Day 3 :
- Track 9: Nano-Therapy
Paul J. Davis
Albany Medical College, USA
Northeastern University, USA
Albany Medical College, USA
Title: Nanotetrac targets the thyroid hormone receptor on integrin αvβ3 on tumor cells to promote apoptosis, disorder cell defense pathways and block angiogenesis
Time : 09:30-09:50
Paul J Davis is Professor of Medicine at Albany Medical College and Professor of Pharmacy at the Albany College of Pharmacy and Health Sciences. He is former Chair of the Department of Medicine at the Medical College. His medical training was obtained at Harvard Medical School, Albert Einstein College of Medicine and the NIH. He is a Past President of the American Thyroid Association, a former Director of the American Board of Internal Medicine and is a Master and former Governor of the American College of Physicians. He is Head, U.S. Endocrinology and Metabolism panel of the Faculty of 1000 Medicine. He has co-authored more than 200 scientific publications, 25 textbook chapters and multiple editorials. Dr. Davis is Associate Editor at Endocrine Research (Informa) and on the Editorial Boards of Hormones and Cancer (Springer Publishing) and of Imunnology, Endocrine and Metabolic Agents in Medicinal Chemistry (Bentham). He is Co-Editor with Dr. Shaker Mousa of Angiogenesis Modulations in Health and Disease (Springer Publishing), a reference manual for angiogenesis drug development published in 2013. His research group was the first to identify a receptor or target for thyroid hormone on the outside of cancer cells that allows the hormone to be a growth factor for certain cancers and for the blood supply of malignancies. He and his colleagues have developed nanoparticulate thyroid hormone derivatives that work exclusively at this cell surface target. Among these derivatives is a novel agent that blocks experimental tumor cell growth and cancer-related blood supplies and that also renders cancer cells more sensitive to radiation therapy. Other nanoparticulate formulations of thyroid hormone work at the receptor in the absence of cancer to generate blood vessels, i.e., angiogenesis applied to wound-healing and ischemia. Dr. Davis has also been involved in the development of drugs to salvage nerve cells at the time of stroke and heart cells in the setting of reduced coronary artery blood flow.
We have described a cell surface receptor for thyroid hormone and hormone analogues on the extracellular domain of integrin αvβ3. The receptor has no structural analogies to nuclear thyroid hormone receptors (TRs) involved in genomic actions of thyroid hormone. The integrin is expressed primarily by cancer cells and rapidly-dividing endothelial cells. Thyroid hormone as L-thyroxine (T4 ) and 3,5,3’-triiodo-L-thyronine (T3) nongenomically supports cancer cell proliferation and anti-apoptosis at this receptor site. Tetraiodothyroacetic acid (tetrac) is a thyroid hormone derivative that has undesirable thyromimetic actions inside cells, but at the cell exterior inhibits binding and actions of T4 and T3 at αvβ3. We covalently bound tetrac to a biodegradable poly(lactic-co-glycolic acid) (PLGA) nanoparticle which restricts action of the hormone analogue to the receptor on αvβ3 and prohibits cell uptake. Nanoparticulate tetrac (Nanotetrac) is ten-fold more potent than unmodified tetrac as an anti-proliferative/anti-angiogenic agent at αvβ3. In addition, Nanotetrac has anti-angiogenic and pro-apoptotic actions lacking in unmodified tetrac. The anti-angiogenic qualities of Nanotetrac include inhibition by several mechanisms of the activities of VEGF, bFGF, PDGF and EGF. Targeted to its receptor on the integrin, Nanotetrac also disorders gene transcription relevant to six cancer cell survival pathways and blocks tumor cell repair of double-stranded DNA breaks that are induced by γ-radiation. The agent has been shown to be highly effective against ten human cancer xenograft models, including pancreas, lung, kidney, prostate and breast. The function of the 200 nm PLGA tail on Nanotetrac is to restrict its tetrac probe to the cell exterior and αvβ3. However, the cassette of tetrac-directed PLGA may be used to deliver a traditional chemotherapeutic agent—adsorbed to the nanoparticle—locally to tumor cells and tumor-relevant endothelial cells. In summary, Nanotetrac is a highly-selective chemotherapeutic/anti-angiogenic agent targeted to a novel small molecule receptor on integrin αvβ3. At this receptor, Nanotetrac is capable of 1) activating pro-apoptotic systems suppressed in cancer cells, 2) suppressing anti-apoptotic mechanisms that are ordinarily activated in such cells, 3) disabling a number of cell survival pathways and 4) inhibiting by several mechanisms the actions of multiple vascular growth factors. Nanotetrac is restricted to the extracellular space and its integrin target is primarily expressed by cancer cells and rapidly-dividing blood vessel cells.
Northeastern University, USA
Title: Stimuli-sensitive combination nanopreparations of siRNA and chemotherapeutic drugs to treat multidrug resistant cancer
Time : 09:50-10:10
Vladimir Torchilin is University Distinguished Professor and Director, Center for Pharmaceutical Biotechnology and Nanomedicine, Northeastern University, Boston. He has published more than 350 original papers, more than 150 reviews and book chapters, wrote and edited 10 books and holds more than 40 patents. He is Editor-in-Chief of Current Drug Discovery Technologies and of Drug Delivery. He is a Member of European Academy of Sciences, Fellow of AIMBE, AAPS and CRS, and received many important national and international awards including the 2013 Blaise Pascal Medal in Biomedicine from EAS. In 2005 he was a President of the CRS. In 2011, Times Higher Education ranked him number 2 among top world scientists in pharmacology for 2001-2010.
Tumor therapy, especially in the case of multidrug resistant cancers, could be significantly enhanced by using siRNA down-regulating the production of proteins, which are involved in cancer cell resistance, such as Pgp or survivin. Even better response could be achieved is such siRNA could be delivered to tumors together with chemotherapeutic agent. This task is complicated by low stability of siRNA in biological surrounding. Thus, the delivery system should simultaneously protect siRNA from degradation. We have developed several types of lipid-core polymeric micelles based on PEG-phospholipid or PEI-phospholipid conjugates, which are biologically inert, demonstrate prolonged circulation in the blood and can firmly bind non-modified or reversibly-modified siRNA. Additionally, these nanopreparations can be loaded into their lipidic core with poorly water soluble chemotherapeutic agents, such as paclitaxel or camptothecin. In experiments with cancer cell monolayers, cancer cell 3D spheroids, and in animals with implanted tumors, it was shown that such co-loaded preparations can significantly down-regulate target proteins in cancer cells, enhance drug activity, and reverse multidrug resistance.
In order to specifically unload such nanopreparations inside tumors, we made them sensitive to local tumor-specific stimuli, such as lowered pH, hypoxia, or overexpressed certain enzymes, such as matrix metalloproteases. Using pH-, hypoxia-, or MMP2-sensitive bonds between different components of nanopreparations co-loaded with siRNA and drugs, we were able to make the systems specifically delivering biologically active agents in tumors, which resulted in significantly improved therapeutic response.
Bio-Synthesis, Inc., USA
Time : 10:10-10:30
Andrei Laikhter graduated from bioorganic chemistry department Moscow Institute of Fine Chemical Technology in 1982. He received his Ph.D. degree in 1987 at the age of 27 years, from Zelinsky Institute of Organic Chemistry, Russian Academy of Sciences. He worked at University of Delaware in 1993 and University of Virginia 1994-1996. Since 1999 he worked at Integrated DNA Technology, Inc., he became the vice president of chemistry research in 2004. From 2007 until beginning of 2014, he was working as chief scientific officer at Chemgenes Corporation. Currently he is working as chief scientific officer at Biosynthesis, Inc. Till now he has published over 40 papers and patents.
Conventional method of RNA synthesis by 3’ - 5’ direction is now well established and currently in use for synthesis and development of vast variety of therapeutic grade RNA and si RNA etc. A number of such synthetic RNA requires a modification or labeling of 3’- end of an oligonucleotide. The synthesis of 3’- end modified RNA requires lipophilic, long chain ligands or chromophores, using 3’ - 5’ synthesis methodology is challenging, requires corresponding solid support and generally results in low coupling efficiency and lower purity of the final oligonucleotide in general because of large amount of truncated sequences containing desired hydrophobic modification. We have approached this problem by developing reverse RNA monomer phosphoramidites for RNA synthesis in 5’- 3’- direction. We demonstrated that a number of lipophilic modifications such as cholesterol, PEG-2000 and PEG 4500 can be introduced at 3’-end with high efficiency. Highly pure desired oligonucleotides have been isolated using reverse-phase HPLC purification. We developed novel solid support that along with reverse RNA and DNA synthesis technology offers convenient synthesis of 5’–adenylated oligonucleotides without any post-synthetic steps.
North Maharashtra University, India
Time : 10:30-10:50
Vinod Jagannathrao Mokale has completed his Ph.D at the age of 30 years from North Maharashtra University, Jalgaon-425001, Maharashtra, India. He is the Head, Department of Pharmaceutical Technology, UICT, NMU, Jalgaon, Maharashtra, India. He visited Thailand, Singapore, Malaysia, Indonesia, London etc. as a resource person in various international conferences, and also deliver keynote with session the chair. He has completed one project and 04 is ongoing who is financially supported by Dept. of Science and Technology, DRDO and AICTE Govt. of India with one Career Award for Young Teacher from AICTE, New Delhi, Govt. of India, he has 02 Indian patents and going to the US Patent with more than 30 publications in reputed international journals and serving as an editorial board member of repute.
We hear a lot about Nanotechnology these days, but what is it actually doing for us, and how will it help in the future?
*Nanotechnology = devices and capabilities that are:
Cheaper than the ones we have today
Better than the ones we have today
Different to the ones we have today - doing things we could never do before…
For patients this also means, technologies that are less invasive and technologies that improve quality of life. For health care professionals, this can mean “doing more with less”. Nanotechnology has an impact on the subject of Global Priorities, which are Health Care, Information Technology, Environment, Chemical & Biodefense, and Energy etc.
We also have current and future health care challenges like obesity, infectious diseases, heart attack, cancer, genetic disorders, aging etc.
To overcome from above all the said problems , we have to move towards nano-therapy which is a future medicine to us by formulating nanomedicine, nanodiagnostics agents & devices, and by the use of various biodegradable/biocompatible nanoparticulate drug delivery systems, because they have been used frequently as drug delivery carrier in various life threatening diseases and due to its better encapsulation capacity, sustained/control release property and less toxicity, one also we can add we can definitely deceases dosing frequency for various dosage forms by the formulation of nanomedicine so, Nano-Therapy is the future medicine which will be helpful for all the human beings.
Theodor Bilharz Research Institute, Egypt
Title: The differentiation potential of human cord blood unrestricted somatic stem cells into hepatocyte-like cells on 3-dimensional nanofibres
Time : 11:05-11:25
Zeinab A Demerdash has completed her MD in 1990 from Cairo Medical School, Egypt, in Clinical Pathology, fine specialty Immunology . She is now a Professor in Immunology at Theodor Bilharz Research Institute, Cairo Egypt. She has been specialized in Monoclonal Antibody production and Stem Cell Research especially Cord Blood Mesenchymal Stem Cells and their clinical applications. She has more than 30 publications in reputed Journals.
Background: Unrestricted somatic stem cells (USSCS), earlier progenitors of cord blood mesenchymal stem cells, hold great promise for liver tissue engineering. Scaffolds are three-dimensional (3D) matrices that provide the initial structural support for cells to attach, proliferate, and differentiate, facilitating the formation of an extracellular matrix (ECM). In vitro culture of USSCs on nano-fibrous scaffold could intiate and enhance their differentiation potential into heptocytes – like cells. Aim of the work: In this study we induced the differentiation of USSCs into hepatocytes in two- and three-dimensional (2D and 3D) culture systems. The effect of nanofibrous scaffold ( 3D) on their differentiation potentiality were evaluated. The more promising culture system are to be adopted in the in vitro- hepatic differentiation of USSCs, to improve their future application in cell-based therapies in patients with chronic liver diseases. Materials and Methods: USSCs were isolated from human cord blood, propagated and characterized. Hepatogenic differentiation of USSCs was performed on both 2D and 3D culture systems. Differentiation potentiality of both systems was evaluated using, indocyanin green (ICG) cellular uptake method, periodic acid schiff (PAS) staining method, immunohisochemistry analysis for albumin and alpha fetoprotein ( AFP), scanning electron microscopy (SEM), and ELISA for albumin and alpha 1 antitrypsin secretion. Results: SEM showed adherence of cells in the nanofiber scaffold during hepatogenic differentiation. Differentiated Hepatocyte- like cells were more abundant, more mature, and hexagonal in shape in the 3D culture system. Both systems were positive for ICG uptake, PAS, albumin and AFP staining. Albumin secretion was significantly higher in 3D culture system while alpha1antitrypsin secretion increased equally in both 3D and 2D cultures Conclusion: In vitro differentiation of USSCs on nanofibrous scaffold, before their use in liver regeneration, could be superior to conventional 2D cultures. This work is extracted from the project 1410 supported by the Science and Technology Development Funds (STDF), Cairo, Egypt.
Harvard School of Public Health, USA
Time : 11:25-11:45
Archana Swami completed her Ph.D. in Biotechnology at Institute of Genomics and Integrative Biology, Council of Scientific and Industrial Research (CSIR), India and was honored the degree of Ph.D. from Department of Biotechnology, University of Pune, India (2009), with a prestigious fellowship for 5 years (Research Fellow, CSIR, India). Dr. Swami Ph.D. research mainly involved in understanding the role of gene therapy and development of multifunctional gene delivery carriers for biomedical applications. Thereafter, Dr. Swami worked as a postdoctoral fellow at Brigham and Women’s Hospital with the groups of Dr. Omid Farokhzad, at Harvard Medical School and, Robert Langer, at The David H. Koch Institute for Integrative Cancer Research, MIT. During her postdoctoral training, Dr. Swami's primary research goals were to develop nanotechnologies for therapeutic applications. Her work lead to several peer-reviewed research articles, patents and reviews. Dr. Swami long term goal is to make use of the skill sets obtained in past, to serve the community and give her part in creating better understanding of disease mechanism, giving novel therapeutic targets. Currently, Dr. Swami is working at Harvard School of Public Health, and is studying to resolve the understanding on how environmental exposure impacts health outcome and this information is translated in the genes.
Our environment (macro- and micro- scale) transmits the signals that integrate at the tissue and cellular level to govern the general health and predisposition to disorders. In cells, the regulation of gene expression by DNA methylation and histone acetylation, is an important example to reflect the significance of environment over genotype. A better understanding of the mechanism of this translation will be advantageous in speculation of disease predisposition and creating better treatment options. For example, studying the impact of noise exposure in occupational setting to induce hearing loss is significant concerning the public health. Furthermore, it is important to understand how at tissue level, the modulation of the cellular microenvironment impacts the disease progression. In another example, we studied the role of bone microenvironment in cancer progression. As tested in multiple myeloma in mice model, where bone targeted nanoparticles strengthened the bone tissue to make the bone tissue resistant to homing of cancer cells.
- Track 10: Bioengineering Therapeutics
Vasiliki E. Kalodimou
IASO Maternity Hospital, Greece
University of Oslo, Norway
Title: Key regulatory junctions stabilizing the osteoblast phenotype: Implications for cell and tissue engineering
Time : 12:15-12:35
Jan O Gordeladze, PhD, holds a triple Professor competence (medical biochemistry, physiology, and pharmacology), and is presently working as a Professor at the Department of Biochemistry, Institute of Basic Medical Science, University of Oslo, Norway. He has previously been employed as the Medical Director of MSD, Norway, serving two years as a Fulbright Scholar at the NIH, Bethesda, Maryland, USA, and from 2006-2009 also being employed as Associate Professor at the University of Montpellier, France. He is a member of the Norwegian Stem Cell Center, and his research has over the past 5-6 years been devoted to differentiation of osteochondral cells from stem cells focusing on the impact of transcription factors and microRNA species constituting regulatory loop interactions with functional target genes. He has published more than 120 scientific articles, reviews or book chapters and presented more than 250 abstracts or posters or talks at conferences worldwide.
MicroRNAs are short, non-coding RNAs, which bind to the 3UTR of messenger RNAs, thus suppressing the translation process. MicroRNAs may be intragenic and positioned more or less adjacent to their targets. However, it has been asserted that transcription factors (TFs) and microRNAs engaged in feed-forward/feed-back loops are stabilizing cell phenotypes. Many researchers, engaged in osteoblast and bone engineering, have focused on Runx2 and Osterix (SP7), being two TFs instrumental in osteoblastogenesis, as well as various microRNA species targeting them. However, when using bioinformatics (the Mir@nt@n algorithm), it appears that other osteoblast-”specific” TFs, like SP1, are involved in regulatory loops. The work presented here indicates that stabilizing the interactions of SP1 with microRNA species targeting it (e.g. mir-204, mir-211, mir-24, -mir-149, and mir-328) may lead to a better differentiation of stem cells to osteoblasts, thus producing a better bone tissue more able to withstand an inflammatory environment compared to over-expressing the TFs Runx2 and Osterix (SP7). The experiments conducted are based on manipulations of TF or microRNA levels using polycistronic constructs of pre-mirs and antago-mirs, up- or down-regulation of TFs using gene-expressing vectors and sh-RNA expressing lentiviral constructs, adipose stem cells grown in an osteoblast-differentiating medium ± cytokines from Th-17 cells, co-cultures with osteoclasts differentiating from PBMCs, as well as manipulated osteoblasts injected into the tibial muscle of SCID mice. Other important transcription factors involved in regulatory loops are SP3, Ets-1, as well as SPI1, JUN, and FOS. Finally, we make an attempt to rank SP1 compared to SP3 and Ets-1 featuring their stabilizing potency.
IASO Maternity Hospital, Greece
Title: The effects of infused mesenchymal stem cells obtained by adipose tissue (ADSCs) to treat chronic scar tissue of the vocal cords in an animal model
Time : 12:35-12:55
Vasiliki E. Kalodimou MSc, PhD is the director of the Flow Cytometry-Research and Regenerative Medicine Department at IASO Hospital in Athens, Greece. She has studied and worked with progenitor cells from placenta, umbilical cord, and adipose tissue along with their applications in regenerative medicine and flow cytometry, with publications in the field, including research fellowships. She has published two books about flow cytometry, the Greek edition was published in 2010 and in 2013 the book was published from AABB Press USA. She is an AABB assessor and her biography is included in Who Is Who in the world 2014 edition.
Numerous studies have provided preclinical data on the safety and efficacy of adipose derived stem cells supporting the use of these cells in current and future clinical applications. This is the first study to our knowledge, which aims to use the cell viability and absolute number of isolate mesenchymal stem cells from ADSCs and their effect to chronic scar tissue of the vocal cords. Adipose tissue (10gr) was taken from the groin of 70 healthy rabbit, New Zealand variety (2.5-3.8Kg). 10 of the samples were used as controls were the remaining 60 rabbits undergo vocal cord injuries under anesthesia and laryngoscopy. The isolated ADSCs were infused directly to the scar tissue of the vocal cords. Multi-parameter flow cytometry with magnetic beats was used to determine the absolute number, viability and the ADSCs isolation. The samples were processed within 1h of collection. The results obtained from the analysis of all samples indicate that the highest absolute number of viable adipose derived stem cells was isolated. Their purity was confirmed by the high expression (> 95%) in the positive markers and low expression (< 2%) in the negative markers. We also studied the ADSCs characteristics after the transplantation at 3, 6 & 12months in order to measure the elasticity of the vocal cords between the groups. Taken together, these data indicate that purified lipoaspirate-derived stem cells maintain their characteristic of staminality, suggesting that they could be applied for cell-based therapy to improve chronic scar tissue of the vocal cords.
Central University of Technology South Africa
Title: Engineering of cytochrome P450 monooxygenases-based novel inhibitors against Mycobacterium tuberculosis
Time : 12:55-13:15
Khajamohiddin Syed completed his Ph.D. at the age of 27 years atSri Krishnadevaraya University, India. His research is focused on P450-enzyme based drug-designing against human pathogens and development of P450-enzyme based novel tools for bioremediation of cancer-causing and endocrine-disrupting chemicals. His research work has been published in highly ranked peer-reviewed scientific journals including Science and PNAS, USA. He is serving as editorial board member and reviewer for reputable international scientific journals. To date he has published 17 articles in reputable scientific journals.
The actinomyceteMycobacterium tuberculosis (Mtb) causes tuberculosis (TB), a chronic lung disease, in humans and continues to be one of the greatest threats to mankind. It is estimated that one-third of the human population are latently infected by Mtb and 9 million cases of active TB occur each year.Development of multi, extensively and totally drug-resistant (MDR, XDR and TDR) Mtb strains,together with the paucity of new drug targets currently being explored, suggests that new basic research is required to delineate novel potential targets. Recent studies suggested that cytochrome P450 monooxygenases(P450s) of Mtb can serve as novel drug targets. P450s are mixed function oxidoreductases well known for their role in essential cellular anabolic and catabolic processes. To identify P450s that can be used as drug targets not only for Mtb but also against all mycobacterial species, we performed comprehensive comparative P450 analysis across 23 mycobacterial species. A total of 730 P450s and fourrtruncated P450s (pseudo) were identified in 23 mycobacterium species. Six P450 families, CYP51, CYP123, CYP125, CYP136, CYP138 and CYP144, were found conserved among 23 mycobacterial species, suggesting their essential role in mycobacterial species physiology and as potential novel drug targets. We successfully constructed reliable 3D-models and mapped the active site cavities for the six P450s. Furthermore, we also identified potential inhibitors for six P450s by virtual screening of a library of chemical compounds. Experiments are in progress to validate the identified inhibitors and to refine/engineer a common inhibitor against six conserved mycobacterial P450sfurther.
University of Leicester, UK
Time : 13:45-14:05
Nawal Helmi has completed B.Sc. in Biochemistry (2004) and Master degree in genetic and pharmacology (2008) from King Abdul Azizi University (Jeddah). Nawal has also undertaken volunteer work at the King Abdul Azizi University Hospital there she saw at first hand the suffering of sickle cell anaemia patients and this contributed to her decision to try to stop the suffering of the sickle cell anaemia patients around the world. For that she get her PhD from University of Leicester (2014) and her dissertationwas about the effect of perfluorocarbon therapy in Streptococcus pneumoniae infected Sickle Cell Mouse, during her PhD study she received for two years in row awards Best 50 research in University of Leicester. Nawal now work as assistant professor in AL-Farabi College in Jeddah.
Sickle Cell Anaemia (SCA) is a chronic haemoglobinopathy. It results from a genetic defect in the globin gene, which leads to a haemoglobin S (HbS). SCA is characterised by a chronic haemolytic anaemia and vaso-occlusive crises. Both result from distorted (sickle) red blood cells (RBCs). Individuals with SCA also have a significantly greater risk of overwhelming pneumococcal infection, which occurs at a much higher rate than for other encapsulated bacteria, suggesting that individuals with SCA are uniquely vulnerable to pneumococci. Streptococcus pneumoniae is a common cause of pneumonia (an inflammatory condition of the lung), which can cause vaso-occlusion in SCA patients and this is due to asplinea. RBC transfusion reduces the risk of further vaso-occclusion but recurrent transfusions risks such as iron overload. An alternative to RBC transfusion is artificial blood. For example Perfluorocarbons (PFC) have an O2 carrying capacity, which is 40 to 50 times higher than haemoglobin. As observed in ex vivo and in vivo models sickle red blood cell -induced vaso-occlusion is often partial, allowing for decreased remnant flow. Hence, if oxygen is delivered to these areas decreased obstruction might be achieved. The hypothesis of this study that treatment with PFC will reverse sickle cell induced pulmonary vascular-occlusion in SCA mice with pneumococcal lung infection. The effect of the perfluorocarbon emulsions was tested in vitro and in vivo on S.pneumoniaeto determine whether it encouraged the pneumonia-causing bacteria to grow. This was done using two different strains of the pneumoniae D39 serotype 2 and TIGR 4 serotype 4. The findings suggest that PFCE may have a role in treating patients with sickle cell anemia
Federal Fluminense University, Brazil
Title: Looking for β3 integrin family selectivity: The use of snake venom disintegrin as a tool for molecular modeling approach
Time : 14:05-14:25
Reinaldo Barros Geraldo is a young scientist, researcher currently postdoc at Fluminense Federal University (Brazil). Graduated in Biomedicine Fluminense Federal University (2007), Master of Neuroimmunology, Fluminense Federal University (2009) and Doctor of Science by Federal University of Rio de Janeiro (2013). Has experience in the areas of biochemistry, with an emphasis on computer modeling, acting on the following subjects: Comparative modeling, structural biochemistry and Molecular Biology. At now, he has published 11 papers in reputed journals.
Disintegrins are a family of modulatory peptides that inhibit the some integrins, such as αIIbβ3 and αvβ3, which are involved in a variety of cellular functions such as platelet aggregation and angiogenesis, physiopathology of arterial thrombosis and cancer. These disintegrins were identified several sources including venom of some snakes. The aims of this study were to analyze using in silico approaches (i.e. molecular modeling and molecular dynamics) this disintegrin sub-family, to observe the selectivity against those integrins. In addition, perform a study of molecular biology with two disintegrins present in the venom of Bothrops jararaca venom, jararacin and jarastatin, to use these disintegrins as a model for study of new prototype of new drugs against those pathologies. The in silico analyses of 70 snake venoms disintegrins showed with the alignment and phylogeny results, the presence of four distinct sub-groups that possess the high similarity among them. From those subgroups, we select two disintegrins of these four sub-groups: sub-group 1 (albolatin and mojastin), sub-group 2 (jarastatin and salmosin) sub-group 3 (trimestatin and flavoridin) group 4 (kistrin and jararacin), and whose were used to construct the complexes with integrins αIIbβ3, αvβ3 and α5β1. These complexes were minimized with the GROMACS program. Moreover, the interaction of them was analyzed with the webservers PISA, protein interaction calculator (PIC) and GROMACS program. The analysis of the complexes with α5β1, the sub-group 1 and sub-group 3 and kistrin present the number of interations is greater than others. The αIIbβ3 complexes show the jararacin, sub-group 1 and sub-group 3 present the number of interactions is greater than the sub-group 2 and kistrin. And the αvβ3 complexes show sub-group 2 with the high interactions values than others followed by sub-group 3 and sub-group 1. The molecular dynamics reveals corroborated the initial in silico results that shows jararacin to αIIbβ3 complex formation and jarastatin possess more stability of αvβ3 complex formation. This study shows the presence of subgroups in the PII-disintegrins, and they present a different selectivity against integrins. We succeeded in producing recombinant disintegrins of Bothrops jararaca with the maintenance of biological activity.
Tehran University of Medical Science, Iran
Time : 14:25-14:45
Seyedmehdi Nourashrafeddin is a recent PhD graduate from Tabriz University of Medical Science, Iran, in the field of Molecular Medicine. He is currently work as an academic research assistant at Magee-Women’s Research Institute, University of Pittsburgh, USA. His research focuses on the molecular mechanisms that govern primate spermatogonial stem cell differentiation. His graduate school research focused on the analysis of gene expression during stem cell-based spermatogenesis in vitro. In 2005, he received his master’s degree in the field of Immunology from Tehran University of Medical Sciences, a premier University in Iran. In addition, from 2004-2006 he worked as a research assistant at the Hematology-Oncology and Stem cell Transplantation Research Center in Tehran, Iran. He is proficient in all standard molecular and cellular biology techniques. He is also an employee as Research Assistant in Tehran University of Medical Science. He has published more than 6 papers in reputed journals.
Approximately 15% of couples are known to have reproductive problems. Embryonic stem cells (ESCs) are pluripotent stem cells derived from the inner cell mass that able to develop to any kind of cell. Recent studies have shown that soluble growth factors are able to govern progression of the mouse ESC into male germ-like cells in the absence of somatic cell support, and these mouse ESC-derived germ-like cells are able to induce normal fertilization and development in vitro. We inserted a fragment of Stra8 gene promoter (-1400 to +7) in ScaI/HindIII multiple cloning site of pEGFP-1 vector. Then electroporation carried out on embryonic stem cells and positive colonies were selected as puromycin-resistant after treatment with puromycin. All-trans retinoic acid (RA) was used for differentiation of mouse ESCs. We applied RT-PCR, Real-Time PCR and immunocytochemistry to analyze the differential expression of spermatogenesis specific molecular markers during different stages of mouse ESC differentiation in vitro parallel with mouse testis development in vivo. GFP-positive mESC colonies were observed after 72 h RA induction. We differentiate more GFP-positive mouse ESC with RA, and after 30 days of differentiation, typical elongated spermatids appeared in culture media. These germ cells expressed specific molecular markers during development stages in vitro as the same as testis development in vivo. The protamin 1 (Prm1) gene, as post meiotic marker, was expressed after 21 days of RA induction. These results leading us to suggest stem cell-based spermatogenesis in vitro using a backbone vector containing the fusion gene Stra8-EGFP as one of the alternative treatment for infertile subjects that suffering from azospermia. Moreover, we suggest the stem cell-based spermatogenesis in vitro as a potent systematic model for male germ cell development and spermatogenesis studies.
- Track 11: Clinical Trials and Research in Cell & Gene Therapies
Track 12: Regulatory & Ethical Issues of Therapies
MD Anderson Cancer Center, USA
MD Anderson Cancer Center, USA
Time : 14:45-15:05
Dr. Vikas Kundra completed his M.D. and Ph.D. from Harvard Medical School and Harvard University. He then did a research and clinical residency followed by a research and clinical fellowship with a concentration on MR at Brigham and Women’s Hospital in Boston. He then joined the faculty at U.T.-M.D. Anderson Cancer Center where he is now Professor of Radiology and Director of Molecular Imaging. He has a joint appointment in the Department of Experimental Diagnostic Imaging. His clinical work and research focuses on body imaging and laboratory research focuses on molecular imaging, including imaging gene expression, and imagable models of disease.
Clinical trials of gene therapy have been hampered by a lack of clinically relevant methods for in vivo detection of gene transfer. Currently in the clinic, evaluating success of gene transfer is primarily limited to analyses of biopsy samples, which provides limited assessment of in vivo gene delivery, is prone to sampling error, has associated morbidity and mortality, and can have problems with patient compliance especially when repeated evaluation or monitoring of multiple sites is needed. Instead, monitoring of exogenous gene expression should be noninvasive and easily repeatable over time in the same patient. This would inform regarding the location, magnitude, and kinetics of gene expression, and, could prove instrumental towards the rational development of innovative formulations designed to selectively target particular tissues, organs, or disease sites. To approach these needs, reporter genes may be used. These often encode enzymes, transport proteins, and receptors that most frequently bind and/or entrap an imaging agent. These may be limited for percutaneous imaging of humans because of scatter, such as light based agents; size; immunogenicity, particularly if not of human origin; quantification; and availability of clinically approved imaging agents. A desirable feature of the reporter is that it does not affect the intracellular milieu by signaling or pump action so that it does not cause untoward effects in expressing cells. We find that human somatostatin receptor type 2 gene-based reporters (SSTR2-based) reporters have desirable features for imaging in animals and for translation to humans. The SSTR2-based systems enable in vitro, in vivo, and ex vivo assessment of the reporter. They can be imaged using clinically approved radiopharmaceuticals and can be designed to be signaling deficient. Using small animal cognates of clinical machines as well as machines designed for patients, we have used a combination of functional and anatomic imaging to quantify in vivo expression of SSTR2-based reporters. We have used these to evaluate methods for improving expression. Imaging and quantification of such reporters has been performed in small animals and as a bridge to translation, in large animals.
Loughborough University, UK
Title: Regulation and development in cell-based therapeutics: The need for comparability and Implications for new business models
Time : 15:05-15:25
Nick Medcalf is a Professor of Regenerative Medicine Manufacture at Loughborough University, UK. He is Director of the EPSRC Centre for Innovative Manufacturing in Regenerative Medicine from 2014. Regulatory science’ i.e. industry-facing, challenge-led research aimed at providing the confidence in process design necessary to satisfy regulatory constraints for advanced therapeutic products, such as cell- and tissue-based products. Thomas Heathman is a PhD student at Loughborough University, UK. He is interested to develop a microcarrier based stirred culture system for scalable expansion of allogeneic bone marrow derived human mesenchymal stem cells to treat disorders of the skeletal system. We will be taking a quality engineering approach to build a comprehensive knowledge base of how various process parameters such as reactor & impeller dimensions, microcarrier type and culture media will affect the product critical-to-quality attributes. From this enhanced process understanding we will develop a control strategy to ensure acceptable overall product quality throughout operational scale up which is critical for regulatory approval and successful commercialisation.
The development and delivery of novel cell-based therapeutic products requires ongoing and improved methods for ensuring consistency of quality, safety and efficacy. Issues that must be addressed for cost-effective, regulatory-compliant supply include the design of suitable control systems, validation of transit arrangements and management of overall costs, especially for indications that are of moderate, rather than life-saving value-release. The introduction of automation is often cited as a solution to these challenges but investment choices for process improvement using automation must takes place within a whole-operation context if effective automation is to take place. This presentation examines the current options for production and supply for new cell therapy businesses and highlights the strengths and weaknesses of these in the light of supply-chain robustness, ease of clinical introduction and cost of manufacture. Some conclusions are presented about the likely direction of new business models and the technology required to enable these to be used while satisfying issues of process comparability.
BioGlobaX Inc., Canada
Anelia Atanassova received BSc in Chemistry, MSc in Biophysics and Ph.D. in Biochemistry from Sofia University, Bulgaria. She was a Postdoctoral Fellow at the University of Toronto, Chemistry Department and Structural Genomics Consortium and Scientist in Faculty of Pharmacy, PharmaGen Ltd., Sofia and Center for Gene Research, Nagoya University, Japan. She is a founder of BioGlobaX Inc, a private service organization established in collaboration with research groups from the University of Toronto and across the globe. She is an author of numerous innovations and provides services as an editorial board member of scientific journals.
Proteins are key biomolecules for cell therapy; however, toxicity and side effects of manufactured proteins arise due to altered properties and factors derived from the complexity of their structures, cellular specificity of action, dynamic folding, and reactivity. They perform diverse functions in the cells and may play dual role, such as enzymatic activity coupled with genes activation, membranes traffic and rearrangements, molecular transport or transmission, among many others. Reactive amino acid residues on the proteins surface participate in their signal transduction pathways via cascades of reversible interactions and redox reactions. In isolated form, proteins lose the native environment, which keeps them in functional states in the cells. The replacement of the cellular compounds by appropriate bio and inorganic molecules is critical for protein production to keep them as close as possible to the wild type. In addition, the formation of homo-oligomers may inactivate proteins, triggering immune responses in the targeted cells. It is challenging to identify the multiple factors that affect the protein properties during the entire cycle of their production from cloning to manufacturing. The fast growth of bioinformatic tools and databases allow statistical evaluation of large variety of those critical factors and their contribution to the manufacturing process. By applying multidisciplinary and Design of Experiments approaches, a rapid decline of the variables is achievable and thus can bring about results that are more consistent to improve the success in clinical trials.
Sime Darby Research, Malaysia
Time : 16:00-16:20
Zahariah obtained Ph.D. in Surfactant Chemistry in 2001 and Master in Business Administration in 2006, major in Techno- entrepreneurship. Her area of academic specialized was in oleochemical. She has worked with Malaysian Palm Board (MPOB) for 23 years. Her involvement was in research & development for rheology of colloid , pharmaceutical, personal care and household products. Now she engaged with Sime Darby Research Sdn Bhd working on the same line. She has published more than 30 papers in reputed journals.
Tocotrienols and tocopherols are members of Vitamin E. They are naturally found in palm oil, barley, wheat germ, oats, rice bran and etc. Palm Tocotrienols is a palm derived Vitamin E consisting of naturally occurring tocotrienols-tocopherol mixture. Others grain consist solely tocopherol. The tocotrienols differs from tocopherol as the former have three double bonds in the carbon side chain. The Tocotrienols differences may result in different efficacy and potential as an antioxidant (Mashima et al 2003 ). The tocotrienols was reported to have higher antioxidant activity compared to tocopherol and has been proven to have anti-aging properties (Sen et al.2007). Many work on other studies on benefit of human health such as DNA damage in older adults, neuroprotective effect, chemo-protective and skin protection have been published (Wan Zurinah WG., et al 2008, Osakada F., et al 2004, Sen CK., et al 2004 and Khana S., et al.2005, Traber MG., et al. 1997, Nesaretnam K, 2004, Sylvester PW. et al.2007, ). The objective of this paper is to confirm the effectiveness of tocotrienols in personal care products. The personal care formulations enriched with tocotrienols, namely nano anti-wrinkle serum, whitening lotion, slimming lotion, and cracked heel cream were prepared. The efficacy/clinical studies were carried out by the third party. The products were measured for skin firmness, skin elasticity, melanin content, fat removal, trans epidermal water loss (TEWL) and crack heel recovery. The result showed that the tocotrienols prevent skin aging by stimulating collagen synthesis and increase skin elasticity in nano anti-wrinkle serum. Skin whitening lotion has shown that the tocotrienols promote skin whitening by reducing tyrosinase activity and decreased melanin content. Tocotrienols has shown in fat removal for slimming lotion. The decrease in skin roughness was found in cracked heel.
San Diego Regenerative Medicine Institute & Xcelthera, USA
Title: Direct conversion of pluripotent human embryonic stem cells into functional human neuronal or cardiomyocyte cell therapy derivatives for regenerative medicine
Time : 16:20-16:40
Xuejun H Parsons received her PhD in Biochemistry/Molecular/Cell Biology from Cornell University, and completed her PostDoc studies as a Leukemia and Lymphoma Society Research Fellow at University of California (UC). She has been supported by grants from NIH to become an independent investigator and leader in human embryonic stem cell research. She is the Founder of San Diego Regenerative Medicine (RM) Institute, Xcelthera, and California Consortium for RM Startup. Previously, she was a stem cell scientist/faculty at UC. She is inventor and corresponding author of more than 20 articles and serves on editorial boards of open access scientific journals.
Given the limited capacity of CNS and heart for self-repair/renewal, cell-based therapy represents a promising therapeutic approach closest to provide a cure to restore normal tissue and function for neurological and cardiovascular disorders. Derivation of human embryonic stem cell (hESCs) from the in vitro fertilization (IVF) leftover embryos has brought a new era of cellular medicine for the damaged CNS and heart. Recent advances and technology breakthroughs in hESC research have overcome some major obstacles in moving stem cell research from animals towards humans trials, including resolving minimal essential human requirements for de novo derivation and long-term maintenance of clinically-suitable stable hESC lines and direct conversion of such pluripotent hESCs into a large supply of clinical-grade functional human neuronal or cardiomyocyte cell therapy products. Such breakthrough stem cell technologies have demonstrated the direct pharmacologic utility and capacity of hESC cell therapy derivatives for human CNS and myocardium regeneration and, thus, have presented the hESC cell therapy derivatives as a powerful pharmacologic agent of cellular entity for CNS and heart repair. The availability of human stem/progenitor/precursor cells in high quality and large commercial scales with adequate cellular neurogenic or cardiogenic capacity will greatly facilitate developing safe and effective cell-based regenerative therapies against a wide range of CNS and heart disorders. Transforming non-functional pluripotent hESCs into fate-restricted functional human cell therapy derivatives dramatically increases the clinical efficacy of graft-dependent repair and safety of hESC-derived cellular products, marking a turning point in cell-based regenerative medicine from current studies in animals towards human trials.
Royan Institute for Stem Cell Biology and Technology, Iran
Title: Repeated injection of autologous mesenchymal stem cells in patients with knee osteoarthritis: Randomized double blind clinical trial royan institute (RIMRO-TriaL)
Time : 16:40-17:00
Nasser Aghdami earned his MD degree from the Urumieh Medical University (1998), and Ph.D. in Immunology from Tarbiat Modares University (2004) in Iran. During (2000- 2006) he was an associate investigator at Hematology and Oncology in Royan Institute and his research focused on Stem Cell Biology. He has published over 40 international papers and editorial boards related to Stem Cell, Clinical Trail, and Cell therapy. What's more, he is the editor of three books in Stem Cell and Regenerative Medical field. He was prestigious award from his research include young investigator in 2012.
Background:Osteoarthritis (OA) is a degenerative joint disorder and is a common condition in elderly. Recent non-randomized studies including our three previous phase I/II clinical trials have shown the safety of mesenchymal stem cells injection in the treatment of osteoarthritis. Study Objects and design: To investigate the effects of intra articular injection of autologous bone marrow derived mesenchymal stem cells (BM-MSC) on the symptoms of moderate to sever knee osteoarthritis we performed a double blind, placebo-controlled study in 46patients. Eligible patients were 18 to 65 years old and had a joint pain visual analog scale of >30 mm (at rest).Patients fulfill criteria for knee OA were randomly assigned into two groups: group one received BM-MSC (20 million cells, twice atday 0 and week12th) and group two received carrier media as placebo. Primary end points were the Western Ontario and McMaster Universities Osteoarthritis Index (WOMAC) and a visual analogue score (VAS) for pain at baseline and at the end of weeks 12, and 24,36. Secondary end points were pain free walking distance and WOMAC stiffness sub score and cartilage thickness, subchondral edema, tumor formation or neoplastic changes at 24 and 36 weeks. Results:The results indicated that the intra articular injection of autologous BM-MSC is very well tolerated during the 9 months trial. Moreover BM-MSC treated group had significantly clinical improvement as compare to placebo group in all clinical end points. In particular, the WOMAC-Total score, WOMAC-Physical Function sub score, WOMAC-pain sub score and pain free walking distance in BM-MSC, were superior compared to the placebo group (P =0.03, p=0.02, p=0.03 and P =0.01, respectively). Other clinical endpoints including the WOMAC-Stiffness, and VAS scores were also improved in both groups as compared to the baseline; but no statistical significance was detectable between two groups. Primary radiologic data indicated that subchondral edema decreased in some patients also thickness of cartilage increased in MSC group. Evaluation of radiographic data is not complete yet. Conclusion:Our short term follow up (9 months) have shown that repeated intra articular injection of BM-MSC is safe and effective in reducing functional impairment and relieving pain in patients with moderate to severe osteoarthritis of the knee. Clinical trial registration: NCT01504464.
Indore University, India
Time : 17:00-17:20
Jyoti Bhojwani, is presently a Faculty of Genetics/Bioinformatics/ Principal Investigator of the M.Tech Research Programs (Bio-Informatics)at University of Indore, India. She obtained her B.Sc.(Bachelors degree) in Biological Sciences/Chemistry/Physics, M.Sc.(Master’s degree) in Life-Sciences, and Doctoral degree (Ph.D.) at School of Life-Sciences, University Of Indore. She pursued her post-doctoral ventures at Max-Planck Institute For Biophysical Chemistry (FRG), University Of California Irvine and University Of Pittsburgh (USA). Currently, her projects mainly focus on translational-research and extrapolation of basic developmental mechanisms from modelsystems like fruitfly (Drosophila) to human. Apart from this, her thrust areas of research interest include Cancer Biology, Stem-Cell Biology and Homeotic-Gene Regulation. She is keen on studying in details the genetic factors, which presumably aid in understanding of mechanism by which cancer stem cells function in transforming a tissue from normal to cancerous states. Her research has a motive to further facilitate the perception of stem cell potential/mechanistic in areas of Regenerative Medicine, Translational Research and Anti-cancer therapy. Being involved in Clinical informatics, her students are also training a Cancer model and a Stem cell model, deploying Systems Biology approach and other Gene-Networking BioInformatics tools. This novel area of research will hopefully lead to further understanding the tipping of balance from a stem cell/normal cell to a transformed cancer cell.
Colorectal cancer (CRC) is the second leading cause of cancer related deaths globally posing a lifetime risk of 80-100% in every individual. Genetics and relevant mechanisms underlying some key signaling pathways like Wnt, TGF, p53, K-ras etc. play a detrimental role in governing the predisposition for CRC. A high percentage of colorectal tumors (adenomas and carcinomas) show activating mutations in beta-catenin or axin, whereas, loss of certain tumor suppressor genes (TSGs), like APC cause the initiation of random polyps in the colon. All of these molecules co-incidentally are critical components of an evolutionarily conserved Wnt signaling pathway, which is instrumental at various time points in the development of this disease. Differences in SNP profiles amongst sample groups in the genomic landscape can be recognized through a smart and efficient use of machine learning techniques. The statistics and pattern analyses of these SNP profiles, interestingly provides us with a concrete and logical platform upon which, relative contribution/s of each unique SNP, ranging from cause to effect can be significantly assessed. The biological relevance of such SNP variations with respect to cancer prediction and predisposition, however, remains unexplored, pending a better understanding of the impact of control design in SNP studies. Our results emerging from critical analyses of significant SNP’s, demonstrates the utility of relevant bioinformatics tools in discriminating diseased populations based on realistic SNP data. In this study, we have primarily targeted members of Wnt signaling pathway, which play important developmental role/s during different stages of colorectal cancer, depicting a classical multigene-multistep nature of cancer. We have identified and related common genetic variants for the early-acting and late-acting members of this pathway, that are most prevalent in patients with CRC disease. Complex relationships and correlations hidden in large data sets have been dug and analyzed here, by deploying various data-mining (bioinformatics) tools. Results will be presented in the light of discussing the scope of such a combinatorial approach, in identifying some potential candidates, in translational research and clinical research interventions.
Centre for Biological Engineering, Loughborough University, UK
Title: Serum-free expansion, harvest and preservation of mesenchymal stem cells from a scalable microcarrier process
Time : 17:20-17:40
Thomas Heathman is a PhD student at Loughborough University’s EPSRC Doctoral Training Centre in Regenerative Medicine, with a focus on the scalable manufacture of cell-based therapies. He has a First Class Master’s Degree in Chemical Engineering from the University of Bath and has previously worked in the oil & gas industry as a process engineer. He has currently published 5 papers in reputable journals, holds positions on 6 committees and leads multiple contract research projects with industry partners.
The use of animal-derived serum is common in MSC culture but has many drawbacks such as limited supply, lot-to-lot variability, possibility of pathogen transmission and reduced scope for process optimisation. These constraints have the potential to impact the development of a consistent large-scale process and therefore must be addressed. In this work, we have demonstrated the successful end-to-end production of MSCs from a potentially scalable serum-free process. Human MSCs have been expanded on fibronectin coated, non-porous plastic P-102L microcarriers (Solohill, USA) in 100 mL stirred spinner flasks at a density of 3x105 cells/mL in serum-free medium with monitoring of key metabolites. MSCs were successfully detached and separated from microcarriers using our recently-developed protocol with a post-harvest viability of 99.63 ± 0.03%, demonstrating full ISCT characterisation and maintaining MSC outgrowth and colony-forming potential. MSCs were held post-harvest for four hours in suspension to simulate a typical pooling time for a scaled expansion process and cryopreserved in a serum-free vehicle solution using our developed controlled-rate freezing process. Post-thaw viability was 75.8 ± 1.4% with a similar three hour attachment efficiency of 75.2 ± 13.1%, demonstrating successful MSC recovery and attachment. MSCs were able to form an F-Actin containing cytoskeleton following attachment and demonstrated five-fold expansion after seven days in culture post-thaw. This study has demonstrated for the first time that MSCs can be expanded, harvested, cryopreserved and recovered from a potentially scalable serum-free microcarrier process. Demonstrating the successful integration of multiple unit operations in a serum-free MSC production process from expansion through to cryopreservation provides an important pilot study in the development of an end-to-end MSC manufacturing process.
Canik Başari University, Turkey
Time : 17:40-18:00
Ferhat Ozturk has completed his PhD at University of Nevada, Reno under the supervision of Christopher Porada and Esmail Zanjani, one of the leading teams of in utero gene therapy. He moved to Creighton University for his Postdoctoral study with Devendra Agrawal in cardiovascular gene therapy research. He then worked with Ali Nawshad at the University of Nebraska Medical Center in the field of cell signaling and bioinformatics of cleft palate. Recently, he moved to Canik Başarı University of Turkey as the founding chair of Molecular Biology and Genetics. He serves as Director of BasGen Cancer Research Laboratory and BalMer Apitherapy Center.
One of the major developments in the field of cancer gene therapy is to use nano-materials as transfer vectors. Nanoparticles serve various advantages to deliver the genetic cargo such as; i) easy penetration into the cell membrane due to their miniscule size and polarity character, ii) targeting of cancer cells using specific molecular labels, and iii) efficient transfer to the cytoplasm and/or nucleus of the host cell. Especially, the gold nanoparticles (AuNP) are being widely used in various biochemical and biomedical applications due to their biocompatibility and surface modification potential. RNA interference (RNAi) agents, such as siRNA and shRNA, are highly specific to inhibit the target genes and are mostly delivered into the cancer cells using the viral vectors. However the viral vectors have some side effects such as immune reaction to the vector itself and low transduction efficiency of the genetic cargo, when used to transduce cancer cells. We have developed a novel method of siRNA transfer into the MCF-7 cells using PEGylated AuNPs, which were also conjugated with either RGD or NLS peptides. Successful and efficient delivery of the AuNP vectors into the cytoplasm and/or nucleus was visualized using darkfield illumination. The activation of the siRNAs and inhibition of the target genes were measured using qRT-PCR and western blotting. Consequently, development of AuNPs equipped with target specific ligands promises to be a critical tool in cancer gene therapy.