Develop novel assays for assessing cellular and gene therapies

细胞和基因治疗产品必须经过无菌性、稳定性、纯度和效力测试。此外，重要的是要测试临床细胞治疗产品的身份，一致性和可比性。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身上收集的，因此可用于测试的材料数量有限。它们通常在生产后立即或不久输入，因此完成检测的时间非常有限。这些疗法中有许多是具有多种功能的复杂细胞。对这些治疗的临床效果至关重要的细胞功能通常不为人所知。传统上，流式细胞术、ELISA、ELISPOT和细胞培养等分析方法已被用于分析细胞和基因治疗。虽然这些分析已被证明是非常有用的，但可以用这些分析分析的因素的数量和类型是有限的。

Cell and gene therapy products must be tested for sterility, stability, purity and potency. In addition, its important to test clinical cell therapy 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 or shortly 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 cytometry, 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 assess the expression of the entire transcriptome. 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. Bone Marrow Stromal Cells (BMSCs) are being use to modulate the immune system and for a number of regenerative medicine applications including bone repair. BMSCs are a heterogeneous population of cells and a small number of BMSCs are skeletal stem cells which are important for regenerative medicine clinical applications. We have been investing BMSCs to determine markers that identify skeletal stem cells. We are using single cell analysis and next generation DNA sequencing to identify markers for the skeletal stem cells. Chimeric Antigen Receptor (CAR) T cells are being used to treat a number of hematologic malignancies, however, clinical outcomes have varied among recipients of these therapies and some of this variability is likely due to variability, and hence, differences in potency among CAR T cell products. We are using gene expression analysis, mRNA analysis, single cell analysis and next generation sequencing to identify factors associated with the clinical potency of these cells.