Blood Cell-Spheres: Growth, Description, and Potential

描述(由申请者提供)：目标和策略：该项目寻求开发个体化(针对患者)的疗法，该疗法将使用抗体和/或免疫细胞(“自然杀伤细胞”、巨噬细胞、细胞毒性T细胞)来攻击致癌的白血病“干细胞”(LSC)，而不损害正常(血液)的造血“干细胞”(HSC)。实验室活动旨在(1)分离和比较同一患者的LSC和正常HSC，(2)确定仅位于致癌LSC上的免疫“靶点”，以及(3)探索根据细胞结构差异提出的免疫治疗方案。成像是至关重要的，它将包括光学显微镜、共焦(3-D)、电子(传输和扫描)和双光子(3-D)。当通过染色技术增强时，细胞显微图像可以识别细胞内和表面结构以及细胞类型和成熟程度。关于细胞的生长、转化和分析，这一建议基本上是对最近(和深刻的)干细胞相关成就的合乎逻辑的延伸。新的见解和技术进步提供了开发免疫疗法所需的所有工具：有用的概念框架、转化细胞的方法以及高科技实验室和成像资源。这些进展激励我们使用从常规患者血液样本中提取的白血病细胞来培育“血细胞球”(BCS)。这些新的白血病BC是令人兴奋的，因为已知其他“癌细胞球体”包含、浓缩或“创造”那些关键的致癌“干细胞”，这些干细胞在患者体内是稀有和难以捉摸的。现在，正常的BCs将被培养出来(令人兴奋的是，像“原始细胞”这样的正常细胞球体也含有“干细胞”--在动物体内，这些细胞可以再生被截肢的肢体)。将对BCS进行成像和分析，以确定(1)LSC是否位于白血病BCS内，以及(2)正常HSC是否位于正常BCS内。如果在相似的细胞球中发现了干细胞-LSC和正常的HSC将使用染色/成像技术进行比较，这种技术可以显示球体和其中的单个细胞的内部和表面细节。如果不存在所需的“干细胞”，则在加入更多促进“干细胞”生成的因子和细胞后，就会培养出“新一代”的BCS。当正常干细胞和癌症干细胞都能被分离出来时，他们将被仔细比较，以确定仅在LSC上发现的潜在“靶点”。最后，将开发出能够攻击这些独特靶点的抗体和/或免疫细胞。对健康的影响：如果能从白血病患者的血液样本中分离出LSC和正常的HSC，就可以开发出针对患者的免疫疗法。个体化治疗将消除由基因变异造成的治疗失败，并将代表可能的最理想和最低调的策略之一。此外，任何使科学家能够从更成熟的细胞中获得“干细胞”或“祖细胞”的实验室程序，都可能帮助科学家再生人体组织和器官。

DESCRIPTION (provided by applicant): Goals and Strategy: This project seeks to develop individualized (patient-specific) therapies that will use antibodies and/or immune cells ("natural killer cells," macrophages, cytotoxic T cells) to attack cancer-causing leukemia "stem" cells (LSC's) without harming normal (blood) hematopoietic "stem" cells (HSC's). Lab activities are designed to (1) isolate and compare LSC's and normal HSC's from the same patient, (2) identify immunologic "targets" located only on the cancer-causing LSC's, and (3) explore immune-therapy options suggested by cell-structure differences. Imaging is vital and will include light microscopy, confocal (3-D), electron (transmission and scanning), and two-photon (3-D). When enhanced by staining techniques, cell micrographs can identify intracellular and surface structures as well as cell type and maturity level. Regarding growing, transforming, and analyzing cells, this proposal is essentially a logical extension of recent (and profound) stem-cell-related achievements. New insights and technical advances provide all the tools needed to develop immune-therapies: a useful conceptual framework, ways to transform cells, and high-tech lab and imaging resources. These advances inspired us to grow "blood cell-spheres" (BCS) using leukemia cells taken from routine patient blood specimens. These new leukemia BCS are exciting because other "cancer cell-spheres" are known to contain, concentrate, or "create" those critical cancer-causing "stem" cells that are rare and elusive inside a patient's body. Now, normal BCS will be grown (exciting since normal-cell spheres like "blastemas" also contain "stem" cells - these regenerate amputated limbs in animals). BCS will be imaged and analyzed to determine if (1) LSC's are inside leukemia BCS and (2) normal HSC's are inside normal BCS. If found - and stem cells have been found in similar cell-spheres - LSC's and normal HSC's will be compared using staining/imaging techniques that can show internal and surface details of spheres and the individual cells inside them. If the desired "stem" cells are not present, "newer generations" of BCS will be grown after adding more factors and cells that promote "stem" cell creation. When both normal and cancer "stem" cells can be isolated, they will be carefully compared to identify potential "targets" found only on LSC's. Finally, antibodies and/or immune cells that can attack these unique targets will be developed. Health-Impact: If LSC's and normal HSC's can be isolated from BCS grown from a leukemia patient's blood specimen, patient-specific immunotherapies can be developed. Individualized therapies would eliminate treatment failures caused by genetic variations and would represent one of the most ideal and unassuming strategies possible. Also, any lab procedure that enables scientists to derive "stem" or "progenitor" cells from more mature cells may help scientists regenerate human tissues and organs.