Stem cell therapy can be useful for treating various diseases, such as spinal cord injury, Parkinson’s disease, Alzheimer’s disease and diabetes. However, the use of human embryos raises ethical as well as supply issues. There is also a major possibility of undesirable rejection of the transplanted stem cells administered to patients who have these diseases. While some recent advances offer promising results, such techniques currently have some critical flaws, hence new innovative approaches are needed to resolve the encountered or anticipated problems. The proposed research contains an effort to lay the ground work for resolving some of the major hurdles using nanotechnology and physical science approaches. To expedite therapeutic applications of stem cells, better understanding of the many factors which influence stem cell behavior, and new methods to cause differentiation of stem cells need to be developed. While there have been significant research investigations of stem cell behavior using cell biology approaches, there have been few studies to explore new ways to direct the fates of stem cells using physical science methods. Therefore, in this proposal of interdisciplinary research, we will introduce forefront nanotechnology to engineer desirable and beneficial changes in the behavior of human embryonic stem cells (hESCs). With recent nanotech advances, nanoscale manipulations (i.e., 1/80,000 the width of a human hair) of very small particles, materials, molecules are possible, inside the cells as well as inside the site where genes are located, the nucleus. Nanotechnology has not yet been seriously applied to stem cell science to advance clinical therapies. The significance of the proposed research lies in the possible control of differentiation pathways (e.g. stem cell to nerve cell), specifically at the level of the nucleus, in order to create therapeutically useful and ethically acceptable stem cells. The specific aims are thus directed toward the demonstration that bioengineered manipulations of stem cells using nanotechnology will accomplish what has not been previously possible with traditional laboratory methods. To validate the nanotech approach, experiments for controlled intracellular and intranuclear stem cell manipulations will be applied, and their effects on stem cell differentiation into a variety of mature cell types (pluripotency), will be investigated. Advances in stem cell manipulations using nanotechnology will provide significant and powerful ways to accelerate realistic therapeutic applications of stem cells.
Statement of Benefit to California:
For some patients with diseases, such as spinal cord injury, Parkinson’s disease, Alzheimer’s disease and diabetes, stem cell therapy offers a great hope for efficient therapeutic treatment of the patients. To expedite therapeutic applications of stem cells, an understanding of the key parameters which influence the stem cell behavior, and efficient technical means to induce differentiation into well-defined lineages are needed. While there have been significant worldwide research efforts for studies of stem cell behavior via biological approaches, with some recent advances offering promising approaches for future stem cell therapeutics, there has been little effort to explore new avenues of possible stem cell controls using physical science technology approaches. The proposed research contains an effort, using nanotechnology and associated innovative approaches, to lay the ground work for resolving some of the major hurdles or anticipated problems in the practical medical application of the current stem cell research advances. The successful outcome of the proposed research will benefit many California residents suffering from Alzheimer’s disease, diabetes, spinal cord injury, Parkinson’s disease. The estimated cost of healthcare in California related to the Alzheimer’s type disease alone is several hundred million dollars every year. There are nearly 1.5 million California residents diagnosed with diabetes with additional a few million people at risk. There is tremendous financial impact on California with the introduction of stem cell therapy for cure of these diseases. The proposed research on nanoscale stem cell manipulations is also substantially dependent on utilization of nanotechnology and materials, devices, processes being employed for semiconductor industries. The State of California is very strong in semiconductor industry including many silicon valley based high-tech companies. The State is also one of the significant leaders in the forefront biotechnology. The success of the proposed stem cell manipulation and control techniques, and their eventual commercialization for stem cell therapeutics, can foster expansion of the business scopes and revenues for some of the biotech as well as nanotech companies in California, thus creating jobs and enabling the State of California to become a world leader in stem cell science, technology, and medical applications. Such blossoming technical and medical advances will have profound positive effects on the vision of young scientists in California. This will also help many of the State’s large medical institutes to become world leaders in their respective therapeutic treatments of various diseases.
SYNOPSIS: This proposal aims to use magnetic nanoparticles to manipulate hESC temperature and intracellular mechanics to effect differentiation. This proposal includes four specific aims. The first aim is an experimental demonstration of introducing local intracellular temperature rise and asymmetric mechanical strain using nanotechnology. The second aim is a study of how the processing and materials parameters for temperature, strain and biomolecule injection can influence and control stem cell differentiation. The third aim proposes to employ engineering manipulation for hESC and somatic cell alignment to generate extremely high efficiency/probability for successful cell fusion and heterokaryon formation. The fourth aim proposes preliminary establishment of novel nanocone/nanopipette manipulation processes for nano/micro surgery and eventual selective inactivation of the human ESC nucleus after somatic nuclear reprogramming in the fused heterokaryon. INNOVATION AND SIGNIFICANCE: The use of physical forces to regulate hESC differentiation is innovative. The specific concept of this proposal - to use use magnetic nanoparticles to manipulate hESC temperature and intracellular mechanics to effect differentiation - is innovative. The use of magnetic particles to control temperature and forces of cells is not new, but its application to ESC differentiation is. It is not clear what the significance of this work would be to the hESC community. Perhaps if successful, it would sensitize hESC biologists to recognize the possibility that physical stimuli can alter ESC differentiation, but most biologists already believe it, and simply don't know what to do with the information if proven. STRENGTHS: The PI is a leader in the field of mechanical engineering who proposes physical science approaches to stem cell control using nanotechnology. The strength of this proposal is the willingness of a talented mechanical and aerospace engineer wanting to contribute to the embryonic stem cell field, and as such is respectable. The investigators have a demonstrated track record in the proposed fields. Magnetic particles delivered to hESCs may be interesting simply from the standpoint of beginning to understand how to deliver materials into these cells. WEAKNESSES: The proposal has many shortcomings, including the following. The investigator doesn’t need stem cells to do the experiments proposed, any cell type would qualify. None of the technology proposed in this grant are likely to be of any use for biologists interested in embryonic stem cells. All of the proposed experiments could be funded by the NIH, NSF, or any other funding agency. More specifically, the proposal is quite detailed (sufficient) in describing the magnetics, but is vague with regard to the hESC studies. What lineages will the investigator quantify? How will the differentiation experiments be carried out? If there is a shift in populations of differentiated cells, so what? What are the appropriate controls to compare various manipulations? There is no reason to believe that temperature or physical forces can regulate hESC's, so why should so much effort be placed into the development of this technology first? It would be more exciting if the investigator were to demonstrate with simple changes in bulk temperature, interesting or important effects on hESC differentiaiton. Thus, it appears that the technology is seeking a problem rather than the other way around. It is not clear what range of forces can be generated with this approach, and whether those forces are in the physiologic range of stresses experienced by cells. It is not clear what stimuli would be appropriate so this seems like a situation where one might be screening thousands of stimuli protocol. It is not clear how the investigator will explore this vast space in a rationale way. The fact that there is no directed goal (say, to increase myogenesis), makes it is very difficult to assess the likelihood that investigators would observe an effect (even if the effect were present - e.g., suppose neutrophilogenesis was enhanced, but investigators only looked at 20 other cell types in their assays). DISCUSSION: There was no discussion following reviewers' comments.