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ASGCT Business Meeting 7:30 AM - 8:00 AM Room: 202 AB
Plenary Session 400 8:00 AM - 8:45 AM Room: Ballroom AB Outstanding Achievement Award Lecture
ChairKatherine P. Ponder, MD
Speakers
David A. Williams, MD 25 Years in Translation: The Hematopoietic Microenvironment and HSC Gene Therapy Initial studies (Williams et al. Nature, 1984) described the use of recombinant retrovirus vectors for transduction of murine engraftable hematopoietic stem cells (HSCs). These studies and those by many other investigators suggested an important role of the hematopoietic microenvironment in HSC function. This talk will summarize the basic and translational studies which sought to understand and exploit the HM in gene therapy, leading to the discovery of the use of fibronectin (Retronectin™) in gene transfer, the discovery of interleukin-11 (Neumega™), the newly defined role of Rho GTPases in HSC engraftment/retention and two newly defined human diseases.
Oral Abstract Session 410 9:15 AM - 11:15 AM Room: 201 C AAV Vector Design & Application
Co-ChairsAravind Asokan, PhD Dirk Grimm, PhD
Oral Abstract Session 411 9:15 AM - 11:15 AM Room: 202 AB Cancer - Immunotherapy: Genetic Modification of T Cells
Co-ChairsBrian C. Beard, PhD Sunil Chada, PhD
Oral Abstract Session 412 9:15 AM - 11:15 AM Room: 203 AB Cancer - Oncolytic Viruses
Co-ChairsGlen Barber, PhD Roberto Cattaneo, PhD
Oral Abstract Session 413 9:15 AM - 11:15 AM Room: 201 AB Hematologic Gene & Cell Therapy
Co-ChairsBernhard Rudolf Gentner, MD Axel Schambach, MD, PhD
Oral Abstract Session 414 9:15 AM - 11:15 AM Room: 204 C Immunologic & Host Responses in Gene & Cell Therapy
Co-ChairsBrad E. Hoffman, PhD Isabelle Riviere, PhD
Oral Abstract Session 415 9:15 AM - 11:15 AM Room: Ballroom AB Stem Cell Biology
Co-ChairsDenise A. Carbonaro Sarracino, PhD Paul Gadue, PhD
Oral Abstract Session 416 9:15 AM - 11:15 AM Room: 204 AB Cell Processing and Vector Manufacture
Co-ChairsMichael Kalos, PhD Johannes C.M. Vanderloo, PhD
Lunch Break (On Own - Not Provided) 11:15 AM - 12:45 PM
Meet the Investigator Lunch Session 420 11:15 AM - 12:45 PM Room: Marriott 303 Immunotherapy for Cancer
Carl H. June, MD
Laura A. Johnson, PhD
Meet the Investigator Lunch Session 421 11:15 AM - 12:45 PM Clinical Vector Core Facility Tour (Children's Hospital of Philadelphia)
J. Fraser Wright, PhD
Meet the Investigator Lunch Session 422 11:15 AM - 12:45 PM Room: Marriott 304 Regulatory Affairs (Approval Process)
Malcolm K. Brenner, MD, PhD
Bambi Grilley, RPh
Plenary Session 430 12:45 PM - 2:45 PM Room: Ballroom AB Outstanding New Investigator Symposium
For more information about ASGCT awards, please go to www.asgct.org.
Nicola Brunetti-Pierri, MD Helper-dependent Adenoviral Vectors for Hepatocyte Gene Therapy Helper-dependent adenoviral (HDAd) vectors are attractive for liver-directed gene therapy because they can provide efficient and long-term transgene expression in the absence of chronic toxicity. However, efficient hepatocyte transduction by HDAd vectors following systemic intravascular injection requires high vector doses which result in dose-dependent activation of innate immunity and acute, potentially lethal toxicity. To address this problem, we have first focused on physical methods to achieve preferential HDAd delivery into the liver. Towards this goal, we have developed in non-human primates a minimally invasive balloon occlusion catheter-based method which resulted in efficient hepatocyte transduction with low vector doses. We have followed a large number of non-human primates injected with HDAd vector and we observed sustained hepatic transgene expression for up to 7 years post-vector administration without chronic toxicity. To further improve the efficiency of HDAd hepatocyte transduction, we next investigated the mechanisms of HDAd vector uptake by non-parenchymal cells of the liver with the goal of evading these important barriers to further improve hepatocyte gene transfer. In most recent work in our laboratory, we have shown that blood-borne HDAd particles interact with scavenger receptor A (SR-A) and scavenger receptor expressed on endothelial cells I (SREC-I), both in Kupffer cells and endothelial cells. We also found that blocking these receptors with monoclonal antibodies increases hepatocyte transduction efficiency. Ultimately, the combination of physical methods with pharmacological agents, such as molecules blocking vector uptake in non-parenchymal cells, may improve the efficiency and the safety of HDAd vectors for clinical applications.
Marco A. Passini, PhD Gene and Antisense Therapies for Neurodegenerative Disorders In collaboration with a consortium of academic and industrial partners, we show that gene- and antisense oligonucleotide-based therapies are highly effective strategies for treating monogenic disorders that affect the CNS, including the neuromuscular disease spinal muscular atrophy (SMA). Although results from proof-of-concept efficacy studies in SMA mice are encouraging, successful translational studies in large animal models are required to address the challenges associated with global spinal cord delivery. Unlike soluble lysosomal enzymes and growth factors, SMN is not a secreted protein and thus motor neurons must be directly targeted by the viral vector or antisense oligonucleotide (ASO) for efficacy to be achieved in SMA. Results in pigs and monkeys show that widespread motor neuron targeting can be achieved with intrathecal delivery of recombinant AAV vectors or ASOs, demonstrating the potential clinical utility of these therapeutics for SMA and other neurodegenerative disorders.
Theresa M. Reineke, PhD Design and Discovery of Glycopolymer Vehicles for the Delivery of Nucleic Acids The Reineke Laboratory has developed a series of polymer-based delivery vehicles that consist of carbohydrate groups copolymerized with cationic moieties that facilitate binding and compaction of nucleic acids into polyplexes. We have examined the delivery efficacy of these vehicles for pDNA, siRNA, and ODN decoy delivery to a variety of tissue types and have examined the intracellular delivery mechanisms of our most promising structures. The synthesis, delivery data, structure-activity relationships in vitro, and preliminary in vivo data will be presented.
Benjamin R. TenOever, PhD Harnessing the Power of Small RNAs in Vector-mediated Therapeutics The discovery that small RNAs could effectively target mRNAs in a sequence-specific manner revolutionized molecular biology and provided a novel approach to therapeutics. A remaining challenge in harnessing the potential of this discovery is the ability to deliver robust levels of functional small RNAs specifically to a tissue of interest. In an effort to address this challenge, we exploit endogenous microRNAs to control the tropism of engineered RNA-based vectors and use the vectors themselves as vehicles to generate specific short interfering RNAs. The biology underlying this technology and vector examples will be discussed.
Scientific Symposium 440 3:15 PM - 5:15 PM Room: 201 C Cancer Theranostics
Co-ChairsBakhos A. Tannous, PhD Margaret E. Black, PhD
Young Jik Kwon, PhD Pinpointed Optical Imaging and Simultaneous Gene Silencing Using Stimuli-Transforming Nanotheragnostics Early, minimally invasive, and accurate diagnosis of cancer is as important as administering efficient and safe therapeutics. In this study, siRNA-carrying nonviral carriers were tethered with gold nanoparticles in order to obtain multiple optical signal changes (diminished light scattering and increased variance of Doppler frequency), in response to a molecular stimulus in cancer, followed by simultaneously enhanced gene silencing. In vitro and preliminary in vivo studies demonstrated the high feasibility of this novel stimuli-transforming nanotheragnostics agent for combined optical diagnosis and gene therapy for cancer.
Bakhos A. Tannous, PhD Imaging and Blood Monitoring in Cancer: Linking Cellular Processes with Therapy The past decades have seen tremondous growth in different molecular imaging technologies for cancer theranostics. These include the development of different reporters which can be tracked with one or more imaging modalities including optical, nuclear and magnetic resonance. During this presentation, we will discuss current trends in reporter systems for pre-clinical cancer theranostics applications. We will focus on multiplex imaging technologies including blood-based reporters which allows simultaneous real-time monitoring of multiple cellular processes. We will then discuss the application of these systems in different gene and cell therapy settings such as tracking of circulating cells, activation of different cellular pathways and monitoring of tumor response to specific therapies.
Vladimir Ponomarev, MD, PhD Cellular Theranostics in Cancer Immunotherapy Adoptive cell transfer therapies rely on ex vivo manipulation and expansion of therapeutic cell populations, which are then transferred back to the patient. Tumor-specific cell infusion into a tumor-bearing host has been used as an experimental approach for studying tumor immunity, as well as an anti-tumor therapy option. The ability to noninvasively monitor the fate and efficacy of adoptively administered cells provides a unique opportunity to assess response early in the course of treatment. Novel imaging technologies allow for spatial and functional assessment of adoptively transferred cells and provide better understanding and evaluation of cellular therapeutics role in treatment outcome.
Mikael J. Pittet, PhD Origins of Tumor-associated Myeloid Cells Cancer progression is affected not only by genetic alterations in cancer cells, but also by diverse populations of seemingly normal host cells that infiltrate the tumor stroma. Tumor-associated macrophages (TAMs) represent one type of host cells that promote adverse outcomes and shorten survival in several cancer types; however, our understanding of the origins and dynamics of these cells remains not fully explored. By using various modalities, including in vivo imaging, we have started to define where, when and how TAM precursors are produced and traffic to the tumor stroma. The findings identify novel contributions of the host response to tumor progression and novel targets for therapeutic intervention.
Scientific Symposium 441 3:15 PM - 5:15 PM Room: 204 C Update on International Clinical Trials and Regulatory Affairs in China, Europe and US
ChairDale Ando, MD
Maria Cristina Galli, PhD Advanced Therapy Medicinal Products in EU: From Clinical Trials to Market Authorization Advanced therapy medicinal products (ATMP) represent a significant emerging field in which new treatment opportunities are offered to patients. The EU regulatory framework for ATMP includes i) Regulation 1394/2007, according to which ATMP are authorized for the EU-wide market by the European Medicine Agency (EMA) in one single (centralized) procedure, and ii) Directive 2001/20/EC according to which clinical trial approval is the responsibility of each EU Member State. According to the EMA centralised procedure, market authorisation application is evaluated by the Committee for Advanced Therapy (CAT), while final opinion to the European Commission is given by CHMP. CAT working parties GTWP and CPWP have issued a number of guidelines describing quality, preclinical and clinical requirements for ATMP to help applicants developing their products. Clinical development takes place at national level, with each EU Member State performing a separate evaluation and authorization procedure. Some EU Member States have issued national guidelines to help applicants in submitting the investigational ATMP dossier, while EMA guidelines addressing requirements for market authorisation also contain guidance for preclinical studies. Procedures and initiatives have been put in place by EMA and national Competent Authorities to facilitate an efficient translation of research discoveries into effective ATMP.
Daniel M. Takefman, PhD International Harmonization of Gene Therapy Regulatory Requirements This presentation will cover some of the international harmonization activities FDA has been involved in. In particular, participation in the International Conference on Harmonization (ICH) will be covered, both regarding harmonization on general and gene therapy specific topics. Additionally, an overview will be given on the ATMP cluster which is a forum for FDA and the European Medicines Agency to have monthly discussions of Cell and Gene Therapies.
Chunming Rao, PhD Regulatory Issues for Gene Therapy Products in China In order to assure the safety and efficiency of gene therapy products,the government has issued a series of Guidelines. In May 1993, the Chinese Ministry of Public Health released An Outline of Quality Controls for Clinical Studies of Human Somatic and Gene Therapy. It was further revised in June 1999 and reissued as Guiding Principles for Human Gene Therapy Clinical Trials by SFDA. In March 2003,the SFDA released Guidelines for Study of Human Gene Therapy and Products Quality Control and Guidelines for Quality Control of Human Recombinant DNA Products. In this presentation the guidelines related to gene therapy products in China will be reviewed, which includes requirements for application of gene therapy clinical study, study protocol format, requirements for the establishment and testing of cell bank and engineered strains, manufacturing process, quality controls, product efficacy and safety tests, foundation of quality control standard, quality standard research of gene therapy products, specifications of gene therapy products in China.
Tae-Gyun Kim, PhD Regulation and Clinical Trials of Gene Therapy Products in Korea Korea Food and Drug Administration (KFDA) established regulations for the ‘Approval of Gene Therapy Product’ in December 2000. Since then, 23 cases of clinical trial protocols have been approved for a wide range of products applicable to cancer, HBV, AIDS, cardiovascular disease, genetic disorder. This presentation is about the KFDA’s regulation on approval of the products after review of safety and efficacy based on risk/benefit assessment and current trends of the product development in Korea.
Scientific Symposium 442 3:15 PM - 5:15 PM Room: 201 AB Gene and Cell Therapy Clinical Trials: Manufacturing Challenges for the Next Horizons
Jointly planned by the Association of Academic Biologics Manufacturers
Co-ChairsJ. Fraser Wright, PhD Isabelle Riviere, PhD
Bruce L. Levine, PhD Challenges in Manufacture of CD19 Redirected T cells from CLL Patients Critical components for generation of cells with non-homologous function include 1) source and isolation of cells, 2) method of activation, 3) method of expansion, 4) characterization and testing. T lymphocytes can be activated, cultured, and endowed with novel functions ex vivo. Ex vivo modified cells can engraft, significantly expand in vivo by >1000 fold, and can survive for a decade. We recently reported the first demonstration of the use of gene transfer therapy to create "serial killer" T cells aimed at cancerous tumors. T cells were modified with a lentivirus to express a chimeric receptor expressing an anti-CD19 antibody to redirect T cells against leukemia cells in three CLL patients. In addition to lentivirus delivery, unique aspects of the ex vivo manipulation included the chimeric receptor using the co-stimulatory domain 4-1BB and the use of T cell expansion technology based on T cell stimulatory antibodies to CD3 and CD28 conjugated to magnetic beads.
Jim Ackland Manufacturing and Control Challenges in the Development of a Gene Therapy Product The development of new biological products is a long and complicated process involving many hurdles between concept and approved product. The early consideration of regulatory guidance and engagement of regulatory agencies is important in avoiding costly reworks or delays due to unanticipated requirements. Issues which need to be considered in transitioning from research depend on the product but generally include the provenance of the source materials (virus, bacteria, genetic material, cell substrate), exposure to adventitious agents, adequate documentation and material controls, establishment of basic consistency in manufacture, adventitious agent testing, qualified/validated release testing, initial product stability, efficacy models and immunological correlates, preclinical safety. Prior to approval for marketing the regulatory requirements increase and all aspects relating to manufacturing and control need to be addressed including GMP compliance, facilities, scale up, process changes, process validation, analytical validation, consistency and stability need to be addressed. Lessons learned from scale up and approval of vaccines will be used as examples.
Adrian P. Gee, PhD Manufacturing for Phase I Clinical Trials Preparation of cellular and gene therapy products presents monetary, scientific, manufacturing and regulatory challenges to academic institutions. The Center for Cell and Gene Therapy (CAGT) at Baylor College of Medicine operates a 22 clean room GMP Facility for manufacture of viral vectors and a variety of cellular therapy products. This presentation will address both how CAGT has tackled these challenges and issues that still require resolution.
Denise Gavin, PhD The Lifecycle Approach to Product Development: Preparation Not Procrastination With the promise of G&CT products at an all time high, it is essential that manufacturers overcome challenges to successfully develop gene and cell therapy products. In addition to establishing that a product is safe and effective, manufacturers must validate control of the manufacturing process by consistently producing a quality product and by meeting CGMPs. It is vital for manufacturers to understand that quality can not be tested into products, but must be built in by design. This requires thorough characterization of product quality attributes and evaluation of the manufacturing process throughout all phases of a product’s lifecycle. Strategies for leveraging the product lifecycle approach to prepare for licensure will be discussed.
Scientific Symposium 443 3:15 PM - 5:15 PM Room: 202 AB New Directions for Non-Viral Delivery of Nucleic Acid Based Drugs
ChairIan MacLachlan, PhD
Vasant Jadhav, PhD Challenges and Opportunities for siRNA Therapeutics RNA interference using siRNAs is one of the most promising approach for treating many diseases. However, to realize the therapeutic potential of siRNA, key issue is the safe and effective intracellular delivery. We are working on number of approaches to tackle this issue. The progress and challenges from this efforts will be presented.
Niren Murthy, PhD New Strategies for Treating and Imaging Inflammation Drug delivery and molecular imaging have tremendous potential for improving human healthcare, however their medical impact has been limited because of a lack of materials. In this presentation I will describe 2 new materials that are either designed to improve the delivery of therapeutics or image disease biomarkers, termed the polythioketal nanoparticles (TKNs) and the metabolic imaging probes. TKNs are nanoparticles formulated from polythioketals, and are designed to target orally delivered therapeutics to intestinal inflammation. TKNs are stable to acid and base, but degrade in the presence of reactive oxygen species (ROS), allowing them to target intestinal inflammation, which has upregulated ROS levels. We demonstrate that orally administered TKNs can deliver TNFa-siRNA, diminish TNFa mRNA in the colon and protect mice from intestinal inflammation. In addition, I will present a new contrast agent for imaging bacteria, termed MIPs, which are designed to target the maltodextrin transport pathway and diagnose bacterial infections at an early stage.
Samir Mitragotri, PhD Technologies to Enhance Delivery of Nucleic Acids into Skin Skin offers an excellent target for nucleic acid-based therapies. However, barrier properties of skin make is very difficult to deliver sufficient quantities of nucleic acids into skin cells. My presentation will provide an overview of challenges and emerging technologies for enhancing uptake of nucleic acids into skin.
Scientific Symposium 444 3:15 PM - 5:15 PM Room: 204 AB Oligonucleotide Therapeutics for Modulation of Splicing
ChairRyszard Kole, PhD
Charles A. Thornton, MD Oligonucleotide Therapeutics for Myotonic Dystrophy The disease process in myotonic dystrophy is a good match for antisense oligonucleotide drugs. In this disease the expansion of a trinucleotide repeat leads to production of a pathogenic RNA. This presentation will focus on different ways in which antisense oligonucleotides can be used to counteract the RNA gain-of-function mechanism. Recent studies suggest that the simplest approach – get rid of the offensive transcript – may be quite effective in animal models.
Mariano Allo, PhD siRNA-Mediated Epigenetic Control of Alternative Splicing Small interfering RNAs (siRNAs) are known to mediate post-transcriptional gene silencing (PTGS) by promoting degradation of target mRNAs. When targeted at promoter regions, siRNAs participate in an alternative pathway known as transcriptional gene silencing (TGS) by promoting histone methylation, heterochromatin formation and inhibition of transcription. We have shown that siRNAs targeting sequences located in the vicinities of a cassette exon are able to regulate its alternative pre-mRNA splicing in human cells. The effect needs both AGO-1 and AGO-2 and is abolished or reduced by factors that favor chromatin relaxation or increase transcriptional elongation. The mechanism involves the presence of facultative heterochromatin epigenetic marks (H3K9me2 and H3K27me3) at the target site and the hetorochromatin associated protein HP1 alpha. The effect is not only triggered by intronic but also by exonic siRNAs. The use of the cancer-related panel for alternative splicing in cells depleted of AGO1 and Dicer shed light on a possible endogenous pathway of this process. Moreover, we found significant differences in the upstream intron size of events affected by AGO1 knockdown relative to non-affected ones.
Daniel A. Norris, PhD Developing Oligonucleotide Therapeutics for the Treatment of Neurodegenerative Diseases
Ryszard Kole, PhD The Use of PMO for Exon Skipping in Duchenne Muscular Dystrophy
Scientific Symposium 445 3:15 PM - 5:15 PM Room: 203 AB Towards Clinical Trials for Genetic and Metabolic Diseases
Co-ChairsValder Arruda, MD, PhD Charles P. Venditti, MD, PhD
Olivier Danos, PhD An Open-label, Single Arm, Monocentric, Phase I/II Clinical Study of Intracerebral Administration of Adeno-associated Viral Vector Serotype 10 Carrying the Human SGSH and SUMF1 cDNAs for the Treatment of Sanfilippo Type A Syndrome Sanfilippo syndrome is a group of mucopolysaccharidoses caused by deficiencies in the catabolism of heparan sulphate and characterized by predominantly neurologic symptoms, progressive mental retardation and death before the age of 15. A Phase I clinical trial was initiated in Sanfillippo type A patients receiving intracerebral injections of SAF-301, an AAV2/rh.10 vector that encodes N-sulfoglycosamine sulphohydrolase (SGSH). Four patients aged 18 months to 6 years are treated with SAF-301 delivered to both sides of the brain through six image-guided tracks in a single neurosurgical session. Patients receive an immunosuppressive treatment and are followed-up during one year. Safety is evaluated on clinical, radiological and biological parameters.
Stephanie Cherqui, PhD HSC Gene Therapy for Cystinosis: New Hope for a Multi-Compartment Lysosomal Storage Disorder Cystinosis is characterized by the accumulation of cystine in all tissues and progressive multi-organ degeneration. We recently showed that syngeneic hematopoietic stem and progenitor cell (HSPC) transplantation from wild-type donors into the mouse model for cystinosis led to significant decreases of cystine in every tissue compartment correlating with the abundant tissue integration of bone marrow-derived cells and resulting in organ preservation. We are now undertaking the first clinical trial of HLA-matched related donor hematopoietic cell transplantation at the University of California, Los Angeles to test the hypothesis that HSPC expressing CTNS gene will improve cystinosis in humans. The long-term objective is to develop the transplantation of autologous hematopoietic cells genetically modified ex vivo using a SIN-lentiviral vector to express a functional CTNS gene in patients. We are also investigating the mechanisms by which transplanted HSPC expressing a functional CTNS gene mediate their long-term therapeutic benefits in the context of a lysosomal transmembrane protein.
Bernhard Rudolf Gentner, MD A microRNA Dimmer Switch for Hematopoietic Stem Cell Gene Therapy Hematopoietic stem cell gene therapy has made significant progress over the last years offering for a variety of genetic disorders the concrete prospect of a less risky and potentially more effective alternative to allogeneic stem cell transplantation. This presentation will outline how microRNA activity can be harnessed to negatively regulate transgene expression in distinct hematopoietic subpopulations and – in combination with appropriate transcriptional control elements – achieve tissue- and lineage restricted expression profiles with unprecedented specificity. We constructed “HSC-off” vectors lacking expression in hematopoietic stem cells as opposed to differentiated cells, thus allowing the delivery of a transgene into HSC without altering their proteome, while benefitting from sustained multi-lineage expression in the progeny. The utility of the “HSC-off” approach will be discussed for 2 genetic diseases: globoid leukodystrophy and chronic granulomatous disease, examples for overt and suspected transgene toxicity upon de novo expression of the corrective cDNAs in HSC, respectively.
Paul Gadue, PhD Generation of Endodermal Stem Cells with the Potential to Form Functional Pancreatic Beta cells as a New Approach for Diabetes
Poster Session III 5:15 PM - 7:15 PM Room: Hall A
