Dual Modality Labels for Macro and Micro Detection and Quantification of Stem Cel

用于干细胞宏观和微观检测及定量的双模态标签

• 批准号: 7909750
• 负责人: Brian David Gray
• 金额: $ 19.62万
• 依托单位: MOLECULAR TARGETING TECHNOLOGIES, INC.
• 依托单位国家: 美国
• 财政年份: 2010
• 资助国家: 美国
• 起止时间: 2010-07-15 至 2012-04-14
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7909750
• 关键词: Affect; Alzheimer' s Disease; Animal Model; Animals; Autoradiography; Binding; Biodistribution; Biological Process; Carbocyanines; Cell Differentiation process; Cell Survival; Cell Therapy; Cell membrane; Cell physiology; Cells; Chelating Agents; Chemicals; Clinical; Clinical Research; Clinical Trials; Controlled Study; Data; Detection; Development; Diabetes Mellitus; Disadvantaged; Disease; Dyes; Engraftment; Evaluation; Family; Fluorescence; Fluorochrome; Future; Generations; Half-Life; Hand; Heart Diseases; High Pressure Liquid Chromatography; Histologic; Histology; Homing; Hydrocarbons; Image; In Vitro; Indium; Indium-111; Individual; Ischemia; Knowledge; Label; Lead; Location; Measurement; Medicine; Membrane; Methods; Microscopic; Microscopy; Modality; Molecular; Monitor; Mus; Noise; Nuclear; Organ; Oxyquinoline; PKH 26; PKH67; Parkinson Disease; Pentetic Acid; Phase; Physiological; Positron-Emission Tomography; Property; Radio; Radioactivity; Radioisotopes; Radiolabeled; Reagent; Reference Standards; Research; Resolution; Signal Transduction; Site; Solubility; Spinal cord injury; Stem cells; Stroke; System; Tail; Time; Tissue Sample; Tissues; Toxic effect; Tracer; Transplantation; Tropolone; analog; cell type; commercialization; cytotoxicity; daughter cell; embryonic stem cell; fluorophore; imaging modality; imaging probe; improved; in vitro Assay; in vivo; insight; interest; killings; meetings; migration; next generation; optical imaging; public health relevance; radiation effect; radiochemical; radiotracer; safety study; single photon emission computed tomography; stem; stem cell biology; stem cell differentiation; stem cell therapy; tool; uptake

DESCRIPTION (provided by applicant): Stem cells hold promise for treatment of a number of disease states such as Parkinson's, Alzheimer's, spinal cord injury, diabetes, ischemia stroke and heart disease since stem cells have the potential under certain physiological conditions to develop into many different specialized cell types with individual functions. There are 2,620 clinical trials involving stem cells that are either on-going or have been completed, however, to-date, no stem cell therapy has received full FDA approval. The potential that stem cells offer remains to be better understood by observing their fate in vivo (e.g. bio-distribution, survival and differentiation) and this requires the means by which to track the cells non-invasively overtime. Methods are available to visualize cells, each having its own advantages and disadvantages, however, at present, no single imaging modality possess all the desired qualities for optimal evaluation of stem cell therapies. Likewise, many currently available direct cell labels have limitations due to cell toxicity, intracellular radiation effects, inefficient uptake and most importantly, rapid elution from the cell. We hypothesize that dual-modality imaging of stem cells using a non-diffusable dual-labeled imaging probe consisting of a far-red fluorophore and a radionuclide can provide complementary information regarding stem cell location longitudinally, thereby providing an accurate global picture of stem cell biodistribution in vivo which may lead to an improved understanding of stem cell biology and guide emerging stem cell therapies. In Phase I, MTTI will synthesize a dual modality probe for stem cell labeling comprising of three components: (i) a chelator (DTPA) for radiolabeling with 111In allowing detection by SPECT; (ii) a far red emitting fluorochrome to permit observation by optical imaging at the macro and micro-levels, and (iii) long hydrocarbon tails to provide stable non-diffusable incorporation of the probe into the plasma membrane. The probe's cytotoxicity, radiotoxicity, signal:noise, membrane retention and effect on various mouse stem cell functions will be characterized using standard in vitro assays. We expect to establish a suitable probe concentration which does not alter cell viability, proliferation or differentiation, and show that the probe is passed on to the next generation of daughter cells; but does not get incorporated into neighboring cells. Finally, utility of the probe to quantify and track stem cell distribution in vivo in a normal mouse using small animal SPECT and optical imaging systems will be evaluated. The fluorochrome present will also permit microscopic evaluation of tissue samples of interest after sacrifice. We expect to demonstrate that stem cells with the dual labeled marker will localize and accumulate in our animal model in a manner consistent with the cell type and be "visible" for several cell generations. Phase II will include studies in larger animal models, synthesis and evaluation of a dual modality probe for PET and optical imaging, commercialization of the dual probes as research tools and initiation of assembly of a data package for eventual clinical use. PUBLIC HEALTH RELEVANCE: Stem cells hold promise for the treatment of a number of disease states such as Parkinson's, Alzheimer's, spinal cord injury, diabetes, ischemia stroke and heart disease, but their true potential remains to be better understood by observing their fate in vivo. We propose to develop a dual modality label for stem cell tracking using nuclear and optical imaging modalities. This label is expected to provide a highly sensitive and accurate global picture of stem cell biodistribution longitudinally, which may lead to an improved understanding of stem cell biology, and in the future guide emerging stem cell therapies.

描述（由申请人提供）：干细胞有望用于治疗许多疾病状态，例如帕金森氏症、阿尔茨海默氏症、脊髓损伤、糖尿病、缺血性中风和心脏病，因为干细胞在某些生理条件下具有发育成具有个体功能的许多不同特化细胞类型的潜力。有2，620项涉及干细胞的临床试验正在进行或已经完成，然而，迄今为止，没有干细胞疗法获得FDA的全面批准。干细胞提供的潜力仍然需要通过观察它们在体内的命运（例如生物分布，存活和分化）来更好地理解，这需要非侵入性地跟踪细胞的方法。方法可用于可视化细胞，每种方法都有其自身的优点和缺点，然而，目前，没有一种成像方式具有用于最佳评估干细胞疗法的所有所需质量。同样，许多目前可用的直接细胞标记由于细胞毒性、细胞内辐射效应、低效率摄取以及最重要的是从细胞中快速洗脱而具有限制。我们假设使用由远红荧光团和放射性核素组成的非扩散性双标记成像探针进行干细胞的双模态成像可以提供关于干细胞纵向位置的互补信息，从而提供干细胞体内生物分布的准确全局图像，这可能会导致对干细胞生物学的更好理解并指导新兴的干细胞治疗。在第一阶段，MTTI将合成一种用于干细胞标记的双模态探针，包括三种组分：（i）螯合剂（DTPA），用于用111 In进行放射性标记，允许通过SPECT进行检测;（ii）远红外发射荧光染料，允许通过宏观和微观水平的光学成像进行观察;以及（iii）长烃尾，以提供探针稳定的非扩散性掺入质膜。探针的细胞毒性、放射毒性、信号：噪声、膜保留和对各种小鼠干细胞功能的影响将使用标准体外测定来表征。我们期望建立不改变细胞活力、增殖或分化的合适的探针浓度，并显示探针被传递到下一代子细胞;但不掺入相邻细胞。最后，使用小动物SPECT和光学成像系统，将评估探针在正常小鼠体内定量和跟踪干细胞分布的实用性。存在的荧光染料还将允许在处死后对感兴趣的组织样本进行显微镜评价。我们期望证明具有双标记标记物的干细胞将以与细胞类型一致的方式在我们的动物模型中定位和积累，并且在几代细胞中是“可见的”。第二阶段将包括在更大的动物模型中进行研究，合成和评估PET和光学成像的双模态探头，将双探头作为研究工具进行商业化，并开始组装数据包以供最终临床使用。 公共卫生关系：干细胞有望用于治疗许多疾病状态，如帕金森氏症，阿尔茨海默氏症，脊髓损伤，糖尿病，缺血性中风和心脏病，但它们的真正潜力仍有待于通过观察它们在体内的命运来更好地了解。我们建议开发一种双重模式的干细胞跟踪使用核和光学成像模式的标签。该标签有望提供干细胞纵向生物分布的高度敏感和准确的全球图片，这可能会导致对干细胞生物学的更好理解，并在未来指导新兴的干细胞疗法。