Develop novel assays for assessing cellular and gene therapies

细胞和基因治疗产品必须进行无菌性、稳定性、纯度和效力测试。此外，对临床细胞治疗产品的同一性、一致性和可比性进行检测也很重要。 测试细胞和基因疗法具有挑战性。这些疗法通常是从一个人那里收集的，因此可用于测试的材料数量有限。它们通常在生产后立即输血，因此完成测定的时间非常有限。这些疗法中的许多是具有多种功能的复杂细胞。对这些疗法的临床有效性至关重要的细胞功能通常是未知的。传统上，诸如流式细胞术、ELISA、ELISPOT和细胞培养的分析测定已用于分析细胞和基因疗法。虽然这些测定已被证明是非常有用的，但可以用这些测定分析的因素的数量和类型是有限的。 我们一直在研究使用基因和微小RNA表达分析细胞疗法。这些检测可能只需要使用少量的细胞，并可用于一次测试多达44，000个因子。我们一直在测试全局基因和微小RNA表达谱的能力，以确定这些测定法用于评估细胞疗法的稳定性、纯度和效力的实用性。我们已经表明，基因表达谱可以检测储存细胞的变化，并检测外周血白细胞（T细胞，B细胞和单核细胞）和造血干细胞之间的差异。基因表达谱分析还能够检测未成熟和成熟树突状细胞（DC）之间的差异，并且可用于比较使用成熟剂的不同组合产生的成熟DC。我们将很快开始分析临床DC产品，以比较基因表达与临床结果，以鉴定可能用于一致性、稳定性、同一性和效价测试的生物标志物。 我们已经评估了给予造血干细胞移植患者以改善干细胞植入和预防疾病复发的T雷帕霉素细胞的全球基因表达谱。这些研究揭示了几种生物标志物，这些生物标志物可能对这些产品的鉴别、一致性和效价检测有用。我们现在正在比较T雷帕霉素产品生物标志物与临床结果测量，以确定效力生物标志物。 我们一直在研究骨髓基质细胞（BMSC）的敏感性，并发现了一个24个基因集，预测BMSC在培养中的时间，通过群体倍增法测量。我们也一直在寻找由肽脉冲的成熟树突状细胞表达的生物标志物，其表达与临床疗效相关。

Cell and gene therapy products must be tested for sterility, stability, purity and potency. In addition, its important to test clinical cell thearpy products for identity, consistency and comparability. Testing cellular and gene therapies is challenging. These therapies are generally collected from a single person so the quantity of material available to test is limited. They are typically transfused immediately after they are produced so there is a very limited amount of time to complete the assays. Many of these therapies are complex cells that have multiple functions. The cell functions that are critical to the clinical effectiveness of these therapies are often not known. Traditionally, analytic assays such as flow cytometery, ELISA, ELISPOT and cell culture have been used to analyze cellular and gene therapies. While these assays have proven to be very useful, the number and types of factors that can be analyzed with these assays is limited. We have been investigating the use of gene and micro RNA expression assays for the analysis of cellular therapies. These assays can require the use of only small quantities of cells and can be used to test up to 44,000 factors at one time. We have been testing the ability of global gene and micro RNA expression profiling to determine the utility of these assays for assessing the stability, purity and potency of cellular therapies. We have shown that gene expression profiling can detect changes in stored cells and detect differences between peripheral blood leukocytes (T cells, B cells and monocytes) and hematopoietic stem cells. Gene expression profiling has also been able to detect differences between immature and mature dendritic cells (DCs) and has been useful for comparing mature DCs produced using different combinations of maturation agents. We will soon begin analyzing clinical DC products to compare gene expression with clinical outcome in order to identify biomarkers that might be useful of consistency, stability, identity and potency testing. We have assessed global gene expression profiles of T Rapamycin cells that are given to hematopoietic stem cell transplant patients to improve stem cell engraftment and prevent disease relapse. The studies have revealed several biomarkers which may be useful for identity, consistency and potency testing of these products. We are now comparing T Rapamycin product biomarkers with clinical outcome measure to identify potency biomarkers. We have been investigating sensecence of bone marrow stromal cells (BMSCs) and have found a 24 gene set that predicts BMSC time in culture as measured by population doublings. We have also been seraching for biomarkers expressed by peptide-plused mature dendrtic cells whose expression coorelates with clinical effectiveness.