Mechanisms of Lineage Commitment in Human Neural Crest Stem Cells

Funding Type: 
Basic Biology I
Grant Number: 
RB1-01398
Investigator: 
ICOC Funds Committed: 
$0
Public Abstract: 
Diseases arising from defects in NC specification, migration and differentiation referred to as neurocristopathies (total ~74) include various skeletal dysmorphology syndromes (e.g. Apert and Beare-Stevenson cutis gyrate syndromes), diseases of the nervous system (neurofibromatosis and Hirschsprung’s disease, peripheral neuropathies such as Familial Dysautonomia) and pigment disorders (Waardenburg syndrome). The defect is often confined to one or two NC lineages, as in pigment abnormalities and the treatment of PNS disorders will require a uniform population of lineage-committed progenitors. In addition, in vitro applications such as initial screening for drug toxicity will require large quantities of pure NC progenitors. In particular, derivation of functional chondrocytes may provide a much-needed alternative for cartilage regeneration. Recently, by means of the support from a CIRM Seed grant, we generated a uniform population of neural crest stem cells (NCSC). These human NCSC can differentiate in vitro producing all neural crest lineages including sensory and autonomic neurons, Schwann cells, smooth muscle cells, melanocytes, adipocytes and chondrocytes. We propose comprehensive in vitro analysis of differentiation in NC for the following two reasons: 1. All previous evidence was obtained in the model organisms. No studies investigating the role of transcription factors or extracellular matrix (ECM) in human neural crest self-renewal and differentiation were reported to date. 2. It is possible that unsuspected conditions will promote directed differentiation of human NCSC compared to physiological stimuli. Specifically we will: 1. Identify transcription factors regulating human NCSC differentiation using a functional genomic approach; 2. Investigate the role of various matrix components and their elasticity in human NCSC differentiation. The deliverables will include: first, a verified set of TFs that regulate human NCSC lineage commitment and differentiation; second, a knowledge of how the nature and stiffness of various ECM matrices affects differentiation into neural crest lineages. The knowledge, tools and reagents will be available to other academic researchers and commercial entities. If successful, the proposed approach will help us to understand mechanisms of NC differentiation along specific NC lineages. This knowledge will allow the development of tools and reagents for diagnostic and therapeutic applications such as screening assays for drugs affecting human sensory neurons, neuronal replacement for peripheral neuropathies, and the generation of functional Schwann cells.
Statement of Benefit to California: 
Diseases arising from defects in NC specification, migration and differentiation referred to as neurocristopathies (total ~74) include various skeletal dysmorphology syndromes (e.g. Apert and Beare-Stevenson cutis gyrate syndromes), diseases of the nervous system (neurofibromatosis and Hirschsprung’s disease, peripheral neuropathies such as Familial Dysautonomia) and pigment disorders (Waardenburg syndrome). The defect is often confined to one or two NC lineages, as in pigment abnormalities and the treatment of PNS disorders will require a uniform population of lineage-committed progenitors. We propose comprehensive in vitro analysis of differentiation in NC for the following two reasons: 1. All previous evidence was obtained in the model organisms. No studies investigating the role of transcription factors or extracellular matrix (ECM) in human neural crest self-renewal and differentiation were reported to date. 2. It is possible that unsuspected conditions will promote directed differentiation of human NCSC compared to physiological stimuli. The deliverables will include: first, a verified set of TFs that regulate human NCSC lineage commitment and differentiation; second, a knowledge of how the nature and stiffness of various ECM matrices affects differentiation into neural crest lineages. The knowledge, tools and reagents will be available to other academic researchers and commercial entities. If successful, the proposed approach will help us to understand mechanisms of NC differentiation along specific NC lineages. This knowledge will allow the development of tools and reagents for diagnostic and therapeutic applications such as screening assays for drugs affecting human sensory neurons, neuronal replacement for peripheral neuropathies, and the generation of functional Schwann cells. An effective, straightforward, and understandable way to describe the benefits to the citizens of the State of California that will flow from the stem cell research we propose to conduct is to couch it in the familiar business concept of “Return on Investment”. The novel therapies and reconstructions that will be developed and accomplished as a result of our research program and the many related programs that will follow will provide direct benefits to the health of California citizens. In addition, this program and its many complementary programs will generate potentially very large, tangible monetary benefits to the citizens of California. These financial benefits will derive directly from two sources. The first source will be the sale and licensing of the intellectual property rights that will accrue to the state and its citizens from this and the many other stem cell research programs that will be financed by the CIRM. The second source will be the many different kinds of tax revenues that will be generated from the increased bio-science and bio-manufacturing businesses that will be attracted to California by the success of the CIRM.

© 2013 California Institute for Regenerative Medicine