Choline Transporter Antibodies as Developmental Biomarkers

胆碱转运蛋白抗体作为发育生物标志物

• 批准号: 7635148
• 负责人: JOY A UMBACH
• 金额: $ 7.7万
• 依托单位: UNIVERSITY OF CALIFORNIA LOS ANGELES
• 依托单位国家: 美国
• 财政年份: 2009
• 资助国家: 美国
• 起止时间: 2009-01-15 至 2010-12-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7635148
• 关键词: Affect; Alzheimer' s Disease; Amyotrophic Lateral Sclerosis; Animals; Antibodies; Antigens; Binding; Biological Markers; Brain; Cell Therapy; Cell surface; Cells; Communities; Development; Disease; Epitopes; Goals; Human; Hybridomas; In Vitro; Label; Life; Monitor; Monoclonal Antibodies; Mus; Neurodegenerative Disorders; Oocytes; Peptides; Protocols documentation; Rana; Replacement Therapy; Research; Sensitivity and Specificity; Source; Stem cells; Surface Antigens; choline transporter; cholinergic; cholinergic neuron; extracellular; human embryonic stem cell; human stem cells; improved; progenitor; public health relevance; success; tool

DESCRIPTION (provided by applicant): This project aims to develop fluorescent monoclonal antibodies as biomarkers for living, human cholinergic neurons and their precursors, in vitro. The most important use of these antibodies will be for identifying human stem cells that are undergoing cholinergic differentiation. Success of this project depends on antibodies that bind to cell surface markers that are preferentially expressed by cholinergic neurons (and their progenitors). Currently, the high affinity choline transporter is the only cell surface antigen that is recognized as being universally expressed by cholinergic neurons. Thus, mice will be immunized with peptide immunogens that correspond to specific extracellular regions of the human, high affinity choline transporter. Hybridoma supernatants from these animals will be screened for their ability to detect the human high affinity choline transporter that is expressed in frog oocytes, HEK-293 cells and in fixed human brain sections. Antibodies that satisfy empirical criteria of specificity and sensitivity will then be fluorescently labeled and evaluated for their utility in detecting the cholinergic differentiation of human stem cells in culture. Antibodies that are viable in this context will be distributed to the stem cell community as tools to improve protocols for obtaining stem cell-derived cholinergic neurons. Such cholinergic neuron precursors are expected to be useful for cell replacement therapies in disorders that involve a loss of cholinergic neurons (like amyotrophic lateral sclerosis and Alzheimer's disease). Thus, this project should serve to catalyze progress toward stem cell-based therapies for disorders that affect cholinergic neurons. PUBLIC HEALTH RELEVANCE: Alzheimer's disease and amyotrophic lateral sclerosis are devastating neurodegenerative disorders that involve a loss of cholinergic neurons. The antibody biomarkers developed by this project will facilitate the identification and characterization of living, human cholinergic neurons (and possibly, their precursors) that are derived from stem cells. The widespread dissemination of these biomarkers to the research community should significantly advance efforts geared to the use of cell-replacement therapies for these diseases.

描述（由申请人提供）：该项目旨在开发荧光单克隆抗体，作为活体人类胆碱能神经元及其前体的体外生物标志物。这些抗体最重要的用途是识别正在进行胆碱能分化的人类干细胞。该项目的成功取决于与胆碱能神经元（及其祖细胞）优先表达的细胞表面标记物结合的抗体。目前，高亲和力胆碱转运蛋白是唯一被认为由胆碱能神经元普遍表达的细胞表面抗原。因此，将用与人类高亲和力胆碱转运蛋白的特定细胞外区域相对应的肽免疫原对小鼠进行免疫。将筛选来自这些动物的杂交瘤上清液，以确定其检测青蛙卵母细胞、HEK-293 细胞和固定人脑切片中表达的人高亲和力胆碱转运蛋白的能力。然后，满足特异性和敏感性经验标准的抗体将被荧光标记，并评估其在检测培养物中人类干细胞的胆碱能分化中的效用。在这种情况下可行的抗体将被分发到干细胞群落，作为改进获得干细胞衍生胆碱能神经元的方案的工具。这种胆碱能神经元前体预计可用于涉及胆碱能神经元丧失的疾病（如肌萎缩侧索硬化症和阿尔茨海默病）的细胞替代疗法。因此，该项目应有助于促进基于干细胞的治疗影响胆碱能神经元疾病的进展。公众健康相关性：阿尔茨海默病和肌萎缩侧索硬化症是破坏性神经退行性疾病，涉及胆碱能神经元的丧失。该项目开发的抗体生物标志物将有助于鉴定和表征源自干细胞的活体人类胆碱能神经元（可能还有它们的前体）。这些生物标志物向研究界的广泛传播应该会显着推进针对这些疾病使用细胞替代疗法的努力。