Bioengineering technology for fast optical control of differentiation and function in stem cells and stem cell progeny
Embryonic stem (ES) cells potentially could provide clinically important replacement tissue for central nervous system (CNS) disease treatment, and regenerative medicine approaches involving ES cells have been suggested for common CNS disorders. But it has been difficult to produce the right kind of replacement tissues from ES cells because the “differentiation”, or cell-type specification process, takes many days to weeks, during which time many different stimuli and signaling molecules need to be physically applied to the stem cells. This process of “stem cell differentiation” is slow, costly, laborious, variable, prone to error and contamination, and ultimately rate-limiting in the long road leading to clinical translation. We propose to develop and apply fast, inexpensive, and robust optical technologies to the fundamental problem of stem cell differentiation and regenerative medicine, with particular focus on CNS disease.
Neuropsychiatric diseases like Parkinson’s disease and major depression are leading causes of disability and death in California and worldwide. They are difficult to treat, poorly understood, and devastating for patients, families, and society as a whole. Our proposed fusion of engineering technology with clinically-inspired stem cell technology represents a unique opportunity, which we anticipate will lead not only to fundamentally new, potent, and specific therapies for diseases representing major burdens for the state, but also to engineering and medical commercial ventures that will add resources, money, and skilled jobs to the robust and growing state economy.
Therapies and diagnostics that depend on differentiating stem cells may be fundamentally accelerated with a fusion of optical bioengineering and stem cell technology that we propose to develop and apply. Indeed, we anticipate that development and dissemination of this technology will lead to 1) in vitro methodology for fast reliable cell production, 2) deeper understanding of the basic science and biology of signaling in stem cells as they proliferate, differentiate, and generate phenotypically complete and stable progeny; and 3) in vivo methodology for noninvasively stimulating new cells in patients with devastating degenerative diseases.
The goal of this work has been to develop optogenetic technology for the study of stem cells. Major aims include optogenetic technology development and dissemination/distribution/collaboration with stem cell scientists. This work has progressed well, and our work (optogenetics) was recently named method of the year across all fields of life science and engineering, by Nature Methods:
Deisseroth K (2011). Optogenetics. Nature Methods, 8:26-9.
Weick JP, Johnson MA, Skroch SP, Williams JC, Deisseroth K, Zhang SC. Functional control of transplantable human ESC-derived neurons via optogenetic targeting. Stem Cells. Epub 2010 Sep 8.
Tønnesen J, Parish CL, Sørensen AT, Andersson A, Lundberg C, Deisseroth K, Arenas E, Lindvall O, Kokaia M. Functional integration of grafted neural stem cell-derived dopaminergic neurons monitored by optogenetics in an in vitro Parkinson model.PLoS One. 2011 Mar 4;6(3):e17560.
Stroh A, Tsai HC, Wang LP, Zhang F, Kressel J, Aravanis A, Santhanam N, Deisseroth K, Konnerth A, Schneider MB. Tracking stem cell differentiation in the setting of automated optogenetic stimulation. Stem Cells. 2011 Jan;29(1):78-88. doi: 10.1002/stem.558.
We have made extensive progress in developing optogenetic tools that can be used for stem cell biology. Optogenetics is a method for using light to control well-defined events within specified cells, even within systems as complex as intact behaving mammals. Broadly our project has included both using optogenetics to control the differentation of stem cells, and for controlling stem cells, their progeny, or surrounding cells in the living mammal to assess and guide function. Over the recent funded period we are pleased to report that we finally achieved the first multiple-color excitation (two separate channels of control) in mammalian brains, which will have far-reaching impact on the study of interactions between stem cells and their environment, as well as on technology used for maintaining, expanding, controlling, and studying these powerful biological tools.
- Neuron (2011) Optogenetics in neural systems. (PubMed: 21745635)
- Nature (2011) Neocortical excitation/inhibition balance in information processing and social dysfunction. (PubMed: 21796121)
- Cell (2011) Dynamics of Retrieval Strategies for Remote Memories. (PubMed: 22019004)
- Nat Neurosci (2011) Optetrode: a multichannel readout for optogenetic control in freely moving mice. (PubMed: 22138641)
- Cell (2011) The microbial opsin family of optogenetic tools. (PubMed: 22196724)
- Neuron (2011) Recombinase-driver rat lines: tools, techniques, and optogenetic application to dopamine-mediated reinforcement. (PubMed: 22153370)
- Stem Cells (2011) Tracking stem cell differentiation in the setting of automated optogenetic stimulation. (PubMed: 21280159)
- Nat Neurosci (2011) An optogenetic toolbox designed for primates. (PubMed: 21278729)
- Nat Neurosci (2010) Ultrafast optogenetic control. (PubMed: 20081849)
- Cell (2010) Molecular and cellular approaches for diversifying and extending optogenetics. (PubMed: 20303157)
- J Neurosci (2010) Dlx5 and Dlx6 regulate the development of parvalbumin-expressing cortical interneurons. (PubMed: 20392955)
- Nature (2010) Global and local fMRI signals driven by neurons defined optogenetically by type and wiring. (PubMed: 20473285)
- Nat Protoc (2010) Optogenetic interrogation of neural circuits: technology for probing mammalian brain structures. (PubMed: 20203662)
- Science (2010) Cholinergic interneurons control local circuit activity and cocaine conditioning. (PubMed: 21164015)
- Science (2009) Optical deconstruction of parkinsonian neural circuitry. (PubMed: 19299587)
- Nature (2009) Temporally precise in vivo control of intracellular signalling. (PubMed: 19295515)
- J Neurosci (2009) Sleep homeostasis modulates hypocretin-mediated sleep-to-wake transitions. (PubMed: 19726652)
- Nature (2009) Parvalbumin neurons and gamma rhythms enhance cortical circuit performance. (PubMed: 19396159)
- Nat Neurosci (2009) Bi-stable neural state switches. (PubMed: 19079251)