City of Hope Shared Resource for Human Embryonic Stem Cell Research in Developmental Biology, Cancer and Regenerative Medicine
Researchers at our institution are increasingly including human embryonic stem cells (hESCs) in investigations of development, regeneration and disease. A hESC shared research resource (hESC Core) will be a valuable and cost effective mechanism to enable researchers to begin work with hESCs, facilitate ongoing efforts, provide a central source for high quality established and newly-derived hESC lines that are not on the list of federally approved lines, and assist researchers with evaluation and selection of hESC lines intended for specific purposes. The facility will also provide access to specialized equipment essential for conducting experiments with hESC lines, which would not otherwise be accessible to researchers working with non-federally approved lines. The pilot projects described in this proposal are examples of efforts employing hESCs and originate in multiple parts of the institution. Projects to understand the origins of genetic abnormalities that accumulate in hESC lines in culture will help ensure that hESC-based therapies are safe. Projects that are developing models of human cancer are creating platforms for basic studies of cancer and for discovery of novel therapies. Projects that are designing strategies for differentiation of hESCs to specific fates will make possible use of hESCs in cell replacement therapies for patients with degenerative diseases, diabetes and cancer. The hESC Core will be responsible for maintenance and propagation of existing hESC lines and derivation of new lines using strict quality assurance. The Core will also be devoted to facilitating studies involving genetic modifications of hESCs and directed differentiation by manipulation of microenvironmental cues. These functions will be required for creation of cancer models and development of cell replacement therapies. In addition, the Core will provide space for investigators constrained by federal funding restrictions or institutional resources. The Core will also provide cells and laboratory space for researchers from neighboring California State University Campuses who have submitted projects that would use this Core and that mesh well with existing programs. Beyond being a starting point for investigators initiating hESC-based projects and an active partner in ongoing projects, this hESC Core will be a shared resource useful to all California scientists interested in hESC research. We anticipate collaboration with other CIRM-funded hESC shared research laboratories, sharing capabilities, techniques and cell lines to maximize leverage of CIRM funds.
Statement of Benefit to California:
The hESC shared resource will play an essential role in the study of hESC biology and investigations of their utility in understanding human disease and developing new therapies. Using hESCs, COH researchers aim to minimize the tumorigenic potential of hESCs and to develop a better understanding of human cancers of the blood and brain, and to develop innovative cell replacement therapies for treatment of diabetes, hematological disorders, skeletal disease and muscular abnormalities. The hESC Core will generate new hESC lines that may be better suited to specific applications compared with existing lines, and that will be derived and maintained under conditions consistent with potential future clinical application. COH, as a leader in hematopoietic and islet cell transplantation, has unique scientific strengths. Our HCT Program is the largest in California and is at the forefront of developing new cellular and biologic treatment for hematological disorders. We also have demonstrated leadership in selection, culture and manipulation of cells under GMP conditions for application to investigator initiated IND-based trials. Our experience and expertise in taking discoveries from bench-to-bedside places us in an outstanding position to do the same for hESC-based discoveries. This experience directly benefits the proposed hESC Core, since its efforts to maintain and derive hESC cells require strict conditions of quality assurance. As the key resource for planned and existing research projects, the Core will facilitate entry of talented new investigators into investigations and applications of hESCs. It is important to note that this facility will serve neighboring institutions (e.g., those in the California State University system), allowing these scientists access to hESC research which is otherwise not possible. We anticipate collaboration with other hESC core facilities—sharing technologies, capabilities and cell lines to maximize the application of CIRM funds.
SHARED LABORATORY SYNOPSIS OF PROPOSAL: This application is for a shared laboratory that will provide economies of scale in maintaining existing stem cell lines of high quality. The budget request is for ~$790,000 to develop a 900 square foot core facility for human embryonic stem cell work. The laboratory will derive new human stem cell lines to replace older lines of diminishing quality and to derive clean lines that have not been exposed to animal products. The core will also support studies that involve genetic modifications of human stem cells to create animal disease models and develop tactics for stem cell replacement therapies. There are no plans for a stem cell course. QUALITY AND IMPACT OF THE SCIENCE: The goals of the proposed research are to understand the growth and nature of human embryonic stem cells (hESC) to create humanized mouse therapy models, and to define conditions to direct differentiation of hESCs for cell replacement. Four pilot projects are described. Timothy O’Connor (Division of Biology) will examine genetic instability in hESCs by testing the hypothesis that DNA repair mechanisms are relatively inefficient in stem cells and that a backlog of unrepaired genetic lesions in cycling stem cells sets the stage for cancer. This is an important aspect of the maintenance of hESCs, and demonstrating genomic stability of hESCs will be critical for future therapeutic applications. Drs. Barish and Bhatia will use stem cells to explore the cancer stem cell hypothesis, which holds that the malignant phenotype of solid tumors is fueled by a relatively quiescent population of developmentally stalled tissue progenitor cells (cancer stem cells). Dr. Michael Barish will examine overexpression of PDGF in combination with tumor suppressor genes in hESCs to develop models for gliomas and other brain tumors. Dr. Ravi Bhatia will express BCR/ABL in HSCs derived from hESCs to better model chronic myelogenous leukemia. As an example of research that will focus on tactics for stem cell replacement therapies, Dr. Carlotta Glackin will manipulate TWIST-1 in hESC derived-mesenchymal stem cells in order to study the mechanisms by which these cells may regenerate bone tissue. The program director (PD) Dr. Barberi will develop methods for converting stem cells into skeletal myoblasts for treatment of muscular dystrophies. Dr. Theresa Ku will attempt to generate transplantable pancreatic beta cells from hESCs. All of the scientific projects are clearly described, and while the quality of experimental approaches and efforts are variable, the studies have applications to important biomedical questions. In general, the science is good, and if successful, the proposed studies will have significant clinical/translational impact. One major concern is that the principal investigators named are very junior and have by and large been unsuccessful in raising support for their research via NIH peer reviewed mechanisms. In addition, the need for newly-derived cell lines seems contrary to the need to expand the hESC-competent research base at City of Hope and the current level of expertise of the staff. The ongoing research efforts with hESCs are modest, with few grants from any major funding agencies, and most hESC projects are currently in the planning stages. However this argument could be turned around and used as justification for a shared research laboratory to leverage the career development of these young scientists. The program director, Dr. Tiziano Barberi, has excellent training. He received a D.Sc. in Cell and Molecular Biology from the State University of Rome, Italy in 1997, and the PhD in Cell Biology from the University of Tubingen, Germany in 2005. He has been an assistant professor in the Division of Neurosciences at the Beckman Research Institute of City of Hope since 2006, and his publications on hESC studies from a postdoctoral career in the Stem Cell and Tumor Biology group at the Sloan-Kettering Institute appear in Nature Medicine, Nature Biotechnology, and other journals. Prior managerial experience is not described. One PI listed has been approved for a CIRM SEED grant to investigate the mechanisms of genetic stability of hESC, and another PI will apply extensive experience directing mouse ES cell-differentiation to hESCs. APPROPRIATENESS OF SPACE AND EQUIPMENT TO SCOPE OF PLAN: The institute apparently does not have NIH-free space already available for this work. However the institution has committed ~ 2,000 square feet of a conventionally designed laboratory wing to stem cell research. This lab would be a boon to the nascent human ES cell group at City of Hope; however, there is no convincing justification for NIH-free lab space, because there is no clear argument that any of the projects require hESC that are not in the NIH registry. Although the need for fresh hESC lines is a useful general argument for NIH-free space, there is no specific need listed in any of the immediate projects, and creating new ones likely would take talent and resources from other projects & blunt the progress of this small group. Importantly, no source of embryos is apparent. The plan is to set aside ~900 square feet for the stem cell core facility described here, and an institutional letter from Dr. Arthur Riggs, Director of the Beckman Research Institute, confirms that if this application is funded it will commit the space, provide matching funds (as required), support the Core Manager, and provide reasonable additional operating funds. Against this backdrop, the space and equipment plans seem appropriate, but the commitment appears to be contingent upon CIRM funding. The shared lab is intended as a means to support the pilot projects of five PIs by providing technical help, supplies, and equipment. The size and scale of the proposed shared lab are more than enough to support the number of users. The lab would maintain and propagate hESC lines and derive new ones, and test for chromosome stability and loss of teratogenicity. The staff will help with genetic modification of hESCs to create cancer models and help develop culture conditions for directed differentiation of cells for potential replacement therapies. The core will be equipped with biosafety cabinets, CO2 incubators, microscopes, flow cytometers, inverted fluorescent microscope, and other equipment necessary for this research. The equipment and personnel should be adequate for the proposed studies. The host lab will acquire the infrastructure and support to enhance greatly hESC work for three hESC researchers and encourage four others to begin. Interested researchers at nearby California State Universities are anxious for access. NIH-free space is assigned for hESC research, but renovations and support for infrastructure are required; thus, CIRM funding is requested to upgrade this space to a functional core facility. QUALITY OF MANAGEMENT PLAN: The key concern is that the program director is quite junior and has no prior administrative experience. The facilities will be made available to qualified investigators with demonstrated productivity (funding and/or publication track record) and a documented inability to use “approved” cell lines for their work. The management plan to support, manage and maintain the lab & equipment, while carefully described, is not adequate because the duties and responsibilities for each staff member are not addressed well. Dr. Barberi, who will devote 20% effort to running the CIRM Shared laboratory, is an experienced investigator in studies that relate to stem cells, but is a newly appointed assistant professor just beginning an independent research laboratory. The PD has no experience or training in directing a multi-user facility, and his fitness to manage this Core are not well described. City of Hope has a track record of several multi-user facilities operating successfully and provides considerable institutional support, although details are not provided here, and the nature and amount of administrative support is not stated. The Director of the Analytical Cytometry shared resource at City of Hope could be an excellent advisor for operations, but the lab manager and technician have not yet been identified thus their qualifications cannot be assessed. In this regard, it is unclear who will provide expertise for the lab services. For example who will train or perform the lentivirus transductions, and who will generate new human lines, much less acquire the embryos? The need for and mechanisms to acquire appropriate institutional approvals are not well addressed. The Oversight Committee will include Dr. Susan Kane, a senior member of the City of Hope faculty; an individual with experience in shared facilities operations; a stem cell biologist; and two ad hoc members. As described, half of the Oversight Committee is comprised of major users of the lab, including the program director. This may compromise decision making since the oversight committee of four investigators chaired by Dr. Barberi will review and prioritize formal research proposals to share the facility and ensure that only qualified investigators from the state of California use the facility. Plans are in place for investigators from Cal State Fullerton and Cal State Los Angeles to access the CIRM lab should it be funded. Some of the potential users outside City of Hope have excellent scientific credentials, and while these investigators from neighboring institutions can gain access through a formal application process, neither the nature of the process nor the assignment of access priority to investigators with prior experience in hESC work are described. It appears that the Beckman Research Institute of the City of Hope is willing to commit adequate laboratory space, the matching funds required by CIRM, salary support for the Lab Manager, and vaguely reasonable operating funds beyond those requested in the application for purposes unspecified. DISCUSSION: In this application, the PD proposes a generic shared lab study plan with two components: 1) a core to derive new ESC lines in order to replace old lines that may have contaminating animal products, and 2) a research center for SC work on DNA repair, replacement strategies for neurodegenerative diseases, etc. The application is carefully prepared and instructive to read in referencing the nuts and bolts of dealing with stem cell problems, but is relatively weak scientifically and has an inadequate management plan. The PD is a very good young scientist who recently completed a post-doctoral fellowship and is now a junior assistant professor. Reviewers believe that it is somewhat unreasonable for a new assistant professor to have the responsibility of this large management task in addition to the pressures of a new faculty position. The co-investigators who will use this facility are similarly junior in that few have a track record of peer-reviewed hESC work; in fact, only one investigator has funding from a major granting agency. There was no convincing justification for the need for NIH free space. This shared lab potentially could have a bigger impact here than other places, but this is an application to fund pilot projects for five investigators with less of a focus on building a shared lab space that would be widely useful to others. Among other concerns, the applicants propose services to be provided by the shared lab including line derivation and lentiviral transduction without making it clear how this will be implemented since no one in the application has done this work previously. Also, there was some question about the institutional commitment. Reviewers note that the space will be provided only if the shared lab is funded. Moreover, in looking at the space plan, the space to be provided appears be superimposed on Dr. Barberi’s own lab. Overall, the panel felt that the lack of experience and lack of experience in ESC work significantly dampens enthusiasm.