The mammalian nervous system is composed of a multitude of distinct neuronal subtypes, each with its own phenotype and differential sensitivity to degenerative disease. Although some specific neuronal types can be isolated from intact rodent embryos or engineered from stem cells for translational studies, these approaches are time-consuming and many neuronal subtypes are inaccessible. Transcription factor-mediated reprogramming might provide a more direct route to the generation of neurons for disease modeling and regenerative medicine, but it is currently unclear if this approach can be used to create cells with translational utility. Here, we propose to identify a set of transcription factors sufficient to convert fibroblasts into functional spinal motor neurons. We will characterize the reprogramming process and examine the molecular and functional properties of the resulting motor neurons in order to determine their therapeutic potential. These studies will provide an accessible source of patient-specific motor neurons for the study of neurodegenerative disease, demonstrate that specific adult cell types can be directly generated from somatic cells using defined factors, and mechanistically dissect the defined-factor reprogramming process.

Public Health Relevance

The mammalian nervous system is composed of a multitude of distinct neuronal subtypes, each with its own phenotype and differential sensitivity to degenerative disease and although some specific neuronal types can be isolated from intact rodent embryos or engineered from stem cells for translational studies, these approaches are time-consuming and many neuronal subtypes are inaccessible. The goal of these studies is to identify a small group of defined transcription factors can convert fibroblasts directly into motor neurons, thereby enabling the rapid generation of these cells for therapeutic use. More broadly, these studies will investigate the potential of using defined-factor reprogramming to create specific adult cell types with translational utility.

Agency
National Institute of Health (NIH)
Institute
National Institute of Neurological Disorders and Stroke (NINDS)
Type
Research Transition Award (R00)
Project #
4R00NS077435-03
Application #
8605244
Study Section
Special Emphasis Panel (NSS)
Program Officer
Owens, David F
Project Start
2013-01-01
Project End
2015-12-31
Budget Start
2013-01-01
Budget End
2013-12-31
Support Year
3
Fiscal Year
2013
Total Cost
$249,000
Indirect Cost
$62,049
Name
University of Southern California
Department
Other Basic Sciences
Type
Schools of Medicine
DUNS #
072933393
City
Los Angeles
State
CA
Country
United States
Zip Code
90089
Gopalakrishnan, Suhasni; Hor, Pooja; Ichida, Justin K (2015) New approaches for direct conversion of patient fibroblasts into neural cells. Brain Res :
Akeju, Oluwaseun; Davis-Dusenbery, Brandi N; Cassel, Seth H et al. (2014) Ketamine exposure in early development impairs specification of the primary germ cell layers. Neurotoxicol Teratol 43:59-68
Carter, Ava C; Davis-Dusenbery, Brandi N; Koszka, Kathryn et al. (2014) Nanog-independent reprogramming to iPSCs with canonical factors. Stem Cell Reports 2:119-26
Kiskinis, Evangelos; Sandoe, Jackson; Williams, Luis A et al. (2014) Pathways disrupted in human ALS motor neurons identified through genetic correction of mutant SOD1. Cell Stem Cell 14:781-95
Okumura, Noriko; Akutsu, Hidenori; Sugawara, Tohru et al. (2013) β-catenin functions pleiotropically in differentiation and tumorigenesis in mouse embryo-derived stem cells. PLoS One 8:e63265
Hanada, Toshikatsu; Weitzer, Stefan; Mair, Barbara et al. (2013) CLP1 links tRNA metabolism to progressive motor-neuron loss. Nature 495:474-80
Navara, Christopher S; Hornecker, Jacey; Grow, Douglas et al. (2013) Derivation of induced pluripotent stem cells from the baboon: a nonhuman primate model for preclinical testing of stem cell therapies. Cell Reprogram 15:495-502
Son, Esther Y; Ichida, Justin K; Wainger, Brian J et al. (2011) Conversion of mouse and human fibroblasts into functional spinal motor neurons. Cell Stem Cell 9:205-18
Egli, Dieter; Chen, Alice E; Saphier, Genevieve et al. (2011) Reprogramming within hours following nuclear transfer into mouse but not human zygotes. Nat Commun 2:488