Embryomics: Commercial Opportunities in the Increasingly Complex Biology of Pluripotency Case Western Reserve University
July 16, 2013
Forward Looking Statements
The matters discussed in this presentation include forward looking statements which are subject to various risks, uncertainties, and other factors that could cause actual results to differ materially from the results anticipated. Such risks and uncertainties include but are not limited to the success of BioTime in developing new stem cell products and technologies; results of clinical trials of BioTime products; the ability of BioTime and its licensees to obtain additional FDA and foreign regulatory approval to market BioTime products; competition from products manufactured and sold or being developed by other companies; the price of and demand for BioTime products; and the ability of BioTime to raise the capital needed to finance its current and planned operations. Any statements that are not historical fact (including, but not limited to statements that contain words such as "will," "believes," "plans," "anticipates," "expects," "estimates") should also be considered to be forward-looking statements. Forward-looking statements involve risks and uncertainties, including, without limitation, risks inherent in the development and/or commercialization of potential products, uncertainty in the results of clinical trials or regulatory approvals, need and ability to obtain future capital, and maintenance of intellectual property rights. As actual results may differ materially from the results anticipated in these forward-looking statements they should be evaluated together with the many uncertainties that affect the business of BioTime and its subsidiaries, particularly those mentioned in the cautionary statements found in BioTime's Securities and Exchange Commission filings. BioTime disclaims any intent or obligation to update these forward-looking statements.
2 The Opportunity in Pluripotency
• Scalable source of all human cell types • Source of embryonic progenitors
3 Challenges of Pluripotency
• Scalable & reproducible product • Purity and identity of cells • A formulation optimizing viability & immobilization of engraftment • Strategies for near-term commercialization
4 Contrasted Scale-Up Strategies
Purification of Problem of impurities Differentiation ES Cells desired cell type
>200-fold diversity Scalable, monoclonally purified progenitors
5 Solving the Challenge of Reproducibility
Embryoid Body
Activin A
FGF2
Wnt 3A
Pluripotent Stem Cells
1,000 + Complexity of Cell Types
6 The Opportunity of hES Cell Scalability
TRF Length Population Doublings (kbp) Germ-Line & Pluripotent SCs 22 34 43 55 65 72 82 90 15 (Telomerase +)
Somatic 10 (Telomerase -)
5 Decreasing Decreasing Telomere with Length Age Neonatal Hayflick Limit Age
7 Solving the Technical Hurdles
8 Solving the Technical Hurdles
9 Solving the Technical Hurdles
10 Solving the Technical Hurdles
11 Manufacturing Technology 2.0
Human embryonic progenitor (hEP) cell lines: > 200 diverse cell types isolated
Kidney
Diversity Smooth Muscle Precise identity Skeletal Muscle Purity Scalability Blood/Brain Patents pending Barrier
12 Purity
Purified Embryonic Progenitor Line E68
CNTN6
CNTN6 Isotype Ab Reg Med 3(3):287 (2008) Direct Scalability
Selection Criteria of Scalability in Rollers hEP Cell Line Scalability
Regen Med 2012 Jul;7(4):481-501 4D20.8 Scalability
Regen Med 2012 Jul;7(4):481-501 4D20.8 Scalability Fate Space Screening
TGFbs Chondrogenesis
FGFs Angiogenesis
BMPs Osteogenesis
RA Neurogenesis
WNTs Myogenesis
>100 Scalable Array of Diverse Approx 4,000 Clonal hEP Differentiation Gene Expression Lines Conditions Microarrays
18 Fate Space Screening
T42 in MM Culture T42 in HyStem Culture
19 Subsidiaries
OncoCyte Corporation Cancer diagnostics and anti-cancer therapies
Cell Cure Neurosciences Ltd. Neurology
OrthoCyte Corporation Orthopedics
ReCyte Therapeutics, Inc. Age-related vascular disorders
ES Cell International PTE Ltd. Research products
LifeMap Sciences, Inc. Online databases
Asterias Biotherapeutics Contribution of Geron and BioTime Assets
20 Osteochondral Differentiation
Micromas s with TGFb3 COL2A1 qPCR Condition7SMOO32
4D20.8 MSCs
SM30 7PEND24 E15 MEL2
SK11 NHAC
21 Diverse Osteochondral hEP Cell Lines Diverse Osteochondral hEP Cell Lines Osteochondral Differentiation
Micromas s with TGFb3 Condition
24 PureStem Line 4D20.8
• HOXB2+ • BARX1+ • LHX8+ • FOXF2-
25 4D20.8 hEP Reproducibility
26 Solving the Manufacturing Challenges
Simplified and more reproducible differentiation
FGF2
Wnt 3A
1,000 + Complexity of Cell Types
27 Osteochondral Differentiation
DPSCs
4D20.8
28 Precise Identity
Foxf1 Lhx8 Barx1
Foxf1 Genes & Dev. 18: 937-951 Lhx8 Science 24:306: 2255-2257 Barx1 Development 136: 637-645 29 Osteochondral Differentiation
30 Osteochondral Differentiation
31 COL2A1 (Cartilage) Induction
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*
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32 Regen Med 2012 Jul;7(4):481-501 Diverse Osteochondral hEP Cell Lines
Effects of GDF5 PureStem Line E15
NNAT+ Chondrogenic Progenitor
E15 Differentiated in GDF5
Positive For: Negative For: NNAT MSX1 HOXA2 FOXF1 HOXB2 FOXF2 PureStem Line SM30
• HOX- • PITX1+ • ZIC2+ • TBX15-
35 PureStem Lines SK11 AND SM30
PITX1 ZIC2 TBX15 SK11 + + + SM30 +PITX1 + ZIC2 TBX15 -
Genepaint.org Diverse Osteochondral hEP Cell Lines Diverse Osteochondral hEP Cell Lines Diverse Osteochondral hEP Cell Lines
E15 Diversity of Defined Osteochondral Cells
40 Limb Bud Markers
We are beginning to understand limb morphogenesis as controlled by site- specific homeobox genes and gradients from morphogen fields. Such lines, if capable of osteochondral differentiation, could have the advantages of homologous use (i.e. not putting mandibular mesenchyme in the knee). First, limb buds are an outgrowth from lateral plate mesoderm (LPM), specifically the somatic mesoderm).
Splanchnic Mesoderm
Lateral Plate Mesoderm
Somatic Mesoderm
41 Limb Bud Markers
The nomenclature is as follows:
42 Limb Bud Markers
Distal LPM, perhaps from distal HOX genes, is thought to trigger PITX1, a dominant factor for conversion of limb bud to hindlimb. PITX1 is also expressed in mandibular mesenchyme as shown below for PureStem SK11 and SM30.
Mandibular Mesenchyme
Forelimb
Hindlimb
Taher L, Collette NM, Murugesh D, Maxwell E, Ovcharenko I, et al. (2011)
43 Limb Bud Markers
HOXB6 as a marker of committed limb bud mesenchyme
44 Limb Bud Markers
HOXA10 expression in limb mesenchyme
45 Limb Bud Markers
NHAC RTPEC
ASCs EN1 RA Series
SK17 MSCs B16 C4ELSR10 SK31 T14 4SKEL20
46 Limb Bud Markers
HOXA13 as a marker of distal limb mesenchyme
47 Limb Bud Markers
NHAC
C4ELSR10 Xgene EN1 RAD20.19
SK17 B16 SK31 T14 MSCs C4ELS5.1
48 Novel Osteogenic Lines E3 & E72
Embryonic Forelimb Mesenchyme (EFM) E3 E72
49 Novel Osteogenic Lines E72 & E75
Embryonic Forelimb Mesenchyme (EFM)
E3 EFM E3 EFM
50 Scalable Human Embryonic Limb Cells
Upper Limb E72 Lower Limb RAD20.5 HOXA10+ HOXA10+ HOXD11 HOXB6+ PITX1- PITX1+
51 Scalable Human Embryonic Limb Cells
Iliac MSCs Lower Limb RAD20.5 HOXA10+ HOXA10+ HOXB6+ HOXB6+ PITX1+ PITX1+ CD74+ CD74-
52 PureStem E69 and T42
53 Potential Uses of PureStem E69 & T42
E69
T42
54 Potential Uses of PureStem E69 & T42
CYP26B1 TTR Basio-Occipital Bone
Choroid Plexus Meninges Of 4th Ventricle Of Upper Medulla
In situ images from Genepaint.org 55 Storage Strategies
Cells can be differentiated and frozen in HyStem
56 LifeMap
57 Orthopedic Applications
> 30 distinct osteochondro-progenitors
BACK PAIN
Degenerative disc disease: Disc repair/regeneration
Spinal fusion: Bone induction Back Pain DDD
KNEE PAIN:
Cartilage injuries, bone defects, osteoarthritis
Meniscus, ligament, Tendon Knee Pain Traumatic injury 58 PureStem Manufacturing Technology
Human embryonic progenitor (hEP) cell lines: > 200 diverse cell types isolated
• Reproducible product Kidney • Purity and identity • Can differ from MSCs in Smooth Muscle Skeletal Muscle being non-hypertrophic • Potential for permanent Blood/Brain engraftment Barrier • Improved survival and translatability with HyStem
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