Day 2 :
Keynote Forum
Cedric Wu
GenScript USA Inc., USA
Keynote: Rapid assembly of multi-gene pathways and combinatorial libraries with semiconductor technologies
Time : 10:00-10:40
Biography:
Cedric Wu obtained his PhD degree in Cellular and Molecular Biology from University of Wisconsin - Madison. He received post-doc training in nucleic acid, surface and analytical chemistry. Dr. Wu possesses more than twenty years of combined experience working in academia, government and industry. He invented and published more than 20 articles and patents, as well as numerous trade secrets, including nucleic acid microarray fabrication technologies and gene assembling methods. He, currently, is leading the R&D department in GenScript focusing on novel DNA synthesis, long and pathway DNA assembly, codon optimization, DNA microarray fabrication, oligo pool synthesis and automation process development
Abstract:
Engineering synthetic biological systems requires the design, build and testing of multi-gene networks. To optimize a synthetic system with the best combination of DNA parts, synthetic biologists will need to screen in a combinatorial design space. But such combinatorial engineering approaches are hindered by the difficulty of simultaneously assembling numerous large, multi-gene constructs from a collection of DNA parts.
Here, we introduce our newly developed combinatorial library synthesis platform, which enables in-parallel construction of thousands of multiplexed gene variants in an efficient and economical way. This platform utilizes our patented semiconductor-based oligonucleotide microarray fabrication technology, high-throughput GenParts® DNA Fragments, and proprietary isothermal GenBuilderTM assembling reagents. Libraries generated using this method are highly customizable and capable of generating over 1x108 variants. As a result, our platform enables faster, easier, and more cost-effective engineering of metabolic pathways and microbial strains in comparison to alternative approaches.
- Molecular Biology | Biomarkers | Next Generation Sequencing | Plant Genomics and Molecular Plant Sciences
Location: Sylt 1-2
Chair
Patrizia Zavattari
Cagliari University, Italy
Session Introduction
Rufina Soomro
Liaquat National Hospital and Medical College, Pakistan
Title: Prevalence of breast cancer intrinsic subtypes and its association with clinico pathological feature
Biography:
Rufina Soomro completed her diploma in medical education (AKU), FACS(USA). She is working as Professor of Surgery at Liquate National Hospital and Medical College Karachi and She is as a chairperson of Postgraduate medical education committee of the same institute. She did Breast surgery training from Dublin, Ireland, and currently running one of the busiest breast services in the country. Has keen interest in research and did a research course from Harvard University. She is serving College and Physicians of Pakistan as an Examiner in G. Surgery, ATLS course Director / Instructor and Secretary of National Surgery faculty.
Abstract:
Introduction: Breast cancer is the commonest cancer in women worldwide and represents a highly heterogeneous group of tumours particularly in terms of molecular features, prognosis and response to therapy. Breast cancer molecular classification can predict the prognosis of breast cancer in terms of recurrence and help and guide us regarding the treatment decision about systemic therapy. Breast carcinomas may be stratified into subtypes similar to those defined by Gene expression profiling using a panel of immune-histochemical (IHC) markers. Routine IHC evaluations of breast cancers may, therefore, provide a reasonable alternative to costly genetic assays especially in under-resourced healthcare systems. The purpose of this study is to investigate the prevalence of molecular subtypes and correlate it to clinic-pathological features.
Methods: From 2005 to 2017 total of 4847 Breast cancer patients, in whom complete information was available to classify them into luminal subtypes were retrieved and classified into intrinsic subtypes and patients information in each type was collected about age, tumour size, stage, grade and nodal status.
Results: In luminal classification, a highly significant difference was found in mean age (p<0.001) tumour size (p<0.001), grade, metastasis and Ki67. The statistical significance of Her 2 positive and triple negative was found with stage, grade, metastasis and Ki67.
Conclusions: IHC assignment into Luminal subtypes is clinically informative in our patients and routinely using this in our practice could identify patients that may need a more aggressive treatment to reduce the likelihood of recurrences.
Waraporn Kraitavin
The University of Tokyo, Japan
Title: Thermal adaptation of yamame (Oncorhynchus masou) in normal condition after undergoing heat stress
Biography:
Waraporn Kraitavin is a Ph.D. student in Aquatic Molecular Biology and Biotechnology at the University of Tokyo, Japan. She worked as a Nutritionist at Cargill Meats (Thailand) Limited for 1 and a half years and she has completed her Marter’s degree and Bachelor’s degree from Kasetsart University, Thailand
Abstract:
Understanding the mechanism of temperature adaptation is crucial for cold-freshwater fish in order to cope with the recent global warming, especially yamame (Oncorhynchus masou), which is an important aquaculture species belonging to the family Salmonidae. The aim of this study is to understand the adaptive response of high-temperature tolerant yamame in normal condition after heat stress. For that, a group of yamame was developed through selective breeding to have high temperature tolerance. Next, we performed a higher-temperature-tolerant test and divided into HT (for the high-temperature-tolerant) group and NT (for the non-high-temperature-tolerant) group. A week later, RNAs were extracted from the gill tissues and analyzed by examining the mRNA expression profiles using Illumina HiSeq 4000 Sequencing System. A total of 2,893 differentially expressed genes (DEGs) from the gill were identified by comparing the HT and the NT groups, then functional analysis were performed to identify associated gene ontology (GO) terms and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway. Several differential biological pathways were detected and we found that the HT showed higher associated gene expression in ECM-receptor interaction, in cell adhesion molecules (CAMs), in cell junction and in adhesion pathway comparing to the gill tissue in NT. Those genes are related to the reparation of the damaged tissues and to the generation of the cytoskeleton of individuals. On this basic, we concluded that the HT may adapt quicker than the NT in normal condition after undergoing the heat stress. These findings can be used to develop high-temperature-tolerant yamame and other Salmonidae.
Chen Jenn-Tzong
Institute of Nuclear Energy Research, Taiwan
Title: Transgenic Mice brain imaging studies of Alzheimer’s disease
Biography:
Jenn-Tzong Chen is a PhD student of Institute of Polymer Science and Engineering, and Chemical Engineering of National Taiwan University. He is also an associate researcher of Isotope Application Department of Institute of Nuclear Energy Research. After graduated from the Chemical Engineering Department of Ming-Chi Institute of Technology, he studied Chemical Engineering in National Taiwan Institute Technology for one year. Then he graduated from Chemical Engineering Department of National Taiwan University with master degree. He has been trained in Forschungszentrum Jülich, Germany and Washington University in St. Louis, US and so on. At present he works for cyclotron produced radioisotope process development and application. He has published some papers and owned some patterns.
Abstract:
Lipophilicity is one of the major brain radiopharmaceutical design criteria. Alzheimer disease PET imaging agents based on lipophilicity modification are [18F]RO6958948 [1] and [18F]Florbetapir, design by replacing with a Nitrogen element either in the aromatic ring of [18F]Flortaucipir or [18F]Florbetaben. The structure of [18F]FEONM (Figure 1) is designed to provide higher lipophilicity than [18F]FDDNP. Structure modification on a certain bioactive molecule in order to increase its lipophilicity will be also possibly increasing the percentage of penetrating blood brain barrier. Increasing the blood brain barrier crossing ratio, the specificity of this active biomolecule targeting effect might be decreased. Therefore, we design an ethyl oxide modified naphthol based Alzheimer disease positron emission tomography imaging agent [18F]FEONM, in order to compare the uptake effect of Tau tangle and Beta amyloid.
PET radiopharmaceiticals for brain imaging are based on very short half-life radionuclides, most of them will be decayed in one day. One of the longest half-life organic radionuclides is fluorine-18, therefore critical step to producing PET radiopharmaceuticals online is radiofluorination reaction. The highest radiofluorination reaction yield can be made from carboxy glass reactor [2]. In carboxy glass reactor, the function of gap area (FG) [3] curve of radiofluorination yield can be approached with Gauss distribution, Gauss or Welch apodization function. After determine the radiofluorination rate constant, the length of microfluidic plug flow reactor can be designed with an analytical form based on Welch apodization function [4].
Brain hippocampus imaging relative specific binding ratio of [18F]FEONM on a Tau tangle P301S/PS19 transgenic mouse model is two time higher than cerebellum [3], Beta amyloid Tg2576 transgenic mouse model is less than two. On a triple transgenic 3xTg mouse model with both Tau tangle and Beta amyloid formed, the uptake ratio of hippocampus is fifty percent higher than cerebellum. Therefore, [18F]FEONM is a new Alzheimer PET imaging agent. Besides, other than transgenic mouse model, streptozotocin induced Tau tangle mouse model [5] also shows higher brain hippocampus [18F]FEONM uptake than control mouse.
From the transgenic mouse model imaging study, we found [18F]FEONM will uptake on both Tau tangle and Beta amyloid transgenic mouse. In comparison to [18F]FDDNP, it shows no Beta amyloid transgenic mice uptake in brain hippocampus [6]. This result represents part of the specific binding of Tau tangle transgenic mouse of [18F]FDDNP has shift to Beta amyloid. Therefore, Tau tangle and Beta amyloid uptake status can be done by [18F]FEONM in the same time for diagnosis Alzheimer disease. Radiation exposure will be half dosage compared to taking both imaging. These findings based on a new design conclude that a new PET radiopharmaceutical design has the same concept like a new radiofluorination microfluidic reactor design. Either a new chemical structure or a new mathematical model contributes an achievement.
Keywords: Alzheimer’s disease, Welch apodization gap area function, [18F]FEONM
Reference
1. Honer M, Gobbi L, Knust H, Kuwabara H, Muri D, Koerner M, Valentine H, Dannals RF, Wong DF, Borroni E (2018) Preclinical Evaluation of 18F-RO6958948, 11C-RO6931643, and 11C-RO6924963 as Novel PET Radiotracers for Imaging Tau Aggregates in Alzheimer Disease. J Nucl Med 59(4):675-681.
2. Hamaker K, Blessing G, Nebeling B (1990) Computer aided synthesis (CAS) of no-carrier-added 2-[18F]fluoro-2-deoxy-D-glucose: an efficient automated system for the aminopolyether-supported nucleophilic fluorination. Appl Radiat Isot 41:49-55.
3. Hsu JP, Chen JT, Lin SH, Chu KY, Tu YH, Chang KW, Huang LY, Huang YC, Farn SS, Hsu SH, Lin JJ, Lin WJ (2016) Radiofluorination process development and tau protein imaging studies of [F-18]FEONM. J Taiwan Inst Chem Eng 68:119-129.
4. Hsu JP, Chen JT, Lin JJ, Li MH, Chang KW, Chen DC, Duh TS, Lo PL, Farn SS, Luo TY, Lin WJ, Shiue CY (2019) Design of PET radiopharmaceuticals for brain imaging. Biomed J Sci & Tech Res 14(2):1-3. Doi: 10.26717.bjstr.2019.14.002527
5. Pradip KK (2015) Streptozotocin induced Alzheimer's disease like changes and the underlying neural degeneration and regeneration mechanism. Neural Regen Res 10(7):1050-1052.
6. Kuntner C, Kesner AL, Bauer M, Kremslehner R, Wanek T, Mandler M, Karch R, Stanek J, Wolf T, Müller M, Langer O (2009) Limitations of small animal PET imaging with [18F]FDDNP and FDG for quantitative studies in a transgenic mouse model of Alzheimer’s disease. Mol Imaging Biol 11:236-240.