Intelligent Systems and Molecular Biology

Richard H. Lathrop Dept. of Computer Science

Univ. of California, Irvine

rickl@uci.edu Donald Bren Hall 4224

949-824-4021

“Computers are to Biology as Mathematics is to Physics.”

--- Harold Morowitz

(spiritual father of BioMatrix, and Intelligent Systems for Molecular Biology Conference)

Goal of talk: The power of information science to influence molecular science and technology

Intelligent Systems and Molecular Biology

Artificial Intelligence for Biology and Medicine Biology is data-rich and knowledge-hungry AI is well suited to biomedical problems

o Examples (omitted for brevity)

o Machine learning -- drug discovery o Rule-based systems – drug-resistant HIV o Heuristic search -- protein structure prediction o Constraints – design of large synthetic genes

o Current Project o Machine learning and p53 cancer rescue mutants

Goal of talk: The power of information science to influence molecular science and technology

Biology has become Data Rich

Massively Parallel Data Generation Genome-scale sequencing High-throughput drug screening Micro-array “gene chips” Combinatorial chemical synthesis “Shotgun” mutagenesis Directed protein evolution Two-hybrid protocols for protein interaction A million biomedical articles per year

“Data Rich” GenBank Genomic Sequence Data

“Data Rich” PDB Protein 3D Structure Data

“Data Rich” PubMed Biomedical Literature

“Data Rich” 10-100K data points per gene chip

Characteristics of Biomedical Data

Noise!! => need robust analysis methods

Little or no theory. => need statistics, probability

Multiple scales, tightly linked. => need cross-scale data integration

Specialized (“boutique”) databases => need heterogeneous data integration

Intelligent Systems are well suited to biology and medicine

Robust in the face of inherent complexity Extract trends and regularities from data Provide models for complex processes Cope with uncertainty and ambiguity Content-based retrieval from literature Ontologies for heterogeneous databases Machine learning and data mining

Intelligent systems handle complexity with grace

Intelligent Systems and Molecular Biology

Artificial Intelligence for Biology and Medicine Biology is data-rich and knowledge-hungry AI is well suited to biomedical problems

o Examples

o Machine learning -- drug discovery o Rule-based systems – drug-resistant HIV o Heuristic search -- protein structure prediction o Constraints – design of large synthetic genes

o Current Project o Machine learning and p53 cancer rescue mutants

Goal of talk: The power of information science to influence molecular science and technology

Cho, Y., Gorina, S., Jeffrey, P.D., Pavletich, N.P. Crystal structure of a p53 tumor suppressor-DNA complex:

understanding tumorigenic mutations. Science v265 pp.346-355, 1994

p53 is a central tumor suppressor protein

“The guardian of the genome”

Controls many tumor suppression functions

Monitors cellular distress

The most-mutated gene in human cancers

All cancers must disable the p53 apoptosis pathway.

p53 core domain bound to DNA

Image generated with UCSF Chimera

p53 and Human Cancers

Consequences of p53 mutations

Cho et al., Science 265, 346-355 (1994)

Loss of DNA contact Disruption of local structure

Denaturation of entire core domain

~250,000 US deaths/year

Over 1/3 of all human cancers express full-length p53 with only one a.a. change

Cancer Mutation

Inactive p53

Anti-Cancer Drug

+ = Active p53

Mutations Rescue Cancerous p53

Cancer Mutation

Inactive p53

Wild Type

Active p53

Cancer+Rescue Mutations

Active p53

Cancer

Cancer

Ultimate Goal

Suppressor Mutations Several second-site mutations restore functionality

to some p53 cancer mutants in vivo.

N C

Core domain for DNA binding Tetramerization

102-292 324-355

Transactivation

1-42

175

245

248 249 273

282

C S

Will not grow.

Will grow.

INACTIVE (-)

ACTIVE (+)

Baroni, T.E., et al., 2004

Class Labels: Active/+ or Inactive/- p53 Transcription Assay

Human p53 consensus

(S) = Strong (W) = Weak (N) = Negative Danziger, S.D., et al., 2009 Baronio, R., et al., 2010

URA−

First measurement Firefly luciferase p53 dependent

Second measurement Renilla luciferase p53 independent

Initial: Yeast Growth Selection, Sequencing

Confirm: Human 1299 Cell-based Luciferase

Theory

Find New Cancer Rescue Mutants

Knowledge

Experiment

Active Machine Learning for Biological Discovery

Spiral Galaxy M101

http://hubblesite.org/

~10^9 stars.

Known Mutants

~312 stars

Known Actives

~1.5 stars

Known Mutants: 31,200 Known Actives: 150

Assuming up to 5 mutations in 200 residues How Many Mutants are There?: ~10^11

Example M

…

Example N+4

Example N+3

Example N+2

Example N+1

Unknown

Example N

…

Example 3

Example 2

Example 1

Known

Training Set

Classifier

Train the Classifier

Add New Examples To Training Set

Choose Examples to Label

Computational Active Learning Pick the Best (= Most Informative) Unknown

Examples to Label

Positive Region: Predicted Active

96-105 (Green)

Negative Region: Predicted Inactive

223-232 (Red)

Expert Region: Predicted Active

114-123 (Blue)

Visualization of Selected Regions

Danziger, et al. (2009)

MIP Positive (96-105)

MIP Negative (223-232)

Expert (114-123)

# Strong Rescue 8 0 (p < 0.008) 6 (not significant)

# Weak Rescue 3 2 (not significant) 7 (not significant)

Total # Rescue 11 2 (p < 0.022) 13 (not significant) p-Values are two-tailed, comparing Positive to Negative and Expert regions. Danziger, et al. (2009)

Novel Single-a.a. Cancer Rescue Mutants

No significant differences between the MIP Positive and Expert regions. Both were statistically significantly better than the MIP Negative region. The Positive region rescued for the first time the cancer mutant P152L. No previous single-a.a. rescue mutants in any region.

Restore p53 function by a drug compound

A Long-held Goal of Anti-cancer Therapy

Restore p53 tumor suppressor pathways in tumor cells

p53 active

inactive cancer mutant

reactivation compound

reactivated

A Serendipitous Discovery (With a Great Deal of Support)

(a) Cys124 (yellow) is occluded in “closed” PDB structure. (b) Cys124 structural “breathing” in “open” MD geometry. (Wassman, et al., 2013)

Other Computational Support c d

(c) Cys 124 (yellow) is surrounded by p53 reactivation (“rescue”) mutations (green) (Wassman, et al., 2013) (d) “Druggable” pockets in p53 from FTMAP (orange) (Brenke, et al., 2009)

Stictic acid docked into open L1/S3 pocket of p53 variants

(a) wt p53; (b) R175H; (c) R273H; (d) G245S. (Wassman, et al., 2013)

14 Actives in first 91 assayed

1

0.8

0.6

0.4

0.2

0

Saos-2 (p53null)

R175H G245S

PRIM

A-1

Stic

tic a

cid

Vehi

cle

35ZW

F 25

KK

L 22

LSV

32C

TM

26R

QZ

27W

T9

33AG

6 33

BAZ

28

NZ6

27

TGR

27

VFS

32LD

E

Soas2, Soas2-p53-R175H or Soas2-G245S cells plated at 10000 per well with the different compounds. Samples are collected after 72 hours and tested for cell viability (Cell-titer Glo, promega). Selective inhibition of

R175H (red) or G245S (blue) cells versus p53null cells (black) identifies a compound that potentially reactivates p53.

Photomicrograph of cell viability (of 91 compounds assayed)

p53-null

R175H

G245S

DMSO 26RQZ 27WT9 33AG6 33BAZ 35ZWF

Compounds induced cell death in cells expressing p53 cancer mutants but not p53null cells. Cells were

cultured with vehicle (DMSO) or the compounds indicated (concentrations as above) for 24 h and

micrographs were taken.

The long road to a future anti-cancer drug

I II III IV V N C C S

I II III IV V N C C S

I II III IV V N C C S

I II III IV V N C C S

I II III IV V N C C S

I II III IV V N C C S

I II III IV V N C C S

I II III IV V N C C S

I II III IV V N C C S

I II III IV V N C C S

I II III IV V N C C S

I II III IV V N C C S

I II III IV V N C C S

I II III IV V N C C S

I II III IV V N C C S

I II III IV V N C C S

I II III IV V N C C S

I II III IV V N C C S

I II III IV V N C C S

drug

Peter Kaiser Rommie Amaro Dick Chamberlin Melanie Cocco Hudel Luecke Wes Hatfield Chris Wassman Roberta Baronio Ozlem Demir Faezeh Salehi Edwin Vargas Da-Wei Lin Scott Rychnovsky Michael Holzwarth Geoff Tucker Feng Qiao

Intelligent Systems and Molecular Biology

Artificial Intelligence for Biology and Medicine Biology is data-rich and knowledge-hungry AI is well suited to biomedical problems

o Examples

o Machine learning -- drug discovery o Rule-based systems – drug-resistant HIV o Heuristic search -- protein structure prediction o Constraints – design of large synthetic genes o DNA nanotechnology and space-filling DNA tetrahedra

o Current Project

o Machine learning and p53 cancer rescue mutants

Goal of talk: The power of information science to influence molecular science and technology

p53 Cancer Rescue Acknowledgments Rainer Brachmann (discovered p53 cancer rescue mutants) Peter Kaiser (co-PI for biology) Rommie Amaro (UCSD, molecular dynamics, virtual screening, docking) Scott Rychnovsky (current synthetic chemistry work) Wes Hatfield (Director, Computational Biology Research Lab) Hartmut (“Hudel”) Luecke (DSF and other structural biology work) Feng Qiao (protein structural biology work)

Chris Wassman (then post-doc, now at Google; L1/S3 pocket) Roberta Baronio (then esearch scientist, now at Oxford; biology work) Ozlem Demir (UCSD, molecular dynamics, virtual screening & docking) Faezeh Salehi (then graduate student, now data science researcher)

Other Colleagues: Linda Hall, Melanie Cocco, Pierre Baldi, Richard Chamberlin, Jonathan Chen, Ray Luo, Edwin Vargas, Geoff Tucker

Funding: UCI Chao Cancer Center, UCI Medical Scientist Training Program, UCI Office of Research and Graduate Studies, UCI Institute for Genomics and Bioinformatics, Harvey Fellowship, US National Science Foundation, US National Institutes of Health (National Cancer Institute)