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CL1-00501-1: CIRM Shared Research Laboratory for Stem Cells and Aging

Laboratory Recommendation: Recommended for funding
Laboratory Score: 75

Course Recommendation: Recommended for funding
Course Score: 72

Public Abstract (provided by applicant)

Age-related diseases of the nervous system are major challenges for biomedicine in the 21st century. These disorders, which include Alzheimer’s disease, Parkinson’s disease, Huntington’s disease, amyotrophic lateral sclerosis and stroke, cause loss of neural tissue and functional impairment. Currently, there is no cure for these devastating neurological disorders. A promising approach to the treatment of age-related neurological disorders is cell therapy, i.e., transplantation of nerve cells into the brain or spinal cord to replace lost cells and restore function. Work in this field has been limited however, due to the limited availability of cells for transplantation. For example, cells from 6-10 human fetuses obtained 6-10 weeks post-conception are required for one patient with Parkinson’s disease to undergo transplantation.

Human embryonic stem cells (hESCs) offer a potentially unlimited source of any cell type that may be required for cell replacement therapy, due to their remarkable ability to self-renew (they can divide indefinitely in culture) and to develop into any cell type in the body. In this proposal, we will build out approximately 3400 square feet of shared laboratory space within our existing research facility for hESC research, as well as approximately 2400 square feet for classroom facilities dedicated to training in hESC culture and manipulation. We seek to understand how hESCs differentiate into authentic, clinically useful nerve cells and will use novel molecular tools to examine the behavior of cells transplanted in animal models of human neurological disease. We will also need to develop a noninvasive method of following cells after transplantation and we propose to develop luciferase-tagged (light-emitting) hESC lines for in vivo animal imaging. In addition, we will use hESC-derived nerve cells to screen drug and chemical libraries for compounds that protect nerve cells from toxicity, and to develop in vitro disease models. We believe that these experiments are critical to enhancing our understanding of neurological diseases and providing the tools that will be necessary to move cell therapy to the clinic.

Before a hESC-based therapy can be developed, it is essential to train scientists to efficiently grow, maintain and manipulate these cells. We propose to teach four 5-day hands-on training courses – two basic and two advanced hESC culture courses per year – to California scientists free of charge. These courses will provide scientists with an understanding of hESC biology and will enable them to set up and conduct hESC research after completion of training.

In summary, the goal of this proposal is to provide over twenty investigators at the home institute and neighboring institutions with the ability to culture, differentiate, and genetically manipulate hESCs – including clinical-grade hESC lines – to develop diagnostic and therapeutic tools.

Statement of Benefit to California (provided by applicant)

We propose to build a Shared Research Laboratory and offer a Stem Cell Techniques Course for over twenty principal investigators at the home institute and neighboring institutes working collaboratively on stem-cell biology and neurological diseases of aging. We propose to: 1) Purify nerve cells at different stages of maturation from human embryonic stem cells and to develop transplantation strategies in animal models that mimic human diseases, including Parkinson’s disease, stroke and spinal cord injuries; 2) Screen drug and chemical libraries for reagents that protect nerve cells from toxicity and develop in vitro disease models using nerve cells generated from human embryonic stem cells; and 3) Assess the long-term integration and differentiation of transplanted cells using a non-invasive imaging system.

We believe these experiments provide not only a blueprint for moving stem-cell transplantation for Parkinson’s disease toward the clinic, but also a generalized plan for how stem-cell therapy can be developed to treat disorders like motor neuron disease (amyotrophic lateral sclerosis, or Lou Gehrig’s disease) and spinal cord injury. As the only stem-cell research facility in California’s 10-12 most northwest counties, we are uniquely positioned to extend the promised benefits of Proposition 71 to this large part of the state. The tools and reagents we develop will be made widely available to California researchers and we will select California-based companies for commercialization of any therapies that may result. We also hope that California-based physicians will be at the forefront of translating this promising avenue of research into clinical applications. Finally, we expect that the money expended on this research will benefit the California research and business communities, and that the tools and reagents we develop will help accelerate stem-cell research in California and worldwide.

Scientific Review Summary of Part One Application

SHARED LABORATORY

SYNOPSIS OF PROPOSAL: The program director (PD) is an assistant professor with strong human embryonic stem cell (hESC) expertise who will head a 3,000 sq ft shared research laboratory (SRL) and another nearly 2,500 sq ft for the hESC techniques course. The PD, who will have major responsibility in the management of the facility, will get support from a senior investigator with experience in managing large projects. A lab manager and technical staff with hESC experience have also been identified. State-of-the-art hESC technology is assured through the close collaboration with a well-known scientist. Considerable space commitments have been made by the institution. Plans are presented for space allocation to intramural and extramural investigators. Access of qualified users will be controlled using an experience-based badge system. Use of National Institutes of Health (NIH) and non-NIH lines including a Good Manufacturing Practice (GMP) line is proposed. The investigators who would use the lab have a focus in neural disease, degeneration and aging.

QUALITY AND IMPACT OF THE SCIENCE: The proposed research goals are very relevant to hESC research with a particular focus on neural differentiation, hESC-based neural disease modeling and in vivo imaging of hESC-derived neural progeny. While the choice of topics is strong and appropriate to the expertise of the investigators, some of the proposed studies and the feasibility thereof are less developed. Many projects seem to be at the early planning stages and the overall success will be heavily dependent on the contribution of the PD. As a recent junior faculty member, the PD may be overwhelmed by the large responsibility and workload.

The focus on neurodegeneration and aging provides the application with clear programmatic strength. Another area of strength is the state-of-the-art hESC technology available particularly in the area of genetic modification through the collaboration and expertise developed originally in one collaborator’s lab. However, there is very little detail provided on how other key goals of the proposal will be pursued. For example, the development of hESC-based high throughput (HTP) assays is discussed but it is unclear how this goal is going to be pursued and implemented. Similarly, the imaging aspects of the proposal need to be better developed. The PD has published extensively in an hESC disease-oriented field and has developed with his/her collaborator strategies for genetic modification of hESCs including site directed mutagenesis and homologous recombination. The PD has also worked with another collaborator on targeted differentiation and in vivo transplantation in disease models. While the PD has published extensively, most of the contributions were rather technical in nature. There is a participating senior investigator who is leader of the stem cell effort at the home institute. This PI has a strong publication/ funding record and has worked on endogenous neural stem cells (NSCs), but as yet does not have direct experience in hESCs. Another PI at the home institute has been approved to receive CIRM funding for studying programmed cell death in hESCs. Other resident investigators are excellent researchers in the area of Parkinson’s Disease and Huntington’s Disease (HD) but without hESC experience yet. Similarly, one investigator with C. elegans experience and another involved in Parkinson’s Disease modeling may become important assets for the project but also have not yet worked with hESCs. A close interaction with one well-known collaborating PI will ensure that the facility is at the forefront of hESC technology. Other outside expertise includes a PI from a neighboring institute with experience in xeno-free derivation of hESC.

In summary: A large group of investigators at the home institution will utilize the planned laboratory. They consist of a mix of senior, mid- and junior level scientists, many of whom are well funded and have significant publication records. While there is a mix of research projects, the majority involve age-related diseases of the brain, the aging process itself, cell death and potential cellular therapies in the neurodegenerative diseases using neurons derived from hESCs. Many of these are important scientifically. They will use the most current techniques in their work on hESCs. One would expect there to be a significant number of publications to be generated from this shared hESC facility, some of which will be high impact.

APPROPRIATENESS OF SPACE AND EQUIPMENT TO SCOPE OF PLAN: It is clear that the applicant institution strongly supports this proposed development, in part as evidenced by the amount of space to be utilized. The applicant proposes a huge amount of space earmarked for the project including > 3,300 sq ft for the main facility and another ~ 2,300 sq ft for use as classroom facilities. There is currently no space for non-NIH lines available in the institute. This is very generous and should be very adequate for a multi-investigator lab. As they have to start from scratch in creating a new hESC lab removed from the rest of the institute which is federally funded, they need to buy all the new equipment to run the new lab. While this is expensive, it will likely be the current cost of establishing a new lab that can handle non-federally approved cell lines. One reviewer felt that starting a facility of such a scale with relatively limited evidence for use by a broad user base seems over-ambitious. A critical strength of the application is the fact that the staff running the facility will already have significant hESC expertise, and that the hESC lines proposed include a broad range of registered and non-registered lines including access to a line apparently derived under GMP conditions, although no details were provided. There is a plan to open the facility to investigators in the neighboring counties. While they plan to open their facility to investigators in other near-by institutions, there may be practical disadvantages given the travel time between them.

QUALITY OF MANAGEMENT PLAN: The PD would devote 25% time to the shared lab; given that the PD is a junior investigator, directing this facility could be quite tough; however a day-to-day lab manager with hESC experience has been identified. The oversight will be the primary responsibility of the PD (25% effort) and a senior investigator from the home institution (5% effort). They have an experienced stem cell scientist in charge of the day-to-day management. The oversight committee is composed of the PD, the senior investigator, the institute’s president, the chair of Institutional Animal Care and Use Committee (IACUC), a member of the imaging core, and the director of compliance. This is a balanced composition though it is unclear how often the committee will meet. Overall the management and oversight plan is good – a clear strength. There is a clear discussion of how space will be allocated among intramural and extramural investigators and the proposal includes details such as discussing proposed bench allocation between intramural (24 spots) versus extramural (12 spots) investigators. On the other hand, there is not much detail on the exact services that are going to be provided beyond the lab space. For example, there is no discussion of generating master banks of hESCs and feeders or formal training sessions for internal investigators. The proposed list of hESC lines that will be available is good. There will be good access to state-of-the-art technology through a collaboration with a PI who is at the forefront of technology development including technology for genetic modification of hESC.

The lab will be run as an independent entity from the institute to assure proper separation of costs with NIH funding. Major equipment is requested including a FACS sorter, electrophysiology equipment, and a molecular imaging system. While this is a rather expensive approach and will duplicate equipment available to investigators in their own labs, this strategy will allow clearer separation of NIH and non-NIH work. There was no discussion in the application on how they will handle other core services such as genomics, proteomics or HTP services (for which the labs do not seem to be setup). Information regarding HTP aspects of the project are lacking.

DISCUSSION: The PD brings together strong expertise in hESC with collaborating investigators, but it might be tough for this relatively junior PD to manage this large (3000 sq ft) lab. The structure seems to be well managed, and the interactions with the PI at the forefront of technology development raise enthusiasm, although there is a question about how much time and effort s/he will be able to contribute. There is a good access plan for this state-of-the-art facility. The body of investigators is quite strong, and the focus on aging and neurodegenerative diseases matches well with their key goals in translational assays and imaging. They have identified a day-to-day manager already, and there is a good description of the oversight plan with IACUC and other committees. This is a key strength. The use of space is also well-described, and there are good extramural investigators listed in the proposal.

Despite the well-organized group, this is a stand-alone institution which may make it harder on them to be successful. There is a good list of cell lines, demonstrated expertise in hESC, and they have been approved for CIRM funding. The main concern is that it will be tough on the PD to run this big facility, which at least initially seems to be over-dimension in scope. That said, this facility likely would benefit other groups. The institute is recognized by the National Institute on Aging of the National Institutes of Health as a Research Center of Excellence. A question was raised regarding the relative isolation of the institute, which is relatively new. One reviewer who did some further background research was impressed by the PD, the investigators, and their enthusiasm, and noted that the people involved have strong CVs. This application should be compared alongside other institutions in terms of how well they can manage the shared facility and the quality of the science.

PROGRAMMATIC REVIEW: A motion was made to recommend this Shared Research Laboratory application for funding. The significant issues were: 1) that the PD is a young investigator and there is concern over taking on this significant responsibility, and 2) the quality of research at the institute. The first concern is valid, but the second is not because the science is strong and focused at this institute. One of the discussants noted that the Institute has prominence in the area of aging – it was pointed out that the institute is recognized by one arm of the National Institutes of Health as a Research Center of Excellence. Also it was noted that the investigators involved in the projects that will use the SRL are outstanding scientists. This application could make or break the institute’s dedication to stem cell research, and there are a lot of very serious, very well-qualified scientists there. There was justification for using non-federal lines because the federally approved lines are not usable for clinical purposes. A comment was made that the proposed staff are experienced with hESC and that much of the day-to-day work is routine and these people could presumably do that. Also this SRL would serve to bring new people into hESC research by providing a place to work at a site that is somewhat distant from the other research centers in the area. Another reviewer was impressed by the dedication and enthusiasm of the PD and believes that s/he will make a contribution and will be able to manage a shared facility. Having the senior co-PD is reassuring and could help with management. Interactions with the well-known collaborating PI are considered a big asset. The motion to recommend this Shared Research Laboratory application for funding passed.

TECHNIQUES COURSE

QUALITY OF THE PROPOSED TECHNIQUES COURSE: The plan is to offer two courses, one is basic and the second advanced. Each course is to be given twice a year with four courses in total. The PD has significant experience in instruction and leadership of ESC courses and has already run a course at the home institute in 2006. Faculty and organization of the course are very strongly influenced by a collaborating PI who is well-known for hESC technology development. The names and participation of additional senior faculty would have strengthened the application. These will be 5-day courses teaching basic hESC handling and differentiation. The course schedule is quite reasonable and based on a similar course that they taught in 2006. Differentiation techniques to be taught are somewhat limited with focus primarily on differentiation of the central nervous system (CNS). There could be a broader focus on other tissue types such as mesendodermal differentiation with the inclusion of appropriate faculty. The-5 day courses seem rather short for learning how to work with hESCs. Plans are provided for advertising the event and for outreach and follow up. Follow up includes continued troubleshooting per phone and e-mail, distribution of a CD containing the information of the courses, and the establishment of a network of former course participants. Courses will be provided free and access will be provided on a first come / first served basis. However, possible selection criteria are discussed especially for the advanced course.

QUALIFICATIONS OF THE INSTITUTION: The institution is qualified to carry out the course and has successfully held such a course in 2006. There is clear commitment to stem cell research and to developing a leadership position in the field of stem cells and regenerative medicine particularly in the area of neurodegeneration and aging. Very generous availability of space is another sign of the commitment of the institution. The location is within reasonable reach to a broad community of potential investigators.

DISCUSSION: The course plan is well-designed, well-planned, and well-written up. This is a 5-day hESC handling and differentiation course with a well-known collaborating PI and other instructors. There are two modules which run from 9am to 5pm. One reviewer would like this to be longer. The institute has done this type of course in the past, and it would be nice to have seen the evaluations of last year’s course. One reviewer would like broader expertise outside of the CNS. Others state that the neurocentric course is good because that is where their expertise lies. There are other courses available, and the instruction can be found elsewhere. The course layout including how it would work, be managed, be advertised and its basic set-up were well described. There was a clear commitment of 2,500 sq ft space allocation.

PROGRAMMATIC REVIEW: A motion was made to recommend that this Techniques Course application not be funded. While there was concern with the PD already being overloaded by his/her responsibilities for the SRL, that is not necessarily the case for the course. A well-known collaborating PI is heavily involved in the course which will take some pressure off the PD. Reviewers felt that s/he has the technical expertise and that the course is good. The main concern was that this collaborating PI resided on the East coast. The motion to recommend this Techniques Course application not be funded failed. A second motion was made to recommend this Techniques Course application for funding. The motion to recommend this Techniques Course application for funding passed.

The following Grants Working Group members had a conflict of interest with this application and were therefore recused from participating in review of, discussion of, and voting on the application:

• Feit, Marcy
• Lansing, Sherry
• Sheehy, Jeff
• Steward, Oswald
• Young, Dr. Wise