For postingNJ SYMP BIOMATLS

DRAFTAI CAPLAN

panel presentation

Adult Mesenchymal Stem Cells (MSCs) in Tissue Engineering

and Inflammation.

October 28,2010NJ SYMP on BIOMATERIALS SCIENCE

ARNOLD I. CAPLAN, PhDSKELETAL RESEACH CENTER

CASE WESTERN RESERVE UNIVERSITY

Disclosure Information.

Founder and was an officer of OSIRIS THERAPUETIC,Inc.

Current status: DIVORCED,NO stock, not a consultant but I do receive royalties thru CWRU.

OTHER CONSULTING activity is unrelated to this lecture.

The SCIENCE and CLINICAL USE of Adult

MSCs:from bench to bedside &

back to the lab.

Adult BONE MARROW

HSC MSCMesenchymal Stem Cells

1988

MSC

Pericytes: cells on capillaries and microvessels.

modified byBRUNO PEAULT

from http://www.geocities.c

o.jp/HeartLand-

Suzuran/9389/kekkan

ALL MSCs are PERICYTES!

INJURY

PERICYTE MSC ACTIVATED

MSCREGENERATIVE

MSC

PROPOSED SEQUENCE OF CHANGE

DUE TO INJURY:

2010

IMMUNO-MODULATORY

Anti-Apoptotic

Anti-Scarring

Angiogenic

Mitotic

MSC=pericyte

natural INJURY RESPONSE

Regenerative Micro-environment

T-cells, B-cells, Dendritic cells,

T-regs,etc

2010

Trophic

MSC = MedicinalSignaling

Cell.(the injury-specific DRUG STORE)

Muscle DerivedStem Cells

AdultMulti-lineage

Stem Cells

Fat Derived Stem Cells

MSC

BRUNO PEAULT

MSC MAPC

Cultured Pericytes Express Markers of hMSCs.

MSCs

Pericytes M Crisan et al, 2008

Adult MSCsHYPOTHESIS:

ALL MSCs are PERICYTESand are, thus, found thru-out the

body.

They function to stabilize blood vessels.They also function at sites of local tissue

damage to facilitate TISSUE REGENERATION.

What do Blood Vessels of different tissues in the organism

have in common?

Perivascular cells

Endothelial cells

ALL BLOOD VESSEL CELLS REFLECT THE TISSUE IN

WHICH THEY FUNCTION. THUS, SINCE EACH TISSUE IS

DIFFERENT, THE CELLS OF THE BLOOD VESSELS ARE

DISTINCTLY DIFFERENT.

11/2/2010

Dr. Bill Futrell, et al – Univ of Pittsburgh 1998“Discovered stem cells in fat that could

create new tissues like bone and cartilage”

Enabling Innovation:

Stem Cells from fat Extracted cells go to bone in vitro Fix fractures

Adipose derived adult Stem Cells(ASCs).

• Adipose tissue is a potential source for stem cells (ASCs) derived from pericytes.

• ~400,000 cells/mL of tissue.• 300-500-fold more cells in fat compared to

the same volume of bone marrow aspirate.• Marrow MSCs require exposure to TGF-β

to be chondrocytes; ASCs require both TGF-β and BMP-6 to be chondrocytes. Therefore, marrow MSCS ≠ ASCs.

Gimble et al. 2003; Guilak et al. 2004; Aust et al. 2004, Oedayrajsingh-Varma et al., 2006.

ALL blood vessels are not equal.Marrow MSCs are not fat MSCs.Marrow MSCs are not muscle MSCs.Marrow MSCs are not pulp MSCs.Marrow MSCs are not X MSCs.Marrow MSCs are not Y MSCs.Marrow MSCs are not Z MSCs.

Centrifugation

Passage Culture

Bone Marrow Aspirate

Adhere to Culture Dish

Primary MSC Culture

ColonyFormation

Plate cells at

Interface

Density Solution

Serum batch dependant

CFU-F1988

hMSCs

Chondrogenesis

ITS+dexamethasone

TGF-βascorbate 2-P

Osteogenesis10% FBS

dexamethasoneascorbate 2-P

β-GP

Adipogenesis

10% FBSdexamethasone

insulinIBMX

indomethacin

MULTI-POTENCY:

For TISSUE ENGINEERING uses.

Aspirate bone marrow

Cartilage Bone Tendon / Ligament

Isolate and culture-expand MSCs

Cryopreservefor future use

Fat

MSCs

TISSUE ENGINEERING

SCAFFOLD

1992, CWRU TRANSFERS MSC TECHNOLOGY TO FORM:

AIC separated in 1997. IPO 2006.

MSCs BIOACTIVE FACTORS

IMMUNO-SUPRESSIVE

TISSUES

REGENERATE

TURNOVER/ MAINTAINANCE

MICROENVIRONMENT/ MILIEU

Tissue Engineering

ALLOGENIC hMSCs: The Business of Today, 2010.

Business Model:allogenic hMSCs.

Donor bone marrow aspirate

Initial isolation and expansion in GMP

process

One Donation

Process Control Testing on

Intermediate

10,000 Doses of Final Product

Osiris Pipeline in Cell Therapy: 2010

GENZYME Corp=WORLDWIDE PARTNER

MSC CLINICAL TRIALS:2009: NIH website

85 WORLDWIDE36 in USA

26 in EUROPE23 in Asia ,

South and Central America, Australia,etc.98 in 2010

200 centers worldwide4 indications in Phase III • 3 with Fast Track

Worldwide Reach

SAFETY and

REGULATORY

Osiris Pipeline in Cell Therapy:

PROCHYMALGVHD Compassionate use:

Now(5/2008) APPROVED for PAYMENT & USE.

MULTIPLE Prochymal infusions(3-21) in 12 pediatric patients (5months to 15 years of age) suffering from treatment resistant, severe( grades

III/IV), end-stage GVHD:•All patients(12/12) showed a clinical response to

therapy with 58%(7/12) achieving COMPLETEresolution of GVHD. Less than 15% who fail

immunosuppression survive to 100days.•Clinical improvements in GI GVHD(75% COMPLETE

REMISSION and 3 with regression to grade I).No infusion toxicity or adverse events.

Dec.10, 2007

allo-MSCs for steroid-resistant, severe, acute GvHD(phaseII):

Katarina LeBlanc et al (Lancet 371:1579-86, 2008)55 patients(30 complete response,9 improved).Dose hMSCs:1.4x106 cells/kg (0.4-9).

27patients(1 dose), 22(2 does), 6(3-5 doses).HLA-identical siblings (=5).Haplo-identical donors (n=18).3rd party HLA-mismatch (=69).POSITIVE EFFECTS IRRESPECTIVE OF allo-DONOR.No side-effects (during or after), Lower transplant-related mortality(1yr), Higher overall survival@2yrs.

MSC

S S Iyer & M Rojas: Expert Opin. Biol. Ther. (2008) 8:569-581.

Immuno-modulation

Osiris Pipeline in Cell Therapy

Efficacy Results– Every patient evaluated had a reduction in disease severity

by day 28

– 105 point improvement in CDAI (p=0.004)

Safety Results– No attributed SAE’s– Well tolerated– Outpatient administration (20-70 min)

Phase II :Clinical Evaluation Prochymal in Treatment-Resistant Crohn’s Disease.

Trial Design— 10 patient, prospective, randomized, open label

trial— Average CDAI was 350— Average length of disease was 14 years— Failed previous treatment with steroids,

methotrexate, and Remicade

Osiris Pipeline in Cell Therapy:

Preclinical data

Swine Ischemia / Reperfusion Model:

TROPHICCAPLAN: CELL PRODUCED,

BIOACTIVE FACTOR MEDIATED:1. ANTI-APOPTOTIC/CYTOPROTECTIVE>

Limits field of ISCHEMIC injury

2. ANTI-FIBROTICANTI-SCARRING

3. ANGIOGENIC> VEGF+ PERICYTE

4. MITOTIC/Regenerative Milieu

PROCHYMALPHASE I: Safety study.

53 patient, double-blind, placebo-controlled and preliminary efficacy of intravenous administration.

• Arrhythmias, 4X less (9% vs 37%,p=.025).• PVCs, premature ventricular contractions >10/hr ,fewer at all

time points,1-6 month (11% vs 24%,p<.001).• Prompt heart rate recovery,<15 min for 6 min walk,

(55% vs 26%, p=.08).• Lung function improved, FEV% predicted values,

(17 point vs 6 point, p<.05).• R. Mills: ”The sicker the patient the better the result”.

MICHAEL CHOPP

MICHAEL CHOPP

MSCs and MSin collaboration

withRobert Miller et al:

Immuno-suppressive & trophic.

MSCs and progression of demyelinating disease:

The EAE-Model.Inject myelin proteins MOG or PLP

to induce disease.

10-20 days: Inject human Mesenchymal Stem Cells.

Do hMSCs alter disease progression?

hMSCs

0 5 10 15 20 25 30 35 40 45 500

1

2

3

4

5 EAE+PBS

EAE+hMSCsC

linic

al s

core

s of

EAE

hMSCs

C57/BL/6J mice/MOG35-55

IMMMUNO-MODULATORY

and TROPHIC

MSC-mediated:1. STROMA: Bone marrow transplant.

2. HEART: Infarct (ischemia).3. BRAIN: Stroke (ischemia).4. SPINAL CORD: Axonal regeneration.

5. LUNG: Asthma,inflammation.6. GI: Inflammatory Bowel Disease( Crohn’s ).

7. KIDNEY: Acute Renal Failure(ischemia).

8. MENISCUS: Tissue Regeneration,OA.

9. TENDON: Horses(Vet-Stem).10. Diabetics: hMSCs into mice.

MSC-mediated:

11. Rheumatoid Arthritus12. SLE13. ALS14. MS15. HIV / AIDS?16. Autism?17. OTHER (poly-trauma)???????

MSCTROPHIC BIOACTIVE FACTORS

IMMUNO-MODULATORY

Anti-Apoptotic

Anti-Scarring

Angiogenic

Mitotic

MSC=pericyte

natural INJURY RESPONSE

Regenerative Micro-environment

T-cells, B-cells, Dendritic cells,

T-regs,etc

2010

Trophic

Mesenchymal Stem Cell (MSC)

SkeletalResearch Center

College of Arts and Sciences

Cleveland,Ohio