Identification and characterization of bladder cancer stem cells

膀胱癌干细胞的鉴定和表征

• 批准号: 7937096
• 负责人: Keith Syson Chan
• 金额: $ 24.9万
• 依托单位: BAYLOR COLLEGE OF MEDICINE
• 依托单位国家: 美国
• 财政年份: 2007
• 资助国家: 美国
• 起止时间: 2007-08-15 至 2012-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7937096
• 关键词: Anchorage-Independent Growth; Basic Science; Biological; Bladder Neoplasm; CD44 gene; Cell Count; Cell surface; Cells; Chemicals; Data; Development; Disease; Disease Progression; Distal; Drug Delivery Systems; Flow Cytometry; Frequencies; Funding; Future; Genes; Goals; Heterogeneity; Histology; Human; Immunocompromised Host; Invaded; Keratin; Malignant Epithelial Cell; Malignant Neoplasms; Malignant neoplasm of urinary bladder; Messenger RNA; Metastatic Lesion; Microfluidics; Mitogen-Activated Protein Kinases; Modeling; Molecular Profiling; Mus; Nature; Nuclear; Oligonucleotide Microarrays; Organ; Pathway interactions; Patients; Plant Roots; Population; Property; Proteins; Reporter; Reporting; Reverse Transcriptase Polymerase Chain Reaction; Role; Sampling; Signal Pathway; Signal Transduction; Site; Specimen; Staging; Stem cells; Technology; Time; Tissue Microarray; Tissues; Transitional Cell Carcinoma; United States; Western Blotting; Xenograft Model; adult stem cell; base; cancer stem cell; cancer therapy; cell type; evidence base; expression vector; genome wide association study; in vivo; inhibitor/antagonist; molecular scale; neoplastic cell; pre-clinical; reconstitution; self-renewal; stem cell population; transcription factor; tumor; tumor xenograft; urologic

DESCRIPTION (provided by applicant): Bladder cancer is the second most common urological malignancy in the United States and is usually not curable at its advanced stage. However, basic research funding geared specifically to diseases in this organ site is relatively meager. It has long been observed that human tumors are heterogenous, as defined by histology or biological properties (i.e. anchorage independent growth property, proliferation and differentiation status). One model hypothesizes the existence of a hierarchy in tumors, which may explain the heterogeneity of tumor cell population within a tumor. This model proposes that rare tumor-initiating cells with stem cell-like properties (cancer stem cells) exist, which can self-renew and differentiate into phenotypically diverse tumor cells that reconstitute the heterogeneity of original tumor. In the current proposal, I describe for the first time the successful isolation and initial characterization of a rare CD44 positive, bladder cancer stem cells (CSCs) from patient tumor. I hypothesize that CSCs exist in all patient bladder tumors and therefore can be prospectively isolated based on their unique biological properties. The primary experimental approaches will be utilizing a combination of cell surface markers (Aim 1A, B&C) and constructs that report on self-renewal signaling pathways (Aim2A). Localization of bladder CSCs and the correlation of their expression level to disease progression will be determined by immunohistochemical analysis in tissue sections and high throughput tissue microarrays (Aim1 D). Further, in order to define a molecular signature" for bladder CSCs, two separate approaches will be taken (Aim3): (I) a small scale molecular screen to look at specific "sternness genes" at mRNA level by quantitative RT-PCR (Aim3Ai), and utilization of cutting edge microfluidics western technologies to look at these candidates at protein level (Aim3Aii); (II) a genome wide screen using HEEBO microarrays (Aim 3B). Preclinical targeting of self- pathways unique to bladder CSCs (Aim2B) in comparison to the MARK pathway (Aim2C) will reveal drug targets in vivo. I hypothesize that these CSCs are in fact the "roots", and therefore ideal targets for future anti-cancer therapies. My long term goal is to enrich for a highly purified population of bladder CSCs, further characterize their biological properties (e.g. ability to invade, and migrate to distal organs), and to develop specific strategies to target such CSCs in attempt to eradicate advanced stage bladder cancers.

描述（由申请人提供）：膀胱癌是美国第二常见的泌尿系统恶性肿瘤，通常在晚期无法治愈。然而，专门针对该器官部位疾病的基础研究资金相对较少。长期以来，人们已经观察到，人类肿瘤是异质性的，如组织学或生物学特性所定义的（即锚定独立生长特性、增殖和分化状态）。一种模型假设肿瘤中存在等级，这可以解释肿瘤内肿瘤细胞群体的异质性。该模型提出，存在具有干细胞样特性的罕见肿瘤起始细胞（癌症干细胞），其可以自我更新并分化成表型多样的肿瘤细胞，这些肿瘤细胞重建原始肿瘤的异质性。在目前的建议中，我描述了第一次成功的分离和初步鉴定的一种罕见的CD 44阳性，膀胱癌干细胞（CSC）从患者的肿瘤。我假设CSC存在于所有患者的膀胱肿瘤中，因此可以基于其独特的生物学特性进行前瞻性分离。主要的实验方法将利用细胞表面标志物（Aim 1A、B&C）和报告自我更新信号传导途径的构建体（Aim 2A）的组合。膀胱CSC的定位及其表达水平与疾病进展的相关性将通过组织切片和高通量组织微阵列（AimlD）中的免疫组织化学分析来确定。此外，为了定义膀胱CSC的"分子标记”，将采取两种单独的方法（Aim 3）：（I）通过定量RT-PCR在mRNA水平上观察特定的“干性基因”的小规模分子筛选（Aim 3Ai），以及利用尖端微流体西方技术在蛋白质水平上观察这些候选物（Aim 3Aii）;（II）使用HEEBO微阵列（Aim 3B）的全基因组筛选。与MARK途径（Aim 2C）相比，膀胱CSC特有的自身途径（Aim 2B）的临床前靶向将揭示体内药物靶标。我假设这些CSC实际上是“根”，因此是未来抗癌治疗的理想靶点。我的长期目标是富集高度纯化的膀胱CSC群体，进一步表征其生物学特性（例如侵入和迁移到远端器官的能力），并开发针对此类CSC的特定策略，以尝试根除晚期膀胱癌。