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Identification and characterization of bladder cancer stem cells

膀胱癌干细胞的鉴定和表征

基本信息

批准号：
8136484
负责人：
Keith Syson Chan
金额：
$ 24.15万
依托单位：
BAYLOR COLLEGE OF MEDICINE
依托单位国家：
美国
项目类别：
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/8136484
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

项目摘要

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(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，专门针对这个器官站点中的疾病的基础研究资金相对较少。长期以来，人们观察到人类肿瘤是异质性的，根据组织学或生物学特性(即与锚定无关的生长特性、增殖和分化状态)来定义。一种模型假设肿瘤中存在一个层次结构，这可能解释了肿瘤内肿瘤细胞群体的异质性。这一模型认为，存在罕见的具有干细胞样特性的肿瘤起始细胞(肿瘤干细胞)，它们可以自我更新并分化为表型多样的肿瘤细胞，重建原始肿瘤的异质性。在目前的提案中，我首次描述了从患者肿瘤中成功分离和初步鉴定一种罕见的CD44阳性的膀胱癌干细胞(CSCs)。我假设CSCs存在于所有患者的膀胱肿瘤中，因此可以根据其独特的生物学特性进行前瞻性的分离。主要的实验方法将是利用细胞表面标记的组合(目标1A、B和C)和报告自我更新信号通路的结构(Aim2A)。膀胱CSCs的定位及其表达水平与疾病进展的关系将通过组织切片和高通量组织芯片(Aim1D)的免疫组织化学分析来确定。此外，为了确定膀胱癌的“分子特征”，将采取两种不同的方法(Aim3)：(I)小型分子筛选，通过定量RT-PCR在mRNA水平上寻找特定的“严厉基因”(Aim3Ai)，并利用尖端微流体西方技术在蛋白质水平上研究这些候选基因(Aim3Aii)；(Ii)使用HEEBO微阵列进行全基因组筛选(Aim3B)。与Mark通路(Aim2C)相比，对膀胱CSCs特有的自身通路(Aim2B)进行临床前靶向治疗将揭示体内的药物靶点。我推测这些CSCs实际上是“根”，因此是未来抗癌治疗的理想靶点。我的长期目标是丰富高纯度的膀胱CSCs群体，进一步鉴定它们的生物学特性(例如侵袭和迁移到远端器官的能力)，并开发针对这类CSCs的特定策略，试图根除晚期膀胱癌。