Speaker: Ryan Hicks
Associate Director, Stem & Primary Cell Group AstraZeneca
November 15, 7:00AM/4:00AM (EST/PST)
Ryan Hicks is the Associate Director of the Stem & Primary Cell Group at AstraZeneca. Ryan’s group is responsible for the generation, engineering and differentiation of iPS cells, adult stem cells and use of primary cells for assay development, target validation, hit finding, hit-to-lead, lead optimization and phenotypic screening activities. Models are generated in 2D and 3D cultures and part of the model development work includes using precise genome modification techniques to introduce gene knock-outs, knock-in mutations or endogenous reporters into iPS cells. The groups work primarily focuses on AstraZeneca’s cardiovascular, metabolic, renal and respiratory disease areas. Ryan Hicks joined AstraZeneca in 2001 and has been based in Sweden for the past 8 years. Prior to working in Sweden, Ryan worked as the Associate Director of the Cell & Protein reagents group at AstraZeneca, Alderley Park, UK, with responsibilities in the cell and protein disciplines. Ryan Hicks received his degree in Medical Biochemistry from the University of Birmingham, UK. Following this Ryan received his PhD in Biochemistry and Molecular Biology from the University of Leeds, UK, in 2001.
Presentation: Generation and application of 3D-organoid cell culture models using human induced pluripotent stem cells (iPSCs) for drug discovery
Development of physiologically relevant cellular models, with strong translatability to human pathophysiology, is critical for identification and validation of novel therapeutic targets. Cell types derived from induced pluripotent stem cells (iPSCs) provide an important resource for modelling native biology in human cells and tissues for drug discovery. Understanding the phenotypic and functional relevance of these iPSC derived cells and tissues and how well they represent human biology is vital and as such we have developed tool iPSC-lines and techniques to optimize and monitor differentiation. Applying CRISPR/Cas9, next-generation sequencing (NGS), functional testing and advanced imaging techniques to iPSC model validation, we have established a platform that enables an accurate assessment of their benefits, maturity, gaps and challenges, as well as application in a drug discovery setting. Data presented will be from a key kidney model system we have developed to examplify our platforms. This will describe the validation and application of our kidney model, using SIX2 and NPHS1 markers non-invasively in real time, to trace early nephron commitment and podocyte-maturation in the same cell. In adition, we will also describe how we use transcriptomic analysis by NGS, in addition to functional assays, to get the most from our iPS-derived 3D cell models.
Speaker: Rhys Macown
Senior Scientist, Cell and Gene Therapy Catapult
November 15, 5:00AM/2:00AM (EST/PST)
Rhys Macown PhD is a senior scientist in the process industrialisation team at Cell and Gene Therapy Catapult UK. Rhys earned a BE(Hons) in chemical and process engineering at University of Canterbury (NZ) before starting his career with a PhD in biochemical engineering in Prof. Nicolas Szita’s group at University College London. There he designed and tested a microfluidic system for the 2D adherent culture of pluripotent stem cells with fine control over the micro-environment. Since joining the Cell and Gene Therapy Catapult in 2014, Rhys has been continually active on fee-for-service, collaborative, and core-funded projects developing pluripotent stem cell based processes and has played a leading role exploring the 3D expansion of pluripotent stem cells as aggregates.
Presentation: Towards commercial manufacture of pluripotent stem cell–derived therapies.
In contrast to autologous cell therapies, allogeneic therapies do not require collection of starting material from patients and could; be manufactured in larger batches, be made available “off-the-shelf”, and potentially utilise established biopharma manufacturing technologies. Despite this, allogeneic therapies constitute half of pre-clinical trials and a third of clinical trials in the UK. In an addition to the potential for an immune response, another major barrier to the adoption of allogeneic therapies is current manufacturing cost which increases when using pluripotent stem cells (PSC) as starting material. The manufacturing process is often open and manual, utilising 2D, adherent culture systems and xeno or serum containing materials. As such, these processes are regulatorily challenging and impractical to scale-up to meet market demands. This talk will cover our work to date and future plans in addressing these barriers and realising the potential of PSC-derived allogeneic therapies.
The Cell and Gene Therapy (CGT) Catapult have been working to address some of the challenges of manufacturing processes for PSC derived therapies, towards improving commercial viability. We used regulatorily amenable starting materials, developed invasive and non-invasive methods for process characterisation, developed exemplar processes for the expansion of pluripotent aggregates in stirred tank reactors, and developed concentration and wash processes for large-scale PSC processing. CGT Catapult’s next development focuses are the scale-up and intensification of PSC expansion in stirred tank reactors, translation of differentiation processes to automated and scalable culture systems, and cost of goods reduction.
Speaker: Alysson Muotri
Professor, Department of Pediatrics/Cellular & Molecular Medicine
Director, UCSD Stem Cell Program University of California, San Diego
November 15, 1:00PM/10:00AM (EST/PST)
Dr. Muotri earned a BSc in Biological Sciences from the State University of Campinas in 1995 and a PhD in Genetics in 2001 from University of Sao Paulo, in Brazil. He moved to the Salk Institute as Pew Latin America Fellow in 2002 for a postdoctoral training in the fields of neuroscience and stem cell biology. He has been a Professor at the School of Medicine, University of California in San Diego since late 2008. His research focuses on modeling neurological diseases, such as Autism Spectrum Disorders, using human induced pluripotent stem cells. His lab has developed several techniques to culture human neurons and glia for basic research and drug-screening platforms. He has received several awards, including the prestigious NIH Director’s New Innovator Award, NARSAD, Emerald Foundation Young Investigator Award, Surugadai Award from Tokyo University, Rock Star of Innovation from CONNECT, NIH EUREKA Award among others.
Presentation: Applications of brain-model technology
The complexity of the human brain, with thousands of neuronal types, permits the development of sophisticated behavioral repertoires, such as language, tool use, self-awareness, symbolic thought, cultural learning and consciousness. Understanding what produces neuronal diversification during brain development has been a longstanding challenge for neuroscientists and may bring insights into the evolution of human cognition. Human pluripotent stem cells have the ability to differentiate in specialized cell types, such as neurons and glia. Moreover, induced pluripotent stem cells can be achieved from living individuals by reprogramming somatic cells that would capture their entire genome in a pluripotent state. From these pluripotent state, it is possible to generate models of the human brain, such as brain organoids. We have been using brain-model technology (BMT) to gain insights on several biological processes, such as human neurodevelopment and evolution. We also applied BMT to measure the impact of genetic variants in autism spectrum disorders and for evolutionary studies. The reconstruction of human synchronized network activity in a dish can help to understand how neural network oscillations might contribute to the social brain. Our findings suggest a potential bridge to the gap between the microscale in vitro neural networks electrophysiology and non-invasive electroencephalogram.
Speaker: Filip Roudnicky
Senior Scientist, Roche
November 15, 10:00AM/7:00AM (EST/PST)
Filip Roudnicky is a senior scientist in the disease relevant cellular assay team in chemical biology at Roche. His is responsible for genome editing for disease modeling and for CRISPR/Cas9 genetic screens. He is also expert on disease relevant cellular assays involving endothelial cells. Filip started his career with a PhD in a lab of Prof. M. Detmar at ETH Zurich. He studied tumor angiogenesis of invasive bladder carcinoma and identified several biomarkers and molecular targets on tumor-associated blood vessels of bladder cancer. As a guest research scientist he has worked, in RIKEN Yokohama, Japan, under Dr. Jay W. Shin, on induced-neuronal stem cells and RNA-sequencing analysis. He has been a postdoctoral fellow in Roche and Harvard with the lab of Prof. C. Cowan developing an in vitro model of retinal endothelial cells.
Presentation: Use of stem-cell derived endothelial cells for disease relevant cell modeling and drug discovery
The use of human pluripotent stem cells (hPSCs) for in vitro disease-modeling is limited by the lack of robust and efficient protocols for the differentiation of relevant adult cell types. Previously, we have reported a method to generate vascular endothelial cells from hPSCs (Patsch et al. Nat Cell Biol. 2015). This novel and robust protocol in conjunction with use of programmed nucleases allows generation of relevant endothelilal cell disease models. We will show examples of modeling high-barrier resistance endothelial cells in vitro, by generation of CLDN5 reporter and use of chemical profiling, and modeling the effect of a severe metabolic genetic disease (AKT2 loss-of-function or dominant active mutation) on endothelial cells.
Speaker: Kris Saha
Assistant Professor, Department of Biomedical Engineering, University of Wisconsin-Madison
November 15, 11:00AM/8:00AM (EST/PST)
Krishanu Saha is an Assistant Professor in the Department of Biomedical Engineering at the University of Wisconsin-Madison. He is also a member of the Wisconsin Institute for Discovery, Carbone Cancer Center, and Stem Cell and Regenerative Medicine Center as well as the National Academies’ Forum on Regenerative Medicine. Prior to his arrival in Madison, Dr. Saha studied Chemical Engineering at Cornell University and at the University of California in Berkeley. He was a Society in Science: Branco-Weiss fellow at the Whitehead Institute for Biomedical Research at MIT and in the Science and Technology Studies program at Harvard University. Major thrusts of his lab involve gene editing and cell engineering of human cells found in the retina, central nervous system and blood.
Speaker: Precise gene editing of human pluripotent stem cells
CRISPR ribonucleoproteins (RNPs) can generate programmable gene edits, however imprecise editing and efficient delivery to human stem cells are key challenges. Here we describe novel biochemical techniques to assemble various biomolecules and coatings with nanoscale precision around a RNP. First, by modifying the sgRNA with a short S1m RNA aptamer, we developed a modular strategy, termed an “S1mplex,” to assemble Cas9 RNPs with biotinylated moieties. Using S1mplexes with biotinylated short oligonucleotides improves the ratio of precise to imprecise editing up to 18-fold over conventional methods approaching a ratio of 4 precise edits to every imprecise mutation, while assembly with fluorescent molecules allows selection and enrichment for cells with multiplexed gene edits. Second, we developed synthetic coatings for nonviral delivery of RNPs to human pluripotent stem cells. Combined, these strategies, which utilize chemically-defined, off-the-shelf reagents, have significant promise for gene editing applications in vitro (e.g., drug discovery, disease modeling) with human stem cells.
Speaker: Ashleigh Schaffer
Assistant Professor, Deparment of Genetics and Genome Sciences Case Western Reserve University
November 15, 3:00PM/12:00PM (EST/PST)
Ashleigh Schaffer graduated with a BS in Genetics from University of Wisconsin-Madison and went on to earn her PhD from the University of California, Irvine under the mentorship of Dr. Maike Sander. She continued her training as a postdoctoral fellow at the University of California, San Diego, and Howard Hughes Medical Institute under the guidance of Dr. Joseph G. Gleeson, focused on elucidating the genetic cause, and molecular mechanisms, underlying recessive pediatric neurological disease. During her time with Dr. Gleeson, Ashleigh discovered over 20 novel genetic causes of disease, many for recessive, syndromic pediatric neurodegenerative disorders. She joined the Department of Genetics and Genomes Sciences at Case Western Reserve University as an Assistant Professor in February 2017.
Presentation: Using human stem cells to model neocortical gyration phenotypes
Neuronal migration defects, including pachygyria, are among the most severe developmental brain defects in humans. Using human genetics approaches, we recently identified bi-allelic truncating mutations in CTNNA2, encoding αN-catenin, in patients with a phenotypically distinct recessive form of pachygyria. Interestingly, two mouse lines harboring nonsense mutations of the ortholog to human CTNNA2 (Catna2) have been characterized, but failed to recapitulate the human cortical lamination phenotype observed in our patients. Therefore, to investigate neuronal phenotypes in a human model, we developed stem cell-derived neuronal assays to assess neuron morphology as well as define the molecular disease mechanism. I will highlight the need for human cell-based models for neocortical gyration disorders as well as discuss the advantages of using stem cell based systems to discover pathogenic mechanisms and novel insight into human brain development, with this case as an example.
Speaker: Takanori Takebe
Associate Director, Center for Stem Cell & Organoid Medicine (CuSTOM)
Assistant Professor, Cincinnati Children’s Hospital
Professor, Tokyo Medical and Dental University
Professor and Principal Investigator, Yokohama City University
November 14, 6:00PM/3:00PM (EST/PST)
Dr. Takebe is an Associate Director of Center for Stem Cell and Organoid Medicine (CuSTOM) at the Cincinnati Children’s Hospital Medical Center and Professor at Yokohama City University, and Tokyo Medical and Dental University, Japan. He serves as Board of Directors at International Society of Stem Cell Research (ISSCR). His lab investigates the mechanisms of human organogenesis, and develops mini-organ technologies from human stem cells—namely organ bud–based approaches. He is applying iPSC liver buds into drug discovery study as well as transplant application—for patients with a rare congenital metabolic disorder, ultimately expanding the clinical indications to diseases like liver cirrhosis.
Presentation: Modeling inflammation and fibrosis in humans with PSC-derived steatohepatitis liver organoids
Human organoid systems recapitulate in vivo organ architecture, yet fail to capture complex pathologies such as inflammation and fibrosis. Here we developed multi-cellular human liver organoids (HLO) from 11 different healthy and diseased PSC lines that display essential features of steatohepatitis. Single-cell-level analysis revealed steatohepatitis organoids exhibited persistent hepatic steatosis, followed by progressive activation of pro-inflammatory and fibrotic lineages. Interestingly, expression of the steatohepatitis phenotype correlates with the presence of clinically significant GWAS factors: PNPLA3, GCKR and TM6SF2. We developed an organoid-level readout with atomic force microscopy, and demonstrated that organoid stiffening reflects the fibrosis severity. Furthermore, organoid model of an iatrogenic parental nutrition associated liver disease (PNALD) identified obeticholic acid as effective in preventing pathology progression, suggesting the potential repurposing for PNALD. The presented key methodology and preliminary results offer a new method for studying human precision for inflammation and fibrosis, facilitating the discovery of effective treatments.
Speaker: Essam Abdelalim
Scientist, Diabetes Research Center, QBRI Assistant Professor College of Science and Engineering, HBKU
Dr. Essam M. Abdelalim is a scientist at QBRI and Assistant Professor at Hamad Bin Khalifa University (HBKU). He received his PhD in Medical Science from Shiga University of Medical Science, Japan, and was later appointed as Assistant Professor in the same university. He was awarded a Postdoctoral Fellowship from the Japan Society for Promotion of Science (JSPS), where he identified novel genes involved in maintaining pluripotency and self-renewal of embryonic stem cells (ESCs). Since April 2014, Dr. Abdelalim has been leading the stem cell team focusing on diabetes at QBRI/HBKU. He has received several prestigious awards, including the State Prize of Encouragement (Egypt) in 2012 and the President’s Award for outstanding PhD student from Shiga University of Medical Science (Japan) in 2007. Dr. Abdelalim is a member of the editorial board of “Stem Cells and Development” and “World Journal of Stem Cells”. He is also a guest editor for “Stem Cell International” and “Frontiers in Cell and Developmental Biology”. His current research at QBRI focuses on using pluripotent stem cell technology to investigate the pathophysiology underlying the development of diabetes and insulin resistance as well as generating stem cell-derived pancreatic beta cells for cellular therapy. He is currently leading several diabetes-related projects funded by QBRI and QNRF. Recently, he has been awarded two NPRP grants (NPRP9 and NPRP10) and two QBRI Internal Research grants to study diabetes and pancreatic beta cell differentiation.
Presentation: Efficient generation of pancreatic β cell precursors from human pluripotent stem cells
Pancreatic β cell replacement therapy is considered as a potential strategy to treat diabetes. To date, transplantation of pancreatic islets from cadavers is the most effective approach for treating diabetic patients, but this approach has limitations in terms of the necessity of strong immunosuppressive drugs and the shortage of donors. Alternatively, human pluripotent stem cells (hPSCs), including human embryonic stem cells (hESCs) and hiPSCs, can be differentiated into insulin-secreting β cells that have a great potential to treat diabetes. However, generation of functional β cells similar to those present naturally in the pancreas is, so far, a work in progress. Indeed, few studies reported the generation of functional β cells from hPSCs, but those cells were low in efficiency and functionality, creating a major obstacle to the use of these cells for cellular therapy. Recent progress showed that patients with diabetes could be transplanted with hPSC-derived pancreatic progenitors co-expressing the two transcription factors (TFs), PDX1 and NKX6.1. Those progenitors are known as the precursors of functional pancreatic β cells.; therefore, after their transplantation, they are converted into functional insulin-secreting β cells inside patient’s body. Recently, we have established an efficient protocol for maximizing generation of PDX1+/NKX6.1+ pancreatic progenitors from hPSCs. We enhanced the generation of PDX1+/NKX6.1+ population, by manipulating in vitro culture conditions. Our optimized protocol dramatically increased the expression of NKX6.1, leading to an increase in the proportion of PDX1+/NKX6.1+ progenitors (~90%) and upregulation of the key TFs controlling pancreatic β cell development. The improved efficiency of pancreatic differentiation was associated with an inhibition in the hepatic lineage and an increase in the proliferation of NKX6.1+ cells. Further differentiation validated the ability of PDX1+/NKX6.1+ progenitors to generate endocrine progenitors (NGN3+ cells). In addition, we were also able to enrich a novel PDX1-/NKX6.1+ population by manipulating the re-plating densities, that appeared in the form of three-dimensional clusters. Our findings provide a novel technique that facilitates appropriate cellular rearrangement in monolayer culture to yield a high proportion of PDX1+/NKX6.1+ pancreatic progenitors with an increased self-replicating capacity, thereby aiding scalable production of functional β cells from hPSCs in vitro. Our innovative method also supports the presence of two distinct NKX6.1+ pancreatic populations during human pancreatic development, which may suggest a new trajectory to generate β cells in humans. In this talk, I will present our recently published work on generating distinct pancreatic progenitor populations from hPSCs.
Speaker: Kapil Bharti
Earl Stadtman Tenure-Track Investigator National Eye Institute Adjunct Group Leader, National Center for Advancing Translational Sciences, National Institutes of Health
Dr. Kapil Bharti holds a bachelor’s degree in Biophysics from the Panjab University, Chandigarh, India, a master’s degree in biotechnology from the MS Rao University, Baroda, India, and a diploma in molecular cell biology from Johann Wolfgang Goethe University, Frankfurt, Germany. He obtained his PhD from the same institution, graduating summa cum laude. His PhD work involved research in the areas of heat stress, chaperones, and epigenetics. He did his postdoc at the National Institutes of Health, where he published numerous papers in the areas of transcription regulation, pigment cell biology, and developmental biology of the eye. He has won several awards, including, most recently, being selected as an Earl Stadtman Tenure-Track Investigator at NIH. His lab was recently awarded two prestigious grants: (1) the only Intramural Common Fund grant to develop a phase I Investigational New Drug (IND) for autologous induced pluripotent stem cell derived retinal pigment epithelium tissue; and (2) a DoD grant to develop a 3D retina tissue to model retinal diseases in vitro. His current work as the head of the Unit on Ocular and Stem Cell Translational Research involves understanding mechanism of retinal degenerative diseases using induced pluripotent stem cell technology, and developing cell-based and drug-based therapies for such diseases.
Speaker: Devon D. Brewer, PhD
Devon Brewer has 25 years of experience in writing successful grant proposals in the health sciences. He has been principal investigator on several grants and a key contributor to many others. Brewer has written funded proposals or reviewed proposals for NIH, NSF, CDC, HRSA, MRC (UK), private foundations, and other agencies. As a scientific advisor and writer with ScienceDocs, he helps researchers develop and compose effective proposals and manuscripts.
Presentation: Strategies for successful grant proposals
I describe several strategies for writing successful grant proposals. Each strategy derives from understanding the implications of two basic facts: funders seek to advance their missions through grants, and proposals are offers to sell research projects to funders. I outline the steps for ensuring proposal content and presentation fit a grant opportunity well. I also highlight logistic and bureaucratic matters, ways to build skill in developing and writing grant proposals, and sources of feedback and help.
Speaker: Phil Greenwood
Senior Lecturer, Weinert Center of Entrepreneurship at the Wisconsin School of Business, University of Wisconsin-Madison, USA
As an educator and trainer in the fields of accounting, finance, and entrepreneurship, Phil has designed and instructed numerous courses for MBA, Executive MBA and executive education customers. Currently, he is a Senior Lecturer for the Weinert Center of Entrepreneurship at the Wisconsin School of Business, University of Wisconsin-Madison (consistently ranked among the top 25 entrepreneurship programs by Financial Times and Entrepreneur magazine). Over the past decade, Phil has taught courses in Entrepreneurial Finance, New Venture Creation, Entrepreneurial Management, and Strategic Management at the undergraduate and full-time MBA levels. Phil is also an adjunct faculty member with the Executive MBA - Kohl's Program, the Evening MBA Program, and Executive Education. Topics include Corporate Finance, Financial Management, Business Valuation, and Financial Analysis. Finally, Dr. Greenwood is a founding faculty member with the Morgridge Entrepreneurial Bootcamp (MEB) and Master's in Biotechnology program sponsored by the UW-Medical School. Phil has extensive experience instructing at other institutions including the Copenhagen Business School MBA, Copenhagen, Denmark; University of St. Thomas in the Twin Cities, Minnesota; MBA program at HEC University in Versailles, France; Executive MBA program at KÖC University based in Istanbul, Turkey; and, the Executive MBA program at the Chinese Academy of Sciences in Beijing, China.
Presentation: Biotechnology entrepreneurship
This presentation provides a brief introduction to biotech entrepreneurship including factors that are pertinent to biotech entrepreneurship and different myths and facts that exist. You will also learn about factors that many investors consider and find attractive for potential investments specifically in biotech. Finally, we will review what to consider after the startup and into the future.
Speaker: Gay Grossman
Co-founder and Patient Advocate, ADCY5.org
Gay started ADCY5.org to provide a place for newly diagnosed families of ADCY5 related-Dyskinesia, an ultra Rare Disease. They currently have one of the largest Rare Disease stem cell projects. Working with biotech partners, almost $3 million has been dedicated to ADCY5 related-Dyskinesia related science through ADCY5.org. Speaking publicly at conferences and to organizations about the first hand experience of Whole Genome Sequencing, personalizes how a diagnosis for her daughter, after 15 years, changed their family’s lives. Sharing this story with a variety of audiences including, but not limited to, precision medicine conferences, patient advocacy conferences, and graduate and medical school seminars, fosters partnerships within our community. We Are All Rare, a patient education book Gay co-authored, together with her daughter, who has a Rare Disease, provides an educational tool for families entering primary school, complete with a list of valuable resources. Meeting with her Congressman to encourage support of The OPEN ACT, Medicaid reimbursement, and access for persons with disabilities, gives her the opportunity to share the patient family perspective with policy makers. She is currently serving on the Global Genes Foundation Alliance Advisory Council, their Summit Planning Committee for the 2018 International RARE Patient Advocacy Summit, and on the developing team for the Genetics 101 class. She is an Ambassador for Illumina; for the Rare Undiagnosed Genetic Disease (RUGD) program. Gay works with key opinion leaders like Rare Science, Global Genes, EveryLife Foundation, and other patient advocacy organizations to engage through social media, conferences, and personal networking. Gay started her career in Sales and Sales Training with Glaxo Inc. where she directed a study on Physician Impressions of Corticosteroids and how to best position them in the marketplace. Gay uses this foundation of science and her own experience to relate the burden of disease, bringing awareness through advocacy and public speaking. She successful advocates in environments including, but not limited to, educational classrooms, private, state, and federal insurance, and collaborating in the Rare Disease space to gain access to medical therapies.
Presentation: Undiagnosed disease to rare disease discovery: Perspectives from a patient family
One would think that getting the diagnosis after fifteen years would end the journey. How does your work at the bench affect patients and how can you change lives through cell therapy? This talk highlights the power of precision medicine with the story of the ADCY5 gene variant and the journey of a family who went from being n of 1 to having fifty stem cell lines. The mother of a patient shares her perspective and determination to find a diagnosis and advancements in science and technology that will help hasten the path towards treatment.
Speaker: William Hendriks
Instructor, Neurology Harvard Medical School Dept. of Neurology, Massachusetts General Hospital
After receiving his PhD in Neuroscience at the VU University Amsterdam in the Netherlands in 2008, Dr. Hendriks joined the lab of Dr. Paola Arlotta at the Center for Regenerative Medicine of Massachusetts General Hospital (MGH) in Boston to study neuronal development focusing on neuronal differentiation of human pluripotent stem cells. In 2011, Dr. Hendriks joined the Harvard Stem Cell Institute (HSCI) iPS Core facility with Dr. Chad Cowan at Harvard University in Cambridge, where initially he worked on developing and implementing foot-print free somatic cell iPSC reprogramming methods. Dr. Hendriks also initiated and managed the hPSC genome editing service for 2 years at HSCI before moving to his current position as a Harvard Medical School Instructor in Neurology at the MGH Collaborative Center for X-Linked Dystonia Parkinsonism in 2014.
Presentation: Human PSC-based disease modeling to study X-linked Dystonia-Parkinsonism
The isolation of human embryonic stem cells (hESCs) and the discovery of human induced pluripotent stem cell (hiPSC) reprogramming have sparked a renaissance in stem cell biology, in vitro disease modeling, and drug discovery. In general, hPSC-based disease models are well-suited to study genetic variation. Studies commonly compare patient-derived hiPSCs, e.g., with a disease-causing genetic mutation, and (age-matched) control subject-derived hiPSCs, typically differentiated to the disease-affected cell type, e.g., neurons. A major caveat of this disease-modeling strategy is the variability of differentiation propensities and phenotypic characteristics, even in hPSCs derived from the same donor. Still, even if the cellular phenotype of a given mutation is strong and highly penetrant, it may be lost due to confounding effects of differences in genetic background of unrelated hPSC lines. A very powerful approach to overcome this hurdle is to use custom-engineered endonucleases, such as CRISPR/Cas9 that enable precise and programmable modification of endogenous hPSC genomic sequences. In our lab we use hPSC-based disease modeling to study the neurological movement disorder dystonia, in particular X-linked Dystonia Parkinsonism (XDP). In this talk I will show how we use hPSC-based disease modeling in combination with CRISPR/Cas gene editing, to elucidate the underlying molecular pathogenesis of XDP. I will also address some of the potential problems one might face using hPSC-based disease modeling in combination with gene editing.
Speaker: Jeanne Loring
Professor, Developmental Neurobiology
Director, Regenerative Medicine The Scripps Research Institute
Jeanne F. Loring is a Professor and the founding Director of the Center for Regenerative Medicine at The Scripps Research Institute in La Jolla. Her laboratory studies human pluripotent stem cells (hPSCs), the remarkable cells that can make every cell type in the body. Her lab is developing a patient-specific cell replacement therapy for Parkinson’s disease and a stem cell-based treatment for multiple sclerosis, and they are studying the underlying causes of autism. With the San Diego Zoo, her lab is developing a "zoo" of induced pluripotent stem cells from endangered species to aid in their conservation. Dr. Loring serves on many scientific and bioethics advisory boards and is frequently quoted in major newspapers, appears on television and in documentary features, and gives many public lectures about science and society. She is particularly concerned with the dangers of unregulated stem cell treatments in the US and other countries (“stem cell tourism”). In 2015, she was designated “Stem Cell Person of the Year” by The Niche blog, and was awarded the Stem Cell Action Advocacy Award by the World Stem Cell Summit.
Presentation: Putting pluripotency to use
Pluripotent stem cells can reproduce indefinitely and develop into any cell type in the body. We are putting pluripotent stem cells to use, for a neuron replacement therapy for Parkinson’s disease, treatment of multiple sclerosis, understanding autism, and rescuing endangered species.
Speaker: Florian T. Merkle
Principal Investigator, University of Cambridge, WT-MRC Institute of Metabolic Science and WT-MRC Cambridge Stem Cell Institute, UK
Florian T. Merkle received his BS in biology from the California Institute of Technology (Caltech), which he attended on full tuition merit scholarship. Florian pursued his PhD in neuroscience at the University of California, San Francisco (UCSF), where he studied adult neurogenesis in the laboratory of Prof. Arturo Alvarez-Buylla. His work revealed the embryonic origin of adult neural stem cells, overturned the widely-held belief that these stem cells are homogeneous and multipotent, and discovered several new types of adult-born cell types. As a postdoctoral fellow at Harvard University, Florian worked with Prof. Alex Schier and Prof. Kevin Eggan to develop gene editing tools in human pluripotent stem cells (hPSCs) and to sequence and bank a large collection of hPSCs to facilitate disease modelling and transplantation studies. He developed novel methods to differentiate hPSCs into hypothalamic neurons that regulate essential physiological processes and are therefore pivotally important for human health, and whose abnormal function causes a range of diseases including obesity. Florian how leads a research group at the University of Cambridge where he is studying obesity using in vitro disease models and pursuing questions relating to the genetic stability of hPSCs. These studies are supported by the Wellcome Trust, Royal Society, and Academy of Medical Sciences.
Presentation: Disease modelling with human pluripotent stem cells: general principles and specific applications to obesity
The capacity to generate disease-relevant cell populations from human pluripotent stem cells has tremendous potential for shedding light on human disease mechanisms. I will discuss basic principles of in vitro disease modelling, including the generation of isogenic models with CRISPR/Cas9, the issue of recurrent culture-acquired mutations and how to assess stem cell genomic integrity, and cell type maturity and phenotyping strategies. To frame these issues, I will discuss my research group's interest in obesity, which is largely a disease of excess food intake, which is in turn regulated by neurons in the hypothalamus. We use human stem cell-derived hypothalamic neurons to gain insight into the environmental and genetic factors that alter neuron function.
Speaker: Rakhi Pal
Chief Technology Scientist Centre for Brain development and Repair, Instem, Bangalore
Dr. Rakhi Pal is the Chief technology scientist at the Centre for Brain Development and Repair, Institute of Stem Cell Biology and Regenerative Medicine, Bangalore, India where she works using stem cell–based technologies to develop platforms for understanding neuro-developmental disorders. A gold medalist in human physiology from the prestigious All India Institute of Medical Sciences, New Delhi and a PhD from Manipal University, Dr Pal has more than 10 years of experience in the field of stem cell biology with particular interest in neurological disorders. She not only has multiple publications and patents to her name, but also possesses an unique blend of both industry and academic experiences.
Presentation: Human stem cells for modelling neurological disease and its therapeutic applications
My talk shall encompass how the stem cell research field has evolved from embryonic stem cells to adult stem cells and currently induced pluripotent stem cells with special reference to the field of neuroscience in a bench-to-bedside approach. Studying neurological disorders has always been a challenge. This is compounded by the lack of predictive pathophysiological models to identify and test potential therapeutic targets. Adult stem cells being multipotent and non-teratogenic have been in the limelight of stem cell clinical research. Multiple clinical trials have been conducted across the globe with limited and inconsistent results. As we progress to understand how stem cells may orchestrate the regeneration process, a whole new avenue of questions have opened up and I will elaborate on this during the course of my talk. While adult stem cells continue to be the focus of clinical trials at the moment, it is essential to note that the discovery of induced pluripotent stem cells has made the field even more exciting. Along with the technological advancements in sequencing and gene manipulation techniques, personalized medicine seems to be the future. Reconstruction of the early developmental stages is now possible by deriving induced human pluripotent stem cells from patient samples using advanced molecular and cellular manipulation methods. For example, we have been successful in reprogramming and differentiating human patient derived–iPSCs into the three common brain cell types: neurons, glial cells and oligodendrocytes of cortical origin, importantly using pathways and trophic factors that can sequentially recapitulate human neural development. This is imperative for accurate understanding of the disease and enhancement of therapeutic strategies. Upon marker analysis and functional characterization, human iPSCs are able to consistently generate cortical neurons that are electrophysiologically active and that have intrinsic electrical properties comparable to adult human neurons capable of generating mature action potentials upon stimulation. However, lack of synaptic activity led us to further explore an astrocyte-neuronal co-culture system which closely mimics the internal milieu of the brain and responds to drugs. This platform is currently being used to understand pathophysiology of various neurological, neurodevelopmental and neurodegenerative disorders. Importantly, it opens up unlimited access to human tissue material in an unprecedented manner, which can be used for drug testing, neurotoxicity studies, epigenetic changes, gene editing studies and personalized medicine.
Speaker: Daniel Paull
VP, Automation Systems and Stem Cell Biology, The New York Stem Cell Foundation Research Institute
Dr. Paull is Vice President, Automation Systems and Stem Cell Biology at The NYSCF Research Institute. Daniel oversees the day-to-day operation of The NYSCF Global Stem Cell Array in both the production of induced pluripotent stem cells and development of novel tools using the robotic platform including such areas as gene editing and differentiation. He also oversees a number of collaborative projects aimed at developing research across a range of disease areas. He received his PhD from University College London, England and followed this with post-doctoral work at the New York Stem Cell Foundation where he helped develop novel approaches for the treatment of mitochondrial disease as well as furthered research into somatic cell nuclear transfer.
Presentation: Beyond automated iPSC reprogramming: Applications for a fully automated cell culture system for pluripotent stem cell research
The use of pluripotent stem cells is dramatically altering the R&D landscape, providing new insights into both the basic biology of disease progression and novel cell types for pharmaceutical drug screening. Additionally, pluripotent stem cells are already in clinical use for a number of disease indications. However, one limitation of the work to date is that in vitro studies have largely focused on highly penetrant point mutations known to directly lead to a disease phenotype. While useful, this approach may not always be applicable to idiopathic versions of the same disease. For the majority of diseases, underwritten by common causal variants, modeling using only a limited number of cell lines is insufficient. To overcome these issues, we have developed a fully automated platform which allows high throughput cell biology to be accomplished: from reprogramming of hundreds of cell line in parallel to high-throughput differentiations, large-scale experiments can now be performed to interrogate these diseases. Here we report on progress in adapting the NYSCF Global Stem Cell Array technology to encompass high-throughput genome editing as well as large-scale differentiations and screens. We will present a number of case studies highlighting the continued development of this automated system for stem cell biology.
Speaker: Stevens Rehen
Head of Research, D'Or Institute for Research and Education (IDOR) & Institute of Biomedical Sciences Professor, Biomedical Sciences Federal University of Rio de Janeiro
Stevens Rehen received his Bachelor’s degree, Master’s degree, and PhD at the Federal University of Rio de Janerio in Brazil. He later completed postdoctoral training at the University of California San Diego and The Scripps Research Institute. Over the past five years, Stevens has published over 76 peer-reviewed publications. Currently, Stevens is a Full Professor at the Federal University of Rio de Janerio. Additionally, he is the Head of Research at D’Or Institute for Research and Education (IDOR) and Regional Committee Member of the Pew Latin American Program in the Biomedical Sciences.
Presentation: Reprogrammed stem cells to study psychedelic substances
For more than four decades, restrictions on research with psychoactive drugs have slowed progress in understanding how such substances impact brain metabolism. Besides the historical restrictions, the impacts of drug exposure in human neural cells have been compromised by limitations of adequate models. I will present the effects of the β-carboline alkaloid harmine, component of the psychoactive plant tea known as “Ayahuasca”, and 5-MeO-DMT (5-methoxy-N,N-dimethyltryptamine), found in the Sonora Desert toad, in cultures of human neural cells and brain organoids derived from induced pluripotent stem cells. Harmine increased the pool of proliferating cells, with DYRK1A (dual specificity tyrosine-phosphorylation-regulated kinase) as a target, which suggests a biological activity possibly associated with the antidepressant effects of Ayahuasca in patients with depressive disorder. Analyzing global protein expression of brain organoids exposed to 5-MeO-DMT, we found proteins broadly distributed on functional activities such as cellular protrusion formation, microtubule dynamics and cytoskeletal reorganization, which are correlated to novel dendritic spine formation. These models offer an exciting new range of opportunities to investigate the impact of psychedelics on human neural cells.
Speaker: BanuPriya Sridharan
Postdoctoral Associate, The Scripps Research Institute, Florida
Banu Priya Sridharan, PhD, is a Postdoctoral Research Associate at the Scripps Research Institute Molecular Screening Center in the department of Molecular Medicine at Scripps in Florida. After a brief stint in MIT-Harvard HST as an undergraduate research assistant, she trained as a tissue engineer in Lawrence, Kansas where she obtained her PhD in 2015. She moved to Medimmune to do a short-term industrial fellowship. She began at Scripps Florida in 2016 to advance her expertise on developing stem-cell based tissue models but now in the context of HTS. Dr. Sridharan pursued her passion for employing cellular models for phenotypic screening and is deeply involved in iPSC-based neuronal differentiation protocols, genome editing for disease modeling and relevant drug screening. Since 2011, she has published numerous peer reviewed articles and conference posters and podium talks and her key expertise includes stem cell differentiation, phenotypic assay development, high-throughput screening and high-content analysis.
Presentation: Addressing the scalability of human iPSC-derived heurons for HTS implementation and phenotypic screening
Traditional high-throughput screening (HTS) assays for neuronal targets employ non-human primary neuronal cells due to the scale necessary for HTS. Isolation of mouse primary neurons can be unreliable and economically demanding. The discovery of new drugs for neuropsychiatric disorders has further been hampered by lack of access to disease-relevant human primary neurons and appropriate disease models. By using human induced pluripotent stem cell (hiPSC) technology, we can address some of the obstacles which affords the generation of human neurons through (1) embryoid body (EB) formation, (2) cultivation on stromal feeder cells, and, (3) employing lineage-specific differentiation factors. Methods exist to reproducibly differentiate hiPSCs into functional cortical induced neurons (iN) in less than two weeks but, they have never been taken to the HTS scale and have been slow and somewhat variable. We have successfully recapitulated the aforementioned technique and leveraged the CRISPR technology to define the path to a plate-compatible format amenable for large-scale HTS implementation. The resulting iN cells exhibit appropriate genetic and fluorescent markers that give confidence of bonafide neuronal differentiation. Imminently, we intend to test the preliminary iN cells for their ability to post-mitotically increase or decrease neurite outgrowth following treatment with LOPAC test compounds via staining and high-content analysis. Ultimately, we will determine reliability and reproducibility over time with industrial-scale robotics. Furthermore, we also intend to leverage the CRISPR technology to create a library of disease-relevant-phenotypes from hiPSC-derived cellular models that will provide more opportunities for all biologists to study epigenetic mechanisms and scale-up screening initiatives with Scripps Research Institute Molecular Screening Center (SRIMSC).
Speaker: Lincon Stamp
ARC DECRA Fellow, University of Melbourne
Dr. Lincon Stamp did his undergrad degree, BSc (Biotechnology) at the University of Newcastle (Australia). Here he developed a strong interest in stem cell biology and so moved to Melbourne to do Honours and then PhD with Prof Martin Pera at Monash University. Here his research involved investigation of the early differentiation of human embryonic stem cells toward hepatopancreatic cell fates. He then joined the lab of Dr. Don Newgreen at the Murdoch Children’s Research Institute where he began working on development of the enteric nervous system, before joining Prof Heather Young’s lab at the University of Melbourne Department of Anatomy and Neuroscience, where he has been focused on developing a stem cell therapy to treat gut motility disorders such as the pediatric enteric neuropathy Hirschprung disease. Lincon is an ARC DECRA Fellow and holds an NHMRC Project Grant as CIB with Prof Young, and has recently published a number of high impact studies in the Gastroenterology, Journal of Clinical Investigation and Stem Cell Reports.
Presentation: A gut feeling about stem cell therapy for enteric neuropathies
The enteric nervous system (ENS) plays an essential role in gut motility. Diseases of the ENS result in bowel motility disorders that are some of the most challenging clinical conditions to manage. Cell therapy offers the potential to treat gastrointestinal motility disorders caused by enteric neuropathies. We have previously shown that following transplantation into the colon of recipient mice, enteric neural progenitors proliferate, migrate and differentiate into a variety of neurochemical types of neurons. However, it was unclear whether graft-derived neurons integrate into the circuitry of the recipient and directly regulate gut motility. We have used optogenetic and electrophysiological approaches to examine whether transplanted enteric neural progenitors generate neurons that functionally innervate the colon. Neural progenitors expressing the light-sensitive ion channel, channelrhodopsin, were isolated from fetal or postnatal bowel and transplanted into the colon of postnatal mice. The responses of recipient colonic smooth muscle cells to light stimulation of graft-derived neurons were examined. Light stimulation of graft-derived cells resulted in excitatory and inhibitory junction potentials, the electrical events underlying contraction and relaxation respectively, in colonic circular muscle cells. The pharmacological properties of the junction potentials evoked by stimulation of graft-derived neurons were identical to those of endogenous excitatory and inhibitory motor neurons. Interneurons were also generated from graft-derived cells, but their pharmacological properties varied with the age of the donors from which the progenitors were obtained. Our data demonstrate that transplanted progenitors generate different functional classes of enteric neurons involved in the control of gut motility.
Speaker: Christina Waters
CEO/Founder, RARE Science Inc.
Senior Vice President/General Manager, Global Rare Disease Program, WuXi NextCODE
Dr. Christina Waters’ broad range of experience in leading medical research in both biotech/large pharmaceutical companies to non-profits converge to specialize in new approaches to personalized medicine and implementation of new innovative research initiatives to accelerate treatments to patients. She serves as SVP and GM of the Global Rare Disease Program at WuXi NextCODE and Founder/CEO of RARE Science, a non-profit research organization that accelerates discovery of therapeutic solutions for kids with rare disease. She serves as a Scientific Advisory board member, for Global Genes, which focuses on rare disease advocacy. Dr. Waters received her PhD in Genetics from UC Davis and was a Postdoctoral Scholar and Associate of the Howard Hughes Medical Institute, California Institute of Technology. Dr. Waters completed an NIH Postdoctoral fellowship at University of California, Berkeley, and received a BS degree from San Diego State University. Dr. Waters received her MBA from UCLA.
Presentation: Accelerating cures for rare childhood diseases by building global patient communities and iPSC collections
Creating inducible Pluripotent Stem Cell (iPSC) collections of rare patients scattered world-wide can lower the barrier of biological discovery of rare disease, provide a platform for potential drug repurposing and seed new drug development programs where there is a great unmet medical need. Currently approximately 8,000 rare diseases have been identified that affect close to half billion people world-wide of which only 5% have available therapies. More than half, hundreds of thousands, of children have rare disease of which 30% die before their 5th birthday. Two main challenges are (1) access to genetic sequencing and (2) if sequencing is available only 30-50% of the time the etiology is identified that leads to clinical intervention. Even with identification of a causal gene, understanding how the gene, gene variants and variants in concert with other genomic mutations change biology and lead to a clinical phenotype is needed. In the first step, there is a need to find a critical number of families with the same rare disease which necessitates searching for them around the globe. Secondly, the creation and use of iPSC lines once families have been identified enable validation of variance of significance through techniques such as gene editing in addition to revealing causal biological pathways. This methodology can accelerate identification of therapeutic options in the immediate addressing the urgency needed in addition to seeding longer-term, new targeted drug development programs.
For Research Use Only. Not for use in diagnostic procedures.