2021 BioMAN Summit

Welcome

10 minutes

2021 BioMAN Summit Introduction

10 minutes

Stacy Springs

Dr. Stacy Springs serves as the Senior Director of Programs at MIT’s Center for Biomedical Innovation and as the Executive Director of Biomanufacturing Initiatives including MIT’s Biomanufacturing Program, (BioMAN), it’s Consortia on Adventitious Agent Contamination in Biomanufacturing, (CAACB), and the BioACCESS initiative.  The objective of BioMAN is to develop knowledge, science, technologies and strategies that […]

Anthony J. Sinskey

Anthony J. Sinskey, ScD, is a Professor of Microbiology at the Massachusetts Institute of Technology. He also holds positions as Co-Director of the Malaysia-MIT Biotechnology Partnership Program and as Faculty Director of the MIT Center for Biomedical Innovation. He conducts interdisciplinary research in metabolic engineering focusing on the fundamental physiology, biochemistry and molecular genetics of […]

2021 BioMAN Summit Framming

10 minutes

Jacqueline Wolfrum

Dr. Wolfrum has been at the MIT Center for Biomedical Innovation (MIT CBI), since 2014. She is Director of the Biomanufacturing Program (BioMAN), a pre-competitive biopharmaceutical industry consortium focused on development of new knowledge, science, technologies, and strategies that advance the manufacture and global delivery of high-quality biologic medicines. She manages sponsored projects and activities […]

Engineering Genetically Enhanced T Cells for Clinical Applications

Since the 1990’s, we have conducted clinical trials of gene modified T cells.  Gene editing has created T cells resistant to HIV infection. Chimeric antigen receptor (CAR) T cells targeting CD19 on B cells leukemias and lymphomas have induced durable complete responses in patients who are relapsed or refractory to all other available treatments.  New designs for genetically modified T cells include switches and potency enhancements that will be required for targeting solid tumors. In one such approach, multiplex gene editing was accompanied by lentiviral transduction of a T Cell Receptor against the cancer antigen NY-ESO-1.  The first use of CRISPR in the US in humans demonstrated that multiplex human genome engineering is safe and feasible.  Translation of these technologies from research bench to clinical application requires knowledge of the critical quality attributes of the engineered cell product and acceptable limits.  Determining dose, potency, and anticipating pharmacokinetics of a living, dividing drug presents unique challenges.  The road forward for wide patient access to engineered cellular therapies depends not only on scientific progress in targeting, gene modification and cellular manipulation methods, but also on meeting automation, engineering, clinical site onboarding, and health policy challenges.

30 minutes

Bruce Levine

Dr. Bruce Levine, Barbara and Edward Netter Professor in Cancer Gene Therapy, is the Founding Director of the Clinical Cell and Vaccine Production Facility (CVPF) in the Department of Pathology and Laboratory Medicine and the Abramson Cancer Center, Perelman School of Medicine, University of Pennsylvania. He received a B.A. (Biology) from Penn and a Ph.D. […]

Advancing the Development of Cell and Gene Therapies

Individualized cell-based therapeutics and gene therapies will become increasingly important to address the variety of molecular defects present in several thousand serious rare diseases.  Some of these therapies may be targeted to as many as a few thousand individuals, but others only may be targeted to one or few individuals. Development of these products poses challenges to the scientific community.  For example, how do we obtain substantial evidence of effectiveness and adequate evidence of safety for products for rare diseases, particularly diseases where there may be as few as one patient with the disease?  How do we ensure manufacturing quality for products intended to treat one or a few patients?  These are some of the questions that must be addressed to facilitate development and access to individualized therapeutics.  Although notable progress has been made since the passage of the 21^st Century Cures Act in the United States, from the regulatory perspective there are still unanswered questions regarding how to best facilitate the development of such individual therapeutics.

30 minutes

Peter Marks

Peter Marks received his graduate degree in cell and molecular biology and his medical degree at New York University and completed Internal Medicine residency and Hematology/Medical Oncology training at Brigham and Women’s Hospital in Boston. He has worked in academic settings teaching and caring for patients and in industry on drug development.  He joined the […]

Rising to the Challenge of Solid Tumors: The Promise of Autologous and Allogeneic TCR T-cell Therapies

The promise of cell therapy for people living with cancer has been realized with several Chimeric Antigen Receptor (CAR) T-cell therapies approved for a range of hematological tumors. However, the challenges presented by solid tumors – including low and heterogenous expression of tumor specific antigen targets and challenging tumor microenvironment – have been associated with limited efficacy, to date, of CAR T-cell therapies for these types of cancer. During this presentation, Dr, Karen Miller, SVP Pipeline Research at Adaptimmune, will talk about the potential of autologous and allogeneic TCR T-cell therapies in clinical trials to treat patients with multiple solid tumor indications. The Company has recently shared data with its most advanced therapy, afamitresgene autoleucel, showing promising efficacy and durability data in synovial sarcoma, which will be used to file a licensing application, and initial data in five solid tumors with a next-generation therapy that will be investigated in Phase 2 trials for people with esophageal, esophagogastric junction and ovarian cancers.

30 minutes

Karen Miller

Karen is an immunologist by training and has 30 years’ experience in drug discovery in a range of therapeutic areas including oncology, inflammatory disease and neurology. She has broad experience in small molecule and antibody drug discovery and in cell and gene therapy.  At AdaptImmune, Karen is the SVP of Pipeline Research, leading the teams […]

Base Editing for the Treatment of Beta-hemoglobinopathies

Base editing is a next-generation, CRISPR-based gene editing technology that makes precise conversion of one nucleobase to another by virtue of a human deaminase enzyme, directly on the gene, without making double-stranded breaks. Using this technology, we are developing two therapeutic products, BEAM-101 and BEAM-102, for the treatment of Sickle Cell Disease (SCD). In the case of BEAM-101, we are using the precision of base editing to reproduce point mutations in the promoter of the human gamma 1 and gamma 2 genes that are normally seen in individuals who have a condition known as Hereditary Persistence of Fetal Hemoglobin (HPFH). These individuals retain high levels of expression of the fetal hemoglobin (HbF) in adulthood and, because of this, they are protected from mutations in the beta globin gene that would otherwise cause sickle cell disease. BEAM-101 has been shown to achieve the highest level of HbF reported pre-clinically so far (65%) and is progressing to clinical studies. BEAM-102 directly corrects the single amino acid Valine that causes the cells to sickle under low oxygen conditions. For BEAM-102, we have engineered a novel base editor, where the active site of the deaminase has been inlaid directly within the CAS protein. Using this editor we are able to convert the Valine at position 6 into an Alanine, thus converting the sickling form of hemoglobin (HbS) to a normal variant known as HbG Makassar, which occurs in ~0.1% of the human population, with greater than 80% efficiency.

30 minutes

Giuseppe Ciaramella

Giuseppe (Pino) Ciaramella, PhD is currently serving as the President and Chief Scientific Officer of Beam Therapeutics. Prior to joining Beam he set-up and led the Infectious Disease unit of Moderna Therapeutics. Dr. Ciaramella has more than 25 years of drug discovery experience, contributing to several clinical candidates and launched products, in a variety of […]

Scalable Production of Recombinant Adeno-Associated Virus Vectors

Despite the rapid growth of recombinant adeno-associated virus (rAAV) gene therapy research, access to high-quality rAAV remains the major limitation to the clinical gene therapy pipeline. Due to the cytoxicity of the Ad helper gene products as well as cytotoxic or cytostatic effects of the AAV Rep proteins, rAAV production is a transient, terminal process. Thus, introducing the essential genetic information into the producer cells remains the major challenge for large-scale rAAV production. The recombinant baculovirus expression vectors (BEVs) and Spodoptera frugiperda (Sf9) cells provides an alternative to transient transfection of human embryonic kidney (HEK) 293 cells. The BEVs replicate and are amplified in the same cell that produces rAAV and the subsequent cell-to-cell spread of the BEVs results in essentially quantitative infection of the culture in any volume bioreactor.   In the two BEV system, the AAV vector genome is on one BEV and the second BEV  provides the Rep and VP ORFs. In the presence of p5 Rep proteins, the AAV vector genome replicates autonomously from baculovirus. Expression of the Rep and capsid proteins are no longer regulated by the native AAV promoters, therefore, optimal AAV gene expression of the Rep and structural proteins involves engineering the coding sequence to achieve stoichiometric expression of Rep 78, Rep 52, VP1, VP2, and VP3. A systematic approach has been developed to increase the likelihood that the capsid composition is near optimal.

30 minutes

Robert Kotin

Robert Kotin has been involved in adeno-associated virus research since 1986 as a post-doctoral fellow at Cornell University Medical College. Most of Kotin’s career was spent as the head of the Laboratory of Molecular Virology and Gene Therapy in the intramural research program in the National Heart, Lung, and Blood Institute of the NIH. During […]

Directed Evolution of New Adeno-Associated Viral Vectors for Clinical Gene Therapy

Gene therapy – the delivery of genetic material to the cells of a patient for therapeutic benefit – has been increasingly successful in human clinical trials over the past decade, and there are numerous FDA-approved gene therapies.  The most successful gene delivery vehicles, or vectors, are based on adeno-associated viruses (AAV); however, vectors based on natural versions of AAV face a number of delivery barriers that limit their efficacy and will thus preclude the extension of these successes to the majority of human diseases.  These delivery limitations arise since the parent viruses upon which these vectors are based were not evolved by nature for our convenience to use as human medicines.  Unfortunately, due to the highly complex mechanisms of virus-host interactions, there is currently insufficient mechanistic knowledge to enable rational design to be sufficiently successful in creating new vectors.  As an alternative, however, we developed the concept of using directed evolution to engineer highly optimized variants of AAV for a broad range of cell and tissue targets.  Directed evolution involves the iterative genetic diversification of a biomolecule to create a gene pool and functional selection to isolate variants with optimal properties.  Using this approach, we have engineered AAV variants with greatly improved delivery efficiency to multiple organs including the retina; lung, and muscle; targeted delivery to specific cell types; and the capacity to evade immune responses.  Our novel AAV variants are currently used in 5 human clinical trials involving delivery to the retina and heart, and two additional clinical trials will be initiated this year. In parallel, the advent of genome editing technologies such as the CRISPR/Cas9 system raises the possibility of using gene delivery not only for gene replacement but for repair or knockout of endogenous genes.  We have thus been combining engineered AAVs with CRISPR/Cas9 for a range of applications.  The integration of these new technologies – AAV delivery or genome editing machinery – can enable a broad range of basic and therapeutic applications.

30 minutes

David Schaffer

David Schaffer is the Hubbard Howe Professor of Chemical and Biomolecular Engineering, Bioengineering, and Molecular and Cell Biology at the University of California, Berkeley, where he also serves as the Director of the California Institute for Quantitative Biosciences (QB3-Berkeley). He received a B.S. from Stanford University in 1993 and a Ph.D. from MIT in 1998, […]

Rational Design of rAAV Production via Mechanistic Modeling

Recombinant adeno-associated virus (rAAV) is one of the most commonly used platforms for in vivo gene therapy treatments. The reduced toxicity, robust and long-term transgene expression, and ability to transduce both dividing and non-dividing cells as well as target a wide range of tissues have made rAAV the most widely used viral vector. However, the standard method of producing rAAV via transient transfection of mammalian cells, specifically human embryonic kidney 293 (HEK293) cells, typically has low yield and generates a high portion of empty particles, laying extra burden on downstream processing. To elucidate the mechanisms of rAAV synthesis in HEK293 suspension-adapted cells, we have developed a mechanistic model based on the published understanding of the underlying biology and existing data. Quantitative analysis suggests the misaligned dynamics of capsid and viral DNA production result in the high ratio of empty particles. Through a model-based strategy, we explored a novel transfection method using low-dose multiple transfections in HEK293 cell culture that successfully increased the ratio of full to empty capsids in the viral harvest without compromising the viral titer. Molecular analysis through a next-generation rAAV production model attributed the improvements to changes in the kinetics of viral protein expression and DNA replication. Here, we demonstrate that the use of multiple transfection times is a practical method for increasing the genome titer and improving the percentage of full capsids for rAAV production. Our results also demonstrated the capability to manipulate product composition from an operational standpoint.

30 minutes

Tam N. T. Nguyen

Tam Nguyen is a Ph.D. student in Chemical Engineering at MIT, where she does research in biopharmaceutical manufacturing systems. Her main research is in mechanistic modeling for AAV-based transfection and its use in improving understanding, optimization, and control of AAV capsid production. Tam received a Bachelor of Science in Chemical Engineering from the University of […]

Richard Braatz

Dr. Richard D. Braatz is the Edwin R. Gilliland Professor of Chemical Engineering at MIT, where he conducts research into advanced biopharmaceutical manufacturing systems. In this role, he leads process data analytics, mechanistic modeling, and control systems for many projects, including on monoclonal antibody, viral vaccine, and gene therapy manufacturing within the Center of Biomedical […]

Overcoming Low Throughout Testing Issues of AAV Using SEC-MALS

Adeno associated virus (AAV) capsids are a leading modality for in vivo gene delivery. Complete and precise characterization of capsid particles, including capsid and vector genome concentration, is necessary to safely and efficaciously dose patients. Size exclusion chromatography (SEC) coupled to multiangle light scattering (MALS) offers a straightforward approach to comprehensively characterize AAV capsids, including, but not limited to aggregation profile, size-distribution, capsid content, capsid molar mass, encapsidated DNA molar mass, and total capsid and vector genome titer. Currently, multiple techniques are required to generate this information, with varying accuracy and precision. SEC-MALS can leverage intrinsic physical properties of the capsids and encapsidated DNA to quantify multiple AAV attributes in one 20-min run with minimal sample manipulation, high accuracy, and high precision. As such, SEC-MALS is a powerful tool for product development and process analytics in future gene therapy programs.

30 minutes

Wade Wang

Wade Wang is a scientist at BioMarin Pharmaceutical. He works in purification process development and specializes in the biophysical characterization of adeno-associated virus. He earned his PhD at MIT under the direction of Prof. Paula Hammond.

Biophysical Approaches for Process Analytical Technology

Previously we developed a method based on the suspended nanochannel resonator (SNR) that is capable of weighing nanometer sized particles with high precision. Here I will describe how we’re using the SNR to quantify the empty:full capsid ratio for a purified Adeno-Associated Virus (AAV) samples in a way that provides a direct readout without the need to calibrate based on serotype.  This approach achieves a resolution near 10 zeptograms (corresponding to 1% of the genome holding capacity of the AAV capsid) in a 10-minute sampling window using only a few μL of sample. I will also present recent progress towards using suspended microchannel resonators to provide nondestructive measurements of single-cell biophysical properties (e.g. mass, density and stiffness) with the goal of predicting transfection and vector production yield.

30 minutes

Scott Manalis

Scott Manalis is the David H. Koch (1962) Professor of Engineering and faculty member in the departments of biological and mechanical engineering at MIT. He is the associate department head of the biological department and a member of the Koch Institute for Integrative Cancer Research and MIT’s Center for Precision Cancer Research. He has been […]

Industrialization of the GMP Manufacture of Exosome Therapeutics and Opportunities for Further Multifold Process Productivity Increase

Exosome therapeutics are rapidly evolving as a promising new modality in various clinical areas, such as oncology, immuno-oncology, neurology and metabolic diseases, among others.  As some of these indications involve large patient populations, the success of exosome therapeutics depends on our ability to manufacture allogenic exosomes at a large scale, with high purity and quality, acceptable CoG, and under GMP conditions.  Due to the complexity and natural heterogeneity of exosomes, the development of such an advanced production technology is not a trivial task.  This presentation will discuss our ability to develop GMP processes to manufacture precisely engineered engEx exosomes from Codiak’s pipeline. With two lead programs in clinical trials in cancer patients, and an IND approved for a third program in immuno-oncology this year, we have significantly scaled our GMP production capacity.  Our large-scale process uses either 2,000L fed-batch or, more recently, 500L high-density perfusion bioreactors culturing HEK cells at high density in chemically defined media.  The bioreactor harvest is processed through a sequence of purification steps yielding exosomes of high potency and purity, and with consistent quality attributes.  Opportunities for a further multifold increase of process productivity and reduction of CoG through implementation of integrated continuous biomanufacturing technology will be outlined.  This presentation will also review the applicability of the above technology to the manufacture of other ATMPs.

45 minutes

Konstantin Konstantinov

During his 30-year career, Dr. Konstantinov has been at the forefront of development and implementation of advanced biomanufacturing technologies. He has been involved in the commercialization of different biotherapeutic modalities, including monoclonal antibodies, blood factors, enzymes, gene therapy constructs, and exosomes produced by mammalian cells. He has published 60 peer-reviewed papers, and has more than […]

Closing Remarks

10 minutes

Stacy Springs

Dr. Stacy Springs serves as the Senior Director of Programs at MIT’s Center for Biomedical Innovation and as the Executive Director of Biomanufacturing Initiatives including MIT’s Biomanufacturing Program, (BioMAN), it’s Consortia on Adventitious Agent Contamination in Biomanufacturing, (CAACB), and the BioACCESS initiative.  The objective of BioMAN is to develop knowledge, science, technologies and strategies that […]