Kök Hücre Anabilim dalının kullanıma açılmış olan laboratuvarlar:
● AREL (Ege Üniversitesi Tıp Fakültesi Eğitim ve Araştırma Laboratuvarı) ● FABAL (Eczacılık Fakültesi Farmasötik Bilimler Araştırma Laboratuvarı ● Ege Üniversitesi Pediatrik Hematoloji Kök Hücre Nakil Ünitesi, ● Ege Üniversitesi Tıp Fakültesi Terapötik Aferez Merkezi ,
● Nevvar – Salih İşgören İç Hastalıkları Kliniği (Kök Hücre İşleme ve Saklama Bölümleri, Bilgi İşlem, Hasta Kabul ve Muayene Odası),
● Doku Tipi laboratuvarı, ● Analitik Kimya Abd., ● Fizyoloji Anabilim Dalı, ● Nükleer Bilimler Enstitüsü ,
● Mühendislik Fak., Biyomühendislik Böl, ● E.Ü. Mühendislik Fak., Biyomühendislik Böl., ● Hücre Kültürü ve Doku Mühendisliği laboratuarı, ● Fen Fakültesi, Biyoloji Bölümü,
● Moleküler Biyoloji Anabilim Dalı, ● Histoloji ve Embriyoloji AD.
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K11Taner Demirer - Ankara Üniversitesi Tıp Fakültesi/Ankara University Faculty of Medicine Türkiye'de klinik kök hücre uygulamaları
Implementation of clinical stem cell therapies in Turkey
Bu konuşmamızda Ülkemizde kök hücre transplantasyon uygulamaları gözden geçirilecektir. Sunumda hem Türk Hematoloji Derneği, hem Sağlık Bakanlığımız, ve hem de EBMT verilerine değinilecektir.
Son 10 yılda Ülkemizde transplantasyon ünitelerinin sayısı 70 e yıllık nakil sayıları ise 3000 lere ulaşmıştır. Ülkemizde kök hücre tedavilerinin kullanımı özellikle son dekatta yaygınlaşmış olup sonuçları ve başarı oranları Avrupa ve Kuzey Amerika ile kıyaslanabilir durumdadır.
Özellikle kordon kanı kök hücresi kullanımı, GVHD tedavisi için mezankimal kök hücrelerinin kullanımı, haploidentik nakillerin artan şekilde kullanımı ve TÜRKÖK Projesi gibi uygulamalar adeta Ülkemizi kök hücre tedavilerinde bölgesinde lider ülke konumuna getirmiştir. Konuşmamızda bu topikler kısaca gözden geçirilecektir.
In this talk, we will review the clinical applications of stem cell therapies in Turkey. For this purpose, we will use the data of Turkish Hematology Society, Ministry of Health and European Group for Blood and marrow Transplantation (EBMT).
During the last 10 years, especially number of transplant units reached to 70 and annual transplants approximately to 3000. It is clear that stem cell transplant therapies are being
increasingly used as times goes by and both success rate and results are comparable with centers in the Europe and USA.
Implementations such as use of cord blood stem cells, mesenchymal stem cells for treatment of graft-versus-host-disease, haploidentical transplants and TÜRKÖK Project in order to establih a donor bank made Turkey a leader country for stem cell therapies in this region. We will review these issues, shortly, in our talk.
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K12Alp Can - Ankara Üniversitesi Tıp Fakültesi / Ankara University Faculty of Medicine İskemik kardiyomiyopatide kök hücre yaklaşımları
Stem Cell Therapy Approaches to Ischemic Cardiomyopathy
Son 15 yılda iskemik kardiyomiyopatide (İKM) gerek otolog gerekse allojeneik olmak üzere çok sayıda kök hücre tedavisi denemesi gerçekleştirilmiştir. Çoğunlukla faz 1 ve 2 düzeyinde ve az sayıda hasta üzerinde yürütülen bu çalışmalardan bir kısmında ümit verici sonuçlar alınmıştır. Dolayısıyla elde edilen sonuçların “klasik bir tedavi yöntemi” olarak kabul edilmeden önce bir “ön bulgu” olarak değerlendirilmesi daha yerinde olur. Yakın zamanda yayınlanan ve akut miyokard enfarktüsü (AMİ) sonrası kök hücre tedavilerinin ele alındığı bir meta analize göre sol ventrikül (SV) ejeksiyon fraksiyonundaki (EF) bir miktar artışın
ötesinde daha fazla bir yararın maalesef elde edilemediği görülmektedir. Bu amaçla kök hücrelerin iskemik kardiyopati olgularında kullanım gerekçelerinin tekrar değerlendirilmesi gerekliliği söz konusudur. Öncelikle İKM olan birçok hastada aşikar AMİ’den önce klinik olarak saptanmış veya saptanamamış (subklinik) bir miyokard enfarktüsü öyküsü bulunmakta ve dolayısıyla bu hastalarda progresif bir ventrikül doku değişikliği ve bir miktar sol ventrikül yetmezliği varlğından söz edilmelidir. Bu durumda her hastanın kök hücre tedavisine
vereceği yanıt birbirinden farklı olacaktır. Bunun yanı sıra İKM’de hemen her hastada belli oranda disfonksiyonel canlı miyokard (DCM) dokusunun varlığı hücresel tedavi yanıtlarını etkilemektedir. AMİ geçiren hastaların bir çoğunda görülen bu doku kök hücre tedavisinin ana hedefinde yer almaktadır ve kardiyak MR ile ayrıntılı olarak saptanmaktadır. Eğer DCM dokusu canlı olmak yerine daha çok skar dokusundan oluşuyorsa kök hücre tedavisi sonrası SVEF’daki artış daha anlamlı olmaktadır. İKM’de ilerleyici SV disfonksiyonuna eşlik eden mekanizmalar düşünüldüğünde temelde dört etkenin bunda yer aldığı görülmektedir; i) inflamatuvar ve immün doku yanıtları, ii) mikrovasküler disfonksiyon, iii) oksidatif stres, ve iv) ekstraselüler matriks üretimi ve artmış apoptoz. Bu konuşmada bu dört temel hücre ve doku yanıtının İKM’deki rolü tartışılacaktır. (Proje no: 0741-STZ-2014)
Over the past 15 years, numerous stem cell trials have been performed in patients with ischemic cardiomyopathy (ICM), using both autologous and allogeneic stem cells. Although many individual studies reported encouraging signals, these were all phase 1 or 2 studies with appropriately small numbers of patients, and their conclusions must therefore be
considered preliminary. In an attempt to increase statistical robustness, a recent meta-analysis assessing the results of all randomized clinical trials of stem cell therapy for patients with acute myocardial infarction (AMI) was performed, demonstrating no net beneficial effects on outcomes, except for a small improvement in ejection fraction. Given these results, a reassessment of the rationale for the use of stem cells in cardiovascular disease is timely. Patients with ICM invariably have usually extensive areas of myocardial scar. ICM patients had areas of myocardial dysfunction due not to scar, but to dysfunctional viable myocardium (DVM). DVM provides a potential target for therapeutic interventions in ICM. If the
dysfunctional tissue consists of viable rather than scarred myocardium, LV function can presumably be improved. The concept of DVM may also help direct which patients may benefit most from stem cell therapy. As the stage of HF that may be considered too late for stem cell therapy is unclear, the presence of DVM may help guide the identification of those patients with the most potential to benefit. The potential of any therapy, including stem cells, to improve outcomes in ICM is related not only to its effects on restoring function to DVM, but also to its capacity to improve processes that contribute to progressive deterioration of LV structure and function. On the basis of this conceptual framework, my aim is to explore the potential of stem cells to exert beneficial effects in ICM by considering the overlap between pathways believed to contribute to disease progression (other than atherosclerotic
disease of the large coronary arteries) and the known activities of stem cells that could favorably influence these pathways. (Project No: 0741-STZ-2014)
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K13Erdal Karaöz - Liv Hospital Rejeneratif Tıp ve Kök Hücre Üretim Merkezi Hastanesi / Liv Hospital Regenerative Medicine Stem Cell Production Center
Nöromuskuler dejeneratif hastalıklarda kök hücre uygulamaları: Laboratuvardan kliniğe
Stem Cell Applications on neuro-muscular degenerative disorders: Bench to bedside
Kök hücreler çoğalma, farklılaşma ve kendini yenileyebilme (self-renewal) özellikleri ile embriyonik dönemde gelişimde önemli bir rol oynamakla birlikte erişkin dönemde de dokularda yenilenmeyi sağlar. Kök hücreleri esas itibarıyla embriyonik ve embriyonik olmayan olmak üzere iki ana başlıkta incelemek olmakla birlikte günümüzde klinikte kullanım potansiyelini dikkate aldığımızda
mezenkimal kök hücre (MKH) olarak adlandırılan gerek fetal gerekse post-natal dönemde birçok doku ve organlardan elde edilebilen hücreler giderek önem kazanmaktadır. MKH’ler embriyonik dönemde gelişimde önemli bir rol oynamakla birlikte erişkin dönemde de dokularda yenilenmeyi sağlar. Hasarlı dokulardaki hücrelerin yerine geçmenin yanı sıra salgıladıkları sinyal moleküller (sitokinler, büyüme faktörleri, anti-apoptotik faktörler ve hücre yapışma molekülleri) aracılığı ile hasarlı bölgedeki hücreleri apoptozdan koruyabilir, dokularda var olan kök hücre potansiyelini uyarabilir, immün düzenlemeyi uyarabilir ve enflamasyonu engelleyici olarak görev alabilir. Bu sunuda erişkin kök hücrelerin pre-klinik ve klinik uygulamalarına ilişkin deneyimlerimizin yanı sıra geleceğe ilişkin projeksiyonlar tartışılacaktır.
Stem cells have the capability of self-renewal and differentiation into a wide range of cell types with various potential clinical and therapeutic applications. Stem cells are providing hope for many diseases that are currently in need of effective therapeutic methods, including neurodegenerative disorders like; stroke, amyotrophic lateral sclerosis, Alzheimer’s disease, Parkinson’s disease and as well as muscular dystrophy disorders like Duchenne Muscular Dystrophy and Facio-scapulo- humeral Muscular Dystrophy.
For this aim, numerous pre-clinical studies have been achieved and\or in progress on different types of stem cells including, induced pluripotent stem cell (IPS), embryonic stem cell (ESC) and neural stem cells. But there are some complications on the clinical utilization of these cells, due to the reason of ethical issues and especially because of the potential of formation of teratomas via IPSs and ESCs. For this reason, as we glance to the clinical trials ongoing nowadays, we see that MSCs are studied intensely on clinical applications.
Due to the ideal characteristics of these cells for regenerative medicine, clinical interest in the use of MSCs has increased significantly over the past few years. Therapies with MSCs have shown promising results on neurodegenerative and muscular degenerative diseases. In addition to their capability of regulating inflammation, they can promote other beneficial effects, such as neuronal growth, decreasing free radicals, reducing apoptosis and releasing different neurotrophic factors which can assist the endogenous regeneration of the injured region.
The results of clinical trials carried out for the treatment of various diseases, especially including neuro-muscular degenerative disorders, via application of MSCs derived from different tissues and manufactured in our own GMP facility, will be introduced in this presentation.
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K14Stuart B. Goodman - Stanford University School of Medicine Osteonecrosis: Etiology to stem cell therapy
Stuart B. Goodman MD PhD
Professor of Orthopaedic Surgery and Bioengineering, Stanford University, Stanford, CA, USA
Osteonecrosis (ON) is a disorder of bone in which the cellular components of the affected bone, including both hematopoietic and mesenchymal cells, die. Some of the conditions/agents associated with ON include trauma, inflammatory arthritis (such as lupus erythematosis,
rheumatoid arthritis etc.), coagulation disorders, altered lipid metabolism (as seen with excessive alcohol intake or genetic disorders of fat metabolism), radiation, drugs (such as corticosteroids), vascular (venous or arterial) insult, and other causes. In load bearing areas, ON leads to progressive collapse of bone, secondary loss of support of the overlying articular cartilage and resultant degenerative arthritis. When considering the hip or knee joint, progressive arthritis with pain and disability unresponsive to conservative management will necessitate total joint
replacement. As joint replacement is a “joint sacrificing” procedure, increasing interest has been focused on methods and interventions to regenerate the bone and save the original natural joint (“joint saving procedures”).
One of the current techniques that attempts to save the host’s natural joint afflicted with ON is core decompression, in which the osteonecrotic area is drilled, supposedly to facilitate an inflammatory response and promote neovascularization and ingrowth of osteoprogenitor cells. More recently, the technique has been expanded to include debridement of the osteonecrotic segment, and the addition of cells harvested from the iliac crest and subsequently concentrated, to enhance bone healing. In the experience of the author and other researchers in this area, this new technique has improved the clinical and radiological outcome of patients with the earlier stages of ON without collapse or arthritis, at intermediate follow-up.
However many questions concerning “joint preserving” operations still remain unanswered. These questions relate to optimal patient selection, preferred cell harvesting, concentrating and delivery techniques, the quantity and quality of cells necessary for a successful outcome, how best to prepare the ON lesion, whether biomechanical stabilization with scaffold or internal fixation is necessary, and appropriate patient rehabilitation protocols.
These questions will only be addressed in the future by hypothesis driven basic science experiments, and continued reporting of well-designed evidence-based outcome studies.
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K15Annelise Bennaceur Griscelli - University Paris Sud XI School of Medicine
Hematopoietic specification of embryonic and induced pluripotent stem cells and their potential use in cell-based therapies
Prof. A Bennaceur Griscelli, MD,PhD. University Paris Sud, Inserm U935, Ingestem Infrastructure. Hematopoiesis consists of two developmental programs, a primitive and a definitive wave
generating hematopoietic multipotent stem cells with long-term engfratment. Understanding how the definitive program develop is essential to design efficient strategies for the generation of mature hematopoietic lineages and for clinical applications.
Human induced pluripotent stem cells (IPSC) by reprogramming adult cells are very close to human embryonic stem cells (ESC) but some inherent differences exist. Description of
hematopoietic hierarchy has allowed to identify in ES and iPSC, the hemangioblast, from which both primitive and definitive programs occurs in vitro, induced by distinct signaling pathways such as, Activin/Nodal for the primitive wave and Wnt, or tenascin C for the definitive hematopoiesis. The induction of long-term hematopoietic potential has been shown to require either a specific micro-environnement such as teratoma injection or inhibition of pathways such as Activin A and notch signaling activation.
We characterized early lymphoid progenitors, CD34+CD45RA+CD7+ cells restricted to NK and TCRαβ+ T lymphocyte and CD34+CD45RA+CD7– cells restricted to myeloid, dendritic and B lymphocytes. Many differentiation protocols has been described to produce hematopoietic cells from PSC with a variable potential between the PSC sources. However, the generation of mature and adult cells still remains challenging and highly inefficient. Moreover, one of a major hurdle to solve is the genomic stress and instability of the hIPCS with the risk of cancer. We have shown that reprogramming process is a source of mutagenesis and culture conditions select genetically
abnormal cell clones conferring growth advantage. By using infrared microspectroscopy (Synchrotron, Saclay), we could track the spectral modification of somatic cells during the
reprogramming process. iPSC adopt a metabolic features entirely different from that of the original somatic cells, and fully programmed iPSC clones were well distinct from the partially programmed clones.
We explored the potential use of IPSC to produce red blood cells in sickle cell disease. A common therapeutic strategy in SCD is the induction of HbF synthesis in order to attenuate the severity of the disease. We generated iPSC from amniotic fluid cells of a SCD HbS-/S-. Erythroid cells, 20-26% of RBC and 74-80% of orthochromatic erythroblasts were produced in vitro, in 25 days. We
provided evidence that IPSC can lead to the production of significant numbers of fetal erythrocytes. In the future, it might be possible using this strategy, to design new autologous fetal blood
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K16Ali Turhan1,2 - University Paris Sud XI School of Medicine Modelling cancer stem cells using iPSC
1Inserm UMR 935 ; « ESTeam Paris Sud » INGESTEM National Infrastructure Institut André Lwoff ;
University Paris Sud 11 ;
2Department of Hematology, University Paris Sud 11, AP--‐HP Hôpital Paul Brousse, Villejuif ,
France
Our ability to reprogram differentiated cells into a pluripotent state has revolutionized our concepts of cell biology and opened major perspectives in regenerative medicine and disease modeling. In the latter context, comparison of primitive “cancer stem cells “ and gene expression patterns of either embryonic stem cells or the reprogrammed cells using iPSC technology revealed similar pattern of expression of pluripotency genes such as Sox2, Lin28 and Oct4. Cell reprogramming also requires a step of EMT ‘Epithelial-Mesenchymal Transition) followed by MET (Mesenchymal
Epithelial Transition), a phenomenon which also takes place during metastatic dissemination of solid tumors. These findings suggest that iPSC technology could be used to model “cancer stem cells” at least in some tumors.
One of the goals in this context is to use this technology to generate a “cancer stem cell in a dish” allowing a large scale drug screening in order to find novel compounds able to target these cells felt to be at the origin of relapses after therapy. The second goal is to use iPSC technology to understand signalling pathways involved in tumor progression. We will describe strategies that have been designed to model primitive malignant stem cells in haematological malignancies and in solid tumors using iPSC methodology. Two types of malignancies are explored in our center using iPSC: 1- Chronic myeloid leukemia 2- Hereditary cancer syndromes, essentially BRCA1-mutated breast cancers. The results that will be presented suggest that iPSC modeling could be a valuable tool for the study of genomic landscape inherent to several diseases and for future cancer drug screening strategies targeting tumor initiating cells.
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K17Tamer Önder - Koç Üniversitesi Tıp Fakültesi / Koc University Faculty of Medicine p300 ve CBP kromatin düzenleyicilerinin yeniden programlamadaki rolleri
A chromatin modifier-based chemical screen identifies a role for p300 and CBP in reprogramming
Fare somatik hücreleri birtakım kimyasallar kullanılarak yeniden programlanıp kimyasal uyarılmış pluripotent kök hücrelere (kUPKH) dönüştürülebilmektedir. Ancak kUPKH'lerin insan somatik hücrelerinden üretilip üretilemeyeceği henüz belli değildir. Fare kUPKH üretiminde kullanılan kimysasalların en önemlilerinden biri H3K9 metil trasnferazı Dot1L'nin inhibitörleridir. Daha evvelki çalışmalarımızda Dot1L inhibisyonunun yeniden programlamanın verimini artırdığını ve Klf4 ve cMyc'nin yerine geçebildiğini göstermiştik. Bu çalışmada Dot1L inhibitörleri ile beraber
kullanıldıklarında insan somatik hücrelerinin uPKH'lere dönüşmesini kolaylaştıracak kimyasallar taranmış ve p300/CBP'nin bromodomain hedefli iki inhibitörü tespit edilmiştir. Bu inhibitörler yeniden programlamanın verimini artırmakta ve süreci hızlandırmaktadırlar. P300 ve CBP'nin shRNA kullanılarak susturulmaları da aynı etkiyi göstermektedir. RNA sekanslama sonucunda söz konusu inhibitörlerin epitel-mezenkimal geçişte rol alan ve pluripotent kök hücrelerde baskılanacak pek çok genin ifadesini azalttığı belirlenmiştir. Bu sonuçlar p300 ve CBP'nin somatik hücre
kimliğinin devamını sağlayan iki kromatin düzenleyicisi olduğu ortaya çıkarmaktadır.
A combination of chemical compounds has been shown to induce pluripotency in mouse somatic cells. Whether chemically induced pluripotent stem cells (ciPSCs) can be generated from human somatic cells remains unknown. One of the key factors that facilitate ciPSC generation is the inhibition of H3K79 methyl-transferase Dot1L, which we had previously shown to increase reprogramming efficiency and substitute for Klf4 and cMyc in reprogramming of human somatic cells. To uncover additional chromatin-related pathways that work in parallel with Dot1L in suppressing reprogramming, we carried out a screen of small molecule inhibitors of chromatin modifiers on human fibroblasts in combination with Dot1L inhibition. Through this screen, we identified two independent acetyl-lysine competitive inhibitors targeting the bromodomains of co- activators P300 and CBP as potent enhancers of reprogramming. These inhibitors significantly increase reprogramming efficiency and substitute for Klf4 and cMyc in conjunction with Dot1L inhibitors as well as the combination of chemicals used in the generation of ciPSCs. Time-course experiments indicate that inhibition of acetylation-dependent p300/CBP bromodomain-mediated interactions has the most effect during early stages of reprogramming and significantly accelerates the emergence of Tra-1-60-positive iPSCs. Simultaneous shRNA-mediated knock-down of P300 and CBP can phenocopy this effect. RNA-sequencing reveals that treatment of fibroblasts with
p300/CBP bromodomain inhibitors results in the downregulation of a large number of genes, among which epithelial-to-mesenchymal transition related genes are highly enriched. A majority of these p300/CBP-regulated genes in fibroblasts are fated to be repressed in pluripotent stem cells. These results suggest that p300 and CBP serve to maintain somatic cell identity by perpetuating cell type-specific enhancer activity.
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K18Kursad Turksen - Editor in Chief, Stem Cell Reviews and Reports What journals are looking for: One editor's perspective
EDITORIAL REFLECTIONS
Kursad Turksen, Ph.D., Editor in Chief, Stem Cell Reviews and Reports
The role of electronic communications and the internet are revolutionizing the dissemination and publishing of information of all sorts. Science and scientific writing and publishing have also been