Growth Factor Regulation of Olfactory Neurospheres

嗅觉神经球的生长因子调节

• 批准号: 8055377
• 负责人: Richard Carl Krolewski
• 金额: $ 4.18万
• 依托单位: TUFTS UNIVERSITY BOSTON
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-05-07 至 2012-05-06
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8055377
• 关键词: 3-Dimensional; Adult; Affect; Animals; Architecture; Back; Basal Cell; Biological Assay; Biological Markers; Cell Communication; Cell Therapy; Cells; Clinical; Complex; Data; Disease; EGF gene; Engraftment; Epithelial; Epithelium; Exhibits; Fluorescence; Goals; Growth; Growth Factor; Harvest; Hematopoietic System; Human; Immunophenotyping; In Vitro; Injury; Investigation; LIF gene; Lesion; Life; Maintenance; Multipotent Stem Cells; Mus; Natural regeneration; Neurodegenerative Disorders; Neurons; Olfactory Epithelium; Olfactory Mucosa; Population; Production; Regulation; Reporter; Resources; Rest; Rodent; Role; Spinal cord injury; Stem Cell Factor; Stem cells; System; Testing; Therapeutic; Time; Tissues; Translating; Transplantation; Tropism; Work; cell type; clinical application; clinical practice; congenic; in vitro testing; in vivo; insight; methyl bromide; neurogenesis; postnatal; progenitor; reconstitution; repaired; self-renewal; stem; stem cell biology; stem cell population; stemness; success; sustentacular cell

DESCRIPTION (provided by applicant): The olfactory mucosa (OM) of both humans and rodents is an accessible tissue that exhibits ongoing neurogenesis and epithelial reconstitution following injury with demonstrated potential for cell-based therapies. To realize the OM's therapeutic potential requires a culture system that can be used to expand, understand the regulation of, and serve as a biomarker for engraftment potential of progenitor populations. The hypothesis will be tested that olfactory neurosphere formation serves as a biomarker for engraftment potential by varying the "state" of the stem and progenitor cells and also the growth factor influences that act on them. A subsidiary goal is to establish the use of neurospheres as a means of evaluating the network of interactions acting on stem and multipotent progenitor cells in vivo. OM-derived neurospheres will be tested as biomarkers for multipotency and stemness using the sphere-forming activity of cells isolated from in vivo conditions that involve different endogenous stem and progenitor cell activation: normal adult, selective neuronal regeneration, full reconstitution of the epithelium after methyl bromide (MeBr)-induced toxic injury, and early postnatal expansion. Resulting spheres will be quantified and their immunophenotype determined. Olfactory neurospheres will then cultured from constitutively expressing GFP mice and transplanted into MeBr lesioned host animals to determine whether the engrafted cells produce OM cell types. To test whether factors that act on globose basal cells (GBCs) or horizontal basal cells (HBCs) specifically enhance sphere formation and engraftment potential, olfactory neurospheres will be treated with a variety of growth factors with both known tropism as well as unknown function in the OM. Reporter mice that allow for live tracking of GBCs and their progeny will be used to obtain readouts of growth factor activity on olfactory neurospheres. Growth factors that produce a significant change in the size distribution and fluorescence intensity of spheres will be tested via the aforementioned transplantation assay. the stem and progenitor cells ofthe OE are accessible in adult humans and undergo neurogenesis throughout adulthood. As in other systems that have translated stem cell biology to clinical practice, such as the hematopoietic system, it is apparent that the ability to expand the progenitor populations will be required for clinical applications. This work will expand on initial studies showing the clinical potential of olfactory mucosal-derived cells for treatment of spinal cord injury and neurodegenerative diseases by studying what factors can be used to enhance the proliferation and propagation of these stem cell populations.

描述（由申请人提供）：人类和啮齿动物的嗅粘膜（OM）是一种可接近的组织，在损伤后表现出持续的神经发生和上皮重建，并具有基于细胞的治疗潜力。为了实现OM的治疗潜力，需要一种培养系统，该培养系统可用于扩增、理解祖细胞群体的调节并用作祖细胞群体的植入潜力的生物标志物。该假设将被测试，嗅觉神经球的形成作为一个生物标志物的移植潜力，通过改变干细胞和祖细胞的“状态”，也对他们的生长因子的影响。一个次要的目标是建立使用神经球作为一种手段，评估网络的相互作用，干细胞和多能祖细胞在体内。将使用从涉及不同内源性干细胞和祖细胞活化的体内条件中分离的细胞的球体形成活性，将OM衍生的神经球作为多能性和干性的生物标志物进行测试：正常成人、选择性神经元再生、溴甲烷（MeBr）诱导的毒性损伤后上皮的完全重建和出生后早期扩增。将对所得球进行定量，并确定其免疫表型。然后从组成型表达GFP的小鼠中培养嗅觉神经球，并移植到MeBr损伤的宿主动物中，以确定移植的细胞是否产生OM细胞类型。为了测试作用于球状基底细胞（GBC）或水平基底细胞（HBC）的因子是否特异性地增强球体形成和植入潜力，将用在OM中具有已知向性以及未知功能的多种生长因子处理嗅觉神经球。允许活体追踪GBC及其后代的报告小鼠将用于获得嗅觉神经球上生长因子活性的读数。将通过上述移植试验检测产生球的尺寸分布和荧光强度显著变化的生长因子。OE的干细胞和祖细胞在成年人中是可获得的，并且在整个成年期经历神经发生。如在已经将干细胞生物学转化为临床实践的其他系统中，例如造血系统，显然临床应用将需要扩增祖细胞群体的能力。这项工作将扩大初步研究，通过研究哪些因素可用于增强这些干细胞群的增殖和繁殖，显示嗅粘膜衍生细胞治疗脊髓损伤和神经退行性疾病的临床潜力。