Growth Factor Regulation of Olfactory Neurospheres

嗅觉神经球的生长因子调节

• 批准号: 7749753
• 负责人: Richard Carl Krolewski
• 金额: $ 4.03万
• 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/7749753
• 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觉粘膜（OM）是一种可访问的组织，在受伤后表现出持续的神经发生和上皮重构，表现出了基于细胞的疗法的潜力。为了实现OM的治疗潜力，需要一种培养系统，该系统可用于扩展，了解对祖细胞种群的植入潜力的生物标志物。该假设将检验，嗅觉神经圈形成是通过改变茎和祖细胞的“状态”的生物标志物的生物标志物，并且生长因子影响对它们的作用。一个子公司的目标是建立神经球作为评估在体内作用于茎和多能祖细胞细胞的相互作用网络的手段。 OM来源的神经球将使用与体内条件中分离的细胞的体形成活性来测试，以实现多能力和干性，这些细胞与体内条件分离，涉及不同的内源性干和祖细胞激活：正常的成人，选择性的神经元再生，甲基溴化物（Mebr Bromide（Mebr Bromide himber）诱导的毒性损伤和早期毒性造成的甲基溴化物术后的全面重新结束，并早期毒性损伤。所得球体将被定量，并确定其免疫表型。然后，嗅觉神经球将从组成型表达GFP小鼠并移植到MEBR病变的宿主动物中，以确定植入的细胞是否产生OM细胞类型。为了测试作用于球体基底细胞（GBC）或水平基底细胞（HBC）的因素是否特异性增强了球体形成和植入潜力，嗅觉神经球将用各种具有已知热带主义以及OM中未知功能的生长因子来处理。允许对GBC进行实时跟踪及其后代的记者小鼠将用于获得嗅觉神经球上生长因子活性的读数。将通过上述移植测定法测试球体尺寸分布和荧光强度的显着变化的生长因子。 OE的茎和祖细胞在成年人中可访问，并且在整个成年期都会经历神经发生。就像在其他将干细胞生物学转化为临床实践的系统中，例如造血系统，显然，扩大祖细胞种群的能力将需要临床应用。这项工作将扩大初步研究，以表明嗅觉粘膜衍生细胞在治疗脊髓损伤和神经退行性疾病方面的临床潜力，通过研究哪些因素可以使用哪些因素来增强这些干细胞群体的增殖和传播。