bio2009 ctc presentation non confidential

Profiling of Circulating Tumor Cells

A novel platform enabling drug response, signaling events, overexpression, mutation profiling, and transcriptional profiling analysis

Allyn Forsyth, Ph.D Paul Rohricht, MSc., MBASenior Director of R&D Vice President Business [email protected] [email protected]

Clinical utility of CTC detection

Prognostic for patients with metastatic disease-progression free and overall survivalPrognostic for early stage disease-assessing relapse riskPredictive for chemotherapy response-early assessment of effectivenessCurrent systems only count the cells that can be detected

What happens when EpCam is not highly expressed (ex., pancreatic cancer) and therefore not counted as a CTC?

Future:Monitoring of disease progression?Disease staging?

ICx platform counts and profiles CTCsdrug response, signaling events, overexpression, mutation profiling, and transcriptional profiling analysis

ICx Purification Process

Sorted cell populations

79921 blood cells

359 Target cells

Labeling

Billions of cells <0.2 million cells

Sorted targets72% recovery

98% purity

Sorted non-targets7 mis-sorted cells out of 80,280

Purification via µFACSDepletion of non target cells1. 2. 3.

Proprietary cell sorting technology

Multiple fluorescence channels enables advanced strategies for positive and negative selection

•Traditional approaches are typically limited to moderate to highly expressed epitopes (usually localized to the cell surface) and validation of targets with other markers

•EpCam+, CK+, Nuclear+, CD45-

•Broad CTC capture Should capture more CTCs from higher percentage of patients•Channel 1- Labeled antibody cocktail to EpCam, CK, ErbB2, EGFR, PSA etc•Channel 2- nuclear•Channel 3- subpopulation specific (IGFR, pHH3?)•Channel 4- counter label; CD45

CK-PEPhospho-H3-A488

Phospho-AKTCD45-A488

ErbB2-PE

CD45-PE CD235a-FITC

ICx Platform enables broad range of CTC measurements

•High purity and yield enables quality molecular analyses

•Enumeration- with ability to use multiple antibodies to ID CTCs

•Immunohistochemistry- via FACS analysis for every cell allows downstream molecular analysis or traditional slide plating

•Drug response •Signaling events •Overexpression

•Mutation profiling

•Transcriptional profiling

•Compatible with marketed preservatives

•ICx preservative enables transcriptional profiling

MagBeadDepletion

~50 cells

µFACspurified

~ 50 cells

Leukocyte RNA

Input to analysis

Cell linePrior to spike-in

~ 50 cells

No TemplateLung cancer RNA

500 pg

Spiked Blood~50 cells

pure

1 in 105

1 in 109

Target dilution

pure

1 in 105

1 in 109

Target dilution

2009 CTC Effort is On ScheduleDemonstrated CTC recovery from clinical samples

Demonstrated stability of process for off-site (shipped) samples

Demonstrated recovery efficiency and molecular analysis on spike-in samples to whole blood

Developing markers relevant to pharma development

PartneringPartnered with Merck to develop a protocol for patient selection of a specific clinical trial

Discussions with other large pharma and large biotech companies

Letter of intent signed with Fred Hutchinson Cancer Research Centercost share clinical research studies

Discussions with other hospital research centerscost share clinical research studies

Contacts:

Paul Rohricht: [email protected]; Allyn Fosyth: [email protected]

bio2009 ctc presentation non confidential

Documents