Funding opportunities

Human Germ-Line Stem Cells As Source for Pluripotent Stem Cells

Funding Type: 
New Cell Lines
Grant Number: 
RL1-00638
Funds requested: 
$1 448 280
Funding Recommendations: 
Not recommended
Grant approved: 
No
Public Abstract: 
We seek to extend our current findings obtained with laboratory mice to human patients. In particular, during the course of the proposed research, we aim to develop routine protocols for isolating and amplifying autologous Germ Line Stem Cells (GLSC’s) from patient-biopsy specimens that retain their potency and are subject to minimal manipulation under sterile technique (amenable to regulation according to cGMP [current Good Manufacturing Practice]). In the future, we expect to translate this technology into clinical application for individualized treatment of male juvenile cancer patients undergoing radiation- and/or chemotherapy-induced treatments that result in infertility, where the freezing of sperm for future use is not yet an available option. Finally, we believe these techniques can be extended to female oocytes as well, where they will play a similar role in treating female infertility as well. Ultimately, these cells, because of their pristine nature compared with other types of stem cells, can be used to treat a wide variety of chronic diseases.
Statement of Benefit to California: 
Basic research into the biology of mammalian adult stem cells based on experimental animals, such as laboratory rodents, is expected to benefit the citizens of California by establishing rigorous protocols for Phase-I human clinical trials in the future for a wide range of chronic diseases from neurodegenerative disorders (Parkinson’s, Alzheimer’s), heart disease, stroke, lung diseases (COPD), Diabetes, traumatic injuries (spinal-cord and head trauma from sports injuries or car accidents), congenital genetic disorders (Huntington’s, Sickle Cell Anemia), post-radiation cancer treatments leading to infertility, and many others. The reason for working with laboratory animal-models of human disease is to establish clinical guidelines for treatment that are not well understood at this time. Some of the clinical procedures that have yet to be resolved are the following: • rigorous characterization and standardization of cultured human stem cells prior to administration; • purity of the cells, e.g., without contamination by mouse feeder cells or oncogenic viruses; • individualizing of doses of cells per administration (from thousands to possibly millions of cells); • frequency of administration (daily, weekly, monthly, annually, or possibly only once); • route of administration (IV perfusion vs. organ-specific injection); • degree of differentiation of the cells (full pluripotent vs. multipotent or organ-specific cells); • the absolute requirement for autologous cells (to prevent acute rejection and/or graft vs. host disease) vs. the sufficiency of non-histocompatible cells, as claimed by some respected scientists; • diagnosis of end points for the successful termination of treatment; • prognosis of systemic side effects or adverse reactions (inflammation, autoimmune disorders, cancer). It is essential for citizens of California that the first human clinical trials conducted in our state be subject to the most rigorous protocol information available before such trials could lead to potentially serious adverse reactions, as we have witnessed with the otherwise promising field of genetic engineering and in the case of at least one profit-making, off-shore clinic where desperate patients, lacking alternatives, suffered the adverse consequences of tissue rejection from incompatible human fetal-cell injections. We are confident that CIRM-funded animal stem-cell research will provide the rigorous foundation for California medical scientists to build the protocols that will be needed for human therapies in the not-too-distant future.
Review Summary: 
Executive Summary This proposal aims to derive new pluripotent cell lines from male germ cells that could be utilized for treatment of male infertility, for fundamental biology studies of male germ cells, or as a tool for drug screening. The applicants propose to initiate studies with an animal model and ultimately to derive human cells under good manufacturing practice (GMP) conditions. Reviewers felt that the significance of exploring and developing a new source for pluripotent cells is great. However, the research plan to achieve this was poorly formulated. Experimental approaches were lacking in critical areas that made it difficult for the reviewers to assess whether meaningful data would be generated. Of particular concern was the fact that the proposed studies have to date only been accomplished in mice and it is unclear if the proposed strategy will translate to larger animal models and humans. This is an important consideration given that culture conditions for mouse and human cells are very different. If the approach does not translate, then the project fails. The applicant will recruit the assistance of several collaborators who are critical in the acquisition of tissue samples and cell sorting facilities. Reviewers indicated that no plans for distribution of the derived cells were presented by the applicant. Reviewer Synopsis The overall goal of this proposal is to isolate male germ cell-derived pluripotent stem cell lines. The applicants propose to do this in three steps. First they will isolate germ-line-stem-cell (GLSC) populations from non-human primate testicular tissues and then will characterize two of the newly derived cell lines. This has been done so far only in the mouse and will be now applied to primates. In the second step, the experience will be used to derive similar cells from human testicular biopsies. In the third step, similar strategies will be used to derive such germ cell stem cells under GMP conditions. Reviewer One Comments Significance: Novel source for pluripotent stem cells, very significant. Feasibility: The plan is logical and straightforward, but it is unclear how likely it is that similar findings will translate from mouse to primates. Characterization of the cells will be the key and the applicant does discuss this in nice detail. Reviewer Two Comments Significance: Deriving additional lines, in this case from male testicular biopsies, is a laudable goal. Feasibility: This is a poorly organized proposal. Parts of the introduction are very non-critical. There is a list of milestones presented for the 3 years but essentially no description of aims or the characterization of cells thereafter. There is no scientific rigor in the preparation of the proposal or in the description of the experiments. Responsiveness to RFA: Responsive Reviewer Three Comments Significance: The studies outlined in this proposal which are directed at the isolation and characterization of human germ line stem cells (GLSC) have the potential to yield a source of stem cells that may have therapeutic application in the autologous treatment of male infertility and as a source of cells for fundamental studies of male germ cell biology and use in drug screening. Feasibility: The PI, Dr Coles, Director of the Supercentenarian Research Foundation in Los Angeles, has enlisted the collaboration of Dr John Sundsmo from PrimeGen Biotech LLC who will provide access to cell sorting facilities and a Senior Research Scientist from the same company, Dr Fariborz Izadyar who has experience in characterization of germ cells from rodent, human and non-human primate sources. Dr Alan Cooperman (Reproductive Medicine Associates, NY) is providing samples of human testicular tissue for the proposed studies. The research plan is not well described or formulated and even basic details of experimental methodologies and approaches are lacking. Without a description of the precise experiments and assays to be performed it is very difficult to assess the scientific merits of this proposal and whether meaningful data will be generated. Responsiveness to RFA: Based on the proposal I am unable to judge whether the studies alluded to by the applicant will yield pluripotent cell lines. I can find no mention of any plans for distribution of any cell lines generated as a consequence of the proposed research included in the application.
Conflicts: 

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