Strategies for Restoring Olfactory Neurogenesis

恢复嗅觉神经发生的策略

• 批准号: 6802765
• 负责人: JAMES E. SCHWOB
• 金额: $ 15.85万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2003
• 资助国家: 美国
• 起止时间: 2003-09-18 至 2006-02-28
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/6802765
• 关键词: cell differentiation; cell transformation; chemoreceptors; flow cytometry; laboratory mouse; neurogenesis; olfactions; oncogenes; receptor expression; respiratory epithelium; stem cell transplantation; stem cells; tissue /cell culture; transforming growth factors

(Revised Abstract) DESCRIPTION (provided by applicant): The maintenance of olfactory function in humans as in other animals depends on the persistence of neurogenesis in the olfactory epithelium throughout life. If the progenitor population is destroyed during the course of an injury to the olfactory epithelium, the tissue undergoes metaplasia and reconstitutes as respiratory epithelium instead. We have shown that the globose basal cells (GBCs), among which are both broadly potent and neuronal-committed progenitors, can be selectively isolated by FACS and will engraft in the epithelium of a new host after transplantation. Accordingly, transplantation may be a viable strategy for restoring the epithelium's capacity for neurogenesis. Progress in implementing that strategy depends on the availability of a sufficiently large population of progenitors. To that end we are proposing experiments to test whether the pool of progenitors can be expanded in vitro and still engraft and differentiate appropriately in vivo. Two Specific Aims will be pursued. First, GBCs will be selectively isolated from normal mice that ubiquitously express GFP and others that have been lesioned with methyl bromide gas shortly before harvest. In the later case, the effect of the lesion is activate olfactory stem cells and to bias the population toward multipotency. The GBCs will be cultured in defined media with a melange of growth factors that have been shown to drive proliferation and/or neurogenesis in heterogeneous explant or dissociated cultures. Second, GBCs will be conditionally immortalized by transfection with a retroviral vector that encodes a temperature sensitive version of the large T antigen oncogene from SV40. The differentiation of the resulting clonal cell lines will be assayed after switching to the non-permissive temperature. Both the expanded primary isolates and cell lines that differentiate well after inactivation of the oncogene will be transplanted in order to define their capacity for differentiation in vivo (including a determination of whether donor-derived neurons express odorant receptors). Successful completion of the Aims will; 1) clarify the nature of growth factor control on olfactory neurogenesis; 2) determine whether cellular replacement is a viable strategy for restoring neurogenesis; 3) generate a set of protocols and cellular reagents that are useful for further studies of progenitor cell capacity and neuronal differentiation.

（修订摘要）描述（由申请人提供）：人类嗅觉功能的维持与其他动物一样，取决于嗅上皮中神经发生的持久性。如果在嗅觉上皮损伤过程中祖细胞群被破坏，则组织发生化生并重建为呼吸上皮。我们已经证明，全球基底细胞（ggbcs），其中既广泛有效又具有神经元承诺的祖细胞，可以通过FACS选择性地分离出来，并在移植后植入新宿主的上皮中。因此，移植可能是恢复上皮神经发生能力的可行策略。执行这一战略的进展取决于是否有足够多的祖细胞。为此，我们建议进行实验，以测试祖细胞池是否可以在体外扩展，并在体内移植和适当分化。