The Role of EPC in Old Age and Atherosclerosis

11Seminar ASPI 25 Oct.2008Seminar ASPI 25 Oct.2008

The Role of EPC in Old The Role of EPC in Old Age and AtherosclerosisAge and Atherosclerosis

Dr.Boenjamin Setiawan, Ph.D.Dr.Boenjamin Setiawan, Ph.D.

25 October 200825 October 2008

Seminar ASPI 25 Oct.2008Seminar ASPI 25 Oct.2008 22

Number of deaths for leading causes of death, USA, 2005: Heart disease: 652,091 Cancer: 559,312 Stroke (cerebrovascular diseases): 143,579 Chronic lower respiratory diseases: 130,933 Accidents (unintentional injuries): 117,809 Diabetes: 75,119 Alzheimer's disease: 71,599 Influenza/Pneumonia: 63,001 Nephritis, nephrotic syndrome, and nephrosis: 43,901 Septicemia: 34,136

TOTAL1.891.470


TEN LEADING CAUSES OF DEATH IN THE U.S.A., 2004TEN LEADING CAUSES OF DEATH IN THE U.S.A., 2004Heart disease is the leading cause of death of Americans 24 years of age and older, 30.3% or Heart disease is the leading cause of death of Americans 24 years of age and older, 30.3% or 724,269 annual deaths are due to heart disease. Sixteen percent of those deaths, 115,883, occur 724,269 annual deaths are due to heart disease. Sixteen percent of those deaths, 115,883, occur in smokers. Cancer is the second leading cause of death causing 23% or 538,947 deaths. in smokers. Cancer is the second leading cause of death causing 23% or 538,947 deaths. Twenty-three percent of those are lung cancer, 83% of lung cancers occur in smokers andTwenty-three percent of those are lung cancer, 83% of lung cancers occur in smokers and

exsmokers (101, 724).exsmokers (101, 724).


Ten leading causes of death, all Ten leading causes of death, all WHO member states, 2000WHO member states, 2000TEN LEADING CAUSES OF DEATH, ALL WHO MEMBER STATES, 2000CAUSE PERCENT OF TOTAL

(1) Ischemic Heart Disease 12.4%

(2) Cerebrovascular Disease 9.2%

(3) Lower Respiratory Infections 6.9%

(4) HIV/AIDS 5.3%

(5) Chronic Obstructive Pulmonary Disease 4.5%

(6) Perinatal conditions 4.4%

(7) Diarrhoeal Diseases 3.8%

(8) Tuberculosis 3.0%

(9) Road Traffic Injuries 2.3%

(10) Trachea, Bronchus, Lung Cancers 2.2%


What is a stem cell?(Sources *STEM CELLS: A PRIMER; *STEM CELLS: NIH)A stem cell is a cell with the ability to self-replicate throughout the life of an organism and can differentiate or give rise to many cell types upon stimulation and given the right conditions. Stem cells have the potential to develop into functioning mature cells characteristic of different tissues or organs. In contrast, most cells in the body are already differentiated, have limited lifespans and cannot give rise to other types of cells


Embryonic stem cells are derived Embryonic stem cells are derived from the Inner Cell Massfrom the Inner Cell Mass


Multiple embryos are produced Multiple embryos are produced in vitroin vitro for clinical for clinical purposes.purposes.Donated surplus embryos are used as a source of ES Donated surplus embryos are used as a source of ES cells.cells.These surplus embryos would otherwise be discarded or These surplus embryos would otherwise be discarded or remain unusedremain unused

How are ES Cells Generated?


Fertilized Embryos Are Allowed to Divide Fertilized Embryos Are Allowed to Divide Several Times in CultureSeveral Times in Culture


Harvesting ES CellsHarvesting ES Cells

The blastocyst is the stage of development (~1,000 cells) at which the embryos inner cell mass (ICM) forms. It is the ICM that harbors ES cells. Stem cells exist only fleetingly at this stage of development


Generating an ES Cell LineGenerating an ES Cell Line

The trophoblast is removed from the embryo, the inner cell mass is isolated and, using a micropipette, ES cells are extracted and placed in tissue culture. These cells are passaged for many generations until some spontaneously form “immortal” lines


Stem Cells Stem Cells are taken are taken from the from the

Inner Cell Inner Cell Mass at Mass at

the the blastocyst blastocyst stage and stage and cultured in cultured in a Petri diska Petri disk





Structure of Animal CellStructure of Animal Cell

Seminar ASPI 25 Oct.2008Seminar ASPI 25 Oct.2008 1616seminar Active Aging 17 Juli seminar Active Aging 17 Juli 20082008 dr.Boenjamin Setiawan, Ph.D.dr.Boenjamin Setiawan, Ph.D. 1616


The Human BodyThe Human Body

• Consist of 12 Organ Systems

• Consist of 220 types of Tissues

• Consist of 50-100 trillion cells

• Consist of 100 billion neurons

• Consist of 46 chromosomes (22 pairs and x and y chromosomes)

• Consist of 30.000 genes

• Consist of 3,2 billion base pairs


• The cardiovascular circulatory system’s main role is to transport nutrients and oxygen to the 50-100 trillion cells that makes up our body and remove the waste material from the body.

• The most prevalent disease that affects the CVS is:

1. Hypertension

2. Acute myocardial infarction

3. Stroke




World’s oldest person celebrates World’s oldest person celebrates 129129thth birthday, died March 2007 birthday, died March 2007

According to national records, Hernandez was born on May 3, 1878, in one of the country's central provinces, where she gave birth to 13 children. She now has 60 grandchildren, 80 great-grandchildren and 25 great-great grandchildren.

2222Seminar ASPI 25 Oct.2008Seminar ASPI 25 Oct.2008

FIGURE 1. Tissue heterogeneity and FIGURE 1. Tissue heterogeneity and stem-cell functionality for homeostasis stem-cell functionality for homeostasis

and repairand repair


FIGURE 2. Variation of maximal lifespan across species.FIGURE 2. Variation of maximal lifespan across species.From the following article:From the following article:

Stem cells, ageing and the quest for immortalityStem cells, ageing and the quest for immortalityThomas A. RandoThomas A. Rando

Nature 441, 1080-1086 (29 June 2006) Nature 441, 1080-1086 (29 June 2006) doi:10.1038/nature04958doi:10.1038/nature04958


Stem cells, ageing and the quest for immortalityStem cells, ageing and the quest for immortalityThomas A. RandoThomas A. Rando

Influence on Stem Cell FunctionalityInfluence on Stem Cell Functionality


FIGURE 4. Ageing of stem-cell FIGURE 4. Ageing of stem-cell functionality functionality


((Circulation.Circulation. 2007;116:3-5.) 2007;116:3-5.)

© 2007 American Heart Association© 2007 American Heart Association, Inc. , Inc.

• Cardiovascular Biomarkers• Added Value With an Integrated

Approach?

• Wolfgang Koenig, MD, FRCP, FESC From the Department of Internal Medicine II, Cardiology, University of Ulm Medical Center, Ulm, Germany.



Figure 1. Origin and differentiation of Figure 1. Origin and differentiation of endothelial progenitor cellsendothelial progenitor cells. .

Scheme depicts the potential origin and differentiation of endothelial progenitor Scheme depicts the potential origin and differentiation of endothelial progenitor


((Circulation.Circulation. 2007;115:1285-1295.) 2007;115:1285-1295.)© 2007 American Heart Association, Inc.© 2007 American Heart Association, Inc.

Contemporary Reviews in Cardiovascular Medicine

Endothelial Function and Dysfunction

Testing and Clinical Relevance John E. Deanfield, MB, BCh, FRCP; Julian P. Halcox, MD, MA, MRCP; Ton J. Rabelink, MD, PhD From the Vascular Physiology Unit (J.E.D., J.P.H.), UCL Institute of Child Health, London, UK; and the Department of Nephrology (T.R.), Leiden University Medical Center, Leiden, The Netherlands.


• EPC facilitates angiogenesis

• EPC mobilization and Migration is stimulated by VEGF, Angiopoetin-1, FGF, GM-CSF, MMP-9 and Pharmacological Agents, (Statins)



• Figure 1. Left, The quiescent state of the endothelium, where NO (green circles) is generated by the endothelial isoform of nitric oxide synthase (eNOS) in its membrane-bound configuration. The released NO targets cysteine groups in key regulator molecules such as NF B (p50/p65) and the mitochondria, which leads to silencing of cellular processes. Right, The state of endothelial activation where reactive oxygen signaling (red circles) predominates. The ROS such as H2O2 are generated from oxidases as well as the uncoupled state of eNOS. Like NO, the molecules target key regulatory proteins, such as NF B and phosphatases, which leads to activation of the endothelial cells. Such activation can occur physiologically in the context of host defense or pathophysiologically in the presence of cardiovascular risk factors. EC indicates endothelial cells


Figure 2. Sustained ROS signaling induces senescence of endothelial Figure 2. Sustained ROS signaling induces senescence of endothelial cells. Left, This is reflected in detachment of endothelial cells or parts of cells. Left, This is reflected in detachment of endothelial cells or parts of the endothelial cell membrane (endothelial microparticles). Right, With the endothelial cell membrane (endothelial microparticles). Right, With increasing age and persisting ROS signaling, the capacity of neighboring increasing age and persisting ROS signaling, the capacity of neighboring endothelial cells to repair the endothelial injury is limited, and vascular endothelial cells to repair the endothelial injury is limited, and vascular

integrity becomes dependent on the incorporation of circulating PCintegrity becomes dependent on the incorporation of circulating PC








• Major Role: The main role of the nervous system is to relay electrical signals through the body. The nervous system directs behaviour and movement and, along with the endocrine system, controls physiological processes such as digestion, circulation, etc.

• The most prevalent diseases that affect the CNS is Stroke, Parkinson’s disease, Alzheimer’s disease, ALS and Autism


• The brain microvascular endothelium constitutes a major exchange area between the blood and the brain: it forms a physiological barrier, known as the blood-brain barrier (BBB), which plays a central role in maintaining cerebral homeostasis.


Spinal Cord InjuriesSpinal Cord InjuriesHwang Mi-Soon: South Korea

Paralyzed 19 years

Multipotent adult stem cells injected into her spinal cord

Currently: debilitating pain

Published in 2005 (Cytotherapy)


J Am Coll Cardiol, 2003; 42:2081-2082, J Am Coll Cardiol, 2003; 42:2081-2082, doi:10.1016/j.jacc.2003.09.016doi:10.1016/j.jacc.2003.09.016

© 2003 by the American College of Cardiology Foundation© 2003 by the American College of Cardiology Foundation

EDITORIAL COMMENT

Aging of progenitor cells: limitation for regenerative capacity? Stefanie Dimmeler, PhD ,* and Mariuca Vasa-Nicotera, MD Molecular Cardiology, Department of Internal Medicine IV, University of Frankfurt, Frankfurt, Germany Department of Cardiovascular Science, Division of Cardiology, University of Leicester, Leicester, United Kingdom


• Journal of Molecular Medicine© Springer-Verlag 200410.1007/s00109-004-0580-xReview

• Vascular repair by circulating endothelial progenitor cells: the missing link in atherosclerosis?

• Stefanie Dimmeler1 and Andreas M. Zeiher1• (1) Molecular Cardiology, Department of Internal

Medicine IV, University of Frankfurt, Theodor-Stern-Kai 7, 60590 Frankfurt, Germany


Figure 1 Possible influences of age on endothelial Figure 1 Possible influences of age on endothelial progenitor levels. VEGF = vascular endothelial progenitor levels. VEGF = vascular endothelial

growth factor.growth factor.


Fig. 1 Regeneration of the Fig. 1 Regeneration of the endothelial monolayer after endothelial monolayer after injury. After induction of injury. After induction of endothelial injury (endothelial injury (11) two ) two possibilities exist to possibilities exist to regenerate the injured regenerate the injured endothelial monolayer (endothelial monolayer (22,, 33).). 2 2 Regeneration by Regeneration by mature endothelial cells mature endothelial cells which migrate and which migrate and proliferate to regenerate proliferate to regenerate the endothelial layer.the endothelial layer. 3 3 Regeneration by bone Regeneration by bone marrow derived endothelialmarrow derived endothelial

progenitor cellsprogenitor cells



Fig. 3 Possible targets of risk factors for coronary artery disease to reduce circulating Fig. 3 Possible targets of risk factors for coronary artery disease to reduce circulating endothelial progenitor cells (endothelial progenitor cells (11––44). Risk factors for coronary artery disease may interfere with ). Risk factors for coronary artery disease may interfere with hematopoietic stem cells in the bone marrow, reducing mobilization or affecting survival and hematopoietic stem cells in the bone marrow, reducing mobilization or affecting survival and differentiation of circulating progenitor cells. Additionally, risk factors such as age may also differentiation of circulating progenitor cells. Additionally, risk factors such as age may also

reduce homing by reducing stimulatory factors such as VEGF and the block receptor reduce homing by reducing stimulatory factors such as VEGF and the block receptor independent kinasesindependent kinases



Competent Marrow compared to Competent Marrow compared to Senescent MarrowSenescent Marrow


Cardiovascular Research 2007 73(2):326-340; Cardiovascular Research 2007 73(2):326-340; doi:10.1016/j.cardiores.2006.06.030doi:10.1016/j.cardiores.2006.06.030 • Nutrition, physical activity, and

cardiovascular disease: An update• Louis J. Ignarroa, Maria Luisa Balestrierib and Claudio

Napolic,* aDepartment of Molecular Pharmacology, David Geffen School of Medicine, UCLA, Los Angeles, CA, United StatesbDepartment of Chemical Biology and Physics, Division of Clinical Pathology and Excellence Research Center on Cardiovascular Diseases, Complesso S. Andrea delle Dame, 1st School of Medicine, II University of Naples, Naples 80138 ItalycDepartment of General Pathology, Division of Clinical Pathology and Excellence Research Center on Cardiovascular Diseases, Complesso S. Andrea delle Dame, 1st School of Medicine, II University of Naples, Naples 80138 Italy

• * Corresponding author. Tel./fax: +39 081 293399. Email address: [email protected]


Fig. 2 Different effects of physical exercise on redox status of Fig. 2 Different effects of physical exercise on redox status of

the cell. Abbreviations: ROS, reactive oxygen species. the cell. Abbreviations: ROS, reactive oxygen species.


Cardiovascular Research Advance Access originally Cardiovascular Research Advance Access originally published online on March 18, 2008 published online on March 18, 2008 Cardiovascular Research 2008 78(3):413-421; Cardiovascular Research 2008 78(3):413-421; doi:10.1093/cvr/cvn081 doi:10.1093/cvr/cvn081

• Vascular repair by endothelial progenitor cells

• Anna Zampetaki, John Paul Kirton and Qingbo Xu* Cardiovascular Division, King's College London, 125 Cold harbour Lane, London SE5 9NU, UK

• * Corresponding author. Tel: +44 20 7848 5322; fax: +44 20 7848 5296. E-mail address: [email protected]


Figure 1 EPC origins. Figure 1 EPC origins. EPCs could be EPCs could be released from bone released from bone marrow, fat tissues, marrow, fat tissues, vessel wall, vessel wall, especially adventitia especially adventitia and spleen, liver, and and spleen, liver, and intestine, where they intestine, where they form a circulating form a circulating EPC pool. They can EPC pool. They can then contribute to then contribute to the repair of the repair of damaged vessels in damaged vessels in pathological pathological

conditions.conditions.


Figure 3 EPC Figure 3 EPC mobilization and mobilization and differentiation.differentiation. Schematic graphics Schematic graphics summarize cytokines, summarize cytokines, growth factors, and other growth factors, and other mediators inducing EPC mediators inducing EPC release from tissues and release from tissues and homing to the surface of homing to the surface of arteries where endothelial arteries where endothelial cells are damaged. After cells are damaged. After attachment to the vessel attachment to the vessel wall, EPCs differentiate wall, EPCs differentiate into endothelial cells into endothelial cells induced by the micro-induced by the micro-

environment.environment.




• EPC facilitates angiogenesis

• EPC mobilization and Migration is stimulated by VEGF, Angiopoetin-1, FGF, GM-CSF, MMP-9 and Pharmacological Agents, (Statins)



Therapeutic Potential of EPC in AMITherapeutic Potential of EPC in AMI

• Figure 2. Therapeutic potential of EPCs. Intracoronary infusion of EPCs in patients with AMI may have the potential to restore myocardial function to the damaged area.

• EPC transplantation increases left ventricular ejection fraction (LVEF), coronary blood flow reserve, and myocardial viability and decreases left ventricular end-systolic volume (LVESV).66


How can we slow down How can we slow down Atherosclerosis and old age??Atherosclerosis and old age??

1. You are what you eat. (CRAN) Consume a healthy diet, Fruits, Vegetables, Fish, lean meat, etc.

2. Healthy LIFE-STYLE, no smoking, no alcohol, no stress, regular exercise, keep mentally active, yoga, meditation etc

3. Prevent atherosclerosis with statins, control hypertension

4. Take ANTI-OXIDANTS (Glisodin) regularly

5. Hypothetically: Infuse YOUNG, HEALTHY EPC regularly to keep the endothelial lining clean


Future PerspectivesFuture Perspectives• EPCs have been extensively studied in

cardiovascular diseases and accumulating evidence highlights their importance in vascular repair and tissue remodelling.

• Despite these encouraging results, there are also several reports on adverse effects in various animal models and there are still many issues concerning the biology of the EPCs that need to be addressed. Numerous studies have underlined the need of increasing EPC numbers to allow sufficient neovascularization and recovery of the ischaemic tissue. Thus, identifying other sources of stem cells and understanding the molecular mechanisms involved in the mobilization, differentiation, and migration of these cells is essential to design effective therapeutic strategies.



Unresolved Questions

1. How to define an endothelial progenitor cell? 2. Origin of endothelial progenitor cells? 3. Definition of subpopulations with different functional

capacities?4. Signals for EPC homing and differentiation in vivo? 5. Optimization of ex vivo culture conditions to enhance the

benefit of cell therapy? 6. Influence of the severity of vascular damage on the

contribution of EPCs to regeneration?7. Mechanisms of action? 8. Transdifferentiation capacity of different progenitor

cells? 9. Importance of paracrine effects?10. Effects of OLD AGE on functionality of EPC


SummarySummary1. Atherosclerosis causes Hypertension, AMI and Stroke.2. Atherosclerosis is still highly prevalent and cost a few

hundred billions dollars to society.3. Atherosclerosis can be prevented by diet, regular

exercise, healthy life style, drugs (statins, estrogens, anti-oxidants, Glisodin etc.) and EPC

4. Glisodin an anti-oxidant stimulating SOD dismutase can reduce atherosclerosis

5. Atherosclerosis can theoretically be prevented by the infusion of young, healthy EPC

6. Endothelial progenitor cells can be used to maintain the smoothness of the intima and prevent cardiovascular aging


Health & Medicine

The Role of EPC in Old Age and Atherosclerosis