Early Translational I
Very few laboratories in the State of California have the capacity to produce stem cells in compliance with GMP regulations, ("Good Manufacturing Practices"). This shortage is critical because it leaves the State deficient in its ability to produce clinical grade stem cells suitable for use in human clinical trials. The shortage of these laboratories and resulting absence of cell lines is a major bottleneck in the advancement of stem cell-based therapies in California. [REDACTED], is expected to offer GMP manufacturing services in early 2009. Working in partnership with the [REDACTED] and the world-famous [REDACTED], [REDACTED] seeks funding under this project to use pluripotent cells to grow and create California based banks of clinical grade stem cell lines. These lines will then be tested for their ability to differentiate into different types of cells as follows: 1) neurons (for brain injury treatment) 2) cardiomyocytes (for heart tissue treatment) and 3) beta cells (for use in the treatment of such patient conditions as diabetes) This project has two main Aims: 1) To develop and validate human embryonic stem cell lines inside [REDACTED] c-GMP compliant facility; and; 2) To determine the capacity of clinical grade stem cell lines to generate functional terminally differentiated cells representing all three germ layers. In the first year of the project, multiple stem cell lines will be created and catalogued. In the second year, characterization of the developed stem cell lines will occur, along with testing their ability to differentiate into functional neurons, cardiomyocytes and beta cells. An interactive web-based system will be established as a repository database for this project, providing detailed information to CIRM affiliated organizations as well as interested scientists and researchers around the world. In year 3, the manufacturing of master cell banks will be completed with the full validation of up to 15 clinical grade embryonic stem cell lines. A comprehensive educational outreach initiative will be implemented with physicians interested in applying these clinical grade cell lines for human trials in California.
Statement of Benefit to California:
This proposal will have a major impact on translational and clinically-oriented stem cell efforts in California. Many protocols differentiating hESCs into various cell types already exist. These protocols are constantly improving and it is certain that other protocols for hESCs differentiation are expected to be developed in the near future. However, according to California’s Sherman Food, Drug and Cosmetic Law all of the hESCs lines or their derivatives produced for human use in California must be manufactured in FDB (Food and Drug Branch) licensed cGMP facilities. Therefore clinical application of any of these protocols critically depend on the availability of such facilities and their capability to manufacture clinical grade hESCs suitable for human clinical trials. The absence of California-produced cell lines is currently a major bottleneck for the advancement of stem cell-based therapies in California, and several disadvantages exist when California stem cell researchers and clinicians contract cGMP manufacturing outside California. 1. In the case of out of country manufacturing (Europe) we will have to deal with importation permits, customs, etc. In addition, the rules outside US, e.g. EU GMP are somewhat different from the FDA GMP. 2. It is difficult for a majority of California stem cell patients to travel due to their medical condition such as stroke or myocardial infarction. Therefore, shipment of live cell products long distance to California clinical sites is problematic and may raise quality issues. 3. Because California FDB inspects all production facilities prior to treatment of the first human patient, cGMP compliance is assured for California production facilities long before any routine FDA inspection. 4. cGMP regulations require auditing of the contract manufacturing facilities and periodic site-visits. Therefore, it is more convenient and cost effective for California biotech companies to use facilities located within the State. By removing the bottleneck through the active support of FDB licensed cGMP facilities in California, this program will accelerate the availability of stem cell therapies for interested physicians and researchers across the State. Such acceleration will lead to the sale and licensing of intellectual property rights which will accrue to the State and its citizens; and, secondarily, generate revenues from increased biotech, bio-manufacturing and health-related treatment business within, and for, California.
This proposal addresses a bottleneck in the advancement of stem cell therapies to the clinic: the generation of well-characterized, clinical-grade human embryonic stem cell (hESC) lines using current Good Manufacturing Practices (cGMP). The applicant proposes to generate 15 such lines, validate their pluripotency and characterize them using a variety of methods, including karyotyping and gene expression profiling. The applicant will also assess the ability of these hESC lines to differentiate into cells representing the three germ layers: neurons and glia (ectoderm), cardiomyocytes (mesoderm) and pancreatic beta cells (endoderm). These differentiated cell types will be evaluated by marker expression and functional assays. Reviewers acknowledged the importance of generating clinical-grade hESCs but raised significant questions about the feasibility of the proposal. They were particularly concerned by the applicant’s admission that “the hESC derivation procedure is new to” the research organization. Reviewers felt that demonstration of previous success in the derivation of hESC lines was crucial for this type of proposal. The learning curve for working with these cells may be longer than anticipated and it is practically difficult to assay “established criteria for purity, identity, strength and potency” as proposed. Although there are accepted cell markers for pluripotency, expression of the full panel does not guarantee the capacity to differentiate into specific cell lines and there is considerable line-to-line and clone-to-clone variability. One reviewer found it surprising that the chosen model for pluripotency testing is in vitro differentiation, instead of more commonly used in vivo teratoma-formation assays. Reviewers also noted the difficulty of differentiating hESCs into insulin-producing pancreatic beta cells, pointing out that even the pioneering company in this field has abandoned their efforts in terminally differentiating these cells in vitro, opting instead to transplant undifferentiated progenitors. Reviewers noted a general lack of detail in the proposal. The applicant does not describe the current state of clinical-grade cGMP hESC production in California, and it was therefore unclear what facilities currently exist and how this proposal fills an important gap. Another reviewer would have also appreciated more details about the preliminary data and the rationale for using a specific kinase inhibitor to establish clonal stem cell lines. In terms of the data, it was unclear to one reviewer whether the mesenchymal stem cells were functionally characterized and whether the hESC differentiation into neurons, cardiomyocytes and beta cells was done using clinical-grade lines. Reviewers found the applicant and research team qualified for cGMP banking but were concerned about their lack of experience working with hESCs. They found the academic collaboration to be a strength but felt it should have been described in greater detail. One reviewer noted that two letters of support mention either technology or investigators that weren’t included in the proposal. This reviewer also pointed out that the majority of the budget is devoted to consultants. Overall, reviewers felt that this proposal lacked sufficient detail to inspire confidence in its feasibility. They were particularly concerned about the applicant’s lack of expertise in hESC derivation.