HIV/NeuroAIDS Patient Derived Induced Pluripotent Stem (iPS) Cells

描述(由申请人提供)：这是SBIR第一阶段的重新提交，以资助从滥用酒精和/或其他物质(如可卡因、鸦片等)的艾滋病毒/神经艾滋病患者群体中开发诱导多能干细胞(IPS)的概念验证研究。酒精和滥用药物是人类免疫缺陷病毒(HIV)引起的疾病进展的重要辅助因素，这一点已被公认。艾滋病毒感染、酗酒和滥用药物严重影响免疫系统以及中枢神经系统(CNS)。在艾滋病毒患者中，异常相互交错，导致一系列被称为神经艾滋病的症状。术语“神经艾滋病”包括无数新的神经疾病，这些疾病是艾滋病毒对中枢和周围神经系统造成损害的主要后果。医疗条件包括感觉神经病、脊髓病、艾滋病毒痴呆症和认知/运动障碍异常。在少数群体中，艾滋病毒感染和神经艾滋病进展的比例不成比例。一个民族内部是否存在可能加剧神经艾滋病转化率的生物差异尚不得而知。必须进行更明确的研究，以了解酒精和药物滥用对神经艾滋病的潜在机制，特别是检查神经免疫、潜在的中枢神经系统储存库、神经元和神经胶质细胞。我们建议从酗酒和/或吸毒的少数艾滋病毒患者身上收集成纤维细胞样本，建立iPS细胞。与人类胚胎干细胞(HESCs)一样，iPS细胞可以无限自我更新，并可以分化为人体大多数细胞，包括中枢神经系统。此外，iPS细胞既可以通过靶向基因破坏进行修饰，也可以从患者身上创建为疾病特异性细胞系。我们已经成功地将iPS细胞和hESCs分化为神经元系，以研究各种药物和已知毒素(如酒精)对神经退行性变的影响。我们已经成功地鉴定出在神经发生过程中受酒精差异调控的基因。我们相信，iPS细胞是一个强大的工具，可以深入了解哪些生物标记物在滥用药物的同时感染艾滋病毒的人群中发展为神经艾滋病的关键。从患者的大脑中获取原代细胞只会加剧他们目前的神经问题。这一提议的成功将创造第一个旨在研究神经艾滋病生物标记物的细胞系。已发表的进入人体临床试验的药物报告估计，根据治疗方法的不同，成本从5亿美元到20亿美元不等。所有进入第一阶段试验的新药和成功完成人类使用的产品许可的成功率约为9%，同时，中枢神经系统候选药物上市治疗的成功率预计为3%。几种药物已经在艾滋病毒相关性痴呆(HAD)的I期和II期临床试验中进行了测试，HAD通常与艾滋病毒/神经艾滋病有关，不幸的是，没有一项研究能够改善健康状况。显然需要采用既鼓励创新又确保药物安全和疗效的战略来提高生产力和降低药物开发成本。HIV/NeuroAIDS患者来源的iPS细胞是研究中枢神经系统发育、功能和疾病的模型，是补充体内实验的有用工具。我们认为，HIV/NeuroAIDS患者来源的iPS细胞的出现可以减少目前与药物开发相关的低效、高磨损率、延长的研发周期和高昂的成本。iPS细胞相对便宜，可以作为生物技术和制药行业第一阶段药物发现和临床前产品开发的高通量体外分析的模型。这一阶段应用的具体目标是：具体目标1：从药物滥用少数民族和患有艾滋病毒/神经艾滋病的非少数民族的成纤维细胞中分离出诱导性多能干细胞。具体目标2：将iPS细胞分化为神经细胞和神经胶质细胞系。这项工作的成功完成将支持第二阶段的申请，目标是包括更大的iPS细胞开发、毒理学筛选和产品获得FDA 510(K)批准的队列。

DESCRIPTION (provided by applicant): This is an SBIR Phase I re-submission to fund proof of concept studies for the development of induced pluripotent stem (iPS) cells from a population of HIV/NeuroAIDS patients who abuse alcohol and/or other substances (i.e. cocaine, opium, etc). It has been accepted that alcohol and drugs of abuse are significant cofactors in the progression of human immunodeficiency virus (HIV)-induced disease. HIV infection, alcohol and drugs of abuse significantly affect the immune system as well as the central nervous system (CNS). In patients with HIV, abnormalities intersect to lead to a constellation of symptoms known as NeuroAIDS. The term "NeuroAIDS" includes a myriad of novel neurologic disorders which are a primary consequence of damage to the central and peripheral nervous system by HIV. The medical conditions include sensory neuropathy, myelopathy, HIV dementia, and cognitive/motor disorder abnormalities. In minority groups HIV infection and NeuroAIDS progression occur at a disproportionate rate. The existence of biological differences within an ethnic group which might exacerbate the rate of conversion to NeuroAIDS is unknown. More definitive research has to be performed to understand the underlying mechanisms of alcohol and drug abuse on NeuroAIDS, specifically examining neuroimmunity, latent CNS reservoirs, neurons and glial cells. We are proposing to create iPS cell from fibroblast samples collected from minority HIV patients who abuse alcohol and/or drugs. Like human embryonic stem cells (hESCs), iPS cells can self-renew indefinitely and can be differentiated into most cells of the human body, including the CNS. Furthermore, iPS cells can either be modified by targeted gene disruption or created from patients as disease specific cell lines. We have successfully differentiated iPS cells and hESCs into neuronal lineages to study the neurodegenerative effect of various drugs and known toxins such as alcohol. We have successfully identified genes that are differentially regulated by alcohol during neurogenesis. We believe iPS cells to be a powerful tool in gaining insight into which biomarkers are essential for the progression to NeuroAIDS in populations infected with HIV while abusing substances. Obtaining primary cells from a patient's brain would only exacerbate their current neurological problem. The success of this proposal will create the first cell line that is designed to look at biomarkers of NeuroAIDS. Published reports for drugs entering human clinical trials estimate costs to vary from $500 million to $2 billion depending on the therapy. The success rate for all new drugs entering phase I trials and the successful completion of product licensing for human use is approximately 9%, meanwhile, the success rate for CNS drug candidates to become marketed therapeutic is projected at 3%. Several drugs have been tested in phase I and II clinical trials for HIV-associated dementia (HAD), which is commonly associated with HIV/NeuroAIDS and unfortunately none of the studies were able to improve health conditions. There is clearly a need to increase productivity and decrease the cost of drug development using strategies that also encourage innovation while ensuring drug safety and efficacy. HIV/NeuroAIDS patient-derived iPS cells represent a model to study CNS development; function and disease and are a useful tool to complement in vivo experimentation. We propose that the current inefficiencies, high attrition rates, protracted R&D cycles and elevated cost associated with drug development can be reduced by the advent of HIV/NeuroAIDS patient derived iPS cells which are relatively inexpensive and can be used as a model for high- throughput in vitro analysis in first phase drug discovery and preclinical product development in the biotechnology and pharmaceutical industry. The Specific Aims of this Phase I application are: Specific Aim #1: To create iPS cells from fibroblast cells taken from substance abusing minorities and non-minorities with HIV/NeuroAIDS. Specific Aim #2: Differentiate iPS cells into neuronal and glial cell lineages. Successful completion of this work will support a Phase II application, with the goals of including a larger cohort for iPS cell development, toxicology screening and the product leading to FDA 510(k) approval.