Day 1 :
Keynote Forum
Joseph J Barchi Jr
National Cancer Institute, USA
Keynote: Glycopeptide conjugates of a specific tumor-associated carbohydrate antigens as anticancer agen
Time : 10:05-10.30
Biography:
Joe Barchi received his Ph.D. in Synthetic/Marine Natural Products Chemistry from the University of Hawaii and was a postdoctoral fellow at Duke University. He then joined the National Cancer Institute as a staff fellow in the Laboratory of Medicinal Chemistry in1988 where he rose to his current position of Senior Scientist/Principal Investigator and NMR Facility Head at the newly formed Chemical Biology Laboratory. His main research interests are in synthetic medicinal chemistry as it relates to carbohydrate-based drug design, the development of novel sugar-conjugated nanoparticles and the high-resolution structural analysis of sugars, glycopeptides and small molecule drug candidates by NMR spectroscopy.
Abstract:
A hallmark of tumorigenesis is the aberrant expression and presentation of cell surface glycans. This modified display of carbohydrate structures has a host of biological consequences, including modified cell adhesion, proliferation and metastasis. Some of these glycans are recognized as “foreign” to the immune system and hence are considered tumor-associated carbohydrate antigens (TACAs). One such antigen, the Thomsen Friedenreich (TF) disaccharide, is directly involved in the metastatic process and has been used as a hapten to for the development of cancer vaccines. Our lab has studied the therapeutic potential of the TF antigen in the context of TF antigen and TF-glycopeptide-coated nanoparticles as vaccine or anti-adhesive platforms. In addition, a glycopeptide with potent antiproliferative and antitumor properties from a bladder disease called interstitial cystitis, also contains the TF antigen, and we have studied its structure activity relationship as an adjunct to develop novel anticancer agents. This lecture will describe some of the results we have obtained on both of these fronts and outline plans for the development of TF antigen-based therapeutics.
Keynote Forum
Michael P Jennings
Deputy Director Institute for Glycomics, Australia
Keynote: Application of Glycan array analysis in the discovery of novel bacterial; host interactions
Time : 10:30-10.55
Biography:
Prof Michael Jennings works in the fields of glycobiology, bacterial genetics and bacterial pathogenesis. His work has focused on bacterial pathogens, in particular the pathogenic Neisseria (meningitis) and Haemophilus influenzae. He was awarded his PhD (1990) from Griffith University. His post-doctoral training was in the laboratory of Prof Richard Moxon at the University of Oxford 1992-1996 funded by the Beit Memorial Fellowship for Medical Research. In 1997 he took up a faculty position at the University of Queensland. He remained at University of Queensland until 2009, until he returned to Griffith University to take up the position of Deputy Director at the Institute for Glycomics.
Abstract:
Glycans are important structures in many host - pathogen interactions. Bacterial lectins such as adhesins and toxins exploit host glycans as targets. Host lectins recognize bacterial glycans in innate immune processes. The molecular details of many bacterial - host interactions remain to be discovered. Understanding these processes is key for the development of novel strategies for prevention and therapeutics. We have applied glycan array to discover novel interactions between bacterial and human cells. Using the Institute for Glycomics glycan arrays, comprising 400 different structures, we have discovered novel glycan targets for the archetypal cholesterol-dependent cytolysin toxins, pneumolysin and streptolysin O. Previously it had been believed that cholesterol rich membrane of host cells were the only receptor of these toxins. We report high affinity binding to glycan structures present on red blood cells. For example, streptolysin binds to lacto-N-neotetraose with a KD <1 nM. Binding to this structure is required for efficient red blood cell lysis by strepolysin O. Using similar approaches we have defined novel classes of bacterial and host lectins.
- Track 1 : Glycans in Diseases and Therapeutics
Track 2: Glycomics and Bioinformatics
Track 3 : Glyco Immunology
Location: Independence C
Chair
Joseph J Barchi Jr
National Cancer Institute, USA
Co-Chair
Cecilia H. Marzabadi
Seton Hall University, USA
Session Introduction
Maria O. Longas
Purdue University Calumet, USA
Title: NaH Inhibits β-Amyloid Fibril Formation in Alzheimer’s Disease Rats
Time : 11:10-11:30
Biography:
Maria Longas has completed her PhD at New York University and done her postdoctoral training at Columbia University School of Medicine with Dr. Karl Meyer. She is a full Professor of Chemistry at Purdue University Calumet in Hammond, IN. She has more than 24 papers published in reputable journals, and has served as a reviewer for several journals.
Abstract:
Alzheimer’s Disease Rats were developed by Taconic Biosciences, MPA; development took 5 weeks. After this time, each of the 6 Alzheimer’s Disease rats was injected with NaH (3 mg/3 mL sterilized phosphate buffer saline, SPBS); 3 grams of NaH/gram of Rat body mass was used for injection. The controls were normal Rats, 6 of which were injected with 3mL of SPBS per gram of Rat body mass. After 5 weeks, all Rats were guillotined; the heads were removed and stored at -35oC. Cells were then cultured following conditions described for human Alzheimer’s Disease cells (Longas, MO, et al, unpublished) and carried out in 6-well-tissue-culture-dishes, each containing 4 mL of the following media: Astrocyte Basal Medium from Lonza adjusted to 15% (w/w) fetal bovine serum and fortified with L-Glutamine (5.0 mL), Ascorbic Acid (0.5 mL), Epidermal Growth Factors (1.25 mL), Insulin (0.5mL) and Gentamicin Sulfate Amphotencin-ß (0.5 mL). Cells grew in these media at 37oC, under 5% CO2 for 7 days. They were then stained with Congo Red. Figure IIIR displays Alzheimer’s Disease Hippocampus cells which show plenty of β-Amyloid tangles, while Figure IVR has cells from the Normal Hippocampus with no tangle formation. NaH is, therefore, effective to inhibit β-Amyloid tangle formation in Alzheimer’s Disease.
George R. Bousfield
Wichita State University, USA
Title: Influence of glycans on biological activities of pituitary gonadotropins, follicle stimulating hormone and luteinizing hormone
Time : 11:30-11:50
Biography:
George Bousfield completed his PhD at Indiana University in 1981. He then pursued postdoctoral studies at the University of Texas, M.D. Anderson Cancer Center with Darrell Ward. He is currently the Dr. L.M. Jones Distinguished Professor in the Fairmount College of Liberal Arts and Sciences at Wichita State University, and director of the WSU Protein Core Facility. He has published more than 56 journal articles.
Abstract:
The gonadotropins, follicle-stimulating hormone (FSH) and luteinizing hormone (LH), play central roles in the control of reproduction by connecting neuroendocrine hypothalamic/pituitary activities to gonadal function. LH and FSH receptor activation is an important part of assisted reproductive technologies, such as in vitro fertilization. Both FSH and LH are heterodimeric glycoproteins, each composed of a non-covalently associated, common ï¡- and a hormone-specific ï¢-subunit. Both subunits are N-glycosylated. Although high affinity binding of both hormones to their cognate receptors occurs exclusively by protein-protein interactions, full gonadotropin receptor activation involves N-linked glycans. However, the activation mechanism remains elusive. Part of the problem is the high degree of oligosaccharide microheterogeneity that challenges even contemporary analysis by high-resolution mass spectrometry. LH possesses three N-glycosylation sites that are always occupied by glycans and can be removed individually, permitting characterization in a site-specific manner. While FSH glycopeptides can be obtained from all 4 potential N-glycosylation sites, only 2 can be thoroughly analyzed by glycopeptide mass spectrometry. Even then, only about a third of the glycan population can be detected. While both FSHï¡ subunit glycosylation sites can be interrogated individually by sequential peptide-N-glycanase digestion, both FSHï¢ glycans are released simultaneously. This is unfortunate, because naturally occurring forms of FSH lacking either one of the 2 FSHï¢ N-glycans bind their receptors more rapidly, occupy more FSH binding sites, and provoke a more robust cellular response than fully-glycosylated FSH. Moreover, the ratio of FSH glycosylation variants may be under physiological regulation. Supported by NIH grant P01 AG029531.
Cecilia H. Marzabadi
Seton Hall University, USA
Title: Novel carbohydrate-based ligands with immunostimulatory properties
Time : 11:50-12:10
Biography:
Professor Marzabadi completed her PhD at the University of Missouri – St. Louis and carried out postdoctoral studies at Hunter College- CUNY. She has been at Seton Hall University for the past 15 years and has been a Visiting Professor at Harvard University Medical School and a Visiting Researcher at Memorial Sloan Kettering Cancer Center. She has published numerous papers in the carbohydrate literature and currently serves as a Member at Large for the ACS Division of Carbohydrate Chemistry.
Abstract:
We are interested in the synthesis and biological evaluation of new carbohydrate structures with the capacity to act as immunomodulators. Towards these ends we have synthesized molecules as mimics of capsular saccharides. These highly simplified saccharide structures have been shown to act as TLR4 agonists and to elicit the secretion of proinflammatory cytokines at concentrations lower than the known agonist, E. coli lipopolysaccharide. In this presentation, we will discuss the synthetic efforts towards a lead compound and the preliminary biological data obtained for this compound. Efforts to elucidate the mechanism of action of this molecule will be discussed.
Myron R Szewczuk
Queen’s University, Canada
Title: Therapeutic targeting neuraminidase-1 in multi-stage of tumorigenesis
Time : 12:10-12:30
Biography:
Myron Szewczuk has completed his PhD 1974 from University of Windsor in Biology and Immunochemistry and his postdoctoral studies from Cornell University Medical College, New York City, U.S.A. in Cellular immunology with Dr. Greg Siskind (1975-1978). He is presently Full Professor of Immunology and Associate Professor of Medicine, Queen’s University, Kingston, Ontario, Canada. He has published more than 100 papers in reputed journals and has been serving as an editorial board member of repute.
Abstract:
Various reports have suggested that receptor glycosylation modiï¬cation may in fact be the invisible link connecting ligand-binding and receptor dimerization. A novel organizational signaling platform linked to the glycosylated receptor tyrosine kinases (RTK) (e.g., EGFR, TrkA, insulin) and TOLL-like receptors induced receptor activation process, all of which are known to play major roles in tumorigenesis. This signaling paradigm proposes that ligand binding to its receptor on the cell surface induces a conformational change of the receptor to initiate MMP-9 activation to induce Neu1. Activated Neu1 hydrolyzes α-2,3-sialyl residues linked to β-galactosides, which are distant from the ligand binding sites. These ï¬ndings predict a prerequisite desialylation process by activated Neu1 enabling the removal of steric hindrance to receptor association. The importance of these findings signify an innovative and promising entirely new targeted therapy for cancer. The role of mammalian neuraminidase-1 (Neu1) in complex with matrix metalloproteinase-9 and G-protein coupled receptor tethered to RTKs and TLRs is identified as a major target in the multi-stage of tumorigenesis. Preclinical studies support an entirely new cancer therapy targeting different growth factor receptors, tumor neovascularization, chemo-resistance of tumors, immune-mediated tumorigenesis and the development of tissue invasion and metastasis. Evidence exposing the link connecting growth factor-binding and immune-mediated tumorigenesis to this novel receptor-signaling paradigm will be presented in its current relationship to pancreatic, ovarian and triple-negative breast cancers.
Francis Jeshira Reynoso
University of Pennsylvania, USA
Title: The clinical phenotype of PIGN deficiency and consequences of defective GPI biogenesis
Time : 12:30-12:50
Biography:
Jeshira Reynoso has completed her Pediatrics Residency at Lincoln Medical Center; Bronx NY affiliated to Weill Cornell Medical College. She is currently a Clinical Genetics Fellow at the Children’s Hospital of Philadelphia and is conducting research under the mentorship of Miao He and her PhD is focused on Congenital Disorders of Glycosylation.
Abstract:
The PIGN gene (MIM606097) encodes glycosylphosphatidylinositol (GPI) ethanolamine phosphate transferase-1 which adds the ethanolamine phosphate to the first mannose on the GPI anchor. GPI anchoring of proteins is a highly conserved process present in most eukaryotic cells. GPI-anchored proteins perform a diverse set of functions including roles in signal transduction, cell adhesion and antigen presentation. To date mutations in 12 genes involved in GPI anchor synthesis have been associated with human disease. Maydan first described mutations in this gene in a consanguineous Israeli family of Palestinian origin; children presented with multiple congenital anomalies, severe neurological involvement and early death. Four additional cases have been reported since then. We report three unrelated patients with mutations in PIGN and diverse clinical characteristics. Patients 1 and 2 presented during their newborn period with hypotonia, intractable epilepsy, facial dysmorphism as well as gastrointestinal, genitourinary and skeletal anomalies including brachytelephalangy. Patient 1 had metopic suture craniosynostosis and left choanal atresia. Patient 2 had an imperforate anus, uterine didelphys and a small splenic cyst. Patient 3 with a milder phenotype manifested as hypotonia, developmental delay, dysmorphic facial features and early onset refractory epilepsy. Fluorescence activated cell sorting analysis was performed on cultured skin fibroblast lines from Patients 2 and 3 which demonstrated decreased cell surface expression of the GPI anchored protein CD59, consistent with deficient GPI anchor biogenesis in these cells. Mechanistic studies of PIGN deficiency are currently underway in our laboratory.
Tony Velkov
Monash University, Australia
Title: Molecular characterization of the hemagglutinin receptor-binding specificity of avian influenza viruses: predicting the jump across the species barrier
Time : 13:35-13:55
Biography:
Dr Tony Velkov completed PhD in 2000 from Monash University. His anti-infective discovery research is at the leading edge globally. He was awarded a NHMRC Research Fellowships in 2006, 2011 and 2014. The quality and impact of his independent research was recognized by the NHMRC with an Excellence Award in 2011. He has published over 50 papers in high-caliber journals, 3 book chapters and 15 conference presentations. The dynamic team he leads consists of 3 postdocs, 3 RAs and 9 PhD students. Over the last 6 years, he has obtained >$9M funding from the NIH, NHMRC and foundations.
Abstract:
Influenza is a constant global burden to human health. Seasonal influenza results in significant infections and death, cycling through both hemispheres. On occasion, a novel avian influenza viruses crosses the species barrier from birds to humans resulting in an influenza pandemic. The threat of pandemic avian influenza continues with H5N1 and H7N9 consistently infecting humans. In order to evolve from its avian form and gain the pandemic potential for increased transmissibility between humans, the hemagglutinin (HA) of avian influenza viruses will need to undergo mutations in its receptor binding site (RBS) that bring about an avian to human receptor preference switch. In order to understand the major determinants of virus transmissibility and the pandemic potential of the novel avian influenza viruses we have determined the crystallographic structure of the novel avian influenza H10N7 A/Turkey/MN/3/79 to 1.96Å and mapped the RBS. The amino acid residues responsible for conferring receptor selectivity were identified by site-direct mutagenesis of recombinant H10 HA proteins. The receptor-binding selectivity of the HAs was determined using sialyl glycan binding assays. Docking models were constructed of the H10 HA in complex with α2,6-sialic acid (human) and α2,3-sialic acid (avian) pentasaccharide receptor analogs to ascertain the correlation between the binding assay data and the interactions within the receptor binding pocket. The presented findings provide a structure-recognition perspective for the receptor binding properties of the novel avian H10 influenza HA.
Albert M. Wu
Chang-Gung University, Taiwan
Title: Roles of Polyvalency in the Mechanism of Glyco Recognition. An Important Direction for the Future Glycosciences
Time : 13:55-14:15
Biography:
Dr. Wu obtained his Ph.D degree with W. Pigman, who is the pioneer in glycoproteins, at New York Medical College; and had his postdoctoral training at E.A. Kabat’s Lab for quantitative immunochemistry, Columbia University Medical Center, New York. He joined as a faculty position at Texas A&M University in 1982; promoted as a full professor at Chang-Gung University since 1989; and as Emeritus Professor in 2011. Dr. Wu published over 120 polyvalent glycotopes related papers. He is the chief editor for three volumes of Molecular Immunology of Complex Carbohydrates 1 to 3 in Adv. Exp. Med. Biol. 228, 451, 705 (Springer Publisher). His major interests are (i) Glycan purification and characterization; (ii) recognition factors of glycans; (iii) combining sites of lectins and antibodies. He received many Outstanding Research Awards (1997-2008) from government agents in Taiwan.
Abstract:
Lectins are an important class of proteins or glycoproteins that specifically or selectively bind to carbohydrates and play many critical roles in life processes. In order to characterize recognition roles of lectins, the following aspects have been taken into consideration: i) lectins affinity to monosaccharides; ii) expression of reactivities toward oligosaccharides (mammalian structural units/recognition units) and finding the most active ligand. However, it is not satisfied, because most lectins with the same mono- or oligosaccharide specificity may demonstrate different specificities in reaction with polyvalent forms – it has even shown a shift of binding specificity of lectin from one type of carbohydrate ligand to another when the density of the specific carbohydrate changed. Therefore, characterization of lectin specificity has been extended to: iii) simple oligovalent or cluster effect; and iv) complex multivalent or cluster effects. Simple oligovalent effect concerns the reactivity of lectins with oligomeric glycoconjugates (e.g. branched oligosaccharides carrying several active disaccharides, glycopeptides with several Tα or Tn glycotopes). A complex multivalent effect applies to interaction with high-molecular or aggregated molecules carrying multiple glycotopes recognized by a lectin. In this talk, we are focusing on the resulting intensities of three basic recognition factors ï€ (ii) essential mammalian structural units, (iii) their clusters and (iv) polyvalency in the recognition processes.
Julian M Menter
Morehouse School of Medicine, USA
Title: Effect of temperature and hyaluronate on photochemical changes in calf skin collagen solutions at physiological PH
Time : 14:15-14:35
Biography:
Dr. Menter received his Ph.D. degree in Chemistry from the George Washington University in 1969. He completed a postdoctoral fellowship with Prof. Dr. Theodor Foerster at the Institut fuer physikalische Chemie der Universtiaet Stuttgart, Germany. Subsequently, he was at the University of Alabama, Birmingham, and the VA Medical Center (Atlanta) He currently serves as Research Professor of Biochemistry at Morehouse School of Medicine. Dr. Menter is recognized internationally for his work in the areas of collagen photochemistry and melanin photobiology as pertaining to redox reactivity.
Abstract:
Mammalian collagens contain several age-related fluorescent chromophores that are unstable to solar UV wavelengths. The consequences of the resulting collagen photo-degradation are not well known, since the direct collagen – UV interactions are poorly described at best. We are studying these interactions by following UV – induced changes in calf skin type I collagen fluorescence as functions of time, temperature, and hyaluronic acid (HA). We have observed that UVC radiation (mainly 254 nm) causes dimerization of tyrosyl residues to dityrosine and the disappearance of an age-related tyrosine oxidation product that is formed in the ground state. Dityrosine formation does not require oxygen and is relatively insensitive to changes in temperature and age of sample. The age-related oxidation product destabilizes the overall collagen supramolecular structure. Added sodium hyaluronate (2:1 ratio) shows little or no effect on these results. This latter result may indicate that there is little physical interaction between the collagen telopeptide and HA domains in vitro.
Xueli Li
University of Pennsylvania, USA
Title: Glycomics profile analysis by MALDI TOF/MS in human CSF
Time : 14:35-14:55
Biography:
Xueli Li has completed her PhD from Leipzig University, Germany. She is a Research Associate in The Michael J Palmieri Metabolic Laboratory, Children’s Hospital of Philadelphia, USA. She has more than 5 publications in the area of Glycomics recently.
Abstract:
Protein glycosylation is important for human brain development and function. The majority of the >100 known subtypes presents with intellectual disability. To explore possible defects in protein glycosylation in the brain, we developed comprehensive glycomics analysis of N-glycome, O-glycome and free glycome by MALDI TOF/MS in cerebrospinal fluid (CSF). 0.4 ml CSF was filtered to separate free oligosaccharides from glycoprotein. N-glycans were released via PNGase F digestion and O-glycans were released separately by reductive beta-elimination reaction. After purified, glycans were permethylated and analyzed by MALDI TOF/MS. Thirty eight different N-glycan species, 85 free oligosaccharide and 25 O-glycan species were identified in 10 control CSF samples. Compared with plasma glycomics data, CSF has much more O-glycan species including O-mannosylated glycans and polysialylated O-glycans. Comparing 166 CSF samples from patients with undiagnosed neurological disease from the NIH Undiagnosed Diseases Program (UDP) with 10 control CSF samples, about 20 UDP patients have CSF glycomics profiles significantly deviated from the profiles of control CSF samples. Four of the UDP patients have known genetic disorders that alter protein glycosylation in human brain, including one patient with putative CAD deficiency, a defect in uridine biosynthesis. Interestingly, one of the known patients has a completely normal plasma glycomics profile and urine oligosaccharide profile, while both his CSF N and O-glycomics profiles are abnormal. Our results suggest that CSF glycomics analysis could be a useful tool to discover new diseases or disease mechanism.
Sourav Sarkar
Lehigh University, USA
Title: MUC1 glycopeptide based anti-cancer vaccines
Time : 14:55-15:15
Biography:
Sourav Sarkar has completed his PhD from University of Toledo (research advisor: Prof. Steven J Sucheck) and postdoctoral studies from Complex Carbohydrate Research Center, University of Georgia (Prof. Geert-Jan Boons). Presently he is working as a Research Scientist III in the Department of Chemistry at Lehigh University. He has published numerous papers in reputed journals and is an inventor of the patent “Xeno-antigenic anti-tumor vaccines”. He is a member of the American Chemical Society and has served as a peer reviewer for Elsevier, Royal Society of Chemistry, Springer etc. journals.
Abstract:
MUC1 variable number tandem repeats (VNTRs) conjugated to tumor-associated carbohydrate antigens (TACAs) have been shown to break self-tolerance in humanized MUC1 transgenic mice. Therefore, we hypothesized that a MUC1 VNTR TACA-conjugate can be successfully formulated into a liposome-based anti-cancer vaccine. The immunogenicity of the vaccine should be further augmented by incorporating surface displayed L-rhamnose (Rha) epitopes onto the liposomes to take advantage of a natural antibody-dependent antigen uptake mechanism. To validate our hypothesis we synthesized a 20-amino acid MUC1 glycopeptide containing a GalNAc-O-Thr (Tn) TACA by SPPS and conjugated it to a functionalized Toll-like receptor ligand (TLRL). An L-Rha-cholesterol conjugate was prepared using tetraethylene glycol (TEG) as a linker. The liposome-based anti-cancer vaccine was formulated by the extrusion method using TLRL-MUC1-Tn conjugate, Rha-TEG-cholesterol and 1,2-dipalmitoyl-sn-glycero-3-phosphocholine (DPPC) in a total lipid concentration of 30 mM. Groups of female BALB/c mice were immunized and boosted with a rhamnose-Ficoll (Rha-Ficoll) conjugate formulated with alum as adjuvant to generate the appropriate concentration of anti-Rha antibodies in the mice. The mice were then immunized with the TLRL-MUC1-Tn liposomal vaccine formulated either with or without the surface displaying Rha epitopes. Sera collected from the groups of mice initially immunized with Rha-Ficoll and later vaccinated with the Rha-displaying TLRL-MUC1-Tn liposomes showed a >8-fold increase in both anti-MUC1-Tn and anti-Tn antibody titers in comparison to the groups of mice that did not receive Rha-Ficoll. The anti-MUC1-Tn antibodies in the vaccinated mice serum also recognized MUC1 on human leukemia U266 cells.
Michael S. Strano
Massachusetts Institute of Technology, USA
Title: New concepts in molecular and energy transport within carbon nanotubes and graphene: optical sensors, resonant ion channels and thermopower waves
Time : 15:30-15:50
Biography:
Professor Michael S. Strano is currently the Carbon P. Dubbs Professor of Chemical Engineering at the Massachusetts Institute of Technology. He received his B.S from Polytechnic University in Brooklyn, NY and Ph.D. from the University of Delaware both in Chemical Engineering. He was a post doctoral research fellow at Rice University in the departments of Chemistry and Physics under the guidance of Nobel Laureate Richard E. Smalley. From 2003 to 2007, Michael was an Assistant Professor in the Department of Chemical and Biomolecular Engineering at the University of Illinois at Urbana-Champaign before moving to MIT. His research focuses on biomolecule/nanoparticle interactions and the surface chemistry of low dimensional systems, nano-electronics, nanoparticle separations, and applications of vibrational spectroscopy to nanotechnology. Michael is the recipient of numerous awards for his work from 2005 to the present.
Abstract:
Our laboratory has been interested in how 1D electronic materials such as carbon nanotubes can be utilized to illustrate new concepts in molecular transport and energy transfer. In the first example, we predict and demonstrate the concept of thermopower waves for energy generation(1). Coupling an exothermic chemical reaction with a thermally conductive CNT creates a self-propagating reactive wave driven along its length. We realize such waves in MWNT and show that they produce concomitant electrical pulses of high specific power >7 kW/kg. Such waves of high power density may find uses as unique energy sources. In the second system, we fabricate and study SWNT ion channels for the first time(2) and show that the longest, highest aspect ratio, and smallest diameter synthetic nanopore examined to date, a 500 μm SWNT, demonstrates oscillations in electro-osmotic current at specific ranges of electric field, that are the signatures of coherence resonance, yielding self-generated rhythmic and frequency locked transport. The observed oscillations in the current occur due to a coupling between stochastic pore blocking and a diffusion limitation that develops at the pore mouth during proton transport. Lastly, our laboratory has been interested in how semiconducting single walled carbon nanotubes (SWNT) can be modified such that their fluorescent emission is modulated in response to specific molecules, hence creating a new class of sensor (3). Such sensors have important advantages, including enhanced optical penetration of tissues in the near infrared, reduced auto-fluorescence, infinite resistance to photobleaching and most recently, single molecule analyte sensitivity. This presentation will review our recent efforts in this space including new platforms for label free protein detection, nitric oxide, H2O2 and the interfacing of sensor arrays to living cells.
Ajiloseda Charles Adeniyi
Adyghe State University, Russia
Title: Cytokine gene polymorphism (IL-17A G197A, TNF-alpha G308A, IL-6 C174G) and human major histocompatibility complex (HLA DRB1) specificities control in bronchial asthma
Time : 15:50-16:10
Biography:
Ajiloseda Charles Adeniyi has completed a Specialist (MSc) and a Professional Education degree at the Adyghe State University, Russia and as a result of his ardent observation and understanding in the hybridoma culture, antibody purification and microscopic usage as well as his keen scientific interest was immediately inducted into the research group at the University’s immune-laboratory working with the lead Professor Tugus. He collaborated with others scientists to determine the importance of medicinal plant on health: A case study of Iris plantae. His personal publication includes but not limited to his recent work on the “Development and integrating of health sciences into foreign languagesâ€.
Abstract:
In the development of non-specific resistance in an immune system, important role belongs to mediators of the said immune system; the pro-inflammatory cytokines which in turn activates expression of class II HLA-DRB1 on antigen presenting cells (APC) that implement specific immunity. The pathologies of many infectious, autoimmune and malignant diseases are influenced by the profiles of cytokine production in pro-inflammatory (TH1) and anti-inflammatory (TH2) T cells. With TLR (Toll-like receptor) on dendritic cells recognizing PAMP (pathogen-associated molecular patterns); antigens that trigger a cascade of specific antibody and cytokine. Many studies have examined the relationship between cytokine gene polymorphism, cytokine gene expression in vitro and the susceptibility to and clinical severity of diseases. Our study here tends to and as extensively as possible expose the relationship of cytokines gene polymorphism and Human Major Histocompatibility complex with respect to bronchial asthma.
Yves St-Pierre
INRS-Institut Armand-Frappier, CANADA
Title: Targeting galectins as a new tumor microenvironment-based therapeutic strategy in cancer
Time : 16:10-16:30
Biography:
Dr. St-Pierre has received his Ph.D. in immunology at the University of Toronto and pursued his postdoctoral training Harvard Medical School, working on the molecular mechanisms controlling trafficking of immune and cancer cells. He was recruited as at the INRS-Institut-Armand-Frappier in 1994 where he has been ever since. He is currently pursuing research aimed at identifying novel molecular targets and markers of cancer using various immunological and molecular approaches.
Abstract:
Carbohydrates are traditionally considered to be an important source of energy for living organisms. In the field of biology, they are defined as organic compounds composed of carbon, hydrogen, and oxygen that are organized into ring structures. The analysis of these structures and their functions has led to a new field of biology called “glycobiology.†In the biomedical sciences, glycobiology is rapidly emerging as an integral part of complex biological processes. Evidence suggesting that the interactions between lectins and their ligands play a major role in the different steps of cancer progression has accumulated at a rapid pace and has gained the attention of several oncologists. This is particularly true for galectin family members because changes in their expression levels correlate with alterations in cancer cell growth, apoptosis, and, due to their cytokine-like properties, they are major activators or inhibitors of immune responses. In mammals, 19 galectins have been identified, of which 13 are expressed in human tissues. While they perform homeostatic functions inside normal cells, under pathological or stress conditions, cytosolic galectins are released either passively from dead cells or actively via non-classical secretion pathways. Once in the extracellular milieu, they bind all glycosylated growth receptors on the surface of normal and cancer cells to set their signaling threshold. Such properties enable galectins to kill of infiltrating immune cells while promoting growth of tumor cells. We hypothesize that galectins play an important role in the tumor microenvironment which undergo profound alterations during cancer. Such alterations remains one of the great obstacle in the successful treatment of cancer is therapeutic resistance and cancer recurrence. This is especially true for cancer with poor outcome, such as triple negative breast cancer (TNBC) or ovarian cancer, for which no therapeutic targets exist. Here, we will discuss the relevance of the galectinome in TNBCs and ovarian cancer.
Peng G. Wang
Georgia State University, USA
Title: Efficient chemoenzymatic synthesis of an n-glycan isomer library
Time : 16:30-16:50
Biography:
Peng George Wang obtained his B.S. Degree in Chemistry from Nankai University, China in 1984 and his PhD. Degree in organic chemistry from University of California, Berkeley in 1990. He then conducted postdoctoral research in Scripps Research Institute and became an Assistant Professor in 1994 in University of Miami. From 1997 to 2003, he was a faculty at Wayne State University. In 2003, he took a position in the Departments of Biochemistry and Chemistry at Ohio State University as Ohio Eminent Scholar in Macromolecular Structure and Function. In 2011 He became Professor and Georgia Research Eminent Scholar in Chemical Glycobiology in the Department of Chemistry at Georgia State University.
Abstract:
Quantification, characterization and biofunctional studies of N-glycans on proteins remain challenging tasks due to complexity, diversity and low abundance of these glycans. The availability of structurally defined N-glycans (especially isomers) libraries is essential to help on solving these tasks. We reported herein an efficient chemoenzymatic strategy, namely Core Synthesis/Enzymatic Extension (CSEE), for rapid production of diverse N-glycan libraries. Starting with 5 chemically prepared building blocks, 8 N-glycan core structures containing one or two terminal N-acetyl-D-glucosamine (GlcNAc) residue(s) were chemically synthesized via consistent use of oligosaccharyl thioethers as glycosylation donors in the convergent fragment coupling strategy. Each of these core structures was then extended to 5 to 15 N-glycan sequences by enzymatic reactions catalyzed by 4 robust glycosyltransferases. Success in synthesizing N-glycans with Neu5Gc and core-fucosylation further expanded the ability of enzymatic extension. High performance liquid chromatography (HPLC) with an amide column enabled rapid and efficient purification (>98% purity) of N-glycans in milligram scales. A total of 73 N-glycans (63 isomers) were successfully prepared and characterized by MS2 and NMR. The CSEE strategy provides a practical approach for “mass production†of structurally defined N-glycans, which are important standards and probes for glycoscience.
Shuxia Wang
University of Kentucky, USA
Title: Thrombospondin1 an important mediator of obesity associated inflammation and insulin resistance
Biography:
Dr. Wang has completed her PhD from Institute of Basic Medical Sciences at Chinese Academy of Medical Sciences and postdoctoral studies from University of Alabama at Birmingham. She is an Associate Professor in the Dept. of Pharmacology and Nutritional Sciences at University of Kentucky and a scientist at VA Medical Center in Lexington. She has published more than 35 papers in reputed journals and has been serving as an editorial board member for several journals.
Abstract:
Thrombospondin 1 (TSP1) is a multifunctional matricellular protein. It is highly expressed in visceral fat tissue (AT) from obese and insulin resistant humans or obese rodents. Recently, both human and rodent data from our lab and others suggest that TSP1 plays an important role in obesity-associated chronic inflammation and insulin resistance (IR). The positive association of adipose tissue TSP1 with AT inflammation and IR has been observed in obese human subjects. By using global TSP1 deficient mice, we revealed a novel role for TSP1 in stimulating macrophage accumulation and activation in AT that promotes inflammation and IR resulting from high fat diet-induced obesity (DIO). Specifically, we found that feeding a high fat diet to wild type and TSP1 deficient mice for 16 weeks caused similar obesity, but only mice with TSP1 deficiency remained insulin-sensitive. The protection of TSP1 deficient mice against IR was associated with reduced ATMs, decreased adipose and systemic inflammation, and reduced AT fibrosis. Moreover, TSP1 deficiency protected mice from obesity-induced hypertension and kidney damage. In vitro data demonstrated that TSP1 deficient monocyte/macrophages had decreased chemotactic activity and a reduced pro-inflammatory phenotype. TSP1 treatment stimulated macrophage migration. In addition, TSP1 stimulated macrophages to produce pro-inflammatory cytokines, which required TLR4 activation and was mediated by interaction between the type 1 repeats of TSP1 (TSR) and its receptor-CD36. Collectively, these data suggest that TSP1 acts as both a chemoattractant and proinflammatory activator for macrophages in inflamed AT, and promotes obesity-induced inflammation and IR.
Adam Kapelner
Queens College, USA
Title: Starvation of cancer via induced ketogenesis and severe hypoglycemia
Biography:
Adam Kapelner is an Assistant Professor of Mathematics at Queens College in New York City. He graduated with a Ph.D. in Statistics from Wharton Business School in 2014. He was previously a software engineer building web applications in San Francisco and a mathematics & computational science undergraduate at Stanford University. He has published in a number of fields: Statistical Methodology, Statistical Software, Economics, Natural Language Processing but has a special love for applications to medicine.
Abstract:
Neoplasms are highly dependent on glucose as their substrate for energy production and are generally not able to catabolize other fuel sources such as ketones and fatty acids. Thus, removing access to glucose has the potential to starve cancer cells and induce apoptosis. Unfortunately, other body tissues are also dependent on glucose for energy under normal conditions. However, in human starvation (or in the setting of diet-induced ketogenesis), the body ‘‘keto-adapts’’ and glucose requirements of most tissues drop to almost nil. Exceptions include the central nervous system (CNS) and various other tissues which have a small but obligatory requirement of glucose. Our hypothesized treatment takes keto-adaptation as a prerequisite. We then propose the induction of severe hypoglycemia by depressing gluconeogenesis while administering glucose to the brain. Although severe hypoglycemia normally produces adverse effects such as seizure and coma, it is relatively safe following keto-adaptation. We hypothesize that our therapeutic hypoglycemia treatment has potential to rapidly induce tumor cell necrosis. This is joint work
Diwakar Jain
New York Medical College, USA
Title: Radiolabeled glucose analogues for myocardial ischemia imaging and for diagnosing coronary artery disease
Biography:
Dr Jain is a Professor of Medicine (Cardiovascular Medicine) and director of nuclear cardiology laboratory at New York Medical College. He attended Medical College, Rohtak (India) and underwent residency and fellowship training in Medicine and Cardiology at Post Graduate Institute of Medical Education and Research, Chandigarh (India). He underwent further training in cardiology at Northwick Park Hospital and Clinical Research Center, Harrow (UK) and Yale University School of Medicine. He has conducted research in the fields of molecular imaging, cardiotoxicity of cancer chemotherapy, and behavior, mental stress and heart disease. He has published over 150 scientific papers, book-chapters, editorials and review articles.
Abstract:
Glucose and free fatty acids (FFA) are the predominant metabolic substrates in heart. Their relative uptake in normal hearts varies with the metabolic milieu (fed/fasting state), plasma levels and insulin levels. Myocardial ischemia results in a profound upregulation of glucose and downregulation of FFA metabolism. Fluorine-18 labeled deoxyglucose (FDG), a glucose analogue is used extensively in clinical practice for imaging glucose metabolism in heart, tumors and other organs. Alternations in glucose metabolism accompanying several disease conditions can be used to diagnose these conditions. Exercise results in ischemia and upregulates glucose uptake in regions perfused by diseased coronary arteries. This can be used for developing a non-invasive diagnostic imaging test for coronary artery disease (CAD). We evaluated the diagnostic potential of exercise FDG imaging for CAD and compared it with exercise-rest myocardial perfusion imaging (MPI), an established and routinely used test. FDG imaging had higher diagnostic sensitivity compared to MPI. Furthermore, increased regional FDG uptake is observed only on exercise images. Persistence of FDG uptake 24 hours later is seen in less that one third of cases and is indicative of more severe CAD. Cardiac imaging using radiolabeled sugars is a highly promising new diagnostic test for CAD and may also provide a powerful tool for studying the pathophysiology of myocardial ischemia.
Yi-Pin Lin
Tufts University School of Medicine, USA
Title: Glycosaminoglycan binding adhesins of Lyme disease spirochetes contributes to tissue colonization and disease manifestations
Biography:
Yi-Pin Lin has completed his Ph.D. from Cornell University currently is a research assistant professor in the laboratory of Dr. John Leong at Tufts University School of Medicine. His research is emphasis on virulence mechanisms of bacterial pathogens that cause infectious diseases in humans and animals and host immunity to bacterial infection. His specific expertise is in pathogenesis of the Lyme Disease agent Borrelia burgdorferi, particularly in the bacterial-cell adhesion and immune evasion mechanisms that promote colonization and disease.
Abstract:
Borrelia burgdorferi is a bacterial spirochete that causes Lyme disease, the most common vector-borne disease in U.S. B. burgdorferi can infect the skin at the site of the tick bite, and subsequently colonize the heart, joints, or brain. The establishment of B. burgdorferi infection at diverse sites may be partly dictated by the attachment of spirochetes to target tissues. Indeed, B. burgdorferi encodes several adhesins that recognize components of the extracellular matrix such as proteoglycans and is often localized in connective tissue during infection. Proteoglycans consist of a core protein and one or more covalently linked long repeating disaccharides glycosaminoglycan (GAG). Upon adaptation to the mammalian host, B. burgdorferi produces surface-localized GAG-binding proteins as known as adhesins. The GAG-binding activity of these adhesins was speculated to mediate the tissues colonization and disease. Recently, spirochetes deficient in specific GAG-binding adhesins, such as DbpA or BBK32, have been shown to display reduced infectivity in a murine model, indicating a critical role for GAG-binding during mammalian infection. We further demonstrated that the GAG-binding activity of DbpA promotes colonization and disease manifestation at all of the tissues while this activity of BBK32 specifically promotes joint colonization. Therefore, GAG-binding activities may play roles in tissue colonization and the clinical manifestations of Lyme disease.
Biography:
Dr. Rosa Peracaula completed her PhD at the University Ramon Llull, Barcelona in 1995. Since 1995 she joined to the group of Biochemistry at the University of Girona and performed postdoctoral stays at MS Anderson Cancer Center in Houston (1997) and at the University of Oxford (2001 and 2002). Since 2003, she is associate Professor of Biochemistry and Molecular Biology at the University of Girona and leads the research group of Glycobiology and Cancer with projects related to find new biomarkers associated to changes of glycosylation in serum proteins and aimed to understand the role of the altered glycosylation in the tumorigenic process. She has three patents and has published over 25 scientific articles in international journals, some of them with more than 150 citations. She has directed five Doctoral Theses and has also given several presentations at national and international conferences. The research performed over these years has a strong translational component, with important collaborations with medical doctors and several national and international groups.
Abstract:
Altered glycosylation is one of the hallmarks of tumor cells and it is involved in each and every aspect of tumor progression. It affects cell surface carbohydrates and cellular and secreted glycoproteins, some of which may reach the bloodstream and be used as tumor markers 1. Our group has focused on the altered glycosylation of serum proteins in prostate and pancreatic cancer as potential tumor markers. We have described glycosylation changes of Prostate Specific Antigen (PSA) glycans, related to sialylation and fucosylation, in prostate cancer compared to Benign Prostate Hyperplasia and seminal plasma from healthy controls 2-5. In pancreatic cancer, we have described glycosylation changes on human pancreatic ribonuclease (RNase 1) and acute-phase proteins. An increase in core fucosylated structures in the N-glycan chains of RNase 1 6 and an increase of sialyl-Lewis x (SLex) and fucosylation of the acute-phase proteins ceruloplasmin and alpha-1-acid glycoprotein, respectively, were described and were found in advanced pancreatic cancer patients 7-9. These tumor associated glycan changes are currently being investigated in larger cohort of patients as cancer diagnostic or prognostic tools. The expression of the glycosyltransferases responsible for the synthesis of the tumor associated carbohydrate antigens such as SLex has been found deregulated in cancer. In particular, our group has focused on the study of sialyltransferases, which have received much attention recently as they are frequently up-regulated in cancer cells. We have described using in vitro and in vivo models the involvement of the α2,3-sialyltransferases ST3Gal III, and ST3Gal IV in key steps of pancreatic tumor progression processes, and have found that they are highly expressed in most pancreatic adenocarcinoma tissues10-12.
Achuth Padmanabhan
Yale University, USA
Title: Tracking cancer progression through monitoring the dynamic serum phospho-proteome
Biography:
Achuth Padmanabhan completed his PhD from University of Maryland Baltimore County and is currently pursuing his postdoctoral studies at Yale University, School of Medicine. His research has helped identify novel signaling modules in prostate cancer that regulates the levels of the key oncogene MYC and the prostate tumor suppressor NKX3.1. These discoveries have opened up new avenues that can be potentially used to target these proteins in prostate cancer. He has published several papers in reputed journals and has been serving as an editorial board member of repute.
Abstract:
Serum proteome has been shown to reflect pathological changes in tissues. Changes in serum proteomic pattern during carcinogenesis have been exploited successfully to serve as diagnostic and prognostic markers. Protein Kinase A catalytic subunit α has been shown to be secreted into the serum as ecto-kinase and the ecto-PKA level is significantly higher in the serum of prostate cancer patients. Here, I describe the utility of the Reverse In-gel Kinase Assay coupled with LC-MS/MS to profile potential serum substrates of PKA and in following their phosphorylation status during disease progression (1-4). The sensitivity and linearity of this assay will help identify potential biomarkers, which can be used to track the progression of prostate cancer. This strategy can be adapted and extended to track the progression of other cancers and pathological conditions.
Robin Polt
University of Arizona, USA
Title: Glycolipids and Glycopeptides-minimally competent lewis acid catalysis produce surfactants for use ex vivo and in vivo
Biography:
Robin Polt developed methods for the synthesis of amino acids in Prof. Martin O’Donnell’s lab at IUPUI. He was then trained in the art of Organic Synthesis in the laboratories of Professors Gilbert J. Stork at Columbia University in New York and Dieter Seebach at the ETH in Zürich. He has continued the development of useful synthetic methods for amino acids, amino alcohols, glycosides and glycopeptides. Polt has mentored undergraduates, Ph.D. students from the USA, Czech Republic, China, India, Iran, Ireland, Kenya, Korea, Mexico and Sri Lanka, as well as post-doctoral students who have taken positions in academia, industry and government.
Abstract:
Use of minimally competent main group Lewis acids such as InBr3 and Bi(OSO2CF3)3 permit the formation of glycosides in high yield and purity from simple sugar per-acetates at room temperature or above with remarkable α/β-selectivity. Classical reactions rely on very reactive glycosyl donors in conjunction with metal promoters (Hg++, Ag+, etc). Newer methods have used trichloroacetimidates and strong Brønsted or Lewis acids, or thioglycosides in conjunction with oxidative or thiophilic activators. Use of the more stable glycosylperacetates in conjunction with stoichiometric amounts of strong Lewis acids (BF3•Et2O, FeBr3, etc) has been explored with limited success. The use of “minimally competent†Lewis acids has allowed us to perform glycosidation reactions with catalytic amounts of InBr3 or Bi(OTfl)3 well above room temperature and without significant decomposition of the glycoside products. We will discuss the exploitation of these methods to produce glycolipid surfactants from renewable resources, as well as glycopeptide drugs that penetrate the blood-brain barrier (BBB). These “biousian glycopeptides†are not subject to “Lipinski’s rules†that would otherwise eliminate them as CNS drug candidates. Glycosylated amphipathic helices, or “address segments†have been used to target G-protein coupled receptors (GPCRs) in the brain after intravenous administration. The addition of glycosides to endogenous peptide neurotransmitters and peptide hormones imparts favorable pharmacokinetic and pharmacodynamics (PK/PD) properties, and enables penetration of the BBB, which is not constrained by molecular weight.