Gene editing, particularly using the CRISPR (Clustered Regularly Interspaced Short Palindromic Repeats)/Cas system, has very rapidly established itself as an important tool in drug discovery and is now being exploited for therapeutic purposes as well. Cambridge Healthtech Institute’s Fifth Annual CRISPR for Precision Medicine symposium will bring together scientists and clinicians to talk about the recent progress made in gene editing and its potential going forward. At the same time, they will also discuss what is being done to overcome some of the inherent challenges that exist in terms of guide RNA design, delivery and off-target effects associated with CRISPR/Cas9, and what are some of the alternatives being developed? Experts from pharma/biotech, academic and government labs, and technology/service companies will share their experiences leveraging the utility of CRISPR-based gene editing for diverse applications such as creating cell lines and disease models for functional in vitro, in vivo and ex vivo screening that will ultimately pave the way for better and safer therapeutics.

Final Agenda

 

Thursday, March 14

7:00 am Registration Open and Morning Coffee (Golden Gate Foyer)

ENGINEERING PRECISION AND ELIMINATING OFF-TARGET EFFECTS
Golden Gate 7

8:25 Chairperson’s Opening Remarks

Kevin Davies, PhD, Executive Editor, The CRISPR Journal, Mary Ann Liebert, Inc.

8:30 Introduction: The CRISPR Craze

Davies_KevinKevin Davies, PhD, Executive Editor, The CRISPR Journal, Mary Ann Liebert, Inc.

From 60 Minutes to Last Week Tonight, CRISPR is well and truly in the public eye. 2018 saw an explosion in the number of CRISPR publications, exciting breakthroughs in CRISPR biology, diagnostic and therapeutic applications of gene editing, and the evolution of base editing technology. But many technical and experimental concerns were also highlighted, leaving no doubt that CRISPR gene editing has many rivers to cross to fully realize its potential. In this brief overview, I will survey the CRISPR landscape, including recent landmark reports and future challenges.

9:00 Engineering Genome-editing Enzymes for Tissue-specific Therapeutic Delivery

Ross WilsonRoss Wilson, PhD, Project Scientist and Principal Investigator, Innovative Genomics Institute, University of California Berkeley

The advent of CRISPR-derived genome-editing enzymes such as Cas9 has transformed biomedical research. Despite the apparent opportunity for an impending revolution in the clinic, several substantial barriers must be overcome before the promise of therapeutic genome editing will become a reality. A primary challenge is that of delivery: cells are exceptionally well-evolved to prevent the entry of macromolecular machinery necessary for editing. To address this need, the Wilson Lab is engineering the Cas9 enzyme itself to allow cellular penetration and editing following molecularly-targeted uptake into cells. 

9:30 Integrating Novel Advances in Gene Delivery and Genome Engineering for Therapeutic Application

Schaffer_DavidDavid V. Schaffer, PhD, Professor, Chemical and Biomolecular Engineering, Bioengineering, Molecular and Cell Biology, Helen Wills Neuroscience Institute; Director, Berkeley Stem Cell Center, University of California, Berkeley

There have been an increasing number of successful human gene therapy clinical trials, but vectors based on natural versions of the AAV viruses used in many trials face a number of delivery challenges that limit their efficacy. We have been developing a high throughput approach termed directed evolution to engineer highly optimized variants of AAV for a broad range of cell and tissue targets, for delivery of CRISPR/Cas9 and other therapeutic cargoes.

Synthego 10:00 Improving Genome Editing in Difficult Cell Types through Guide Design and Automation

Wronski_AnnaAnia Wronski, PhD, Product Manager: Engineered Cells, Synthego Corporation


10:30 Coffee Break in the Exhibit Hall with Poster Viewing

11:15 Technologies Enabling Detection and Precision of Gene Editing

Dresios_JohnJohn Dresios, PhD, Senior Biology Director and Chief Scientist, Advanced Solutions Group, Leidos

In an effort toward agnostic detection of genome editing, we undertook a comprehensive approach for sample interrogation at multiple levels of genome organization, ranging from primary DNA sequence to higher structural features, including DNA-protein interactions, chromatin accessibility, and tertiary chromatin folding. The results showed that multidimensional DNA organization analysis provides a sensitive approach for identifying genomic perturbations arising from genome editing and can be utilized for biomedical and biosecurity applications.

11:45 Broad Spectrum and Personalized HIV Excision/Inactivation Therapeutics

Wigdahl_BrianBrian Wigdahl, PhD, Professor and Chair, Department of Microbiology and Immunology, College of Medicine; Director, Institute for Molecular Medicine & Infectious Disease; Director, Center for Molecular Virology & Translational Neuroscience, Drexel University

HIV persistence during therapy is a major hurdle to a cure. Genomic editing techniques, like the CRISPR/Cas9/gRNA system, hold promise to excise or inactivate the integrated provirus. However, due to the infidelity of the viral reverse transcriptase, the virus in patients exists in reservoirs as a collection of distinct genomic variants, termed quasispecies. The evaluation of newly designed broad-spectrum gRNAs will be discussed with respect to targeting the HIV quasispecies.

12:15 pm Enjoy Lunch on Your Own

1:15 Session Break

BUILDING CRISPR-DERIVED DISEASE MODELS
Golden Gate 7

1:55 Chairperson’s Remarks

Lukas Dow, PhD, Assistant Professor of Biochemistry in Medicine, Department of Medicine, Hematology and Medical Oncology, Sandra and Edward Meyer Cancer Center, Weill Cornell Medicine

2:00 Redefining Animal Genome Engineering with CRISPR-Cas9 Technology

Guru_ChannabasaviahC. B. Gurumurthy (Guru), PhD, Associate Professor, Developmental Neuroscience, Munroe Meyer Institute for Genetics and Rehabilitation, Director, Mouse Genome Engineering Core Facility, University of Nebraska Medical Center

CRISPR-Cas9 has radically changed how the decades-old traditional transgenic technologies are practiced. Our lab has pioneered some breakthrough CRISPR based technologies to create complex animal models including long cassette knock-ins and conditional knockout mouse models. Our technological contributions, particularly Easi (Efficient additions with ssDNA inserts)-CRISPR and Genome editing via Oviductal Nucleic Acids Delivery (GONAD) have been widely adapted in the animal transgenesis field. I will discuss how the latest technological advances have redefined the traditional transgenic methods.

2:30 Building Disease Models with Single Base Pair Resolution

Dow_LukasLukas Dow, PhD, Assistant Professor of Biochemistry in Medicine, Department of Medicine, Hematology and Medical Oncology, Sandra and Edward Meyer Cancer Center, Weill Cornell Medicine

Comprehensive tumor sequencing has provided a comprehensive map of cancer-associated mutations. For instance, more than 350 distinct mutations have been identified in p53, while KRAS shows up to 9 different alterations within a single codon. But, are all mutations created equal? To begin to understand the consequences of distinct mutational variants we have built new animal models and optimized genetic tools that enable a detailed interrogation of cancer-associated SNVs.

3:00 Efficient Mouse Genome Engineering by CRISPR-EZ Technology

Lin HeLin He, PhD, Thomas and Stacey Siebel Distinguished Chair Professor, Molecular Cell Biology Department, University of California at Berkeley

CRISPR/Cas9 technology has transformed mouse genome editing with unprecedented precision, efficiency, and ease; however, the current practice of microinjecting CRISPR reagents into pronuclear-stage embryos remains rate-limiting. We thus developed CRISPR ribonucleoprotein (RNP) electroporation of zygotes (CRISPR-EZ), an electroporation-based technology that outperforms pronuclear and cytoplasmic microinjection in efficiency, simplicity, cost, and throughput. In C57BL/6J and C57BL/6N mouse strains, CRISPR-EZ achieves 100% delivery of Cas9/single-guide RNA (sgRNA) RNPs, facilitating indel mutations (insertions or deletions), exon deletions, point mutations, and small insertions. In a side-by-side comparison in the high-throughput KnockOut Mouse Project (KOMP) pipeline, CRISPR-EZ consistently outperformed microinjection. Here, we provide an optimized protocol covering sgRNA synthesis, embryo collection, RNP electroporation, mouse generation, and genotyping strategies. Using CRISPR-EZ, a graduate-level researcher with basic embryo-manipulation skills can obtain genetically modified mice in 6 weeks. Altogether, CRISPR-EZ is a simple, economic, efficient, and high-throughput technology that is potentially applicable to other mammalian species. 

3:30 Refreshment Break and Poster Competition Winner Announced in the Exhibit Hall

4:15 CRISPR/Cas9-Based Precision Medicine Approaches in Lung Cancer Research

Thales_PThales Papagiannakopoulos, PhD, Assistant Professor, Department of Pathology, New York University School of Medicine

Our laboratory investigates how genetic alterations in metabolic pathways, which are mutated in a large subset of lung cancers, promote tumor initiation and progression by rewiring cell autonomous and non-cell autonomous metabolic pathways and enable cancer cells to overcome metabolic bottlenecks. We use a combination of genetically-engineered mouse models, an accelerated CRISPR/Cas9-based experimental platform and biochemical approaches to identify metabolic liabilities that can be exploited using novel targeted therapies.

4:45 TECHNOLOGY PANEL: Latest in CRISPR Tools and Strategies

Symposium speakers and service providers come together to address current gaps in know-how and technology for CRISPR applications. They discuss current challenges, share best practices and their experiences using various CRISPR reagents and tools for research and therapeutic use.

Moderator: Kevin Davies, PhD, Executive Editor, The CRISPR Journal, Mary Ann Liebert, Inc.

Panelists: Paul Diehl, PhD, COO, Cellecta

Lukas Dow, PhD, Assistant Professor of Biochemistry in Medicine, Department of Medicine, Hematology and Medical Oncology, Sandra and Edward Meyer Cancer Center, Weill Cornell Medicine

C. B. Gurumurthy (Guru), PhD, Associate Professor, Developmental Neuroscience, Munroe Meyer Institute for Genetics and Rehabilitation, Director, Mouse Genome Engineering Core Facility, University of Nebraska Medical Center

Kevin Holden, PhD, Head of Science, Synthego

Thales Papagiannakopoulos, PhD, Assistant Professor, Department of Pathology, New York University School of Medicine

 

5:45 Reception in the Exhibit Hall with Poster Viewing

6:45 Close of Day

 

Friday, March 15

8:00 am Registration Open and Morning Coffee (Continental Foyer)

CRISPR FOR IDENTIFYING DRUG TARGETS & DISEASE PATHWAYS
Continental 9

8:25 Chairperson’s Remarks

Daniel P. Dever, PhD, Instructor, Laboratory of Dr. Matthew Porteus, Department of Pediatrics, Division of Stem Cell Transplantation and Regenerative Medicine, Stanford University Medical Center

8:30 CRISPR Gene Editing for Drug Discovery

John Feder, PhD, Associate Director, Leads Discovery and Optimization, Bristol-Myers Squibb

While much attention has been given to the therapeutic aspects of CRISPR genome engineering, over the past several years CRISPR has become a research tool that is now embedded into the drug discovery process. From new target discovery/validation to dissecting biological pathways, CRISPR allows for the creation of cellular reagents that are having an impact on drug discovery. An overview of how CRISPR is being used at Bristol-Myers Squibb will be presented.

9:00 Using Base Editing to Model and Treat Laminopathies in Patient-Specific iPSCs and Monkey Embryos

Chengzu_LongChengzu Long, PhD, Assistant Professor, The Helen and Martin Kimmel Center for Stem Cell Biology and the Leon H. Charney Division of Cardiology, Department of Medicine, New York University School of Medicine

CRISPR/Cas-derived base editing systems represent a powerful means of modeling and treatment of genetic disease. Here, we show that the base editing mediate efficient generation and correction of disease-causing LMNA mutations which are associated with several diseases, including Hutchinson-Gilford progeria syndrome and dilated cardiomyopathy in human stem cell models. Furthermore, we efficiently achieve base editing in cynomolgus monkey embryos, demonstrating the potential of this technology in non-human primate models.

9:30 Epigenome Engineering Using Targeted CpG Island Methylation Editing

Takahashi_YutaYuta Takahashi, PhD, Postdoctoral Fellow, Laboratory of Dr. Juan Carlos Izpisua Belmonte, The Salk Institute for Biological Sciences

DNA methylation plays major roles in defining cell identity and function during both embryogenesis and adult physiology. Aberrant DNA methylation is deeply involved in human diseases. However, a detailed understanding of this process has been impeded by a lack of robust tools for DNA methylation editing technologies. In this talk, I will present our technologies that allow for targeted and stable DNA methylation editing.

10:00 Combining CRISPR Editing and Cell Barcoding to Elucidate Gene Function and Advance Target Discovery

Paul Diehl, PhD, COO, Cellecta

Labeling of target cells with lentiviral barcode libraries offers an effective approach for monitoring cell phenotype in time-course experiments iusing single-cell molecular analysis. Further, cell barcodes can be incorporated with CRISPR sgRNA libraries to identify phenotypic changes in progeny cells.

10:30 Coffee Break in the Exhibit Hall with Poster Viewing

CRISPR-BASED ENGINEERING OF T CELLS & STEM CELLS
Continental 9

11:15 Gene Correction in Hematopoietic Stem Cells for the Treatment of Blood and Immune System Diseases

Dever_DanielDaniel P. Dever, PhD, Instructor, Laboratory of Dr. Matthew Porteus, Department of Pediatrics, Division of Stem Cell Transplantation and Regenerative Medicine, Stanford University Medical Center

Hematopoietic stem cell (HSC) transplantation has the power to cure diseases of the blood and immune system. The CRISPR-Cas9 system now enables targeted manipulations of the HSC genome that have revolutionized the concept of correcting disease-causing genetic mutations in patient-derived autologous HSCs. We outline therapeutic programs focused on gene correction in autologous HSCs from the discovery stage to the preclinical stage with the ultimate goal of developing novel cell and gene therapies for serious genetic blood disorders.

11:45 Therapeutic Non-Viral Genetic Engineering of Human T Cells

Roth_TheodoreTheodore Roth, MD/PhD Student, Laboratory of Dr. Alexander Marson, University of California, San Francisco

Decades of work have aimed to genetically reprogram T cells for therapeutic purposes. The need for viral vectors has slowed down research and clinical use. We have developed a CRISPR-Cas9 genome-targeting system that does not require viral vectors, allowing rapid and efficient insertion of large DNA sequences (greater than one kilobase) at specific sites in the genomes of primary human T cells. We have applied non-viral genome targeting to enable rapid and flexible therapeutic engineering of primary human immune cells for autoimmune and cancer cellular therapies.

12:15 pm Engineering the Blood to Treat Complex Multi-Systemic Diseases: Correction of Lysosomal Storage Diseases Using Human Genome-Edited Hematopoietic Stem Cells

Natalia-Gomez-OspinaNatalia Gomez-Ospina, PhD, Assistant Professor, Department of Pediatrics, Stanford University

Lysosomal storage diseases (LSDs) are a large group of genetic disorders, many of which lack effective treatments. Most are caused by deficiencies in enzymes resulting in progressive multi-systemic disorders marked by neurologic and musculoskeletal deterioration. A potential treatment approach is to engineer the patient’s own hematopoietic system to secrete the enzyme via autologous transplantation of genome-edited cells. Additionally, a genetic strategy where the enzymes are targeted to safe harbor loci, constitutes an adaptable “one-size-fits-many” approach.

12:45 Close of Symposium

Register