop a cost-effective approach for efficiently
prioritizing the toxicity testing of thousands of chemicals
 BioSeek has been a contractor since 2007
 Three Phases
 Phase I – Proof-of-concept
 Phase II – Verification / Extension
 Phase III – Reduction to Practice
 http://www.epa.gov/ncct/toxcast/
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3. ToxCastTM Program Timeline
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4. ToxCastTM Program Details
Number of Chemical Number of Cost per Target
Phase Purpose
Chemicals Criteria Assays Chemical Date
Data Rich Signature
I 320 552 $20k FY08
(pesticides) Development
Ib 15 Nanomaterials Pilot 166 $10K FY09
Data Rich
IIa >300 Validation >400 ~$20-25k FY09
Chemicals
Known Human
IIb >100 Extrapolation >400 ~$20-25k FY09
Toxicants
Expanded Structure
IIc >300 Extension >400 ~$20-25k FY10
and Use Diversity
IId >12 Nanomaterials PMN >200 ~$15-20K FY09-10
Prediction and
III Thousands Data poor >300 ~$15-20k FY11-12
Prioritization
7/23/2012 CONFIDENTIAL
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5. BioSeek ToxCastTM Data
Now Posted on EPA’s ACToR Public Database
http://actor.epa.gov/
ACToR
Aggregated Computational Toxicology Resource
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6. BioMAP® Analysis of ToxCastTM Phase I Chemicals
Diversity of Mechanisms Distinguished
NFB
cAMP Elevation
DNA Alkylation Mitochondrial Dysfunction
Reference: Houck, K.A., D.J. Dix, R.S. Judson, R.J. Kavlock, J.
Yang and E.L. Berg. 2009. Profiling bioactivity of the ToxCast Tubulin Inhibition
chemical library using BioMAP primary human cell systems. J.
Biomol. Screen, 2009, 14:1054-66.
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7. BioMAP Endpoints (BSK) in Predictive Signatures
Rat & Rabbit Developmental Toxicity
Knudsen, T.B., Computational Toxicology and Prenatal Development, presentation for
the EPA's Computational Toxicology Community of Practice (8/25/2011)
7 Toxicol Sci. 2011, 124:109-27

8. BioMAP® Analysis of Nanomaterials
ToxCast Phase II Chemicals
• Nanomaterials are active and distinguishable by composition
• Reference: Wang, A., E.L. Berg, M. Polokoff, J. Yang; D. Reif, N. Kleinstreuer, S. Marinakos, A.R. Badireddy,
S. Gangwal, C. Matson, M. Wiesner, and K. Houck. Nanomaterial (NM) bioactivity profiling by ToxCast high-
throughput screening (HTS). Society of Toxicology Annual Meeting, March 12, 2012, San Francisco, CA.
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9. BioMAP® Profile of Titanium Oxide Nanoparticles
Similarity to Paclitaxel
• Paclitaxel – binds b-tubulin and causes microtubule stabilization
• TiO2 nanoparticles interfere with microtubules (PMID: 18776989)
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10. BioMAP® Analysis of Failed Pharma Compounds
 Nicole Kleinstreuer, Society of Toxicology Annual Meeting, March 12,
2012, San Francisco, CA
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11. Publications with BioSeek ToxCastTM Data
 Sipes NS, Martin MT, Reif DM, Kleinstreuer NC, Judson RS, Singh AV, Chandler KJ, Dix DJ, Kavlock
RJ, Knudsen TB. Predictive models of prenatal developmental toxicity from ToxCast high-throughput
screening data. Toxicol Sci. 2011, 124:109-27
 Knudsen, T. B., K. A. Houck, N. Sipes, A. V. Singh, R. Judson, M. T. Martin, A. Weissman, N.
Kleinstreuer, H. M. Mortensen, D. Reif, J. R. Rabinowitz, R. W. Setzer, A. M. Richard, D. J. Dix, And R.
J. Kavlock. Activity profiles of 309 ToxCast™ chemicals evaluated across 292 biochemical targets.
TOXICOLOGY. Elsevier Ireland Limited, Limerick, Ireland, 282(1-2):1-15, (2011).
 Kleinstreuer, N. C., R. S. Judson, D. M. Reif, N. S. Sipes, A. V. Singh, K. J. Chandler, R. Dewoskin, D.
J. Dix, R. J. Kavlock, And T. B. Knudsen. Environmental Impact on Vascular Development Predicted by
High Throughput Screening. ENVIRONMENTAL HEALTH PERSPECTIVES. National Institute of
Environmental Health Sciences (NIEHS), Research Triangle Park, NC, Online Edition:1-34, (2011).
doi:10.1289/ehp.1103412
 Houck, K.A., D.J. Dix, R.S. Judson, R.J. Kavlock, J. Yang and E.L. Berg. 2009. Profiling bioactivity of
the ToxCast chemical library using BioMAP primary human cell systems. J. Biomol. Screen, 2009,
14:1054-66.
 Judson RS, Houck KA, Kavlock RJ, Knudsen TB, Martin MT, et al. 2009, In Vitro Screening of
Environmental Chemicals for Targeted Testing Prioritization: The ToxCast Project. Environ Health
Perspect 118(4): doi:10.1289/ehp.0901392
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12. Presentations with BioSeek ToxCastTM Data
 Kleinstreuer, N. C., vEmbryo in silico models:Predicting vascular developmental toxicity, Wiley-
Blackwell Symposium on Computational Embryology: Integration and Modeling of Developmental
Mechanisms, Teratology Society 2012, June 25, 2012, Baltimore, MD.
 Wang, A., E.L. Berg, M. Polokoff, J. Yang; D. Reif, N. Kleinstreuer, S. Marinakos, A.R. Badireddy, S.
Gangwal, C. Matson, M. Wiesner, and K. Houck. Nanomaterial (NM) bioactivity profiling by ToxCast
high-throughput screening (HTS). Society of Toxicology Annual Meeting, March 12, 2012, San
Francisco, CA.
 Kleinstreuer, N. C., K. Houck, R. Judson, D. Reif, P. Kothiya, M. Martin, T. Knudsen, A. Richard, M.
Polokoff, J. Yang, E.L. Berg, R. Kavlock, and D. Dix. Biological profiling of the ToxCast Phase II
Chemical Library in Primary Human Cell CoCulture Systems. Society of Toxicology Annual Meeting,
March 12, 2012, San Francisco, CA.
 Houck, K., Use of primary human cell systems for creating predictive toxicology profiles. SBS Annual
Meeting, 2009, Lille, France.
 Houck, K. Evaluation of the ToxCast™ suite of cellular and molecular assay for prediction of in vivo
toxicity. GlobalChem, 2009, Baltimore, MD.
 Houck, K., Characteristics of the ToxCast™ In Vitro Datasets from Biochemical and Cellular Assays
(EPA/ORD/NCCT). ToxCast™ Data Analysis Summit, May 14, 2009, RTP, NC.
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13. Contacts
BioSeek, LLC
310 Utah, Suite 100
South San Francisco, CA 94030
650-416-7600
www.bioseekinc.com
www.BioMAPSystems.com
Bridging the Gap From In Vitro to In Vivo
BioSeek, LLC
310 Utah, #100
South San Francisco, CA 94080