How to Make a Heart:Towards Pluripotent Cell-Based Heart Stem Cell Therapeutics

ACI 2011 BCIS Meeting

London Hilton MetropoleLondon, UK

January 28, 2011

Kenneth R. Chien MD, PhD

MGH Cardiovascular Research Center Department of Stem Cell and Regenerative

BiologyHarvard University

Harvard Stem Cell Institute

• Islet-1 heart progenitor story

• Heart Parts: Engineering patch of mature heart muscle tissue from ES cells

• Human master Islet-1 progenitors: Human cardiogenesis, congenital heart disease , and regenerative therapeutics

• Laser technology devices for cell based therapy

2

The Unmet Clinical Need: Growing Heart Muscle

22 57

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2,107 2,1632,199

2,363

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500

1,000

1,500

2,000

2,500

75 80 85 90 95 00 08

Years

Num

ber o

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ants

Hansson, Lindsay, and Chien, Cell Stem Cell, In Press, September, 2009

Regeneration Next: Towards Heart Progenitor Derived Heart Parts

The Meltonian Stem Cell Era: Directed Differentiation of Pluripotent Stem Cells

J Embryol Exp Morphol. 1985

The in vitro development of blastocyst-derived embryonic stem cell lines: formation of visceral yolk sac, blood islands and myocardium.Doetschman TC, Eistetter H, Katz M, Schmidt W, Kemler R.

• Diversification• Scalability• Maturation• Coordination• Durability

5

How to Make a Heart

ES/iPS Pluripotent Stem Cells

Committed Ventricular Progenitor

Differentiated Ventricular Cell

Mature Ventriuclar Cell

Durable VentricularMuscle Tissue

Multipotent Iwlet Heart Progenitor

Ventricular Cardiac Muscle Cell Fate Map:Towards Human Heart Stem Cell Models and Therapeutics

Laugwitz et al, Nature, 2005 Moretti et al Cell, 2006 Qyang et al Cell Stem Cell, 2007 Martin-Puig et al, Cell Stem Cell, 2008 Chien, Nature, 2008 Chien, et al, Science, 2008 Bu et al, Nature, 2009 Domian et al, Science, 2009 Hansson et al, Cell Stem Cell, 2009

Mesodermal Precursor

Multipotent Isl1 Cardiovascular Progenitor

BipotentProgenitor

Differentiated Progeny

Endothelial Cell

Smooth Muscle Cell

Right Ventricular

Cardiomyocyte

AtrialCardiomyocyte

Smooth Muscle Cell

Isl-1Flk-1Nkx2.5

Isl-1Nkx2.5

BryFlk-1

Cardiac Mesodermal Precursor

Primary Heart FieldCardiovascular Progenitor

Nkx2.5Others ?

Left Ventricular Cardiomyocyte

Mlc2-vNkx2.5MHC

Mlc2-vMHC

SLNMHC

CD31vWf

SM-MHC

SM-MHC

Bry

Wnt/-cat

Wnt/-cat

Common PrimordialCardiovascular Progenitor

?

Atrioventricular Nodal Cell

Sinoatrial Nodal Cell

§

Towards defining the heart cell lineage map

?

Coordinated Contractility of a Syncitium of Murine ES cell

derived Islet-1 Progenitors Driven into the Cardiac Lineage

Cardiac Progenitor Cells Form a Thin Film of Beating Cardiac Muscle

Making a Ventricular Heart Muscle Patch:Towards Regenerative Cardiovascular Medicine

Domian et al, Science, 2009

10

Islet Heart Progenitors in the Human Heart

Hansson, Lindsay, and Chien, Cell Stem Cell, October, 2009 Bu et al, Nature, 2009

11

Master Human Islet Heart Progenitors from ES Cells Can Make Any Cardiovascular Heart Cell Type

Master ISL1 heart progenitors

A Stem Cell Paradigm for Human Cardiogenesis & Congenital Heart Disease

Hansson, Lindsay, and Chien, Cell Stem Cell, October, 2009

• TBX1• NKX2.5• TBX20

Malformation of several organs, including the cardiovascular system

- arterial tortuosity - diffuse aneurysms - rupture / death young age - congenital heart disease

(PDA, BAV, ASD)

Caused by mutations in TGFRB1 (ALK5) or TGFRB2

Loeys et al., Nature Genetics, 2005

Loeys-Dietz syndrome

iPS derivation EB differentiation

+ / - Alk5 inhibitorPlate & culture underISL1+ promoting conditions

LD syndrome iPS cells display a >4 fold increase in the induction of ISL+ progenitors

+/- Alk5 inhibitor vs control

Analysis of SHF progenitor cells carryingTGFBR mutations causing Loeys-Dietz syndrome

Mesodermal precursors

Multipotent Isl1+ cardiovascular

progenitors

Intermediates committed

to CV lineages

Cardiac mesenchymal cells

prespecification

Islet/Wnt3a

Wnt3a

differentiation

Endothelial cells

Cardiomyocytes

Smooth muscle

TGF-beta

• Diversification (Islet)• Scalability (Wnt)• Maturation (TGF beta Pathway)• Coordination (?)

A Islet Progenitor Fate Map for Regenerative Cardiovascular Medicine

-

CM Progenitors

?

Heart Muscle Tissue Engineering

In vivo delivery

Vascularization

Grafting/Rejection

Side effects/arrhythmogenesis

Efficacy endpoints

The “Apollo” Mission of Cardiac Regenerative Medicine

General Pathway for Cell-based Regenerative Medicine

Lasers in MedicineOphthalmology

Plastic Surgery

Dermatology

Cardiology

Oncology

Radiology

Vascular Medicine

Advantages of Lasers in Medicine

Finely focused laser beams offer precision over scalpels

Hemostasis Miniaturization of laser Adaptability to different needs

Many types of lasers with properties tunable to specific needs

Liver

Day 0 Day 1

Liver

Day 4Day 7

Skeletal muscle regeneration

• Lasers can create finely focused areas of injury

• Skeletal muscle regenerates quickly with a minimum of fibrosis

CD-1 Mice

Human, Human, and Human: Towards Human Heart Parts and Heart Models

• The checkpoints that control the expansion of ventricular progenitors?

• Promoting the in vivo survival and grafting of CVP derived heart patches and in the in vivo context

• In vivo delivery of CVPs—miniaturized endoscopic delivery systems

• Engineering other “Heart Parts”---Coronary Arteriogenesis

• Drug discovery/safety in human models systems

•CHD and adult monogenic CV human disease models