Using human IPS cells to study fate, function and neurodegenerative disease

使用人类 IPS 细胞研究命运、功能和神经退行性疾病

基本信息

批准号：
10207780
负责人：
LORRAINE IACOVITTI
金额：
$ 37.82万
依托单位：
THOMAS JEFFERSON UNIVERSITY
依托单位国家：
美国
项目类别：
财政年份：
2012
资助国家：
美国
起止时间：
2012-04-01 至 2024-06-30
项目状态：
已结题

来源：
https://reporter.nih.gov/project-details/10207780
关键词：
1-Methyl-4-phenylpyridinium Affect Aftercare Amyotrophic Lateral Sclerosis Animal Model Autologous Binding Cell Line Cell surface Cells Chromatin Chromatin Structure Clinic Cues DNA DNA biosynthesis Data Development Disease Disease model Dopamine Epigenetic Process Genetic study Goals Grant Harvest Heterogeneity High Pressure Liquid Chromatography Histones Human Implant In Vitro Investigational Therapies LRRK2 gene Midbrain structure Mind Modeling Molecular Conformation Motor Motor Neuron Disease Motor Neurons Muscle Names Neurodegenerative Disorders Neurons Oxidopamine Parkinson Disease Pathogenicity Pathway interactions Patients Phenotype Physiological Population Positron-Emission Tomography Process Rattus Specific qualifier value Technology Testing Therapeutic Therapeutic Uses Time Toxic Environmental Substances Translations Transplantation Undifferentiated Work alpha synuclein cell replacement therapy cell type dopaminergic neuron drug discovery human embryonic stem cell in vivo induced pluripotent stem cell insight morphogens motor behavior nerve stem cell progenitor recruit stem cell differentiation stem cells synaptic function tool transcription factor

项目摘要

Human induced pluripotent stem cells (iPSCs), with their potential to generate autologous patient-derived cells, hold great promise for the study and treatment of a host of devastating diseases, including Parkinson's disease (PD) and amyotrophic lateral sclerosis (ALS), to name a few. One of the major obstacles slowing the translation of this powerful technology to the clinic is the heterogeneity of both desired and unwanted cell types generated in grafts of iPSCs, even after cells have been directed down specific differentiation pathways. Similarly in culture, a multitude of cell types are generated after treatment of iPSCs with lineage-specifying cocktails. These realities, combined with the current lack of suitable cell surface markers for the selection of specific desired cell types, has significantly impacted the field, hampering our ability to develop cell replacement therapies or to accurately model diseases in the dish. One plausible explanation for the observed cell heterogeneity is that presumptive undifferentiated pluripotent cells sometimes spontaneously initiate the process of differentiation after encountering lineage- specifying cues in culture, thereby precluding their subsequent directed differentiation by exogenously added differentiation cocktails. Currently, there are no assured ways to know if iPSCs have begun to spontaneously differentiate. However, an exciting new discovery made during our previous grant cycle suggests that the epigenetic state of chromatin shortly after DNA replication serves as a reliable and very early indicator of the state of differentiation of a stem cell. Our results suggest that it may be possible to uniformly direct the differentiation of all iPSCs toward a specific cell fate if chromatin can be kept closed until incubation with exogenous fate-specifying differentiation factors. If these insights are indicative of a more generalized principle, then it should be possible to generate pure populations of neural progenitors (NPs) of different subtypes which can give rise to homogeneous populations of various neurons for studies in culture and in animal models of multiple diseases. With these goals in mind, our Specific Aims for this proposal are: 1) to assess chromatin status during commitment to a motor neuron phenotype; 2) to generate pure populations of midbrain dopamine (mDA) and motor NPs and neurons which will be characterized for phenotype and synaptic function in culture and 3) to further determine whether homogeneous mDA-committed NPs/neurons can accurately model PD in the dish and be used in transplants to therapeutically treat PD rat models.

人类诱导的多能干细胞（IPSC），其潜力产生自体患者衍生细胞，对研究和治疗多种毁灭性疾病的巨大希望，包括帕金森氏病（PD）和肌萎缩性侧索硬化症（ALS）仅举几例。专业之一减慢这项强大技术向诊所的翻译的障碍是两者的异质性即使在细胞被定向后，在IPSC的移植物中产生的期望和不需要的细胞类型特定的分化途径。同样，在培养中，处理后会产生多种细胞类型具有谱系指定鸡尾酒的IPSC。这些现实，结合当前缺乏合适的单元格选择特定所需细胞类型的表面标记已显着影响该领域，阻碍了我们开发细胞替代疗法或准确对菜肴中的疾病进行建模的能力。观察到的细胞异质性的一个合理解释是推定未分化多能细胞有时会自发启动分化过程，遇到谱系 - 在培养中指定线索，从而排除了其随后的定向分化增加了分化鸡尾酒。目前，尚无保证的方法知道IPSC是否已经开始自发区分。但是，在我们以前的赠款周期中提出的令人兴奋的新发现表明DNA复制后不久的染色质的表观遗传状态可作为可靠且非常非常早期指标的干细胞分化状态。我们的结果表明，可能有可能如果可以关闭染色质，将所有IPSC的分化均匀地指向特定的细胞命运直到与外源性命运指定因子一起孵育。如果这些见解表明更广泛的原理，那么应该有可能产生神经祖细胞的纯种群不同亚型的（NP）可以引起各种神经元的同质种群进行研究在文化和多种疾病的动物模型中。考虑到这些目标，我们的具体目标是建议是：1）评估对运动神经元表型的承诺期间的染色质状态； 2）到产生中脑多巴胺（MDA）和运动NP和神经元的纯种群培养中的表型和突触功能的特征和3）进一步确定是否是否同质的MDA承诺的NP/神经元可以在菜肴中准确建模PD，并用于移植到治疗方法治疗PD大鼠模型。

项目成果

期刊论文数量（4）

专著数量（0）

科研奖励数量（0）

会议论文数量（0）

专利数量（0）

BMP and TGF-β pathway mediators are critical upstream regulators of Wnt signaling during midbrain dopamine differentiation in human pluripotent stem cells.

DOI：
10.1016/j.ydbio.2013.01.012
发表时间：
2013-04-01
期刊：
DEVELOPMENTAL BIOLOGY
影响因子：
2.7
作者：
Cai, Jingli;Schleidt, Stephanie;Pelta-Heller, Joshua;Hutchings, Danielle;Cannarsa, Gregory;Iacovitti, Lorraine
通讯作者：
Iacovitti, Lorraine

Cell-to-Cell Transmission of Dipeptide Repeat Proteins Linked to C9orf72-ALS/FTD.