BioMAN Summit - November 18-19, 2015

November 18-19, 2015

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Poster Competition Winners | Poster Competition Submissions | Poster Abstracts

The following abstracts have been accepted to compete in the 2015 BioMAN Summit's Poster Competition:

1.	B-Cell Receptor-Based Genetic Sensors for Extracellular Biomarker Detection
	Lyla Atta^1, Kathryn Brink^1, Christian Richardson^1*, Brian Teague1 and Ron Weiss¹
	¹Synthetic Biology Center, Massachusetts Institute of Technology, Cambridge, Massachusetts, USA. *These authors contributed equally to this work.
	Engineering cells to convert extracellular cues into a desired intracellular response is a key component of creating successful cellular therapies. Current approaches rely on repurposing naturally-occurring receptors that specifically detect a molecule of interest. However, these methods are limited by the existence of a suitable receptor and require re-engineering of each receptor depending on its structure and mechanism of action. We have engineered a modular solution to the extracellular detection problem by creating a versatile receptor platform based on the architecture of the B-cell receptor (BCR). In our system, binding of an extracellular antigen to the B-cell receptor’s antibody domain recruits an intracellular signaling protein. Binding of this protein to the receptor releases a transcriptional activator, which then induces an engineered therapeutic response. By exchanging the antibody variable region, this platform can be easily modified to detect any disease-relevant antigen with a corresponding sequenced antibody. This modularity gives our system a wide range of applicability relative to existing methods. In addition, its ability to dynamically respond to extracellular biomarkers makes it specifically relevant to dysregulation-related diseases, such as Alzheimer’s Disease and diabetes.
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2.	Consortium on Adventitious Agent Contaminations in Biomanufacturing (CAACB): A resource for the manufacturers of cell and gene therapy products
	Paul W. Barone^1, James C. Leung^1, Stacy Springs^1, and Michael E. Wiebe^1
	¹Center for Biomedical Innovation, Massachusetts Institute of Technology, Cambridge, MA, 02139, USA *Equal contribution first authors
	CAACB, a member-funded consortium at the Center of Biomedical Innovation (CBI) of MIT, was founded in 2011. We have 25 companies as members. They are manufacturers of biotherapeutics - recombinant proteins and vaccines, and technology and service providers to the manufacturers. Some of these members are involved in the development of cell and gene therapies. The consortium goal is to identify and to promote best practices for risk mitigation of adventitious agent contaminations in biomanufacturing with the final aim to safe guard patient safety. The approach of the consortium is to create and coordinate a neutral forum for the sharing of pre-competitive experience, knowledge and practices in the industry, to facilitate discussions and debates on important issues, and to promote networking for community building for concerned stakeholders. Important lessons on strategic and tactical considerations of risk mitigation plans and their implementations have been accumulating through consortium workshops and member-defined projects. We highlight here selected topics and issues tackled by CAACB to illustrate their value to the biotherapeutic manufacturers and their commonality and applicability to cell and gene therapy products.
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3.	Multiplexing Metabolomic-based Disease Diagnosis by Surface Enhanced Raman Spectroscopy (SERS) platform
	Ying Chen¹, W. Ranjith Premasiri¹ and Lawrence D. Ziegler¹
	¹The Photonic Center and Department of Chemistry, Boston University, Boston, MA
	SERS-based platform has the potential to be successful point-of-care diagnostic tool addressing various human health concerns because it’s information-rich, multiplexing, rapid and ease-to-use. As a signal enhancement method, SERS arises from the well-known Raman cross-section enhancement effect on molecules close to (<5 nm) the surface of nanostructured metal substrates. Since Raman vibration features are uniquely dependent to the molecular structure, SERS platform is a powerful method for identifying biomarkers at near cell membranes and extracellular regions during pathological progression. These attributes allow SERS platform to be a rapid, cell-growth free diagnostic option in urinary tract infections (UTI), sexually transmitted diseases (STD) chlamydia and gonorrhoea, and cancer identification. With both species and strain specificity we will demonstrate the ability of SERS platform to distinguish different bacteria strains, as well as human cancer from normal cells. Combining with multivariate data analysis techniques, expendable reference library and portable instrument, decision can be made with our SERS platform in the clinic within 1-2 hours. Moreover, since we had discovered that the molecular contributors of our SERS vibrational features to be metabolite molecules secreted by the cells in response to environmental change, our SES platform can be a powerful bioanalytical method studying metabolomics in bacteria and human cells more generally.
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4.	Does Adult Gene Therapy in Shank3 Mutant Model Rescue Autistic-Like Phenotypes?
	Yuan Mei^1, Patricia Monteiro^1,2, Yang Zhou¹, Jin-Ah Kim¹, Xian Gao^1,4, Zhanyan Fu^1,3, and Guoping Feng^1,3
	¹McGovern Institute for Brain Research, Department of Brain and Cognitive Sciences, Massachusetts Institute of Technology, Cambridge, MA, USA ²PhD Programme in Experimental Biology and Biomedicine (PDBEB), Center for Neuroscience and Cell Biology, University of Coimbra, Coimbra, Portugal ³Stanley Center for Psychiatric Research, Broad Institute of MIT and Harvard, Cambridge, MA, USA ⁴Key Laboratory of Brain Functional Genomics (Ministry of Education & Science and Technology Commission of Shanghai Municipality), Institute of Cognitive Neuroscience, School of Psychology and Cognitive Science, East China Normal University, Shanghai 200062, China *These authors contributed equally to this work.
	Because autism is a neurodevelopmental disorder and patients typically display symptoms before the age of three, one of the key questions in autism research is whether the pathology is reversible in adults. Here we investigate the developmental requirement of Shank3, one of the most prominent monogenic autism genes. SHANK3 is a postsynaptic scaffold protein that regulates synaptic development, function and plasticity by orchestrating the assembly of postsynaptic density (PSD) macromolecular signaling complex. Disruptions of the Shank3 gene in mouse models have resulted in synaptic defects and autistic-like behaviors including anxiety, social interaction deficits, and repetitive/stereotyped behavior. Here we have generated a novel Shank3 conditional knock-in mouse model and used it to demonstrate that re-expression of the Shank3 gene in adult leads to improvements in synaptic protein composition, spine density and neural function in the striatum. We also provide behavioral evidence that certain behavioral abnormalities including social interaction deficit and repetitive grooming behavior can be rescued, while anxiety and motor coordination deficit cannot be recovered in adulthood. Together, these results elucidate the profound impact of post-developmental activation of Shank3 expression on neural function and demonstrate certain degree of continued plasticity in the adult diseased brain.
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5.	Using the Shake Flask for the Expansion of Adipose-derived Mesenchymal Stem Cells
	Paul Schwein^1,2, Kimberly Ramos¹, Rachel Yao¹, Shriram K. Chandramohan¹, and Jean-Franois P. Hamel¹
	¹MIT Department of Chemical Engineering ²MIT Department of Biology
	Mesenchymal stem cells have a number of applications in medicine and biological research; however, the extent to which they can be employed for cell therapy is limited by their large-scale production. Here, we demonstrate a method for expanding adipose-derived mesenchymal stem cells (Ad-MSCs) in the shake flask and producing a viable inoculum for scale-up in the bioreactor. Experiments were carried out with either Cytodex™ 1 or Solohill® microcarriers, using Ad-MSCs at passage less than 7. In a comparison experiment, equal concentrations of Ad-MSCs at passage 7 were seeded onto Cytodex™ 1 microcarriers in either a 250-mL shake flask or 125-mL spinner flask, each with 75-mL cell culture medium. The shake flask produced higher cell counts than the spinner flask (3.0x104 cells/mL vs. <5x103 cells/mL), and ammonium concentration per cell in the shake flask was one order of magnitude lower than that in the spinner flask. Ad-MSCs grown in the shake flask are also shown to preserve their pluripotency, able to differentiate into osteocytes or adipocytes.
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6.	Elastic Net with Monte Carlo Sampling for Data-based Modeling in Biopharmaceutical Manufacturing Systems
	Kristen Severson¹, Jeremy G. VanAntwerp^1,2, Venkatesh Natarajan³, Chris Antoniou³, Jörg Thömmes³, and Richard D. Braatz¹
	¹Department of Chemical Engineering, Massachusetts Institute of Technology, 77 Massachusetts Avenue, Cambridge, MA 02139 ²Engineering Department, Calvin College, 3201 Burton SE, Grand Rapids, MI 49546 ³Biogen, 14 Cambridge Center, Cambridge, MA 02142
	Biopharmaceutical manufacturing involves multiple process steps that can be challenging to model. Oftentimes, operating conditions are studied in bench-scale experiments and then fixed to specific values during full-scale operations. This procedure limits the opportunity to tune process variables to correct for the effects of disturbances. Generating process models has the potential to increase the flexibility and controllability of the biomanufacturing processes. This work proposes a statistical modeling methodology to predict the outputs of biopharmaceutical operations. This methodology addresses two important challenging characteristics typical of data collected in the biopharmaceutical industry: limited data availability and data heterogeneity. Motivated by the final aim of control, regularization methods, specifically the elastic net, are combined with sampling techniques similar to the bootstrap to develop mathematical models that use only a small number of input variables. This methodology is evaluated on an antibody manufacturing dataset.
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7.	Development of a customizable microwell array for controlled study of heterogeneous cell populations
	Marianna Sofman^1, Natasha Arora^1, Elijah Karvelis², Rachel R. Katz¹, Paula T. Hammond³ and Linda G. Griffith¹
	¹Bioengineering Department, Massachusetts Institute of Technology ²Department of Chemical and Biomolecular Engineering, University of Illinois Urbana-Champaign ³Chemical Engineering Department, Massachusetts Institute of Technology *Co-presenters
	In developing cell and gene therapies, it is important to recognize and understand the heterogeneity of cellular populations. To date, photolithography and soft lithography methods for fabricating micromolding tools have allowed for the development of micropatterned cell culture substrates. These methods facilitate the isolation of small cell populations which provide a high-throughput imaging and testing platform for cell and gene therapies. However, current micropatterning techniques using PDMS molds have substantial drawbacks including delamination of the cured microwell, poor sealing with the culture substrate, and a resolution limitation due to the inherent swelling properties of the elastomeric stamp. We developed a patterned hydrogel microwell array that was UV cured to a functionalized glass coverslip via a photomask. We have successfully cultured small populations of human induced pluripotent stem cells (hiPSCs) staining positive for pluripotency markers Oct4 and Sox2, as well as differentiated hiPSCs into Cdx2+ hindgut in these patterned microwells. This hydrogel microwell platform holds great potential for culturing, manipulating, and testing complex heterogenous populations in a controlled setting.
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8.	Intracellular delivery by cell squeezing is affected by perturbation of actin cytoskeleton and lipid rafts
	Martin P. Stewart^1,2, Xiaoyun Ding^1,2, Armon Sharei^1,2, Robert S. Langer^1,2, and Klavs F. Jensen^1,2
	¹Koch Institute for Integrative Cancer Research, Massachusetts Institute of Technology, Cambridge, MA ²Department of Chemical Engineering, Massachusetts Institute of Technology, Cambridge, MA
	Cell squeezing is a new intracellular delivery modality based on rapid mechanical deformation of cells passaging through precisely fabricated constrictions in a microfluidic device. Delivery of molecules is achieved by diffusion through membrane disruptions, however, little is known about how the properties of the cell surface affect the membrane disruption and recovery behavior. Using specific inhibitors in conjunction with flow cytometry to assess delivery efficiency, we uncover roles for actin and lipid rafts in the cell response. While perturbation of rafts decreased cell robustness and recovery, inhibitors of actin dynamics exhibited variable effects, with some even improving membrane recovery. We anticipate continued studies on these underlying mechanisms should yield insight into the mechanical response to rapid cell deformations and lead to improved efficiency in delivery applications.
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9.	Comprehensive characterization of human growth hormone degradation products by integration of a top-down and bottom mass spectrometry approach
	Annie Y. Wang¹, Jared R. Auclair¹, Joseph P. Salisbury¹, Di Wu1, Shiaw-lin Wu^1,2, Jeffery N. Agar¹ and William S. Hancock¹
	¹Barnett Institute and Department of Chemistry and Chemical Biology, Northeastern University, Boston, MA ²BioAnalytix Inc, Cambridge, MA
	With the advent of biosimilars to the US market, it is important to have better analytical tools to assure quality of the follow on products. In addition, the popularity of using a continuous process for production of biopharmaceuticals, traditional bottom-up proteomics, alone, for product characterization and quality analysis is no-longer sufficient. On the other hand, top-down proteomics requires minimal quantities of sample and allows for analysis of the intact protein and cleavage peptides. However, fragmentation usually occurs at the N-terminal and C-terminal of protein and less fragmentation occurs with the inner region of the protein, thus requiring methods to produce new internal fragmentations. Herein, we combine the use of the complementary techniques, top-down and bottom-up proteomics, for the characterization of human growth hormone variants as an example of our integrated approach. Notably, our method requires little sample, which is a requirement due to sample limitations for the development of continuous manufacturing. We were able to characterize various protein variants, containing degraded, and cleaved forms, which represents a typical sample generated in a stability study. Thus we were able to highlight the complementary nature of top down and bottom up protein analysis resulting in consistent results as well as unique observations.
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10.	Biopharmaceutical Stability Studies on Therapeutic Proteins with off-line Tandem Mass Spectrometry and Real Time Raman Techniques
	Di Wu¹, William Herrington², Gajendra P Singh², Yu Wang¹, Rajeev J. Ram², and William Hancock¹
	¹Barnett Institute and Dept. of Chemistry and Chemical Biology, Northeastern University, Boston, MA 02115 ²Massachusetts Institute of Technology, 77 Massachusetts Ave, Cambridge, MA 02139
	Biosimilar therapeutics are marketed worldwide nowadays and stability assessment is necessary to validate the long term safety, purity and potency of the products. In this study, a range of controlled stressed conditions, including temperature, humidity and pH, have been applied to the therapeutic proteins. The resulting degradations from the accelerated stability study are monitored with LC-MS-MS. With the high efficiency, accuracy, and sensitivity of mass spectrometry, deamidation and oxidation variants were detected. The differences in structural integrity and conformational stability profiles were monitored and potential degradations were evaluated by the MS analysis. In addition, real time Raman techniques were utilized in this study, which demonstrated a powerful ability to monitor degradation reactions as well as identify biosimilar therapeutic products including hGH, IFN, IgG as well as the variants. Data obtained from Raman spectrometry are comparable with mass spectrometry, and was achieved by the development of novel Raman methods to improve sensitivity and robustness of the measurement. The study indicates that Raman spectroscopy has potential to provide an alternative to conventional analytical methods for the monitoring of protein therapeutics.
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Three poster prizes:
1st place will receive $1000 (plus travel grant*)
2nd place will receive $500 (plus travel grant*)
3rd place will receive a travel grant*
*Travel grants include $1000 travel reimbursement and waived registration
to a CASSS-sponsored meeting (i.e. WCBP 2016 in Washington, DC).