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(申请人提供)：干细胞有望治疗多种疾病，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有可能发展为多种不同的具有个别功能的特殊细胞类型。有2620项涉及干细胞的临床试验正在进行中或已经完成，然而，到目前为止，还没有干细胞疗法获得FDA的完全批准。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)来更好地了解，这需要通过非侵入性的超时跟踪细胞的方法。细胞可视化的方法是可用的，每种方法都有自己的优缺点，然而，目前还没有一种成像方式拥有最佳干细胞治疗评估所需的所有特性。同样，目前许多可用的直接细胞标记由于细胞毒性、细胞内辐射效应、摄取效率低以及最重要的是从细胞中快速洗脱而具有局限性。我们假设，使用由远红外线荧光团和放射性核素组成的不可扩散的双标记成像探针对干细胞进行双模式成像可以提供关于干细胞纵向位置的补充信息，从而提供干细胞在体内生物分布的准确全球图像，这可能有助于加深对干细胞生物学的理解，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探针，由三种成分组成：(I)用于111In放射标记的螯合剂(DTPA)，以便通过SPECT进行检测；(Ii)能够通过光学成像在宏观和微观水平上观察的远红荧光染料；(Iii)长碳氢化合物尾巴，用于稳定、不可扩散地将探针结合到质膜中。该探针的细胞毒性、放射毒性、信号噪声、膜滞留以及对各种小鼠干细胞功能的影响将使用标准的体外检测方法进行表征。我们期望建立一个合适的探针浓度，该浓度不会改变细胞的存活、增殖或分化，并表明该探针可传递给下一代子代细胞；但不会整合到邻近细胞中。最后，将评估该探针在使用小动物SPECT和光学成像系统对正常小鼠体内干细胞分布进行量化和跟踪的有效性。目前存在的荧光色素还将允许在牺牲后对感兴趣的组织样本进行显微镜评估。我们希望证明，带有双标记标记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，并在几代细胞中都是“可见的”。第二阶段将包括更大动物模型的研究，用于正电子发射计算机断层扫描和光学成像的双模式探头的合成和评估，作为研究工具的双探头的商业化，以及开始组装最终用于临床的数据包。 与公共健康相关：干细胞有望治疗多种疾病，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。我们建议开发一种使用核和光学成像模式进行干细胞追踪的双重模式标记。这一标签有望提供干细胞纵向分布的高度敏感和准确的全球图景，这可能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。