Using Stem Cells as Drug Discovery Tools

Presentation for XenoTech LLCLenexa, KS May 1st, 2008

By: Jim Hardy President & CEO Gahaga Biosciences, Inc.

Background Wittenberg University (BA) University of Rochester BRL/Life Technologies, Inc. BP Solar In Vitro Technologies Gahaga, APE-BridgePath Scientific @

FITCI

Stem Cell Time Line20

05

2006

2007

2008

2009

2010

2011

2012

2013

2014

2015

2016

$- $1,000 $2,000 $3,000 $4,000 $5,000 $6,000 $7,000 $8,000 $9,000 1961: Canadians Jim Till & Ernest

McCulloch prove the existence of stem cells (Nature 1963)

1978: Hematopoietic SC discovered in UCB

1981: mouse ES cells isolated

1988: HSC isolated from blood

1992: Adult neural SC discovered

1994: first Cancer SC discovered

1996: Dolly the Sheep

1998: first human ES line

RegMed 2.0

USD (m

illions)

Stem Cells for Drug Discovery

ES cells: › pluripotent › Limited number of cells and strains available› Regulatory environment› Political/ethical issues

“Adult” SC: › lineage dependant› Not easy to isolate or proliferate in sufficient

quantities› Most widely used/accepted in clinical

applications

Stem Cells for Drug Discovery continued…

IPS (induced Pluripotent SCs)› Latest thing› Still a lot of uncertainty

Perinatal tissue: UCB & Afterbirth› Ephemeral organ› Not really “adult”› Pluripotent (?)› Easy to obtain in large volume, various

donor demographics

Uses of Stem Cells in Drug Discovery

As a replacement for Primary Cells› Hepatocytes› Renal cells› Circulatory› Cardiomyocytes› Neurons› Pulmonary cells› Bone, cartilage› Skin: Wound healing & absorption models

In vitro models for metabolic homeostasis & Organogenesis› Demonstrate a “stimulation” of injury & repair mechanisms› Inhibition of necrosis/apoptosis› Tissue specific developmental pathways› Cancer Stem Cells

The Players

Why Neuro-Tox? The path of least resistance Neurodegenerative diseases

› Alzheimer's› Parkinson's› MS› ALS/Lou Gehrig's disease

Acute Injuries› Spinal cord injury› Stroke› Head injury› Cerebral Palsy

Abnormal Neural function› Depression› Epilepsy› Autism

HIV

Alzheimer's Disease and neurodegeneration:

The modular systems biology approach to investigate the control of apoptosis in Alzheimer's disease neurodegeneration, BMC Neurosci. 2006; 7(Suppl 1): S2.

Gahaga Cells From term amniotic membrane tissue Primary, adherent cultures &

cryopreserved Mesenchymal morphology and

Immunophenotype Assessed differentiation side-by-side vs.

Lonza bm-MSC Assessed growth in proprietary GHG

medium vs. Invitrogen MesenPRO® medium Developed two-step differentiation protocol

Nestin: G

HG

vs IVGN

m

edium

Gahaga medium

Invitrogen MesenPRO

©

ß-Tubulin III G

HG

vs IVGN

medium

Gahaga medium

Invitrogen MesenPRO

©

Neuronal Differentiation Grown to confluence in 96 well plates, with

proprietary placental basement membrane extract

Differentiation for 4 days 7 days Neuronal maintenance in IVGN

Neurobasal medium

Challenges Establish clinical significance, IVIVC Choosing the right target: pick one or two

target assays most relevant in the field Optimize isolation, differentiation

protocols Collaborate with one or more existing

ADMET/DMPK companies Always the issue of IP and litigation in this

emerging field

Conclusions A readily obtainable & reproducible

source of neuronal precursors and neurons for the study of neuronal function, dysfunction, and development

A culture system for neuronal differentiation

Potential for similar systems for other stromal cell types

Questions ?