Day 2 :
IMB A*STAR, Singapore
Victor Nurcombe obtained his PhD in Developmental Neurobiology from the University of Sydney in 1984. He took both CJ Martin and Humboldt Fellowships to initiate his Postdoctoral training at the Max-Planck Institute for Biochemistry in Munich, and then worked in Oxford, Paris and New York, before returning to Australia in 1990 first to the Walter & Eliza Hall Institute and hence to the University of Melbourne in 1992 as a Senior Lecturer. In 1998, he became a Reader and Associate Professor at the University of Queensland as Head of Developmental Biology, before being headhunted to the IMCB in Singapore as a Principal Investigator in Stem Cell Biology in 2003. In 2008, he became Senior Principal Investigator within the IMB. He is also Adjunct Professor at the LKC School of Medicine, Imperial College London-NTU, Singapore, as well as the University of Lille in France.
Statement of the Problem: Bone marrow-derived mesenchymal stem cells (hMSCs) are a valuable resource for cell-based therapy. However, their abundance is low, necessitating ex vivo expansion to reach useful numbers; such expansion compromises their “stemness”. Fibroblast Growth Factor-2 (FGF2), now in widespread use, increases hMSC proliferation, but triggers premature differentiation. Certain heparan sulfate (HS) glycosaminoglycan variants, abundant in stem cell Extracellular Matrix (ECM), are known to regulate the activity of many growth factors, including FGF2. Heparin has been used extensively to support human stem cell expansion as an analogue of the more physiologically relevant HS, although it adversely affects hMSCs, resulting in senescence. To obviate the need for heparin, we have isolated HS variants better targeted to the growth factor FGF2. An affinity chromatographic approach was utilized to extract an HS variant (HS8), using peptide sequences derived from the heparin-binding domains of FGF2 as bait. ELISA assays demonstrated that HS8 binds to FGF2 with much higher affinity than to other FGFs, as well as to other heparin-binding factors such as PDGF or VEGF. The melting temperature of FGF2 was markedly increased by HS8, indicating it acts to stabilize FGF2, so prolonging its activity. Both FGF2-stimulated ERK signaling and proliferation were amplified by HS8 in hMSCs. Crucially, hMSC cultures expanded with HS8 supplementation yielded a subpopulation of cells enriched for the early marker Stro-1+ as well as displaying greater CFU-F capacity. When applied into critical-sized calvarial defects in rats, HS8 significantly accelerated bone healing. Our work demonstrates that affinity-selection of HS is able to enrich for HS variants that can trigger faster hMSC ex vivo expansion without adversely changing their biological properties or potential. Such HS preparations are components for the scale-up technologies required to meet the expanding clinical need for adult stem cells.
- Tissue Science | Regenerative Medicine | Cell Science | Stem Cell Research | Clinical trials in cell and gene therapy |Regulatory & Safety Aspects of Cell & Gene Therapy
University of Wisconsin School of Medicine and Public Health, USA
María Jesús del Olmo Barros
Autonomous University from Madrid, Spain Session
Revitacell Clinic, Spain
Miguel Guillermo Garber has over 30 years of experience in Internal Medicine and Cardiology, with expertise in regenerative medicine, training and education, research, product development and senior management. He has more than 12 years of working experience with stem cells, including building and managing the stem cell evaluation, exploring and developing stem cell therapies for cardiomyopathies, osteoarthritis and regenerative medicine at Stem cell Therapeutics Department of American Medical Information Group and Clinica Quirurgica Quantum. He is currently serving as Medical Director of Revitacell Clinic and Clinical Director of Regenerative Medicine Madrid. He is also a Professor of Regenerative Medicine and Editor of scholarly journals.
Chronic kidney diseases affect thousands of people worldwide and the final consequence is a renal failure (RF) becoming an extremely important public health issue, hemodialysis alleviates the situation by filtering the patient's blood, but it does not replace other kidney functions such as hormone release or homeostasis regulation. Transplantation of donor organs is the ultimate treatment for patients suffering from end-stage renal failure. Chronic kidney disease is a state of endothelial dysfunction, accelerated progression of atherosclerosis and high cardiovascular risk. As a consequence, cardiovascular disorders are the main cause of death in end-stage renal disease (ESRD). Actually, seeking for novel therapies, Kidney regeneration is a challenging but promising strategy aimed at reducing the progression to end-stage renal disease (ESRD) and improving the quality of life of patients with ESRD. Adult stem cells are multipotent stem cells that reside in various tissues, such as bone marrow and adipose tissue. Nowadays, there is growing evidence indicating that, under pathophysiological conditions, stem cells (SCs) derived from adipose tissue are able to migrate to the injured kidney, and they seem to play a role in glomerular and tubular regeneration. Moreover, adipose derivative stem cells can promote structural and functional repair, including hematopoietic stem cells and mesenchymal stem cells, which can also participate in the repair process by proliferation and differentiation into renal lineages. Mesenchymal SCs have been shown to decrease inflammation and enhance renal regeneration.
Cancer cells can acquire pluripotent character by forced expression of reprogramming factors, leading to the deletion of epigenetic memory and resulting in decreased tumorigenicity. However, studies on cancer cell reprogramming are limited and contradictory. In this study, the main aim is to investigate the effect of cellular reprogramming on the tumorigenicity of cancer cells. MDA-MB-435, G361 human melanoma cells and human primary melanocytes were transfected with Sendai viral vectors encoding the OKSM factors. After culturing, colonies have emerged in the reprogrammed melanocytes whereas; cluster of cells with distinctive morphology were observed in the transfected cancer cells. OKSM factors were upregulated in both cancer cells and primary melanocytes. All pluripotency markers showed increased expression in the reprogrammed melanocytes. However, only Nanog expression was observed in the transfected cancer cell lines. When embryoid bodies were formed, reprogrammed melanocytes showed expression of all lineage markers. Transfected MDA-MB-435 overexpressed FGF-5 and downregulated Brachyury; whereas G361 overexpressed both FGF-5 and Brachyury, compared to naive cancer cells; indicating that there were changes in epithelial to mesenchymal transitions. In order to understand the tumorigenicity of these resulting cells, sensitivity of chemotherapeutic agents, response to oxidative stress and invasion potential were compared. MDA-MB-435 cells were shown to decrease tumorigenicity after transfection with OKSM factors; whereas G361 increased tumorigenicity. These results suggest that even though transfection of cancer cells with OKSM factors were not able to fully reprogram cells towards pluripotency, it changed the tumorigenicity and modified cancer related pathways.
Autonomous University from Madrid, Spain
María J Del Olmo is Graduated in Musical Pedagogy, Royal Conservatory (Madrid). Degree as Music Therapist, Center for Music Therapy Research MI-CIM (Bilbao). Member of the Atelier de Musicotherapie de Bordeaux A.M.Bx, France. Founding member of the program RBL (rhythm, breathing and lullabies) Albert Einstein College of Medicine. The Louis Armstrong Center for Music & Medicine. Beth Israel Hospital, NY.
Professor in the Music Department at Universidad Autonoma, Madrid. Director of the Music Therapy Master Degree at Universidad Autonoma, Madrid. Collaborates with different training programs in several Spanish and foreign universities. Director of the Music Therapy Program at La Paz Hospital, Madrid. Chairperson of the Music Therapy and Health Foundation, Spain.
In pure biological terms there are many examples of perfectly synchronized musical actions organized in a musical form as structured and coherent as that of a symphony.
The most basic musical element we find in biology is rhythm. The heartbeat, breathing, the baby's sucking rhythm, the brain waves, the walking movement, all of them are rhythmic actions.
There are other musical elements, like melody, that are also present from birth. Crying is its first emotional expression.
Music can play a very important therapeutic role in early development with children at risk.
The physiological results as a consequence of carrying out a musical intervention for a therapeutic purpose show the importance of the use of music therapy in the hospital setting.
Study data: babies from 0 to 6 months of age hospitalized in Pediatric Intensive Care Unit (PICU) until reaching a number of 100 interventions. The study involved 87 subjects.
The predominant pathologies at these ages were: Bronchiolitis
38.98%, cardiac patients (with or without surgery) 13.56%, solid organ transplant 1.69% and various pathologies 45.76%.
Method: In this study the physiological and psychological responses can be observed as well as the results of the Comfort Behavior Scale test given to babies from 0 to 6 months of age hospitalized in a Pediatric Intensive Care Unit before, during and after the interaction with the responsible adult visiting the Unit, WITH and WITHOUT music.
The observation of the above-mentioned responses of the babies WITHOUT music was considered a control group.
Results: The decrease in RF (the rhythm of breathing ?) at the end of the adult interaction compared to basal breathing is significant when there is music (p = 0.016), whereas it is not significant if there is no music.
The decrease in the baby’s heartbeat during and after the adult interaction with respect to the time before is significant WITH music (p = 0.01 and 0.00 respectively), while it is not WITHOUT music. The increase produced in the SAT (?) O2 at the end of the adult interaction compared to the basal rate is significant WITH music (p = 0.00), whereas it is not WITHOUT music.
Musical improvisations of songs known to the child or proposed by the music therapist, immediately change the environment, which in the words of Wallon (1951/1985) will have a direct impact on biology
The Hebrew University-Hadassah Medical School, Israel
Nayef Jarrous is currently working at The Hebrew University of Jerusalem, Israel. His research interest is based on Human nuclear RNase P ribonucleoprotein in tRNA processing. He has published many articles in reputed journals.
Akershus University Hospital, Norway
Title: Relationship between cytochrome P450 polymorphisms and prescribed medication in elderly haemodialysis patients
Time : 12:00-12:30
Krystina Parker is a Specialist in Internal Medicine with ten years of experience. In the last five years, she has been a Consultant in the Nephrology department of Akershus University Hospital. She is currently working on her PhD at the University of Oslo which includes teaching medical students at various stages of their study. She is a Co-author in various projects including “Increased levels of inflammatory mediators and proinflammatory monocytes in patients with type I diabetes mellitus and nephrology” and “Glucarpidase (Carboxypeptidase G2) intervention in adult and elderly cancer patients with renal dysfunction and delayed methotrexate elimination after high-dose methotrexate therapy”.
Background: Elderly patients on haemodialysis treatment have polypharmacy. The drug response can be influenced by age, smoking, as well as through the genetic variations in the cytochrome P450 (CYP) enzymes system. More than 80% of all prescribed drugs are metabolized by CYP enzymes. Aims of this study were to describe the prevalence of polymorphism in 3 CYP isoenzymes and the relationship between CYP polymorphism and prescribed drugs.
Methods: 51 elderly haemodialysis patients aged ≥65 years were included. CYP-genotyping was carried out in whole blood by a real-time PCR method for detecting common variant alleles in CYP2C9, CYP2C19 and CYP2D6. The allele frequencies were calculated using the Hardy-Weinberg equation.
Results: The overall prevalence of CYP polymorphisms (heterozygous and homozygous) was 77%. The prevalence of heterozygous carriers of variant alleles coding for defective CYP2D6, CYP2C9 and CYP2C19 was 64%, 22% and 55%, respectively. The prevalence of homozygous carriers was 6% for each of the CYP2D6, CYP2C9 and CYP2C19 enzymes. The prevalence of CYP2D6*6, CYP2D6*9 and CYP2D6*41 variant alleles did not differ from that in a European Caucasian reference population (p=0.31). 23 patients (45%) had at least one CYP mutation and used drugs that are metabolized by CYP isoenzymes. Metoprolol and proton-pump inhibitors were the most commonly used drugs that could be affected by a heterozygous or homozygous mutation.
Conclusions: Polymorphisms of CYP2C9, CYP2C19 and CYP2D6 are common in elderly haemodialysis patients. Many of these patients have a phenotype with altered CYP enzyme activity and could benefit from close drug monitoring.
Chinese PLA General Hospital, China
The clinical successes of CD19-targeted chimeric antigen receptor (CAR) T-cell treatment in B-cell lineage hematopoietic malignancies in recent years, especially in acute lymphocytic leukemia (ALL), have shown that tumour cells can be precisely and efficiently targeted and eliminated by tumour-associated antigen-redirected immune cells. However, consideration of the significant differences between liquid and solid tumours is needed to ascertain rational, feasible, and efficient CAR T-cell–based therapeutic modalities in solid tumours. How to precisely instruct transfused CAR T cells to attack tumour cells growing outside of normal circulation is still a great challenge. Herein, we would like to briefly introduce CD20-, CD30-directed CAR T trials in patients with refractory lymphomas, and will emphatically address the results from our clinical trial data in patients with relapsed/refractory solid tumours treated by autoglous EGFR-, HER2-, and CD133-directed CAR T-based therapeutic modality.
Southeast University, China
Mesenchymal Stem Cells (MSCs) are attractive sources for cell therapy. However, MSCs have limited expansion capacity. Cellular senescence induced by long-term culture results in the loss of self-renewal property and disrupts their therapeutic functions. MicroRNAs (miRNAs) have been demonstrated as crucial regulators of senescence. In this study, we identified the core miRNA that regulated self-renewal of MSCs from human Umbilical Cord (UC). By a high-throughput deep sequencing, we first investigated the miRNA expression changes of UCMSCs, after a long-term ex vivo expansion. We found the expression of miR-18a was markedly down-regulated in late passaged UCMSCs. To verify miR-18a’s role in preventing senescence, we performed a knock down and overexpression analysis. Inhibition of miR-18a blocked the cell cycle process, repressed expression of “stem genes” including Oct4 and Nanog, and induced expression of senescence marker. Moreover, enforced expression of miR-18a promoted expression of Oct4 and Nanog and prevented senescence. Bioinformatics analysis identified CTDSPL and VEGFB as putative targets of miR-18a. Western blot and reporter assay confirmed miR-18a inhibited CTDSPL expression by binding its 3’UTR site. Furthermore, miR-18a improved VEGFB expression by targeting its promoter region. Finally, we designed a lentivirus vector to stably express miR-18a in MSCs. MSCs with stably expressed miR-18a had improved proliferation ability and differentiation potential. Taken together, these findings suggested that the expression of miR-18a is crucial for maintaining self-renewal of human MSCs.
Hamadan University of Medical Sciences, Iran
Sara Malih has completed her MSc in Medical Biotechnology from Hamadan University of Medical Sciences. She has published more than 3 papers in reputed journals. She has 2 research articles under review. She is the Reviewer of Tumor Biology-Springer and has participated in more than 3 international congresses.
Mesenchymal stem cells (MSCs) are currently the most convenient primary source of stem cells applicable in cell therapy and regenerative medicine. Due to low number of viable MSCs after cell transplantation novel approaches are required to improve viability, robust migration and proper homing of the cells. Recent evidence demonstrated that AdipoRon, a small-molecule AdipoR agonist has cytoprotective effect on post-ischemic cardiomyocytes. Herein we investigated the effect of AdipoRon on major factors involved in survival, migration and angiogenesis of rat bone marrow-derived MSCs cells. The expression level of cyclooxygenase-2 (COX-2), hypoxia-inducible factor-1 (HIF-1), C-X-C chemokine receptor type 4 (CXCR4), C-C chemokine receptor type 2 (CCR2), vascular endothelial growth factor (VEGF), matrix metalloproteinase-2 (MMP-2) and MMP-9 were assessed by Real-Time PCR, compared to untreated MSCs. Prostaglandin E2 (PGE2) production was investigated using ELISA assay. Measurement of Caspase-3 activity was carried out by an enzymatic assay. The migration potential of MSCs was observed by scratch test. AdipoRon significantly promoted the MSCs viability. The Real-Time PCR results indicated that the expression of COX-2, HIF-1, CXCR4, CCR2, VEGF, MMP-2 and MMP-9 were higher in AdipoRon- treated MSCs compared to control groups. The PGE2 level was enhanced by AdipoRon preconditioning; in contrast the caspase-3 activity was attenuated in these cells compared to the control group. Scratch test showed that AdipoRon could promote the migration ability of MSCs. Preconditioning of MSCs with AdipoRon prior to transplantation could enhance cell survival, angiogenesis and migration via activating the COX-2/ PGE2/ HIF-1 pathway and other contributing factors.