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CL1-00510-1: Shared facilities for the preclinical testing of embryonic stem cell therapies

Laboratory Recommendation: Not recommended for funding

Public Abstract (provided by applicant)

Blood Systems Research Institute (BSRI), located in San Francisco, has a 45-year history of research in transfusion medicine and related fields. BSRI has undergone a recent expansion in investigators, laboratory space and equipment to address the growing number of opportunities that exist in the broad field of cellular therapeutics, particularly in the area of human embryonic stem cell (hESC) technology. The notable strength of BSRI lies in the expertise of its investigators in clinical and translational research owing to its long history with cellular therapy in the form of blood cell transfusion.

Laboratory space at BSRI will be remodeled to support our program in hESC research. We request funds to improve two tissue culture rooms to support basic research and pre-clinical studies on hESC lines. A 3rd laboratory will be converted into a cell isolation and analysis laboratory housing equipment used in the isolation and analysis of hESC-derived cells and tissues. A 4th room will be converted into a vivarium to study stem cell transplantation.

The research facility will be used by scientists at BSRI and neighboring institutions in the San Francisco Bay Area to perform basic research and to develop methodologies appropriate for the clinical of application of hESC-based therapies. Therapies under investigations include developing blood and liver stem cells as a new source of tissues to treat birth defects and disease. A training program for research technicians is also included which will help train new workers in the stem cell field.

Statement of Benefit to California (provided by applicant)

The shared stem cell facility will help develop a new cell therapies such as those based on blood and liver cells. Blood diseases such as sickle cell anemia and thalassemia as well as liver diseases caused by viral infection, drugs or inherited disease affect many thousands of Californians. Often, transplanting healthy cells offers treatment or a cure for many of these diseases, but a lack of available or suitable donor tissue prevents such therapy in many cases. Embryonic stem cells offer the hope of generating a sufficient supply of tissues for cellular therapy. The successful outcome of this work will offer new hope to many Californians suffering from blood or liver diseases. This will improve lives and save money on long-term health care costs associated with these diseases.

Additionally, a training program has been included to help train research technicians for work in the stem cell field, therby helping to creat high paying jobs and provide the workforce needed for California to be the leader in stem cell research.

Scientific Review Summary of Part One Application

SHARED LABORATORY

SYNOPSIS OF PROPOSAL: This is an application by an institution to develop space for use in human embryonic stem cell (hESC) research with a focus on translational and clinical applications. The proposal is for renovation of laboratory space for basic and preclinical studies on hESC lines; for isolation and analysis of hESC lines; and for a vivarium to house NOD/SCID mice for in vivo analysis of these lines. The total renovated space would include ~1300 square feet and would be equipped with standard analysis and manipulation equipment. A total of 14 faculty are listed as potential users from the institution as well as from four neighboring research and/or educational institutions.

QUALITY AND IMPACT OF THE SCIENCE: The institution is a research entity of the parent organization. Much of the research support for the investigators at the institution is from the foundation of the parent organization, but all PIs have adjunct appointments at a neighboring research/educational institution. The proposed faculty have modest track records and much of the publication list includes a focus on hematopoietic cells and transfusion medicine. There are also multiple faculty listed without NIH grants or listed as co-investigators on other grants. The research focus of the proposed shared research laboratory is on hESC differentiation into hematopoietic cells and into hepatocytes which would then be tested for engraftment and reconstitution in immunodeficient mice. Much of this work is NIH funded; thus, while the proposed research is not unique, it is clearly important to the field. There are 8 PIs listed from the applicant institution but there is very limited hESC research currently ongoing and there is not much justification for the need of non-federally funded hESCs. The program director (PD) and an associate program director are doing an NIH-funded comparison of differentiation of erythroid cells from hESCs and adult and fetal erythropoiesis; the PD is working on fetal hepatic progenitors and stem cells and will extend this to differentiation of hESCs to hepatocytes. The PD is also collaborating on a pilot project using human fetal ovarian tissue as a source of germ cells from which hESCs could be derived by the growth of this human tissue in immunocompromised mice. One investigator’s interest is in optimizing the methods of viral transduction of hESCs (without need of federally funded free hESCs). Based on the publication record of the key personnel listed on this grant, it is not clear that the work they will do with hESCs will be of high impact, and the brief nature of the description of their projects does not suggest that this research will be at the cutting-edge.

The two strengths of the shared laboratory would be 1) an animal facility free of NIH funding which might draw several investigators from neighboring institutions (although there are some issues, see below) and the interest and expertise of an associate program director and one of the principal investigators on facilitating the translation of small scale research methods to clinically feasible methods that could be scaled up in a GMP facility. Another interesting aspect of this proposal is an internship program with a local community college in which students in their stem cell certification course would have 14 week half time internships at the applicant institution. If this provided hands on experience with stem cells, it would be mutually beneficial, but that is unclear and it is even suggested that these interns might “run samples on FACS, [perform] daily checks and maintenance of mouse colony, perform cell counts, prepare media, etc”.

The predicted quality and impact of the science is thus related mainly to the unique expertise of this group in the potential translation of basic hESC work into the clinical setting. However, it is not clear from the included biosketches to what extent any of the intended users has experience with hESC. This point is underscored by the statement that investigators of the institution have “been promised access to hESC lines” derived from other institutions. In addition, the experience and expertise of this group in xeno-transplantation is not evident from their published work.

APPROPRIATENESS OF SPACE AND EQUIPMENT TO SCOPE OF PLAN: Currently there is no facility at the applicant institution for using non-federally approved hESCs; indeed the level of current hESC research activity appears minimal. The space to be renovated had been for a contracted facility that was closed with consolidation of storage sites. The proposed space includes a tissue culture laboratory to maintain hESC stocks, a second tissue culture room for routine work on these cells and derivative cells, an equipment room for analysis equipment (flow cytometry and microscopy) and a vivarium to house immunodeficient NOD/SCID mice for transplantation studies. Currently, the institution’s investigators utilize the the animal facilities at a neighboring institution. While the vivarium would enhance the possible studies and is currently not available, it is unclear how the one 306 sq ft room would be used for 504 cages and the necessarily separated area for surgery and dissection. Additionally there is no indication of how the appropriate animal care will be handled (licensing, daily care and cleaning, cage cleaning, vet supervision, etc). Thus, the proposed facility will be small and the monies requested will be utilized primarily for renovation of space and equipment purchases.

Given the size of their group, the space that will be dedicated to hESC research is appropriate. The proposed equipment includes the usual cadre of microscopes, hoods and centrifuges. The proposed purchase of a WAVE bioreactor is for large scale closed system expansion of hESC. The requested equipment includes mouse caging and cryostat for analysis of tissues. The equipment requests are justified and reasonable. How this lab will fit into the overall programs of hESC research in the area seems less certain. There would seem to be competition from neighboring institutions as recognized by some of the collaborators. It would appear that the applicant institution has a weaker track record.

QUALITY OF MANAGEMENT PLAN: The management plan is simple as expected for such a small operation. The stem cell facility will be managed by the Program Director (PD) and two principal investigators with the PD taking the lead role. The PD(10% effort) will have overall responsibility for development and management of the laboratory. The PD will also be responsible for maintaining the mouse vivarium, which will apparently fall under the regulation of the animal regulatory domain (IACUC review) of a neighboring institution. One of the investigators will be an associate director (25% effort) with responsibility for maintenance, quality control, banking and distribution of hESCs. He/she has been trained in a noted hESC investigator’s laboratory. Another investigator will be associate director (5% effort) with responsibility for maintenance and function of the Stem Cell Laboratory. This Associate Director will be the primary contact point for investigators and will work with those investigators wanting to take hESC systems to clinical applications. An individual has been named(10% effort) who will be responsible for day-to-day operations and for running and maintaining the equipment for cell analysis (FACS, microscope). A research associate (100% effort) was budgeted to help maintain stocks. Thus, while there is a clear delineation of responsibilities in the written document, the formal management plan is somewhat weak. There is a list of guidelines that includes time-share for the facility, with priority given to “users that sign-up over casual drop-ins”. There is a plan for one of the four managers to be on site during usage by any investigators and a plan to ensure users have undergone basic safety training, but it is not clear what this training is to include. A supply budget of $30,000 is requested along with $10,000 in equipment maintenance. Letters of support are included from several institutions.

The oversight committee will consist of the PD and the two associate directors plus a representative from the stem cell program at the community college. Thus the oversight committee appears to be made up primarily by the management team, rather than by informed, outside investigators. The oversight committee will be responsible for planning, making equipment purchases and oversight of the renovation process.

The application includes funding for an internship program to be carried out with a local community college. This program includes 24 interns over 36 months with each intern rotating on 3 month blocks. There will be 2 interns at a time on site with a commitment of 20 hours/week. The interns will receive hands-on training and participate in the research activities of the hESC program. They will be assigned a mentor from the applicant institution. This program will be carried out in collaboration with the head of the community college’s stem cell program (letter included).

DISCUSSION: This is a research institute that also participates in clinical studies. They propose to renovate lab space for the purpose of hESC culture and differentiation into hematopoietic stem cells and hepatocytes; the space would also include a vivarium for NOD-SCID mice. Based on the information provided, most of the users have limited/no experience in hESC or in xenotransplantation. Many of these users lack NIH grants and no CIRM grants have been approved for award to this institution. One reviewer noted that one of the strengths of this application could have been the vivrium, however what they are proposing is not reasonable. A concern was that 300 square feet for a animal facility and procedure room may not be allowable because of inadequate space; this was not well thought through. Also, the institution currently uses the vivarium at a neighboring institution and even if this award were to be approved, IACUC and SCRO approvals would still have to go through the neighboring institution – so it unclear to at least one reviewer as to why they would change the current animal care and use arrangements. The management plan includes an oversight committee that is comprised primarily of the shared lab management team – at least one reviewer felt that the oversight team should have had more outside input. There are letters of support but they are vague. The application includes funding for an internship program with the community college with a commitment of 20 hr /wk of lab training. This seems reasonable except there does not seem to be a lot of on site expertise in hESC. Given the group’s expertise in clinical trials, there is some interesting potential in the proposed clinical trial and cellular products work, but this is outweighed by the negatives of the proposal.

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