［主催］公益財団法人先端医療振興財団 臨床研究情報センター（TRI）［共催］Caladrius Biosciences, Inc.
［協賛］ミルテニーバイオテク株式会社、日立化成株式会社［後援］文部科学省、厚生労働省、国立研究開発法人日本医療研究開発機構（AMED）

36

抄録集
37

Opening Remarks

Opening Remarks

Contents

Opening Remarks

Contents
Opening Remarks

Atsuhiko Kawamoto（Translational Research Informatics Center）

Atsuhiko Kawamoto (Translational Research Informatics Center)…………………………………………………………… 2
Session I

CD34+ Cell Therapy for Ischemic Tissue Repair: Two Decades of Evidence

Session I

Douglas W Losordo (Caladrius Biosciences, Inc.)……………………………………………………………………………………… 6
CD34+ Cell Therapy for Chronic Heart Failure: Clinical Experience in More than 300 Cases

Bojan Vrtovec (Ljubljana University Medical Centre)…………………………………………………………………………………… 8

Gustav Steinhoff (Rostock Medical University)………………………………………………………………………………………… 10
CD34+ stem cell therapy for Ischemic Stroke

Soma Banerjee (Imperial College Healthcare NHS Trust, UK)…………………………………………………………………… 12[Discussant]Session II

Nobuyuki Ohara (Kobe City Medical Center General Hospital)…………………………………………………………………… 14

Session II

CD34+ Cell Therapy for Critical Limb Ischemia

Atsuhiko Kawamoto (Translational Research Informatics Center)…………………………………………………………… 18
Highly Effective CD34 Cell Therapy on Critical Limb Ischemia in Hemodialysis Patients

Shuzo Kobayashi (Shonan Kamakura General Hospital)…………………………………………………………………………… 20
The use of adipose tissue stem cells in the treatment of patients with critical limb ischemia (CLI)

Andrei Kalinin (Central Clinical Hospital with Outpatient Health Centerof the Business Administration for the President of the Russian Federation)………………………………………………… 22

Session III

Stem cell therapy has been attracting attention for its regenerative potency. Bone marrow-derived CD34
antigen-positive (CD34+) cells are a well-characterized population of stem cells which have traditionallybeen used in clinic to reconstitute the hematopoietic system after radiation or chemotherapy. In 1997,
Asahara et al first identified human endothelial progenitor cells (vascular stem cells) in peripheral blood
CD34+ cell fraction. In animal models of ischemic vascular diseases, CD34+ cells induce therapeutic
angiogenesis by direct incorporation of the cells into the expanding vasculature and paracrine secretion ofangiogenic growth factors developing microvasculature. Recent research also revealed that enhancement of
microcirculation by CD34+ cell therapy contributes to tissue regeneration in non-vascular diseases.
Based on these preclinical achievements, phase I/II clinical trials have explored the usefulness of CD34+
cells in the treatment of patients with ischemic conditions such as peripheral, coronary and cerebral
artery diseases and those with non-vascular diseases including dilated cardiomyopathy, unhealing fracture(nonunion) and liver cirrhosis. Some of the clinical projects for CD34+ cell therapy are already at the pivotal
stage aiming pharmaceutical approval by global regulatory agencies.
This international symposium is held to introduce such clinical development of CD34+ cell therapy invarious fields of medicine. Distinguished researchers from USA, UK, Germany, Slovenia and Russia as wellas domestic renowned researchers will demonstrate the latest achievements and the future perspectivesof this therapeutic modality for intractable diseases. We hope all the participants would share theexact knowledge about this regenerative medicine product awaiting fast provision to the Japanese andinternational patients.

Induction of Cardiac Regeneration is Linked to a Response Signature of CD133 and SH2B3– Outcome Analysis of the Randomized Phase III PERFECT Trial

Session III

CD34+ Cell Therapy for Liver Cirrhosis

Toru Nakamura (Kurume University School of Medicine)………………………………………………………………………… 26
Autologous Transplantation of CD34+ Cells for Patients with Fracture Nonunion

Tomoyuki Matsumoto (Kobe University School of Medicine)………………………………………………………………… 26
Closing Remarks

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Closing Remarks

Masanori Fukushima (Translational Research Informatics Center)………………………………………………………… 30

Opening Remarks

International Symposium

7th World Centenarian Initiative

Clinical Application of

CD34-Positive Cells

Session I

for Cardiac, Cerebral, Vascular, Bone and Liver Regeneration

CD34+/CD133+ Cells for Cardiac and Cerebrovascular Diseases
Session I

CD34 陽性細胞が変える再生医療の世界
─治験実施による早期実用化を目指して
日時

場所

2017.9.9

13:00-18:00（Sat）（受付開始12:30）

虎ノ門ヒルズフォーラム 4F［Hall B］〒105-6305 東京都港区虎ノ門1-23-3（銀座線「虎ノ門駅」
1番出口 徒歩約5分）

13:00 - 13:05

Opening Remarks▲

川本 篤彦（臨床研究情報センター）

13:05 - 15:25

Session I

CD34陽性細胞、CD133陽性細胞による心血管・脳血管再生治療［座長］川本 篤彦（臨床研究情報センター）
▲

		

▲

CD34+ Cell Therapy for Chronic Heart Failure: Clinical Experience in More than 300 Cases
Bojan Vrtovec (Ljubljana University Medical Centre, Slovenia)

Session II

Moderator : Atsuhiko Kawamoto

CD34+ Cell Therapy for Ischemic Tissue Repair: Two Decades of Evidence
Douglas W Losordo (Caladrius Biosciences, Inc., USA)

(Translational Research Informatics Center)▲

Induction of Cardiac Regeneration is Linked to a Response Signature of CD133 and SH2B3– Outcome Analysis of the Randomized Phase III PERFECT Trial
Gustav Steinhoff (Rostock Medical University, Germany)▲

CD34+ stem cell therapy for Ischemic Stroke
Soma Banerjee (Imperial College Healthcare NHS Trust, UK)［Discussant］ 尾原 信行（神戸市立医療センター中央市民病院）

15:35 - 16:50

Session II

CD34陽性細胞、脂肪幹細胞による末梢血管再生治療［座長］横井 宏佳（福岡山王病院）▲

慢性重症下肢虚血に対するCD34陽性細胞治療

川本 篤彦（臨床研究情報センター）▲

血液透析患者における重症下肢虚血に対するCD34陽性細胞治療の高い有効性

Session III

小林 修三（湘南鎌倉総合病院）▲

The use of adipose tissue stem cells in the treatment of patients with critical limb ischemia (CLI)
Andrei Kalinin (Central Clinical Hospital of the Presidential Administration of Russia, Russia)

Session III

CD34陽性細胞による組織再生治療［座長］小林 修三（湘南鎌倉総合病院）

定▲中村 徹（久留米大学）▲

松本 知之（神戸大学）

日英同時通訳あり

Closing Remarks▲

福島 雅典（臨床研究情報センター）350名［参加無料］

偽関節患者に対する自家末梢血CD34陽性細胞移植

17:50 - 18:00

員〈事前登録が必要です〉

肝硬変に対するCD34陽性細胞療法※プログラムは予告なく変更となる場合がございます。

［主催］公益財団法人先端医療振興財団 臨床研究情報センター（TRI）［共催］Caladrius Biosciences, Inc.
［協賛］ミルテニーバイオテク株式会社、日立化成株式会社（AMED）［後援］文部科学省、厚生労働省、国立研究開発法人日本医療研究開発機構［お問い合わせ］

4

オンライン参加登録事務局

E-Mail： tri.workshop@tri-kobe.org

参加申込

▲

〒650-0047 神戸市中央区港島南町1-5-4

http://www.tri-kobe.org
5

Closing Remarks

17:00 - 17:50

Opening Remarks

Session I

CD34+ Cell Therapy for Ischemic Tissue Repair:
Two Decades of Evidence

Presentation slides

Douglas W Losordo
Caladrius Biosciences, Inc.

USA

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Session II

Dr. Losordo is the Chief Medical Officer and Senior Vice President of Clinical, Medical and Regulatory Affairs of Caladrius
Biosciences, Inc, Clinical Professor of Medicine at the New York University Langone Medical Center and Adjunct
Professor of Medicine at the Northwestern University Feinberg School of Medicine in Chicago, Illinois.
Dr. Losordo’s career has been dedicated to patient care and to the development of novel therapeutics aimed at the reversal
or repair of chronic conditions such as heart failure, critical limb ischemia, cancer, diabetes and autoimmune disease.
A native of Brooklyn, NY, he received his medical degree from the University of Vermont. Dr. Losordo completed aninternship, residency and fellowship at St. Elizabeth’s Medical, Boston, Massachusetts, where he subsequently joined thefaculty, working with the late Jeff Isner to develop a program in gene therapy and cell-based tissue repair. Dr. Losordo’sgroup has executed the full“translational medicine”paradigm: identifying potential therapeutic approaches in thelaboratory, investigating these strategies in pre-clinical/IND-enabling models and designing and executing first-in-humanand proof-of-concept clinical trials as the study sponsor/IND-holder. His work has included developing VEGF gene therapy
for myocardial ischemia and diabetic neuropathy, CD34+ cell therapy for refractory angina, critical limb ischemia, severe
claudication and coronary microvascular dysfunction and regulatory T cell therapy for autoimmune disease. Two of thesecandidates advanced to phase 3. At Caladrius Dr. Losordo has initiated a phase 2 study of autologous regulatory T celltherapy for new onset type 1 diabetes in children and recently received Japanese PMDA agreement on a study of CD34
cell therapy for critical limb ischemia targeting conditional approval under the new Japanese regulatory rules governing
regenerative therapies. In addition to his own work Dr. Losordo has also mentored numerous scientists and physicianscientists from around the world who now have their own independent programs in translational research.

Session I

ProfileAs the population ages and the acute mortality from cardiovascular disease decreases, a large populationof patients is emerging who have symptomatic chronic ischemic cardiac and vascular disease, many ofwhom remain severely symptomatic despite exhausting conventional medical therapy and mechanicalrevascularization. Mounting evidence suggests that microvascular insufficiency plays a significant role in thepathophysiology of ischemia. Recognizing this problem, investigators have worked to understand the biology of thevascular system and harness this information to develop new treatments for patients with ischemic diseases.
With this end in mind, the science of therapeutic angiogenesis has been evolving for over two decades.
Pre-clinical and early clinical data provide evidence that CD34+ cells may be employed therapeutically forrepair of ischemic tissue.
Preclinical studies have provided evidence for safety and the potential therapeutic potency of freshly isolated
CD34+ cells. Clinical trials involving over 700 patients have been completed providing data supporting
the feasibility, safety and efficacy of CD34+ cell therapy for treatment of advanced cardiovascular disease.
Accordingly, the goal of ischemic tissue repair appears within reach and is entering a pivotal clinical trial in
Japan for patients with critical limb ischemia.
The evolution of the strategy of CD34+ cell based ischemic tissue repair will require an ongoing dialogue
between clinicians, scientists, regulators, industry and payors to take full advantage of advances in our
understanding of the biology of these processes and thereby drive their appropriate application to patients who
are disabled with their condition and have exhausted conventional medical and surgical therapies.

6

7

Closing Remarks

Chief Medical Officer, Senior Vice President Clinical, Medical and Regulatory Affairs, Caladrius Biosciences,
Inc., New York, NY, USA, Adjunct Professor of Medicine, Northwestern University Feinberg School of Medicine,
Chicago, IL USA, Clinical Professor of Medicine, New York University Langone Medical Center, New York, NY

Session III

Abstract

Opening Remarks

Session I

CD34+ Cell Therapy for Chronic Heart Failure:
Clinical Experience in More than 300 Cases

Presentation slides

Bojan Vrtovec
Ljubljana University Medical Centre

ProfileBojan Vrtovec received his MD and PhD at Univeristy of Ljubljana School of Medicine. He completed his
postgraduate training in the field of advanced heart failure and heart transplantation at Texas Heart Institute,
Stanford University and Columbia University. Currently he is serving as a medical director of Advanced Heart
Failure and Transplantation Programme at University Medical Center Ljubljana, and as a full professor of
cardiovascular medicine at University of Ljubljana School of Medicine. His research interests include clinical aspectsof stem cell therapy for heart failure, heart transplantation and mechanical circulatory support.

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2

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5

6

Session I

Slovenia

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Session III
Closing Remarks

Chronic heart failure represents a major healthcare burden with troubling morbidity and mortality rates. The
most frequent cause of advanced chronic heart failure is non-ischemic dilated cardiomyopathy (DCM), which
accounts for 50% of all heart transplantations. There is increasing evidence that heart failure progression in
DCM is not only a consequence of injury to the myocytes, but also results from decreased myocardial perfusion.
In these patients, coronary flow reserve is often impaired both globally and regionally due to defective
vascularization and impaired vasculogenesis and angiogenesis. In our clinical studies in non-ischemic DCM
we were able to demonstrate that that CD34+ cell therapy results in significant improvement in myocardial
perfusion as assessed by SPECT, which is associated with improved left ventricular function. Furthermore, wehave shown that transplantation of CD34+ cells may reverse heart failure progression in DCM, which resultsin improved 5-year survival rate. These effects demonstrate a significant dose-dependency and appear to beparticularly prominent when CD34+ cells are administered via transendocardial route. Thus, the focus of ourcurrent research is to investigate the effects of repetitive transendocardial CD34+ cell therapy in patients with
non-ischemic DCM.

Session II

Abstract

Opening Remarks

Session I

Induction of Cardiac Regeneration is Linked to a Response
Signature of CD133 and SH2B3– Outcome Analysis of the Randomized Phase III PERFECT Trial

Presentation slides

Gustav Steinhoff
Rostock Medical University

Germany

Gustav Steinhoff, M.D., is a professor of cardiac surgery and head of the German Reference and Translation Centerfor cardiac stem cell therapy at the University of Rostock, Germany. Born 1958, graduated for medicine at Erasmus
University Rotterdam in 1984, trained for residency in general surgery, vascular surgery, thoracic, and cardiac
surgery, Prof. And head cardiac surgery in Rostock since 2000. He performed the worldwide first intramyocardial
stem cell therapy in 2001 and is a leading expert and pioneer in regenerative medicine. Publications 300, H-F. 48.
He installed and completed The Phase III multicentre randomized GCP trial PERFECT as a pioneering project todevelop highly standardarized molecular clinical research and advanced stem cell transplantation trials.

Session I

Profile

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2

3

4

5

6

Conclusion:
1. Heart function improvement is dependent on circulating endothelial progenitor cells.
2. Suppression of bone marrow response is associated to SH2B3 gene expression.
3. Peripheral blood angiogenesis response can be predicted by a biomarker signature.

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Session III
Closing Remarks

Objective: The phase III clinical trial PERFECT was designed to assess clinical safety and efficacy of
intramyocardial CD133+ bone marrow stem cell treatment combined with CABG for induction of cardiac
regeneration.
Design: Multicentre, double-blinded, randomised placebo controlled trial.
Setting: The study was conducted across six centres in Germany October 2009 through March 2016 andstopped due slow recruitment after positive interim analysis in March 2015.
Participants: Post-infarction patients with chronic ischemia and reduced LVEF (25-50%). Interventions: Eightytwo patients were randomised to two groups receiving intramyocardial application of 5mL placebo or asuspension of 0·5-5x106 CD133 +.
Outcome: Primary endpoint was delta (∆) LVEF at 180 days (d) compared to baseline measured in MRI.
Findings (prespecified): Safety (n=77): 180d survival was 100%, MACE n=2, SAE n=49, without difference
between placebo and CD133 +. Efficacy (n=58): The LVEF improved from baseline LVEF 33·5% by +9·6% at
180d, p=0·001 (n=58). Treatment groups were not different in ∆LVEF (ANCOVA: Placebo +8·8% vs. CD133 ++10·4%, ∆CD133 + vs placebo +2·6 %, p=0·4).
Findings (posthoc): Responders (R) classified by ∆LVEF >5% after 180d were sixty percent of the patients (35/58)
in both treatment groups. ∆LVEF in ANCOVA was +17·1% in (R) vs. non-responders (NR) (∆LVEF 0%, n=23).
NR were characterized by a preoperative response signature in peripheral blood with reduced CD133 + EPC(RvsNR: p=0·005) and thrombocytes (p=0·004) in contrast to increased Erythropoeitin (p=0·02), and SH2B3mRNA expression (p=0·073). Actuarial computed mean survival time was 76.9 ± 3.32 months (R) vs. +72.3± 5.0 months (NR), HR 0.3 [Cl 0.07-1.2]; p=0.067. Using a machine learning algorithm, 20 biomarker response
parameters were identified allowing preoperative discrimination with an accuracy of 80% (R) and 84% (NR)after 10-fold cross-validation.
Interpretation: The PERFECT trial analysis demonstrates that the regulation of induced cardiac repair islinked to the circulating pool of CD133 + EPC and thrombocytes, associated with SH2B3 gene expression.
Based on these findings, responders to cardiac functional improvement may be identified by a peripheral bloodbiomarker signature. Heart function improvement is dependent on circulating endothelial progenitor cells.

Session II

Abstract

Opening Remarks

Session I

CD34+ stem cell therapy for Ischemic Stroke
Presentation slides
Soma Banerjee

My Background

Imperial College Healthcare NHS Trust, UK

UK

CD34+ stem cell therapy for
Ischemic Stroke

Dr. Soma Banerjee is a Consultant Physician and Specialist in Stroke Medicine at Imperial College Healthcare NHS
Trust, where she is the Head of Department. She was elected to become a Fellow of the Royal College of Physiciansin 2017.
Her clinical practice involves all aspects of the hyperacute treatment of stroke and early rehabilitation. Her
research interest lies in stem cell therapy for the treatment of stroke, and she completed an MD Research degree
investigating the use of CD34+ stem cells for acute ischemic stroke. She will be presenting on this aspect at the
symposium.

• Interested in the translaBon of Stem cell therapy to the bedside, forstroke paBents

Session I

Profile• Consultant in Stroke Medicine• Imperial College group: Completed 1st UK-based clinical trial ofstem cells for the treatment of acute, severe stroke paBentsDr Soma Banerjee
Consultant Physician in Stroke Medicine
Charing Cross and St Mary’s Hospitals
Imperial College Healthcare NHS Trust

Financial Disclosures•

I have received honoraria from BMS/Pﬁzer as an invited speaker•No other disclosures

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Abstract• Classiﬁed by Source & Potency• Stem Cells– Embryonic Stem Cells (pluripotent)– Adult Stem Cells (mulBpotent)– Induced Pluripotent Stem Cells• RaBonale in Stroke• Evidence & RaBonale for CD34+ stem cells• Imperial trial– Autologous - ‘self to self’
Vs Allogeneic – ‘one-to-many’• Challenges & Future DirecBons

CD34+ cells are a subset of bone marrow mononuclear cells which act as the main endogenous source of
haematopoietic and endothelial cell precursors. There are several lines of evidence suggesting that they mightact as a potentially useful therapy in acute stroke. In this talk, I will be concentrating on CD34+ stem cell use,and discussing the evidence from basic science and human studies, to date.

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Ra9onale in Stroke

Embryonic Stem Cells

Adult Stem Cells

Advantages• Pluripotent• Immortal in vitro• Highly expandable

Advantages• MulBpotent• PotenBal for Induced
Pluripotency• Suitable for autologous andallogeneic use

Disadvantages• Ethical issues• Insuﬃcient availability• Unknown risk of tumour
formaBon

Session II

Types of Stem Cells

Disadvantages• Immunosuppression
required for allograZing• Certain types diﬃcult toobtain eg neural stem cells• Expansion & Potency issues• Evidence of
Neurogenesis in
Human Brain• Neurogenesis in
StrokeHirsch MC. interBRAIN: Springer 2004

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Session III

Cell-based therapies have the potential to open up new avenues of treatment in this arena. Targets for stem celltherapy include neuroprotective approaches, which are aimed at protecting the at-risk tissue during the acutephase of stroke, as well as neuroreparative approaches, which may involve direct replacement of damaged brain
tissue or, alternatively, promote endogenous repair processes of the brain.

Presenta9on Outline

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Closing Remarks

There is a major unmet need for treatments that can reduce tissue injury progression, and enhance functional
recovery in stroke. Stem cell therapy offers the promise of a novel reparative strategy for acute brain injury.
Its basis stems from the observation that certain parts of the adult brain are capable of regeneration. However,whilst the regenerative capacity of certain parts of the brain has been demonstrated, it is clear that thisendogenous neuroreparative process is unable to overcome the devastating damage to the brain tissue that
occurs after acute, severe stroke.

Opening Remarks

Session I[Discussant]Translational Research Informatics Center
Foundation for Biomedical Reseach and Innovation

Nobuyuki Ohara
Kobe City Medical Center General HospitalJapan

Session I

ProfileSession II

Education and Work Experience
2017-present Chief, Department of Neurology, Comprehensive Stroke Center,
Kobe City Medical Center General Hospital
Attending, Department of Vascular Neurology
2014-2016
National Hospital Organization, Osaka Minami Medical Center
2013-2014
Visiting scholar, Department of Neurology, Stroke Center,
David Geffen School of Medicine at University of California, Los Angeles
2010-2013
Chief, Department of Neurology and Cerebrovascular Center,
Osaka Neurological InstituteAttending, Department of Neurology and Stroke Center,2008-2010
Osaka University Hospital
Senior Residency, Department of Neurology,
2005-2008
Kobe City Medical Center General Hospital
Residency, Kobe City Medical Center General Hospital
2003-2005

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Closing Remarks

Session III

Licensures and Certificates
1) Unrestricted Physician’s License in Japan (#436732)
2) Board Certified Member of the Japanese Society of Internal Medicine (#32526)
3) Fellow of the Japanese Society of Internal Medicine (#21599)
4) Board Certified in Clinical Neurology (#4382)
5) Japanese Society of Neuro-endovascular Therapy, Specialist Certificate (#572)
6) Board Certified in Stroke Medicine (#20110077)

Opening Remarks

Translational Research Informatics Center
Foundation for Biomedical Reseach and Innovation

Session II
Session I

CD34+/Adipose Stem Cells for Peripheral Arterial Disease (PAD）

Session II

Moderator : Hiroyoshi Yokoi(Fukuoka Sanno Hospital)

ProfileAppointments and Positions
CURRENT POSITION: Director, Cardiovascular Medicine Center, Fukuoka Sanno Hospital
Director, Cardiovascular Medicine Center, Takagi Hospital
Professor, International University of Health and Welfare
PRIOR POSITION:
Director, Department of Cardiology, Kokura Memorial Hospital
Memberships in Professional and Scientific Societies
Member The Society of Japanese Circulation
Member The Japanese Society of Internal Medicine
Member The Japanese College of Cardiology
Fellow
The Japanese Association of Cardiac Rehabilitation
Fellow
Japanese Association of Cardiovascular Intervention and Therapeutics
Japanese Society of Limb Salvage and Podiatric Medicine
Fellow
Japanese Society for Foot Care
Fellow
Fellow
Japan Endovascular Treatment Conference
Fellow
Japan Society for Clinical Pathway
Kyushu Regional Office of Japanese Circulation Society
Fellow
Alliance for Revolution and Interventional Cardiology Advancement
Fellow
Editorial Board
2008-present Journal of Cardiology Cases

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Session III

M.D., Kanazawa University, Ishikawa, Japan
Internship: 1st Department of Internal Medicine
Kanazawa University Hospital, Ishikawa
Residency: 1st Department of Internal Medicine
Kanazawa University Hospital, Ishikawa

Closing Remarks

Education and Training
GRADUATE:
1980-1986
1986-1987
POST GRADUATE:		
1987-1991
		Opening Remarks

Session II

CD34+ Cell Therapy for Critical Limb Ischemia
Presentation slides
Atsuhiko Kawamoto
Translational Research Informatics Center

Japan

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Session II

Current Position Title
Vice Director, Translational Research Informatics Center (TRI)
Unit Director, Unit of Regenerative Medicine, Institute of Biomedical Research and Innovation (IBRI) HospitalGroup Leader, Vascular Regeneration Research Group, IBRI Research Center
Foundation for Biomedical Research and Innovation (FBRI)
Research and Professional Experience
1987:		M.D., Nara Medical University, Kashihara, Japan
1992:		Ph.D., Nara Medical University Graduate School
1995-1998:
Assistant Professor, Coronary Care Unit, Nara Medical University
1998-1999:
Assistant Professor, Department of Cardiology, Nara Medical University
Postdoctoral Research Fellow, Division of Cardiovascular Research, St. Elizabeth’
1999-2003:
s Medical Center,
Boston, MA, USA
Senior Researcher, Vascular Regeneration Research Group, IBRI Research Center, FBRI, Kobe, Japan
2003-2010:2010-present: Unit Director, Unit of Regenerative Medicine, IBRI Hospital, FBRI
2013-present: Group Leader, Vascular Regeneration Research Group, IBRI Research Center, FBRI
2015-present: Vice Director, TRI, FBRI
Research Interest
Cardiology, Cell Therapy, Clinical Trial, Regenerative Medicine, Vascular Medicine

Session I

Profile18

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Closing Remarks

CD34+ stem cell is an endothelial progenitor cell-enriched fraction in bone marrow (BM) and peripheral blood(PB) mononuclear cells. CD34+ cells can be mobilized from BM into PB by administration of granulocyte colony
stimulating factor (G-CSF). Based on these findings, since 1990s, transplantation of BM, G-CSF-mobilized PB or
non-mobilized PB mononuclear cells has been applied in patients with critical limb ischemia (CLI) for vascularregeneration. Although these therapies have been approved as the Advanced Medical Treatments in Japan,
they have never been pharmaceutically approved or covered by insurance due to lack of validated clinical data
complying with good clinical practice (GCP).
We have performed a series of clinical trials regarding intramuscular transplantation of G-CSF-mobilized
CD34+ cells in patients with CLI. Initial phase I/IIa study was performed in 17 no-option patients with CLI. All
patients were free from death and major amputation for 1 year. CLI-free ratio serially increased up to 88% at 1
year. Rest pain scale, total and pain-free walking distances, toe brachial pressure index, transcutaneous partial
oxygen pressure and ulcer size significantly improved by week 12 compared with baseline. Long-term study
revealed the sustained efficacy for 4 years. Next, we performed a phase II study of G-CSF-mobilized CD34+
cell therapy in 11 CLI patients, which was the first investigator-led, GCP-complied clinical trial in the fieldof regenerative medicine in Japan. Promising results in the initial pilot study were almost reproduced in thisvalidated clinical trial.
Since 2014, in addition to the pharmaceuticals and medical devices, a novel review category,“regenerative
medicine products”was created in the amended Pharmaceutical Affairs Law in Japan. Under this law,
conditional pharmaceutical approval may be considered after relatively small-sized clinical trial for“regenerative
medicine products”. We are preparing for a phase II/III, multicenter and randomized clinical trial under GCP,
aiming the conditional approval. We have already agreed with the Pharmaceuticals and Medical Devices
Agency (PMDA) regarding the overall study design and endpoints. Hopefully, the study will start in the fourth
quarter of 2017.

Session III

Abstract

Opening Remarks

Session II

Presentation slides

ProfileAbstract
Background: Peripheral arterial disease (PAD) is highly prevalent in hemodialysis (HD) patients. Furthermore,critical limb ischemia (CLI) has strong impact on mortality in HD patients. However, satisfactory effect has notbeen achieved by several kinds of therapies including revascularization treatment in HD patients with CLI.
Method: Prospective phase 1/2 interventional clinical trial of autologous granulocyte colony stimulating factor(G-CSF)-mobilized CD34+ cell transplantation in HD patients with no optional CLI was conducted in our
hospital. We used peripheral blood (PB)-derived CD34+ cells instead of whole PBMNCs. This was the firstclinical trial in which selected CD34+ cells were used in terms of pre-specified subjects only for HD patientswith CLI. After 5 days’
G-CSF 5 µg/kg/day subcutaneous injection, peripheral blood-derived mononuclear
cells were collected. The CD34+ cells were separated using anti-CD34 monoclonal antibody labeled magnetic
beads, and were injected intramuscularly in ischemic lower limbs. Primary endpoint was safety, and secondary
endpoint was the efficacy of CD34 cell therapy.
Result: Six patients (4 in Rutherford category 5, and 2 in Rutherford category 4) were registered and treated.
The number of injected CD34+ cells were 0.78-14.1 × 105/kg/limb. There was no serious adverse event related
to cell therapy. As to the efficacy, resting pain in 2 patients (Rutherford category 4) completely resolved, and3 out of 5 intractable ulcers (ulcer size ≤ 30 mm) completely healed within 3 months after treatment. Limb
ischemia represented by skin perfusion pressure (SPP) improved at one month after treatment (38.0 ± 16.5mmHg to 55.1 ± 7.5 mmHg, p<0.05), and remained to be above 40 mmHg during 12 months follow-up period.
Six minutes’walking distance also significantly improved. At the end of 12 months’follow-up period, the rateof changes from CLI stage to non-CLI stage was 71% (5 out of 7 limbs). Furthermore, 1-year amputation-freesurvival rate was 100%.
Conclusion: Autologous G-CSF-mobilized CD34+ cell transplantation was safe, feasible, and extremely effectivefor HD patients with CLI.

1year
survival

5year
survival

Normalrenalfunction

75.4%

42.2%

HD

51.9%

14.4%

Uremia is the major risk.

Ulcer

Apheresis
Product(CD34+ cell)

Climimax
Injected
product
CD34+ Cells(CD34+ cell)

1

79

4

0

1.19x107

8.9X106

1.6

2

77

5

15 (Rt)9.63x106

5.44x106

0.95

3

70

5

6 (Rt)

3.43x107

2.61x107

42

4

57

5

22 (Rt)1.12x1088.68x107

13.3

5

68

4

0

1.70x107

1.31x107

2.0

6

74

5

20 (Rt)2.88x107

9.58x1060.72

535 (Lt)

0.72

(mm)

(x105/kg/Limb)

6’(Aulivola B et al. Arch Surg 2004, 139; 395-399)

1
Peripheral blood WBC count and CD34+ cell countbefore and after 5 days’ G-CSF S.C. injection in HD patients.
WBC
(/μL)

CD34+ cell(/μL)

Change of ulcer size (Rutherford 5)（mm）
Case6 Lt
EVT

4

40000
30000

20

2.80

3

26480

2

20000

10000

2

30

5700

1

baseline

10

0.37

baseline

5th day

Case 4

Case6 Rt
EVT

Case 2

Case 35th daypre

G-CSF 5 μg/kgBW was injected s.c. for 5 days.
Data were expressed as mean±SD.

1

3

6

12

(months)

3（m） Change of 6 minutes’ total walking distance
400

350

100% (7 out of 7 limbs)

300

*

*

250
200

4

1 year amputation-free rate (AFS)**
Rate of Changes from CLI stage to non-CLI stage（mean±SD, *p<0.05 vs. pre）

71% (5 out of 7 limb)
pre

1

3

6

12

(months)

5

20

Rutherford
category

Session I

(yr)

Japan

I am very active as a councilor for several scientific and medical professional societies: Japanese Society of Nephrology,
Japanese Society of Internal Medicine, Japanese Society for Dialysis Therapy, Japanese Society of Acute Blood
Purification, and Japanese Society of Hypertension. Also, I am on the Board of Directors of the Japanese Societyof Artificial Organs and Engineering and President of the Japanese Society of Foot Care Medicine. I obtained my
MD degree in 1980 and postgraduate training that led to a PhD degree in 1986 from Hamamatsu University Schoolof Medicine in Hamamatsu, Japan. I was working as visiting assistant professor in the Department of Pathology,
University of Texas. Currently, I am Chairman of Internal Medicine, Chief of Kidney & Dialysis Center and Deputy
Executive Director of the Shonan Kamakura General Hospital, Kamakura, Japan. I am author of more than 200
research papers and I continue to be active in clinical research.
Recently, we are qualified for the 2nd Class cell therapy by the Ministry of Health, Labor and Welfare, aiming totreat limb ischemia with autologous CD34. Moreover, we obtained the certification of the eligibility of state-of–theart medicine with LDL-apheresis for the patients with diabetic nephropathy with massive proteinuria. By establishingthese innovative clinical medicines, we have been appointed as outstanding reviewer of the committee of PMDA
(Pharmaceuticals and Medical Devices Agency).
Since June the 1st 2017, I have worked as Standing Director of Tokusyukai Medical Group (the third biggest medicalgroup in the world), Representative for Foreign Medical Affairs & Cutting-Edge Technology, and Acting Executive
Director of Shonan Kamakura General Hospital.

age

Prognosis according to renal function

Session II

Shonan Kamakura General Hospitalcases

Prognosis after major amputation

Session III

Shuzo Kobayashi

6

21

Closing Remarks

Highly Effective CD34 Cell Therapy on Critical Limb
Ischemia in Hemodialysis PatientsOpening Remarks

Session II

The use of adipose tissue stem cells in the treatment ofpatients with critical limb ischemia (CLI)

Presentation slides

Andrei Kalinin
Central Clinical Hospital with Outpatient Health Center of the Business
Administration for the President of the Russian Federation

My name is Andrei Kalinin. I was born in 1973. I graduated from Russian State Medical University in 1997. After graduation, I was a
resident in the Cardiovascular Surgery program at the A.V. Vishnevsky Institute of Surgery. I gained my post-graduate degree at thesame Institute during the next three years. I wrote my dissertation in 2002 which centered on Cardiovascular Surgery.
From 2002 to 2012 I worked as a surgeon in the division of Vascular Surgery at the A.V. Vishnevsky Institute of Surgery under theguidance of academician A.V. Pokrovsky.
In 2012 I obtained the qualification of Cardio-Vascular Surgeon of the Highest qualification category.
I have been working in the Central Clinical Hospital of the Presidential Administration of Russia as Head of the Department of Vascular
Surgery since 2012.
I specialize in all types of reconstructive vascular surgery for diseases of the abdominal aorta, arteries of upper and lower extremities,
brachiocephalic arteries, and all kinds of operations in venous pathology.
In my surgical operations we use advanced technology, including minimally invasive techniques: EVAR for aneurysms of the aorta, hybrid
surgery for lesions of lower limb arteries and aorta.
I actively participate in research work. I have more than 20 scientific articles in leading publications. I also take an active part in Russianand foreign conferences specifically devoted to problems of vascular diseases and present results of my work at conferences.

1

2

3

4

5

6

Abstract
Critical limb ischemia is caused by a severe damage of lower extremity arteries and is characterized by rest pain and trophic changes.
According to some authors, critical ischemia leads to amputation in 25-30% of cases (North Shore Medical Center, Salem, MA, USA).
The effectiveness of conducting reconstructive surgery is achieved only in 40-50% of cases. (J. Vascular and Endovascular Surgery)

Session II

ProfileSession I

RussiaIn spite of the amount of research done on the use of stem cells in various human pathology, including critical ischemia, cell therapy has
not been widely applied yet.

Adipose tissue derived stromal cells (ATDSC) are produced as a result of special processing of adipose tissue. ATDSC contains various
populations of stem cells-predecessors.
The present clinical study presents the experience of using adipose tissue derived stromal cells in eight patients with critical ischemiaof the lower extremities.
The inclusion criteria was the inability to perform revascularization in patients with critical lower limb ischemia in atherosclerotic
lesions (atherosclerotic peripheral artery disease) (three patients with rest pain, and five patients with trophic changes in the limbs).
All patients had a complex examination, including contrast angiography, duplex ultrasound (DU) arteries / extremities with thedimension of ABI in the dynamics of Transcutaneous oxygen tension (TcPo2) lower extremities (in dynamics), as well, a questionnaire
was used to assess quality of life (SF-36).
Contrast angiography of the extremities was performed to determine the possibility of a reconstructive operation. In the absence of
the ability to perform open surgery or endovascular procedures, these patients were included in the study. The effectiveness of thetreatment was controlled by means of the ABI, TcPo2, as well as evaluation of quality of life.
The results of the study were observed from 6 months to 12 months. There were no adverse reactions after ATDSC administration.

Session III

Adipose tissue is one of the effective sources enriched by progenitor cells, the number of which is much higher than the number of theprogenitor cells in the bone marrow and other tissues.

The progression of critical ischemia was indicated in one patient within one month after inclusion in the study. The patient underwent
amputation of the limb at the hip level. Analysis of the progression of ischemia in this patient showed initially low rates of TcPo2, ABI,
and the presence of the expressed trophic disorders on the foot.
The remaining seven patients saved their limbs. Rest pain was relieved, existing trophic disorders significantly decreased. The complete
epithelialization was indicated in two patients.

BeforeAfter 1 m

After 6 m

ABI0,11±0,21

0,24±0,14

0,38±0,18

After 12 m
0,41±0,26

TcPO2 (mmHg)

6,2±0,21

12,7±0,34

30,14±0,22

32,08±0,19

Conclusions: the results can be considered satisfactory, despite the limited experience of the use of autologous stem cells in criticalischemia. The use of ATDSC in critical limb ischemia is a safe and effective method. A significant increase in indicators of ABI and
TcPo2 in the first 6 months, and clinical symptoms demonstrate the positive effect of this method.
It is important to continue the research for more reliable results.

22

7

23

Closing Remarks

The dynamics of changes in ABI and TcPo2 in seven patients are presented in the table below:

Opening Remarks

Translational Research Informatics Center
Foundation for Biomedical Reseach and Innovation

Session III
Session I

CD34+ Cells for Tissue Regeneration(Shonan Kamakura General Hospital)

24

25

Closing Remarks

Session III

		

Session II

Moderator : Shuzo Kobayashi

Opening Remarks

Session III

CD34+ Cell Therapy for Liver Cirrhosis
Presentation slides
Toru Nakamura
Kurume University School of Medicine

Japan

Ph.D. Kurume University School of Medicine
M.D. Kurume University School of Medicine

1

2

3

4

5

6

Session II

Membership in Medical Societie
The Japanese Society of Internal Medicine
The Japanese Society of Gastroenterology
The Japanese Society of Hepatology

Abstract
Background: Liver cirrhosis (LC) is a leading cause of morbidity and mortality worldwide. In patients withdecompensated disease, ascites decreases quality of life (QOL) and is a poor prognostic factor. The only curative
treatment for end-stage liver cirrhotic patients is liver transplantation. However, there are only a limited
number of donor livers available for patients. Preclinical studies in rodent models of chronic liver fibrosis
have shown that transplantation of peripheral blood (PB)-CD34+ cells leads to hepatic regeneration and areduction of liver fibrosis by suppressing hepatic stellate cell activity and increasing matrix metalloproteinase
activity. Based on these findings, we conducted a prospective clinical trial to evaluate the safety and efficacy
of autologous granulocyte colony-stimulating factor (G-CSF)-mobilized PB-CD34+ cell transplant in patientswith decompensated LC. Methods: The inclusion criteria were the following: Patients with decompensatedLC, aged from 20 to 75 years, serum albumin of less than 3.0 g/dL, total bilirubin of less than 5.0 mg/dL, andno viable hepatocellular carcinoma on CT or MRI. The exclusion criteria were the following: Patients withcreatinine of more than 2.0 mg/dL, PT-INR of more than 2.4, and liver tumors or other cancer. Patients wereadmitted for liver function tests, full blood count, coagulation profile, tumor marker levels, and ultrasound (US).
Seventeen patients were included and ten patients were treated. Patients included in the study received a daily
subcutaneous injection of G-CSF 10 µg/kg/day for 5 days, to mobilized bone marrow derived CD34+ stem
cells. Leukapheresis was performed on day 4 and 5. In the treated patients, CD34+ cells were infused via thehepatic artery. Patients were divided into three groups according to the amount of CD34+ cell collection. All
the patients were followed up every 4 weeks, and laboratory data were analyzed for 24 weeks. Results: G-CSF
treatment and leukapheresis were well tolerated, and no serious adverse events were observed. After 24 weeks,
serum albumin level was significantly increased in patients who had received middle (1×106cells/kg) or high (2×106cells/kg) doses of CD34+ cells compared with baseline. Doppler US showed a significant increase in hepatic
blood flow velocity and blood flow volume after CD34+ cell therapy. The average Child–Pugh score, which wereused to evaluate the hepatic reserve function, of all treated patients at week 24 was not significantly different
from baseline. Conclusions: CD34+ cell therapy is feasible, safe and effective in slowing the decline of hepatic
reserve function.

26

27

Session III

Professional Training and Employment
2003-present Assistant, Division of Gastroenterology, Department of Medicine, Kurume University School of Medicine
2001-2003
Resident in Internal Medicine, Kurume University
1997-2001
Kurume University Graduate School of Medicine
1997		
Passed the examination of National Board

Closing Remarks

Education
2001		
1997		

Session I

ProfileOpening Remarks

Session III

Kobe University School of Medicine

Japan

Autologous Transplantation of CD34+ Cells
for Patients with Fracture Nonunion

Methods of experimental study

Tomoyuki MASTUMOTO, Takahiro NIIKURA,
Ryosuke KURODA

Transplantation

Department of Orthopaedic Surgery,
Kobe University Graduste School of Medicine

ProfileMedical School
4/1994–3/2000

Immunodeficient rat unfracture modelref) Kokubu T, J Orthop Res, 2003

Human peripheral blood CD34+ cells

Osteocalcin

Isolectin B4

Medical Doctor, Kobe University School of Medicine

Clinical Training
6/2000–5/2001
Kobe University Hospital
6/2001–11/2006 Wadayama Hospital, Hyogo Rehabilitation Center Hospital, Hyogo Emergency Medical Center,
Rokko Hospital
Graduate
4/2003–9/2006(6/2004–3/2006)

Session I

Tomoyuki Matsumoto

Presentation slides

Doctoral, Kobe University School of Medicine
Stem Cell Translational Research, Institute of Biomedical Research and Innovation/RIKEN Center
for Developmental Biology

Postgraduate
12/2006–11/2008 Postdoctoral and Fellowship, Department of Orthopaedic Surgery University of Pittsburgh Medical
Center/Stem Cell Research Center
Clinical Staff, Kobe University Hospital
12/2008-7/2010
8/2010-3/2017
Assistant Professor, Kobe University Hospital
4/2017- present
Associate Professor/Lecture, Kobe University Hospital

Truidine blue
Laser Doppler
Xray
Clinical Application of CD34-Positive Cells(September 9, 2017)、Tokyo

Clinical Application of CD34-Positive Cells(September 9, 2017)、Tokyo

KOBE

KOBE
UNIVERSITY

UNIVERSITY

1

2
Series of experiments (2)Series of experiments (1)Locally transplanted CD34+ cells were dose-dependently
contributed to fracture healing via enhancing
angiogenesis and osteogenesis.

Human peripheral blood CD34+ cells, which weresystemically transplanted, recruited to the fracture siteand contribute to local environment by enhancing
angiogenesis and osteogenesis.

Mifune Y, Matsumoto T, Kuroda R, et al. Stem Cells 2008

Matsumoto T, Mifune Y, Kuroda R, et al. Am J Pathol 2006

More than 104 cells / ratbone marrow

hCD31/SMA

Session II

Autologous Transplantation of CD34+ Cells for Patientswith Fracture Nonunion

Local administration

cortical bone28

osteoblast

Osteoblast

CD34+

103

VEGF

105

capillary

Endothelial cell

GM-PB CD34+ cellCD34+ cell

Blood vessel

CD34+ cell

Clinical Application of CD34-Positive Cells
Osteogenesis (September
Vasculogenesis
9, 2017)、Tokyo

104
Clinical Application of CD34-Positive Cells(September 9, 2017)、Tokyo

KOBE
hOsteocalcin

KOBE

UNIVERSITY

UNIVERSITY

3
Autologous transplantation of peripheral blood
CD34+ cells for patients with nonunion:PhaseⅠ/Ⅱ Clinical trial

4
Comparison with Historical control
CD34+ cells

Historical control

number7

11

Gender（male/female）

6/1

9/2

Age

34 (20-45)

37 (21~56)

Femur/Tibia

2/5

4/7

Healing rate at 12
weeks (%)
Duration to heal
(week)

5/7 (71.1%)

2/11 (18.1%)

16.1 (8~38)

29.1 (12~44)

Kuroda R, Matsumoto T, et al. Stem Cell Transl Med 2015

Clinical Application of CD34-Positive Cells(September 9, 2017)、Tokyo

KOBE
UNIVERSITY

5

Clinical Application of CD34-Positive Cells(September 9, 2017)、Tokyo

Session III

Failures in fracture healing are caused due to many factors, in which severe skeletal injuries consisting of fractureswith a compromised blood supply result in either delayed unions or established nonunions. An essential requirement
for these fractures is to restore the local blood flow. One emerging strategy in the regeneration and repair of boneand surround tissue is the use of stem cells. We have proved the first proof-of-principle that elucidate the healing
potential of CD34+ cells for fracture nonunion. As a series of preclinical studies, we first demonstrated that systemic
infusion of human circulating CD34+ cells into immunodeficient rats with non-healing fracture contributes tofracture healing. Next, we attempted local transplantation of CD34+ cells with atelocollagen gel, a bio-absorbablescaffold, in the same animal model and demonstrated the similar effect at the lower dose compared with thesystemic administration. Based on the in vitro experiments and pre-clinical study using in vivo animal experiments,
we started clinical trial of autologous transplantation of granulocyte colony-stimulating factor (G-CSF) mobilized
peripheral blood CD34+ cells for patients of tibial or femoral nonunion.
Based on these scientific backgrounds, we started phase I/IIa clinical trial: autologous local transplantation of
G-CSF mobilized peripheral blood CD34+ cell for patients with the tibial or femoral nonunion. Inclusion criteria are (1)patient with tibial or femoral fracture, (2) patient with non-infectious nonunion, (3) 20 to 69 year-old patient, (4) patientwith informed consent by writing. After leukoapheresis following 5 days G-CSF injection, patient receives magnet
sorting of CD34+ cells. Treatment is conventional surgery for nonunion combined with autologous transplantationof G-CSF mobilized peripheral blood CD34+ cells suspended with atelocollagen gel. Radiological fracture healing
at week 12, the primary endpoint of this study, was achieved in 5 cases out of 7 patients (71.4%). Both of 2 cases
without fracture healing at week 12 had femoral fractures in which cases ultimately radiological healing was finallyobserved at week 19 and 36. The interval between the cell transplantation and union, the secondary endpoint, was12.6 ± 5.4 (range 8 to 24) weeks for clinical healing and 16.1±10.2 (range 8 to 36) weeks for radiological healing.
Promising outcomes in this phase I/IIa clinical trial encourage the application of the CD34+ cells for nonunion asa novel therapeutic modality. To further elucidate the safety and efficacy of CD34+ cell transplantation for bone
fracture healing, multicenter clinical trial has been started.

bridging callus
callusendothelial cell

KOBE
UNIVERSITY

6

29

Closing Remarks

Abstract

Opening Remarks

Closing Remarks
Translational Research Informatics Center

Closing Remarks

Foundation for Biomedical Reseach and Innovation

Masanori Fukushima

Session I(Translational Research Informatics Center)

Session II

Twelve years have passed since we organized the consensus meeting for vascular regenerative therapy inMarch 2005 at Yokohama. However vascular regenerative therapy is not yet put into daily clinical practice
to date. The major reason is that the clinical trial focusing on responsible cellular fraction of regenerating
blood vessel has not been conducted with appropriate endpoint. This is likewise in the clinical trial using
growth factor other than cell preparations, such as HGF producing vector, bFGF, etc. In latter cases,
cellular process induced by such factor has not well been understood and thereby appropriate clinical trial
was not designed.
Nevertheless, due to steady efforts by researchers, the crucial effect of CD34 cell therapy has been provenby well-designed clinical trials, and finally we could forecast its cell therapy to be disseminated. Today you
should find specific data supporting its outcome, and should feel confident that lower limb amputation ordeath caused by critical limb ischemia (CLI) should be dramatically reduced hereafter. Furthermore, CD34
indication is not limited to CLI but also to patients with non-union, liver cirrhosis, stroke, etc. They will also
receive clinical benefit from CD34 cellular therapy in the very near future.

ProfileWith comprehensive experience spanning over the past quarter-century as a medical oncologist at Aichi Cancer
Center and Kyoto University Hospital, Dr. Fukushima has engaged in the practice and dissemination of standard
cancer treatment and reform of Japan’s medical care system, and is active to date contributing to building up theinfrastructure of clinical trial, particularly focusing on translational research. He was in charge of supervising thenational translational research promotion programs conducted by the government of Japan, and currently, assuming
the Board of Director for CDISC, and Program Officer of Rare / Intractable Disease Project of Japan, aiming tocontribute to the standardization and harmonization of global data sharing in clinical trials; ultimately for the
disease control over the world.

30

31

Closing Remarks

Session III• Director and Chairman, Translational Research Informatics Center (TRI), Foundation for Biomedical Researchand Innovation (FBRI)• Professor Emeritus, Kyoto University• Program Officer, Rare / Intractable Disease Project of Japan, AMED (Japan Agency for Medical Research andDevelopment)• Board of Director, CDISC (Clinical Data Interchange Standards Consortium)