Blood Cell-Spheres: Growth, Description, and Potential

目标和策略：该项目旨在开发个体化（患者特异性）疗法，利用抗体和/或免疫细胞（“自然杀伤细胞”、巨噬细胞、细胞毒性T细胞）攻击致癌白血病“干细胞”（LSC），而不损害正常（血液）造血“干细胞”（HSC）。实验活动旨在(1)分离和比较来自同一患者的LSC和正常HSC，(2)确定仅位于致癌LSC上的免疫“靶点”，以及(3)探索细胞结构差异所建议的免疫治疗方案。成像是至关重要的，将包括光学显微镜，共聚焦（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，细胞显微照片可以识别细胞内和表面结构以及细胞类型和成熟度。关于细胞的生长、转化和分析，这一建议本质上是对最近（和深刻的）干细胞相关成就的逻辑延伸。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、高科技实验室和成像资源。这些进步激励我们利用从患者常规血液标本中提取的白血病细胞来培养“血细胞球”（BCS）。这些新的白血病BCS令人兴奋，因为已知其他“癌细胞球”含有、浓缩或“制造”那些在患者体内罕见且难以捉摸的关键致癌“干细胞”。现在，正常的BCS将被培养出来（令人兴奋的是，像“胚”这样的正常细胞球也含有“干细胞”——这些干细胞可以在动物身上再生截肢）。将对BCS进行成像和分析，以确定(1)LSC是否位于白血病BCS内，(2)正常HSC是否位于正常BCS内。如果在类似的细胞球中发现了干细胞，将使用染色/成像技术对LSC和正常HSC进行比较，这些技术可以显示球的内部和表面细节以及其中的单个细胞。如果没有所需的“干细胞”，在添加更多促进“干细胞”生成的因子和细胞后，将培养“新一代”的BCS。当正常细胞和癌症“干细胞”都可以被分离出来时，它们将被仔细地比较，以确定仅在LSC上发现的潜在“目标”。最后，可以攻击这些独特目标的抗体和/或免疫细胞将被开发出来。健康影响：如果能够从白血病患者血液标本中培养的BCS中分离出LSC和正常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遗传变异引起的治疗失败，并将成为最理想和最谦逊的可能策略之一。此外，任何能够使科学家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.