Day 1 :
AmpTec GmbH, Germany
Keynote: Synthetic mRNAs as optimized tools for stem cell generation and for manipulating cellular phenotypes
Time : 10:00-10:45
Dr. Jean A. Boutin graduated (Thèse d’Etat en Sciences Biologiques) from Nancy University (France) on drug metabolism. He did his postdoctoral training at Johns Hopkins (Baltimore) and at the Karolinska Institutet (Stockholm, Sweden). He was hired as protein chemist in Les Laboratoires SERVIER (LLS) in 1986. During the 30ish last years, Dr. Boutin moved from oncology to peptide research and then molecular & cellular pharmacology. Recently, LLS created a drug discovery platform that Dr. Boutin leads. This structure involves all the aspects of drug discovery, from molecular modeling to ligand/protein biophysical interaction measurements, including protein chemistry, stem cells, structural biology, chemogenetics, HTS, biophysics, Biologics…The main interests of Dr. Boutin are N-myristoyltransferase, melatonin, quinone reductase 2, MCH and autotaxin. In relation with Biologics, our group more particularly explores all the areas related to the possibilities to incorporate exotic amino acids into proteins, especially enzymes.
Synthetic biology is a growing field in which the contribution brought by chemical synthesis of proteins and particularly enzymes is fundamental. Despite this fact, the chemical synthesis of catalytic active proteins remains poorly documented, essentially because it is hard to obtain enough material to use it in biochemical experiments. Chemical synthesis of proteins could permit to have access to the incorporation of unnatural (exotic) amino acids into catalytic active proteins, a feature amenable by recombinant technologies, but that requires delicate manipulation of the bacterial machinery. The developments brought by this approach, include but are not limited to the measure of the influence of unnatural (exotic) amino acids on the 3D structure of enzyme, its activity as well as their recognition of substrate, co-substrate or regulator. The main limitation remains a quantitative problem: How to progress from microgram of proteins produced nowadays with essentially recombinant techniques to tens of milligrams? We chose to present as a model the synthesis and thorough characterization of calstabin, a short protein proline isomerase of 107 amino acids. The protein was synthesized using the native chemical ligation approach. Several tens of milligrams were obtained. Therefore, we were able to refold the polypeptide properly, to characterize its biophysical properties, to measure its catalytic activity and finally to crystallize it in order to obtain its tridimensional structure after X-ray diffraction. Further to it and as a first step of validation of the whole process, we incorporated exotic amino acids in the easiest reachable part of the protein N-terminus. Avenues are now open to obtain further proteins modified with exotic amino acids in a way that is only barely accessible by molecular biology. We hope that these approaches will permit to gain detailed information on the structure-function relationship of proteins as long as they are reachable by complete chemical synthesis (below 300 amino acids).
Seattle Children’s Research Institute, USA
Time : 10:45-11:30
Michael C Jensen was graduated from the University of Pennsylvania; School of Medicine and completed training in Pediatric Hematology and Oncology at the University of Washington, Fred Hutchinson Cancer Research Center. He is the Sinegal Endowed Professor of Pediatric Hematology-Oncology and Bioengineering at the University of Washington, School of Medicine. He serves as the Founding Director of the Ben Towne Center for Childhood Cancer Research at Seattle Children’s Research Institute. He has been a pioneer in the area of cancer immunotherapy focusing on the genetic engineering of immune system T cells. He is an Inventor on a broad portfolio of patents in this area and leads first-in-man clinical trials of chimeric antigen receptor redirected T-cell immunotherapy. He is also a Scientific Founder of Juno Therapeutics, Inc.
Recent conceptual as well as technological advances in the areas of molecular immunology, gene transfer and cell processing have fostered increasingly sophisticated translational applications of adoptive T-cell therapy for oncologic disease employing genetically-modified T-lymphocytes. My laboratory’s work focuses on T-cell genetic modification for re-directing antigen specificity to tumors utilizing recent advances not only in the composition and specificity of receptor antigen recognition domains but also the evolution of multifunctional cytoplasmic signaling domains developed for these chimeric antigen receptors (CARs) that provide dual activation and co-stimulatory signaling. My group is also investigating the context of adoptive transfer with respect to the conditioning of the recipient for enhanced T-cell engraftment and expansion, the grafting of CARs on to central memory T-cells having endogenous TCR specificities for viral epitopes to which the host has robust immunity, and the provision tumor microenvironment survival capabilities. The increasingly broad array of genetic manipulations including not only transgene insertion but targeted gene knock out using engineered targeted nucleases such as TALEN’s and ZFN as well as expression regulatory constructs provides for the creation of synthetic biology of orthogonal immune responses based on gene modified T-cell adoptive transfer. The next decade of advances in this arena will depend on iterative bench-to-bedside back-to-the-bench translational studies capable of sustaining the evolution of these technologies in the context of clinical parameters relevant to the pediatric oncology patient population.