Scientific Program

Conference Series Ltd invites all the participants across the globe to attend 3rd International Conference and Exhibition on Cell & Gene Therapy Embassy Suites Las Vegas, USA.

Day 1 :

  • Track 1: Plant Stem Cell Rejuvenation
    Track 2: Plant Stem Cells: Human Therapeutics
    Track 3: Cell & Gene Therapy- Potential Applications
Speaker

Chair

Paul L. Hermonat

Central Arkansas Veterans Healthcare System, USA

Speaker

Co-Chair

Noah Weisleder

The Ohio State University, USA

Session Introduction

Paul L. Hermonat

Central Arkansas Veterans Healthcare System, USA

Title: The new genetics of adeno-associated virus

Time : 11:30-11:50

Speaker
Biography:

Paul L. Hermonat completed his Ph.D. at the University of Florida in 1984, and postdoctoral studies at the National Institutes of Health. He is a professor of Internal Medicine and OB/GYN at the University of Arkansas for Medical Sciences, and Research Career Scientist at the Central Arkansas Veterans Healthcare System. He mapped AAV’s genes, determined their functions, was the first to generate recombinant adeno-associated virus (AAV/NeoR) and to transfer genes into cells via this method, and has over 140 manuscripts published. From his first study on AAV-based gene therapy in 1984 there are now over 2,400 papers on the topic. He now continues studies on AAV genetics and helper genes, and on gene therapy for cardiovascular disease and cancer.

Abstract:

Adeno-associated virus (AAV) (type 2) is now a very popular human gene therapy vector with a long active transgene expression period and has had no reported vector-induced adverse reactions. Yet the basic molecular biology of this virus has not been fully addressed. For thirty years AAV2 genetics has been comprised of two genes: rep, needed for replication, and lip-cap needed for capsid formation. However, recently Jurgen Kleinschmidts group has identified a new gene and protein, called assembly activating protein (AAP) which is involved in AAV2 particle maturation. We have also recently analyzed another potential gene at the far 3 end of the AAV2 genome, previously referred to as X (nt 3929 to 4393), overlapping the 3 end of the cap gene, has never been characterized, although we did previously identify a promoter just up-stream (p81). Computer analysis suggested that X was involved in replication and transcription. The X protein was identified during active AAV2 replication using a polyclonal antibody against a peptide starting at amino acid 38. Reagents for the study of X included an AAV2 deletion mutant (dl78-91) a triple nucleotide substitution mutant, destroying all three of the 5 AUG-initiation products of X, with no effect on the cap coding sequence, and X-positive-293 cell lines. Here, we found that X up-regulated AAV2 DNA replication in differentiating keratinocytes (without helper virus, autonomous replication) and in various forms of 293 cell-based assays with help from wild type adenovirus type 5 (wt Ad5) or Ad5 helper plasmid (pHelper). The strongest contribution by X was seen in increasing wt AAV2 DNA replication in keratinocytes and dl78-91 in Ad5-infected X-positive-293 cell lines (both having multi-fold effects). Mutating the X gene in pAAV-RC (pAAV-RC-3Xneg) yielded approximately a ~33% reduction in recombinant AAV vector DNA replication and virion production, but a larger effect was seen when using this same X-knockout AAV helper plasmid in X-positive-293 cell lines versus normal 293 cells (again, multi-fold). Taken together these data strongly suggest that AAV2 X encodes a protein involved in the AAV life cycle, particularly in increasing AAV2 DNA replication, and suggests that further studies are warranted.

Speaker
Biography:

Noah Weisleder received his PhD in cell biology from Baylor College of Medicine and conducted postdoctoral studies in physiology at Robert Wood Johnson Medical School. Currently, Dr. Weisleder is an associate professor of Physiology and Cell Biology at The Ohio State University and an Investigator in the Davis Heart and Lung Research Institute. He has authored numerous manuscripts on muscle physiology, cardiovascular disease, membrane repair and calcium homeostasis. He is an inventor on multiple U.S. and international patents. These inventions led to formation of TRIM-edicine, a biotechnology company developing protein therapeutics where he is a Founder and Chief Scientific Officer.

Abstract:

Regenerative medicine approaches utilizing stem cells to increase heart performance represent an important potential therapeutic approach for the treatment of myocardial infarction (MI). Two challenges in the use of embryonic stem cells (ESCs) in cardiovascular regenerative medicine is the poor differentiation of these cells into cardiomyocytes and additional concerns with the formation of tumors following implantation. Here we use an innovative encapsulation technology coupled with a predifferentiation approach that increases the differentiation of ESCs into cardiomyocytes and reduces tumor formation following direct injection into the infract zone of mouse hearts. Mouse ESCs were encapsulated in alginate microcapsules with an aqueous liquid core using coaxial electrospray and cultured for 7 days to form pluripotent ES cell aggregates that were then treated with BMP-4 and bFGF for three days to induce early differentiation into the cardiac lineage. Mice were subjected to permanent surgical occlusion of the left ascending artery and 2x105 of the encapsulated aggregated cells were injected in to the infarct zone. Encapsulated ESCs increase survival time as well as cardiac structure and function compared to mice treated with ESCs alone. Labeling of ESC cells showed the appearance of new cells in the infarct zone that express cardiomyocyte markers such as troponin. Additionally, while injection of ESC alone would produce tumor formation at the injection site, no tumor formation was observed with encapsulated ESC. We conclude that direct injection of encapsulated ESC in an infarct zone allows for improved pathology and reduced tumorgenesis following experimental induction of MI in mice.

Denis Ivanov

Scientific Research Institute for New Medical Technologies, Russia

Title: Cell therapy patients with cardiomyopathy

Time : 12:10-12:30

Speaker
Biography:

Denis Ivanov completed his MD and PhD from Medical Institute Tula State University. Dr. Denis Ivanov has received many patents and scientific discovery. His is a professor on faculty of biomedical sciences Medical Institute, Tula State University and serving as an editorial board member journal of New Medical Technologies.

Abstract:

Aim: To evaluate the efficacy of systemic allograft multipotent mesenchymal stromal cells (MMSC) for the treatment of dilated cardiomyopathy (DCMP) of viral origin. A group of patients (n = 32) with DCMP after viral infection, aged 18 to 46 years under surveillance 24 months. One of the main factors for patient selection was a parameter of ejection fraction by echocardiography, which was less than 30%. All patients underwent ECG, biochemical blood tests, 6-minute walk test, tested by questionnaire - before to the injection, in 14 days after the initial injection, 1, 3, 6, 12, 24 months after the injection of MMSC. All procedures were carried out with the written informed consent of patients. The injection cells are characterized by the absence of contamination viruses, bacteria, fungi, viability of 97-100%, the expression of CDmarkers. The injection was carried out in the hospital after a few days of pharmacological correction of the general condition. All patients performed course injection MMSC. We have perfected an algorithm of the procedure. Allergic reactions, as the nearest and distant period, was not noted. During the observation period the mortality was the main group 9,4% in the control group, 37,5%. The estimate of the results of allogenic MMSC in patients with DCMP showed an increase in life expectancy, increased EF, reduction of heart failure, reducing the number and duration of rehospitalization, compared with the control group. The obtained positive results show the safety and efficacy of injection of allogeneic MMSC patients with DCMP after viral myocardial damage.

Jan H. Spaas

Global Stem cell Technology, Belgium

Title: Wound healing capacities of allogenic skin-derived stem cells in a large animal model

Time : 12:30-12:50

Speaker
Biography:

Jan H. Spaas, veterinarian, graduated from the Faculty of Veterinary Medicine, Ghent University (Belgium) in 2010. In that year he also won the price for young authors of the Flemish Veterinary Journal. He completed his Ph.D. at the Department of Comparative Physiology and Biometrics of the Faculty of Veterinary Medicine, Ghent University. Since 2012, he became the laboratory director of Global Stem cell Technology, an organization that is specialized in regenerative therapies for horses. Thanks to external research funding, he is the industrial promotor of Ph.D. and master students who actively participate in the companys research program.

Abstract:

In recent years, different classes of stem cells have been investigated for their ability to regenerate organs and tissues after injury. Consequently, the isolation and characterization of stem cells from different adult tissues such as bone marrow, adipose tissue, peripheral blood, or umbilical cord (blood) have been investigated. However, ongoing regeneration of the skin is achieved through somatic stem cell differentiation within the epidermis and the hair follicle. The skin may, hence, serve as an excellent source of epithelial stem cells (EpSCs). Accordingly, it has been proposed that EpSCs might be useful in the treatment of several diseases, such as burn wounds, chronic wounds, and ulcers. Nevertheless, only limited information concerning EpSCs is available to date. Nonetheless, this stem cell type could prove itself useful in skin reconstitution after injury. After harvesting from equine skin, the purified cells were characterized as EpSCs by means of positive expression for CD29, CD44, CD49f, CD90, Casein Kinase 2b, p63, and Ki67, low expression for cytokeratin (CK)14 and negative expression for CD105, CK18, Wide CK, and Pan CK. Moreover, the isolated cells were differentiated toward keratinocytes and adipocytes. To assess the regenerative capacities of allogenic EpSCs, a double-blinded clinical study was performed. This revealed significant macroscopic as well as microscopic improvements in different wound-healing parameters in the EpSC treated group.

Tae Suk Ro-Choi

University College of Medicine, USA

Title: Dynamics of RNA string and hnRNP compactions

Time : 12:50-13:10

Speaker
Biography:

Tae Suk Ro-Choi is Professor at University College of Medicine, USA. She has vast experience in the field of DNA and RNA dynamics. She is interested in analyzing chemical approaches for structure and function of RNA.

Abstract:

The dynamics of RNA transcription involves in protein binding and RNA compaction. Co-transcriptional mechanisms involve with hnRNPs and other processing factors such as capping enzyme, and components of spliceosomal complexes. And transport of the complexes are well established. The hnRNP formations are observed in perichromatin fibrils, perichromatin granules, interchromatin granules and granulses at the nuclear pore. These granulse are ~22-24 nm in diameter and composed of hnRNA and hnRNPs. Larger spliceosomal particles are also observed. In vitro reconstitution of hnRNP revealed the stoichiometry of ~700-800 nucleotide RNA and 48 hnRNPs (A112, A212, B14, B24, C112, and C24). Observation of the reconstituted 40S particels showed a diameter ranging 22-24 nm. The RNA of 700-800s nucleotide may measure 400-500 nm in length. It is interesting to observe a model, where a simple enclosing accommodation of the RNA length for the circunference of the 40S particle would force to undergo 6-7 times of wrapping around. Therefore, a logical requirment of the particle model would besecondary foldings to satisfy the compact nature of hnRNP particles. It has been known that single stranded RNA tends to make small stem-loop structures and their free energy can be readily evaluated for thermodynamic applications. In order to find a best fitting molding pattern, various model studies were made to evaluate configuration of hnRNPs using 25-hydroxyvitamin D3 1αhydroxylase, insulin, FMR1 and ovomucoid transcrips by Q-fold program. The hnRNA folding patterns are uniform. Initial compaction ratio is 3-9 and 1 stem-loop is formed every 15-18 nucleotides. It is in good agreement with calculated chain of 15-17 nucleotides per hnRNP protein in reconstituted particles.

Break: Lunch Break 13:10-13:50 @ Atrium
Speaker
Biography:

Yan Li has completed her Ph.D. from the Fourth Military Medical University (China) in 2009 and she used to be a visiting scholar for post-doctoral research in Purdue University (USA). Now she is a lecturer of the department of Biochemistry and Molecular Biology, the Fourth Military Medical University (China). She has published more than 30 papers in reputed journals, including J BiolChem, MolTher,Breast Cancer Res, and Cell Death Dis.

Abstract:

Xerostomia is a subjective symptom and prevalent in more than one-third of postmenopausal women usually complainof seriouslyinterferedchewing, deglutition, and speaking. Saliva production is subjected to the processes of a plasma-like, isotonic primary secretion insalivary acini and subsequent reabsorption ofNa+ and Cl- in the collecting ducts, leavinghypotonic saliva into the oralcavity.Some xerostomia patients actually do not suffer from decreasedsaliva secretionbut from the increasedionic osmotic pressure orviscidity of saliva.Although the feeling of oral dryness can be ameliorated with estrogen supplementation administration, the side effects of estrogen greatly restrict its application.We found that N-myc downstreamregulated gene 2 (NDRG2) is involved in estrogen-mediated Na+ and Cl−transport in salivary ductal cells. In addition, an ovariectomized rat model was used to mimic xerostomia in postmenopausal women and adenovirus vectors bearing NDRG2were constructed to validate their therapeutic potential. Ovariectomized rats exhibited severe sialadenhypofunction, including decreased saliva secretion and ion reabsorption, as well as increased water consumption.We also found that the expression of NDRG2, Na+/K+-ATPase, and epithelial sodium channels (EnaC) decreased in ovariectomized rat salivary glands, which are all involved inNa+reabsorption. Retrograde ductal delivery ofNDRG2adenovirusin ovariectomized ratthroughsubmandibular glandcan efficient lyimprove the dysfunction of Na+ and Cl− reabsorption, and promote saliva flow rate and water intakereturn to normal levels. This study elucidates theunderling mechanismof estrogen deficiency-mediated xerostomia and provides an attractive strategy for therapeutic intervention.

Sonal Malhotra

University of Tennessee-Health Science Center, USA

Title: An examination of the role of disulfide bond isomerase a protein in bacterial virulence

Time : 14:10-14:30

Speaker
Biography:

Sonal Malhotra is a Biotechnology Professional with over 15 Plus years of Experience in biotech Industry and Academia (India & USA). She completed her PhD in University of Tennessee-Health Science Center, USA. She is interested in Translational research, Genomic and Proteomic approaches to understanding disease pathogenesis and management. Infectious Disease Process: In particular the role of Disulfide Isomerases in Infectious disease.

Abstract:

In the last two decades there has been a tremendous increase in number of bacterial genome sequencing projects. This effort has yielded a large collection of data that is freely available to researchers. The role of this study to perform selective and informed literature searches to establish the role of disulfide isomerases from different organisms in virulence in their respective hosts. DsbA and DsbA-like proteins have been implicated in bacterial virulence for a long time. Recently DsbA proteins have been implicated in biofilm formation or maturation in several bacterial species. Biofilms are relevant to the medical community. In particular, Bacterial cells in biofilm exhibit altered resistance to antibiotics. Furthermore biofilms- comprising mixed populations of different bacterial species-have been demonstrated on medical implant devices. Therapeutic intervention on such medical implants has been attempted. Disruption of dsbA gene in P. aeruginosa leads to a reduction in virulence in an animal infection model. We have recently shown that a dsbA disruption in P. aeruginosa PAO1 exhibits enhanced biofilm formation with respect to the wild type strain. This result is similar to results with P. putida: where a dsbA disruption exhibits enhanced exopolysachharide and biofilm formation. In our simple biofilm assay examining gross biofilm formation: the kinetics of biofilm formation on plastic (petri dish) and aluminum are higher in the dsbA mutant with respect to the wild type strain. The similarity in findings between P. aeruginosa and P. putida support the notion of conservation in the mechanisms of biofilm formation amongst different bacterial species. This lends credence to examining biofilm formation pathways in different bacterial species. Such a study will likely reveal conservation in mechanisms of biofilm formation and maturation. It is likely to pave the way for potential avenues for further research and therapeutic intervention. We will report here in this study some some key players in biofilm formation and any likely conserved mechanisms of biofilm formation. Further studies are likely to support a comparison of biofilm formation in a plant infection model (Wt and dsbA disruption mutant) and evaluation of biofilm formation on different materials.

Tahsin Shoala

Misr University for Science and Technology, Egypt

Title: Oxi1 mutant plays an important role in Arabidopsis resistance against aphid (Myzus persicae)

Time : 14:30-14:50

Speaker
Biography:

Tahsin Shoala has completed his PhD from Newcastle University and he is currently an assistant professor at College of Biotechnology, Misr University for Science and Technology. He is the chief executive of MEDIC, Misr Entrepreneur Development Innovation Centre. He has published more than 5 papers in reputed journals and running number of research projects.

Abstract:

The interactions between plants and insect herbivores comprise a complex, co-evolved natural system. Arabidopsis mutants provide a powerful tool for studying plant-insect interactions as the phenotype of individual gene deletions can be studied with relative ease. Oxidative signal inducible (Oxi1) is a serine/ threonine kinase required for oxidative burst-mediated signalling in Arabidopsis roots and full activation of MAPKs cascades. The expression of β-1,3-glucanase, important in the response to aphid feeding, are also investigated through the use of Gns1, Gns2 and Gns3 mutants.
Oxi1 mutants showed resistance to aphids and delay the developmental rate in both adults and nymphs compared to the Col-0 wild type. β-1,3-glucanase genes Gns1, Gns2, Gns3 and Gns5 were not expressed in Oxi1 mutant. However, Gns2 was expressed to high levels in Col-0 in response to aphid feeding. Also, Gns2 was up-regulated in both Oxi1 null mutant in WS2 and WS2 background. Callose synthase GSL5 was down-regulated in both Oxi1 null mutant in WS2 and WS2 background. β-1,3-glucanase mutants Gns1, Gns2 and Gns3 were resistant to aphid feeding and aphid development in both adults and nymphs.
Callose synthase is important for plant resistance especially callose synthase GSL5. Expression of β-1,3-glucanase genes, especially Gns2, play an important role in plant susceptibility in response to aphid feeding. Oxi1 mutants showed resistance in response to aphid feeding. Potentially through the induction of callose deposition via MAPKs resulting in inducing ROS as an early response and signal transduction improves the resistance level of the plant.

Speaker
Biography:

A.S. Singh has is presently working as postdoctoral fellow in Dr. Axel Brockmanns Lab at NCBS. He completed his Ph.D. research in May 2013 in the University of Calcutta under the supervision of Dr. Usha Rajamma. His main research interest is to understand the mechanism of how genes regulate behaviors through certain neural circuitry systems.

Abstract:

Autism spectrum disorder (ASD) is one of the most common genetically predominant neurodevelopmental disorders and the prevalence rate is more than one percent. The pathophysiogy of the disorder is highly complex and several genes are believed to involve with each having minor contribution leading to the final ASD phenotype. In the last two decades, various risk variants of different genes have been identified but none of the individual variants account for more than 1% of ASD cases in average. Recently, identification of genes that contribute to specific autism endophenotype is highly appreciated. So far two most common finding in ASD research are transient increased of head circumference and platelet hyperserotonemia. And platelet hyperserptonemia has been considered as autism endophenotype. Moreover, abnormal serotonin synthesis capacity has been implicated in the brain of ASD individuals, thereby the involvement of serotonergic system abnormality both in the brain as well as periphery in the pathophysiology of ASD. Investigation of few selected single nucleotide polymorphisms from three potential serotonergic candidate genes, TPH1, TPH2 and ITGB3 via population and family based approaches, revealed that these genes are moderately risk to the disorder. Gene-gene interaction analysis further suggests their interactive role towards the cause of ASD. However, as we do not have the functional experimental data, the present finding provides only preliminary evidence. However, these data highlight the need of extensive research both in genetic and functional level for clear understating the role of these genes in the etiology of ASD.

Hussain Mustatab Wahedi

Gachon University College of Pharmacy, South Korea

Title: Co-regulation of Sirt1 and Pin1 contributes to UV A/B-induced skin photo ageing

Time : 15:10-15:30

Speaker
Biography:

Hussain Mustatab Wahedi finished his Masters in Biosciences at the age of 25 from COMSATS Institute of Information Technology, Islamabad. He has been working in well reputed educational and research institutions over the last few years. He has published 7 papers in international journals and is doing PhD from Gachon University, South Korea.

Abstract:

Cellular senescence is the hallmark of the ageing. Many characteristic features causing cells to age or senesce are observed during the process of ageing. Factors contributing to ageing may be intracellular or external. One of the most common factors causing cells to age and depict morphological changes characteristic to the ageing is ultraviolet (UVRs) light. In the present study we demonstrated that exposure to UVA and/or UVB light decreased the viability of skin cells (NHDFs and HaCaT cells) in vitro and caused the surviving cells to show symptoms of ageing like increased ROS production, high NO secretion and apoptotic DNA ladder. Expression patterns of two proteins Sirt1 and Pin1 which are critically involved in ageing were also checked in vitro and in vivo. It was found that upon treatment of UVA and/or UVB light, expression of Sirt1 was decreased and Pin1 was increased. However, it was surprising to find that treatment of cells with the compound KSY-PH1, which is presumably a Pin1 inhibitor, caused the expression of Sirt1 to increase significantly. This increase in Sirt1 expression was even more noticeable than treatment with Resveratrol, an established Sirt1 activator. This finding is thus promising for the reason that protective and rejuvenating role of Sirt1 in the process of ageing (especially UV light induced ageing) can be strengthened by the treatment of KSY-PH1. So we assume that KSY-PH1 may delay the ageing and senescence induced by UV irradiation. This work was supported by a grant from the Next-Generation BioGreen 21 Program (No.PJ009511), Rural Development Administration, Republic of Korea.

Maryam Eslami

Harvard Medical School, USA

Title: Fiber reinforced hydro gel scaffolds for heart valve tissue engineering

Time : 15:30-15:50

Speaker
Biography:

Eslami graduated with MD (2010), PhD (2014) degrees. She has joined in Harvard Medical School (Harvard-MIT Division of Health Sciences and Technology) as a part of her Ph.D. dissertation in the field of Heart valve. She has carried out some broad research on Orthopedic fractures and has published a book and papers in this field. Her U.S and PCT patent achieved the rank of “Best 2008 Invention” from WIPO (World Intellectual Property Organization of the United Nations) and she has received the title of "Best 2008 Women inventor" from WIPO and 6 Gold Medals and 6 Honorary Diplomas in the Contests and Fair of the Inventors in Geneva and South Korea. International and national awards for her research were one of the fundamental achievements that she has received.

Abstract:

Heart valve related disorders are among the major causes of death worldwide. Although prosthetic valves are widely used to treat this pathology, current prosthetic grafts cannot grow with the patient while maintaining normal valve mechanical and hemodynamic properties. Tissue engineering may provide a possible solution to this issue through using biodegradable scaffolds and patients own cells. Despite their similarity to heart valve tissue, most hydrogel scaffolds are not mechanically suitable for the dynamic stresses of the heart valve microenvironment. In this study, we integrated electrospun poly(glycerol sebacate) (PGS)-poly(ε-caprolactone) (PCL) microfiber scaffolds, which possess enhanced mechanical properties for heart valve engineering, within a hybrid hydrogel made from methacrylated hyaluronic acid (HAMA) and methacrylated gelatin (GelMA). Sheep mitral valvular interstitial cells (MVICs) were encapsulated in the hydrogel and evaluated in hydrogel-only, PGS-PCL scaffold-only and composite scaffold conditions. Although the cellular viability and metabolic activity were similar among all scaffold types, the presence of the hydrogel improved the 3D distribution of MVICs. As seen by similar values in both the Young's modulus and the ultimate tensile strength (UTS) between the PGS/PCL scaffolds and the composites, microfibrous scaffolds preserved their mechanical properties in the presence of the hydrogels. Compared to electrospun or hydrogel scaffolds alone, this combined system may provide a more suitable 3D structure for generating scaffolds for heart valve tissue engineering.

Break: Coffee Break 15:50-16:05 @ Atrium

Ebtesam M. Al-Ali

Kuwait institute for scientific research, Kuwait

Title: Important multiple plant viruses present on vegetable crops in Kuwait

Time : 16:05-16:25

Speaker
Biography:

Ebtesam M. Al-Ali has completed her BSC Studies at 1993 from Kuwait University and joined the work as scientific researcher at Kuwait institute for scientific research with the molecular genetics group, she led 5 projects, 3 completed and 2 ongoing, she has published more than 5 papers in reputed journals and international conferences.

Abstract:

Farmers complained to the Kuwait Institute for Scientific Research (KISR) about diseases affecting their vegetable crops, leading to subsequent heavy economical losses. Initial field visits indicated that the diseases might be caused by viruses. Four plant families, including Cucurbitaceae (cucumber, squash, melon, zucchini), Solanaceae (tomato, potato, pepper, eggplant), Liliaceae (onion) and Leguminosaea (bean) were surveyed throughout the growing seasons for viruses by ELISA from December through May. The survey was conducted on six farms located in the two main agricultural districts areas, Wafra (South) and Abdally (North). Each farm was visited four times during the production season (planting, pre-flowering, post-flowering and fruiting). This preliminary survey constitutes the first report in Kuwait of fifteen out of eighteen viruses detected, including Cucumber mosaic virus, Garlic common latent virus, Iris yellow spot virus, Onion yellow dwarf virus, Melon necrotic spot virus, Papaya ringspot virus, Pepino mosaic virus, Pepper mild mottle virus, Potato moptop virus, Potato virus M, Potato virus X, Potato virus Y, Squash mosaic virus, Tomato mosaic virus, Tomato spotted wilt virus, Tomato yellow leaf curl virus, Watermelon mosaic virus, and Zucchini yellow mosaic virus. Virus incidence was close to 100% on some crops, including cucurbit and onions and double or triple infections were common. Several important viruses were not found, including Potato virus A, Potato virus S, Potato leafroll virus, and Potato virus V. TYLCV was also detected by PCR, and partial sequence of the amplicon was done.
Keywords: ELISA (Enzyme Linked Immunosorbant Assay), Cucurbitaceae, Solanaceae, Liliaceae, Leguminosaea.

Ghazala Rubi

University of Health Sciences, Pakistan

Title: Genetic Susceptibility of HCV RNA and its genotypic distribution in the Punjab, Pakistan

Time : 16:25-16:45

Speaker
Biography:

Ghazala Rubi U.K trained Clinical Molecular Biologist with M.Phil degree in Human Genetics & Molecular Biology and PhD Thesis in Human Genetics and Molecular Biology having more than 15 years of experience in molecular biology Pathology Laboratory services, as HCV RNA, HBV DNA PCR, REAL TIME, HCV & HBV Genotyping. Four years of experience of teaching to M.Phil graduates of a medical students. I did my services at University of Health Sciences as a Senior Research/Teaching Faculty for four years in Human Genetics and molecular biology department. Her Ph.D. dissertation submitted and she did a research project on Susceptibility of HCV RNA in our isonym group". Used to work in mixed community as already worked in U.K. and had done Molecular Biological trainings in UK, Germany and Italy.

Abstract:

Introduction and rationale: Hepatitis C, a widespread infectious disease targeting circa 130 million people worldwide, is caused by the Hepatitis C Virus (HCV). HCV genome, consisting of 9,600 nucleotides, is fully sequenced. Population specific high variation in HCV genome exists. HCV is classified into 7 different genotypes with several subtypes. Genotypes 1, 2 and 3 are found worldwide.
Objective: To find out the ecology and genetics of susceptibility of HCV RNA in various isonym groups of the Punjab population
Subjects and Methods: A sample of 349 chronic HCV hospital patients was studied who were already taking the treatment of standard therapy of Interferon+Ribavirin thrice a week. The sample was naturally divisible into three groups: Responder, the patients who received the standard therapy and recovered/cured; Relapser, the patients who after a course of therapy became negative for HCV RNA but after sometimes (6-18 months) became HCV positive again; Non-responder, the patients who did not show positive response to therapy.
Results It was found that HCV genotype 3a is very common (84.0%) among responders group while genotype 1a is more common in relapser (66.2%) and non-responders (54.0%). Five of the Six main genotypes, namely, 1a (61.40%), 2a (0.50%), 2b (20.00%), 3a (13.70%) and an Untypeable (4.40%) were found among the 12 different castes/tribes/isonym ethnic groups. Genotype 4 was not found. The HCV frequency in 12 isonym groups is as follows: Arain (15.26%), Gujjar (10.02%), Jutt (18.91%), Kashmiri (10.02%), Malik (10.44%), Mughal (3.21%), Pathan (17.19%), Rajput (11.46%), Sheikh (3.43%), and Sayyed (4.87%). Jutt caste was found to have the maximum infection of HCV, while the minimum was found in Mughals. Genotype 3a among responder was most common in Rajput caste. Among the relapser 1a is most prevalent in Jutts. Pathans top the list of non-responders having 1a and 2b. Genotype 2a was found only in one sample of Rajput, who was non-responder.
Gene-polymorphism in IL-10 and IL-28B genes to ascertain the genetic susceptibility among various isonym groups revealed six SNPs. In IL-10, SNP at 1082 position, AA (14.5%), GA (80.30%) and GG (5.20%); SNP at 819, AA (3.2%), AC (84.7%) and CC (12.0%); and SNP at 592 position, AA (6.0%), CA (69.9%) and CC (24.1%). CA was in high frequency than CC and AA homologous gene polymorphism. In IL-28B SNP at location a, GG (4.8%), TG (40.6%), TT (54.6%); SNP at location b, CC (34.9%), CT (58.2%), TT (6.8%) and CC (40.2%), CT (43.8%), TT (16.1%) was found. Frequency of TT homologous high at one position, CT hetrozygous polymorphism was frequent at second and third position.
Conclusion: Human genetic susceptibility to HCV genotypes appears to be of importance in getting the infection. The study suggests that IL-10 and IL-28B interleukin genes are common in two major caste of the Punjab. A cohort study needs to be done for better understanding of human susceptibility to HCV infection and its management.

Break: Break out @ Riviera
  • Track 4: Stem Cell Therapies
Speaker

Chair

Steven Gitt

The Phoenix Stem Cell Treatment Center, USA

Speaker

Co-Chair

Rafael Gonzalez

DaVinci Biosciences, LLC USA

Session Introduction

James Wang

StemBios Technologies, Inc., USA

Title: The application for the SB small stem cell

Time : 11:30-11:50

Speaker
Biography:

Wang brings extensive experiences in stem cell research. He was the Senior Scientist/Director of MorphoGen Pharmaceuticals which is a pioneer stem cell company focusing on muscle-derived stem cells. Prior to MorphoGen, Dr. Wang was at VitaGen which is a cell-therapy company dedicated to treating fulminant hepatic failure using bioartificial liver devices. Dr. Wang received his Ph.D. in Molecular Biology at Vanderbilt University. He completed his postdoctoral work at The Salk and Burnham Institute in La Jolla, California. Before joining StemBios, Dr. Wang was an assistant research professor at City of Hope Medical Center. Current, Dr. Wang is a Chief Scientific Officer in StemBios to lead the stem cell research. He is the co-inventor of several patents and numerous scientific publications.

Abstract:

Small stem cells, such as spore-like cells, blastomere-like stem cells (BLSCs), and very-small embryonic-like stem cells (VSELs) have been described and debated in recent studies, although their multipotency in human tissues has not yet been confirmed. Here, we report the discovery of adult multipotent stem cells derived from human bone marrow, which we call StemBios (SB) cells. These isolated SB cells are smaller than 6 micron and are DAPI+ and Lgr5+ (Leucine-Rich Repeat Containing G Protein-Coupled Receptor 5). Here, we discuss the SB cells and its application.

Speaker
Biography:

Steven Gitt, MD, FACS, graduated college at UCLA and medical school at Wayne State University. He completed his General Surgery residency at Good Samaritan Hospital in Phoenix, Arizona and his Plastic Surgery Residency at the University of Michigan Hospitals in Ann Arbor. Dr. Gitt is a Clinical Assistant Professor of Plastic Surgery at the University of Arizona, Phoenix School of Medicine and holds faculty appointments at several other medical schools. He is the founder and Medical director of North Valley Plastic Surgery and The Phoenix Stem Cell Treatment center. Dr. Gitt is on the editorial advisory boards of several scientific publications. He has spoken at both national and international meetings on a variety of topics and is a clinical researcher for RTI, inc., Ulthera Inc., Cynosure/Palomar inc., and Medi Khan, inc.. Dr. Gitt is a clinical spokesperson for Allergan inc., Ulthera inc., RTI inc., and Cynosure/Palomar inc.. He has spoken on Stem Cell/SVF/Regenerative medicine topics dozens of times.

Abstract:

Regenerative medicine promotes the repair or replacement of lost tissue or organ function due to damage or congenital defects, thereby creating living, functional tissues, and/or stimulating previously irreparable organs to heal themselves. Bone Marrow and embryonic stem cell therapies have presented great difficulties while autologous SVF is readily bioavailable, easily harvested, processed in a closed surgical setting with only minimal manipulation, and can be immediately utilized. Patients presented with a variety of orthopedic (peripheral and spinal), rheumatologic, neurologic, cardio-pulmonary, autoimmune, genitourinary, ophthalmic, gastrointestinal, degenerative, and deteriorative conditions. Nearly all patients received intravenous deployment, while intra-articular, intra-lesional, nebulized pulmonary, and direct injection (with radiologic guidance as needed) techniques were employed as dictated by protocols. Clinical efficacy showed that between 60%-80% of patients enjoyed positive responses. Global complication rate (generally minor incidents related to the liposuction required) was below 2% and there were NO serious or major adverse events. Our results show that this procedure is extremely safe, ethical, versatile and appears to have a consistently high efficacy and therapeutic yield.

Biography:

Julie Murrell is a Senior R&D Manager for Stem Cell Biology and Collaborations at EMD Millipore. Dr. Murrell has led an early technology assessment group for the past 7 years and has been part of the Stem Cell group for 3 years. Through that time, she has led the efforts to establish robust assays and identify new targets as key quality attributes for large scale stem cell manufacturing, with a special focus on hMSCs. Dr. Murrell’s background is in Cell and Molecular Biology. Her multi-disciplinary background has led to innovative team-driven approaches in the field of stem cell production.

Abstract:

Human mesenchymal stromal/stem cells (hMSCs) are used extensively in clinical studies for therapeutic agents and as drug discovery tools. Current in vitro culture methods are proving cumbersome to scale. We have previously demonstrated an expansion paradigm that uses a scalable, single use, stirred tank bioreactor with microcarrier scaffold for hMSC expansion. A bioreactor system enables direct monitoring for the specific characteristics of hMSCs at any point during the expansion, thus assuring product quality and consistency. There is a desire desire to remove animal-derived materials from the production process to minimize risk to the patient for a product that is minimally processed. We are evaluating alternatives to serum including human-derived material and other reduced serum formulations. Additionally, we have identified reagents that support robust growth in traditional 2D and more advanced suspension culture while maintaining cellular phenotype and functional potential. These materials can be used in place of currently used reagents to support xeno-free cell therapy production with confirmed consistency, quality and reproducibility in large scale in vitro systems.

Juliann G. Kiang

Uniformed Services University of The Health Sciences, USA

Title: Bone marrow mesenchymal stem cells increases survival after ionizing irradiation combined with wound trauma

Time : 12:30-12:50

Speaker
Biography:

Juliann G. Kiang completed her PhD and postdoctoral studies at the University of California at Berkeley. She is a professor of Radiation Biology at the Uniformed Services University of the Health Sciences, Bethesda, MD and Principal Investigator at Armed Forces Radiobiology Research Institute. She is an inventor and editorial members of journals. She has over 140 publications. Among all awards, she receives the Research and Development Achievement Award from the US Department of Army. She is a US DoD STEM model. She is the first to describe the skin-wound amplifies iNOS activation, cytokine concentrations, and sepsis after ionizing irradiation.

Abstract:

Ionizing radiation combined with wound trauma (CI) induces more mortality than radiation alone (RI). The increase is mediated partly by activation of inducible nitric oxide synthase (iNOS) pathway, cytokines increases, bacterial infection, and ATP loss. A promising therapeutic regimen for managing CI is transfusion with bone marrow-derived mesenchymal stem cells (BMSCs). The aim of this study was to evaluate efficacy of BMSCs on survival after CI. BMSCs were collected from the femur bone marrow of B6D2F1/J mice, expanded and cultivated in hypoxic conditions for 28 days. The cell phenotype was identified by BMSC-positive markers: CD44, STRO1, and SCA1. B6D2F1/J mice were exposed to 60Co-γ photon radiation followed by a 15% total body skin-wound trauma. Twenty-four hours later, these mice received a single injection of BMSCs. As a result, BMSCs treatment significantly improved mouse 30-day survival by more than 30% above the control group. To understand mechanisms underlying BMSC action in septic conditions because of CI-induced bacterial infection, BMSCs were treated with LPS in vitro. LPS significantly increased gene expression of IL-1α, IL-1β, and iNOS in a dose- and time-dependent manner. At 24 h after exposure to 0.5 µg/ml LPS, expression of NF-kB-p65, IL-1β, and iNOS proteins, and NO production in the cells were increased 2-, 11-, 4-, and 6-fold, respectively. These data are consistent with the idea that LPS-induced alterations in BMSCs are mediated by the TRL4/NF-κB axes, which in turn modulates the host adaptive responses during septic conditions. (Supported by NIH/NIAID YI-AI-5045-04. The views expressed do not necessarily represent NIH/AFRRI/USUHS/US DoD).

Neelam Yadav

Albert Einstein College of Medicine, USA

Title: Bone marrow-stem cell therapy for hemophilia A disease

Time : 12:50-13:10

Speaker
Biography:

Neelam Yadav has completed her PhD from Industrial Toxicology Research Centre, Lucknow and postdoctoral studies from Stem Cell Biology Laboratory of National Institute of Immunology (NII) New Delhi, India. She is Assistant Professor in Department of Biochemistry, at Dr. R.M.L. Avadh University, Faizabad, India. Dr. Neelam has published many papers in reputed international journals and filed one patent. She is Visiting Scientist in Department of Medicine, Albert Einstein College of Medicine, New York, USA, under Raman Fellowship Programme of University Grant Commission (UGC) New Delhi, Government of India.

Abstract:

Hemophilia A (HA) is caused by mutations within the Factor VIII (FVIII) gene, which leads to depleted protein production and inefficient blood clotting. The incidence of HA is 1 in 5,000 males, constituting about 80% of all hemophilia cases, and manifests in mild to severe disease, depending on the relative expression of functional FVIII. Current therapies include fixes-dose FVIII prophylaxis, factor VIII replacement therapies, and most recently, gene therapy. Several attempts at gene therapy have failed for various reasons-including immune rejection. Liver is the primary site of FVIII synthesis; however, the specific cell types responsible for its synthesis remain controversial. Several reports have demonstrated the capacity of bone marrow stem cells (BMSCs) to transdifferentiate in to hepatocytes and liver sinusoidal endothelial cells (LSECs). These finding created enormous interest because they uncovered a new property of BMCs and opened the possibility that these cells could be used in the treatment of liver injury and acute or chronic liver failure. We propose that the severity of the bleeding disorder could be ameliorated by partial replacement of mutated liver cells by healthy cells in HA mice. Our study showed that BM-derived hepatocytes and endothelial cells can synthesize FVIII in liver and correct bleeding phenotype in HA mice. Thus, BM- stem cell therapy is potential alternative approach to managing HA.

Break: Lunch Break 13:10-13:50 @ Atrium

Nady Golestaneh

Georgetown University Medical Center, USA

Title: In vitro disease model of dry AMD

Time : 13:50-14:10

Speaker
Biography:

Golestaneh has earned her PhD in 2000, from University of Paris VI, Pierre et Marie Curie, Paris France, her Master Degree from University of Paris VI, Pierre et Marie Curie, Paris France and her Bachelor Degree from University of Paris VII, Jussieu, Paris France. She performed her postdoctoral training at NEI/NIH, Johns Hopkins University and Georgetown University. She is Director of Research at the Department of Ophthalmology at Georgetown University Medical Center and Assistant Professor at Departments of Ophthalmology, Neurology, Biochemistry and Molecular & Cellular Biology at Georgetown University School of Medicine. She has published over 29 peer reviewed papers and has received several national and international awards. She serves as Editorial Board Member of Human Genetics & Embryology, International Journal of Bioengineering and Technology, and CellR4.

Abstract:

Age-related macular degeneration (AMD) is the leading cause of blindness in people over age 55 in the U.S. and the developed world. AMD is a multifactorial disease and there is no adequate animal model that recapitulates key characteristics of dry AMD. The lack of a robust model for AMD is a major impediment for mechanistic studies and drug development. Consequently, effective treatments for AMD are not available, although vitamin supplementation is recommended and is modestly beneficial for a small population of patients. Therefore, the need for novel therapeutic and preventive strategies is pressing.
Recent generation of RPE cells from embryonic stem cells (ESCs) and induced pluripotent stem cells (iPSCs), offers new promise for cell replacement therapy and disease modeling in AMD.
Previously, we showed that the induced pluripotent stem cell-derived retinal pigment epithelium (RPE), (iPSC-RPE) are phenotypically and functionally identical to the native RPE, offering promise for cell replacement therapy in AMD.
We recently generated an iPSC-derived disease model for dry AMD. AMD Patients’ skin cell were collected and cultured under an IRB protocol and induced to pluripotency using the Yamanaka vectors, prior to differentiation into RPE. Striking disease phenotypes and impaired functions were identified in AMD iPSC-RPE compared to normal iPSC-RPE. The mechanisms underlying the disease phenotypes have been investigated. Our research elucidates whether iPSC-RPE from skin of AMD patients are a reliable and safe source for autologous cell-based therapy in AMD, and proposes important new information for development of novel drugs for treatment of dry AMD.

Speaker
Biography:

Alain Chapel is scientific investigator at IRSN. He has been developing cell therapy to protect against the effects of radiation. He collaborates with clinicians to strategies for treatment of patients after radiation. He has participated in the establishment of proof of concept of the therapeutic efficacy of MSCs. He is a member of national and international societies. He is associate editor of World Journal of Stem Cells, World Journal of Gastrointestinal Surgery, World Journal of Radiology, The Open Gene Therapy Journal and Journal of Clinical Rehabilitative Tissue Engineering Research. He has participated in the scientific organization of international conferences.

Abstract:

The IRSN (Institute for radiation and nuclear safety) is the French National Agency responsible for prevention and treatment of irradiation victims. IRSN group has in the past five years contributed to (i) the understanding of the deleterious effects of therapeutic/accidental radiation on healthy tissues and (ii) their regeneration by MSC therapy. We have developed and tested cell therapy for protection against radiation side effects in several animal models, and we proposed mechanisms to explain the benefit brought by this new therapeutic approach. We established the proof of concept that MSC migrate to damaged tissues in the NOD/SCID immunotolerant mice model and in non-human primates (J Gen Med 2003, Blood 2004, Stem Cells 2006, Br J Radiol 2007). In the NOD/SCID mouse model, we showed that the intravenous injection of MSC (i) sustains haematopoiesis after total body irradiation (Blood 2004), (ii) improves wound healing after radiodermatitis (Annals of Hematology 2006) and (iii) protects gut function from irradiation damages (Adv Exp Med Biol. 2006). Thanks to a tight collaboration with clinicians from several French hospitals (Clinical trials…), we have been the only ones so far able to report successful treatments of therapeutic/accidental radiation damages in several victims with MSC injection. Haematopoiesis correction: In collaboration with Saint-Antoine Hospital (Paris, France), we first reported the haematopoiesis recovery in two patients with Bone Marrow failure (graft failure post grafting and Aplastic Anemia) after intravenous injection of MSC which restored the BM micro-environment, mandatory to sustain haematopoiesis after total body irradiation (Leukemia 2004, 2007). We also treated with the clinical team the first patients over irradiated in Epinal with infusion of MSC, following a specific mission form the ministry of health. Radio-induced burns: Cutaneous reactions are major actors in radiation accidents and a limitation for radiotherapy. In collaboration with Percy hospital (Clamart, France) we have shown for the first time the efficiency of MSC therapy in five patients with acute cutaneous and muscle damages following accidental irradiation delivered at doses and to fields higher than initially planned (Regen Med 2007, Health Phys 2010). Gastrointestinal disorder management: We demonstrated in the rat model that MSC restore gut functions after radiation damages (Cell Death and Differ 2010), through regulation of endogenous epithelial cell homeostasis (Methods Mol Biol, 2012). Four patients were successfully treated for pelvic overdose exposure (IJROBP in press, Cytotherapy in press). Cell therapy combining different sources of adult stem cells (endothelial progenitor, gingival fibroblast) is under investigation and is being tested in preclinical models of radio induced damage (Art Thromb Vasc Biol 2009, Haematologica 2012, Plos One 2012). In parallel, we started analyzing potential side effects after injection (Tissue Cell 2010).

Speaker
Biography:

Glavaski-Joksimovic is an Assistant Professor at the Department of Neurosurgery, Medical College of Wisconsin. She received her Ph.D. degree in Neuroscience and Veterinary Anatomy at the Iowa State University. Following graduation, she conducted a postdoctoral training at the Iowa State University, Karolinska Institute, and Northwestern University. From 2009 to 2011, Dr. Glavaski- Joksimovic worked as a Research Assistant Professor at the Department of Pediatrics, Feinberg School of Medicine, Northwestern University. In May 2011, she joined the Medical College of Wisconsin. Dr. Glavaski-Joksimovics studies are focused on the therapeutic potential of adult stem cells in a brain injury and disease.

Abstract:

Aleksandra Glavaski-Joksimovic, Anna P. Miller, Alok Shah, Brandy V. Aperi, Matthew D. Budde, Shekar Kurpad, Frank A. Pintar, Brian D. Stemper

Blast traumatic brain injury (bTBI) has high incidence among military personnel. Unfortunately,available therapies cannot stop or reverse the blast-evoked neurodegenerative cascade. However, an increasing body of evidence indicates that bone marrow-derived mesenchymal stem cells (BMSC) have therapeutic effects for non-bTBI, although they have not been tested following bTBI. Our group developed an in vitro bTBI model to study mechanisms of blast-evoked neurodegeneration and determine therapeutic potential of BMSC for bTBI. Organotypic hippocampal cultures (OHCs) were exposed to a blast overpressures of 150 or 270 kPa using a helium-driven open-ended shock tube. OHCs were exposed off-axis from the shock tube to isolate effects of shockwave overpressure. Blast-evoked cell damage was analyzed by propidium iodide (PI) uptake at different time points following exposure. Both overpressures induced OHCs damage, with higher overpressures causing significantly more cell death. Half of the OHCs were transferred to wells with BMSC at 2 hrs following exposure to 150 kPa and following confirmation of similar damage levels in all OHCs. OHCs and BMSC co-cultures were grown for 7 days. The BMSC-treated group exhibited a decrease in cell death (i.e., decreased PI uptake) compared to the untreated group at 24 hrs post-injury. By 72 hrs, BMSC-treated OHCs were statistically indistinguishable from the sham-injury group, highlighting BMSC rapid protective effects. Since OHCs and BMSC were grown without direct cell-cell contact, these experiments demonstrate that BMSC protective effects were achieved through paracrine factors released in the culture medium. BMSC therapeutic potential for bTBI and underlying mechanisms will be further explored. Supported by the Department of Neurosurgery, Medical College of Wisconsin and VA Medical Research.

Speaker
Biography:

Martin Lehmann has completed his Ph.D in 2014 from the University of Cologne. He currently works as a postdoctoral Marie-Curie fellow for the Institute of Neurophysiology of the University Hospital of the University of Cologne on a EU-granted project, performing parts of his research in a Biotechnology Company in Budapest (Hungary).

Abstract:

Embryonic stem (ES) cells are pluripotent cells derived from the inner cell mass of the blastocyst. These cells possess the ability of infinite self-renewal and to differentiate into all cell types of the three germ-layers. In defined conditions ES cells are committed to the mesodermal lineage and differentiate, amongst other cell types, into cardiomyocytes (CMs). The processes underlying mesodermal and subsequent cardiac differentiation are yet only partially understood. Catecholamine release is well known to modulate heart rate and force in adult mammals. Despite first evidence, only little is known about an involvement of catecholamines during embryonic heart development. Therefore our work aimed to investigate in more detail whether catecholamines are involved in the process of ES cell cardiac differentiation in vitro. Effects of catecholamine depletion induced by reserpine were investigated during murine D3 aPIG44 ES cell differentiation. Reserpine is a drug blocking vesicular storage of monoamines, and as a result depletes cells of the catecholamines norepinephrine and epinephrine. Cardiac differentiation was assessed by quantification of beating clusters, immunocytochemistry, molecular biology, flowcytometry and pharmacological approaches. Proliferation and cytotoxicity was evaluated by embryoid body cross-section measurements and impedance monitoring, while functional characterization of CMs was performed using extracellular field potential (FP) recordings with microelectrode arrays (MEAs). Involvement of b-adrenoceptor signaling was studied differentiating ES cells in the presence of reserpine and isoproterenol. To further discriminate between drug-specific effects of reserpine and catecholamine action via adrenergic receptors we applied the unspecific α- and β-receptor antagonists phentolamine and propranolol during differentiation. Reserpine treatment led to a remarkable reduction of beating cardiac clusters, downregulation of cardiac proteins α-actin and troponinT and delayed mesodermal and cardiac gene expression. In more detail, the average ratio of ~40% spontaneously beating control clusters was significantly reduced by 100%, 91.1% and 20.0% on days 10, 12, and 14, respectively. In line, significant reduction by 71.6% (n=11) of eGFP expressing CMs after reserpine treatment was revealed by flowcytometry. Reserpine neither reduced EB size nor acted cytotoxic on CMs, while increased numbers of neuronal cells were observed. MEA measurements with reserpine-treated EBs showed lower FP frequencies and weak responsiveness to adrenergic and muscarinic stimulation. Co-application of isoproterenol and reserpine during differentiation partially rescued cardiac development. The developmental inhibition after α- and β-adrenergic blocker application mimicked developmental changes with reserpine and proved an involvement of adrenergic receptors in the process. We therefore conclude that catecholamines and adrenergic signaling play a critical role during cardiac development in ES cells.

Speaker
Biography:

Linda Peltier is a registered nurse who recently completed a Ph.D. in Experimental Medicine at McGill University. Practicing for more than 30 years in different health care fields, she became an authority in clinical and laboratory quality assurance. The past 10 years, she specialized in cord blood banking and stem cell processing. Working as Clinical Research Associate with Dr Pierre Laneuville at the McGill University Health Centre, she aspires to collaborate in the translation from bench to bedside of cellular therapies.

Abstract:

Cord blood unit (CBU) is a valuable alternative source of stem cells for patients without allogeneic stem cell donor. Few banked CBUs contain sufficient hematopoietic stem cells (HSC) to transplant adult patients. Different approaches have been used to increase the number of infused HSC, however most approaches have the disadvantage of increasing the cost of graft procurement or relying on a related haploidentical donor that may not be available. A new developed alternative approach consists of creating a third-party unit of enriched CD34+ cells from a pool of multiple HLA-blind cryopreserved CBUs. These pooled units were rejected by the public cord blood bank, due to small volume and/or low nucleated cell count. Seven recipients with hematological cancers received myeloablative conditioning followed by the co-infused of a ≥ 4/6 HLA compatible CBU and a third-party composed of the pool CD34-selected cryopreserved CBUs. Six patients engrafted with a median neutrophil engraftment time of 19.5 (15-29) days. All engrafted patient showed a 100% HLA-matched CBU chimerism on day +14. All recipients had grade I-III acute graft-versus-host-disease (GVHD) that responded promptly to treatment and no patients developed chronic GVHD. Two patients died, one at day +28 of disease relapse and one at day +360 of multisystem organ failure. Not only this new method support the importance of increasing the number of HSCs to ensure rapid engraftment, but also demonstrate that clinical grade rejected CBUs can be used to create a third-party of enriched CD34+ cells to support a small ≥ 4/6 HLA compatible CBU.

Speaker
Biography:

Jan H. Spaas, veterinarian, graduated from the Faculty of Veterinary Medicine, Ghent University (Belgium) in 2010. In that year he also won the price for young authors of the Flemish Veterinary Journal. He completed his Ph.D. at the Department of Comparative Physiology and Biometrics of the Faculty of Veterinary Medicine, Ghent University. Since 2012, he became the laboratory director of Global Stem cell Technology, an organization that is specialized in regenerative therapies for horses. Thanks to external research funding, he is the industrial promotor of Ph.D. and master students who actively participate in the company’s research program.

Abstract:

Degenerative joint disease (DJD) is a major cause of reduced athletic function and retirement in human and equine performers. Medical treatment for DJD may include anti-inflammatory and analgesic drugs to reduce inflammation and pain, and so-called disease-modifying drugs. Nevertheless, the aforementioned therapies are merely aimed at alleviating the symptoms or enhancing clinical recovery, without inducing an actual regeneration of the affected joint. The field of equine regenerative medicine is drawing increasing attention in the scientific community for its treatment strategies of joint pathologies. Mesenchymal stem cells (MSCs) are of specific therapeutic interest as they can differentiate in vitro towards cells with a chondrocyte morphology and produce cartilage-specific components such as collagen type II and glycosaminoglycans. Moreover, horses may serve as a valuable large animal model for the evaluation of new human therapies concerning in vivo efficiency and safety, due to interspecies similarities in thickness of the non-calcified cartilage of the stifle joint.
Twenty horses with naturally occurring DJD in the fetlock joint were divided in 4 groups and injected with: 1) platelet-rich plasma (PRP); 2) MSCs; 3) MSCs and PRP; or 4) chondrogenic induced MSCs and PRP. Evaluated was performed after 6 weeks (T1), 12 weeks (T2), 6 months (T3) and 12 months (T4). Subsequently, 30 horses with the same medical background were randomly assigned to one of the two combination therapies and evaluated at T1. The combined use of PRP and MSCs significantly improved the functionality and sustainability of damaged joints from 6 weeks until 12 months after treatment, compared to PRP treatment alone. The highest short-term clinical evolution scores were obtained with chondrogenic induced MSCs and PRP.

Break: Coffee Break 15:50-16:05 @ Atrium