Stem cells and their aberrant self-renewal pathways in DCIS malignant progression

干细胞及其在 DCIS 恶性进展中的异常自我更新途径

• 批准号: 8550781
• 负责人: Kelli E Valdez
• 金额: $ 12.63万
• 依托单位: UNIVERSITY OF KANSAS MEDICAL CENTER
• 依托单位国家: 美国
• 财政年份: 2012
• 资助国家: 美国
• 起止时间: 2012-09-25 至 2015-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8550781
• 关键词: Applications Grants; Area; Automobile Driving; Award; Biological Assay; Breast; Breast Cancer Prevention; Cell Count; Cell Proliferation; Cell Separation; Cell surface; Cells; Clinical; Clinical Investigator; Conditioned Culture Media; Conflict (Psychology); Data; Development; Diagnosis; Duct (organ) structure; Ductal Epithelial Cell; ERBB2 gene; Epithelial; Estradiol; Faculty; Fluorescence; Focus Groups; Foundations; Funding; Funding Opportunities; Future; Genes; Genetic Heterogeneity; Goals; Growth; Heterogeneity; Histopathology; Hormonal; Hormones; Human; Human Resources; Immunoassay; Immunodeficient Mouse; In Situ Lesion; In Vitro; K-Series Research Career Programs; Laboratory mice; Lead; Leadership; Left; Lesion; Longitudinal Studies; Malignant - descriptor; Malignant Neoplasms; Mammary gland; Mastectomy; Mentors; Mesenchymal; Methods; Modeling; Molecular; Molecular Profiling; Mus; Neoplasm Metastasis; Non-Invasive Lesion; Noninfiltrating Intraductal Carcinoma; Ovarian Steroid Hormone; Paracrine Communication; Pathway interactions; Patient Care; Patients; Phenotype; Population; Positioning Attribute; Premalignant; Prevention strategy; Progesterone; Property; Public Health; Radiation; Research; Research Personnel; Research Training; Risk; Risk Assessment; Role; Sampling; Scientist; Secure; Series; Signal Pathway; Signal Transduction; Solid; Sorting - Cell Movement; Specimen; Staging; Stem cells; Surface; TACSTD2 gene; Techniques; Testing; Therapeutic; Time; Training; Translational Research; Variant; Woman; Xenograft Model; Xenograft procedure; anticancer research; base; breast lesion; breast lumpectomy; cancer stem cell; carcinogenesis; cell growth; design; human disease; human stem cells; improved; in vitro Assay; in vivo; malignant breast neoplasm; meetings; novel; paracrine; progenitor; programs; research study; response; self-renewal; skills; stem; stem cell division; stem cell population; steroid hormone; therapeutic target; tumor; tumor progression; working group

DESCRIPTION (provided by applicant): My goal is to lead a translational research group focused on breast cancer prevention. The Career Development Award to Promote Diversity (K22) will allow me to gain additional research training in the areas of human breast pre-malignancy and to gather the critical preliminary data that will be the foundation of my future proposals as I pursue an independent research position. Upon receipt of this prestigious award, I anticipate securing a position within two years and pursuing independent funding such as an RO1. During the first year of the award period, I will focus on my training as a translational researcher by taking the Introduction to Clinical and Translational Cancer Research Course and by participating in the Breast Cancer Working group, a meeting designed to the bring bench scientists, clinical investigators, patient care personnel, and public health researchers together to determine collaborative research directions and to obtain a major goal - the eradication of breast cancer. To develop my leadership and management skills as I prepare to lead my own independent research group, I will participate in the Professional Development Series offered throughout the year. I will meet weekly with my mentors to discuss my research progress, potential funding opportunities, and my search for an independent position. During my second year I plan to pursue independent funding and apply for faculty positions. I anticipate having gathered sufficient data from the experiments proposed below to provide a solid foundation for my own research program. This, along with the funding already in place by the K22, should place me in a strong position as I begin applying for faculty positions. Ductal carcinoma in situ (DCIS) is a non-invasive cancer lesion that progresses to invasive cancer approximately half the time. The ability to predict DCIS malignant progression could significantly improve patient treatment and survival. However, we are not currently able to distinguish those DCIS lesions with invasion potential. Studies have identified a subpopulation of cells within breast cancer lesions which appear to drive metastasis and epithelial-mesenchymal transition (EMT). These tumor initiating or cancer stem cells share many of the properties of their normal stem cell counterparts such as long term self-renewal potential. Additionally, several groups recently demonstrated that ovarian steroid hormones indirectly influencing mammary stem cell numbers and cancer progression. The objective of this application is to characterize the steroid hormone paracrine signaling pathways involved in aberrant stem cell self-renewal and invasion which have potential for development of subtype-specific therapies. Our hypothesis is that the risk for DCIS malignant progression is determined by the aberrant self-renewal pathways in DCIS tumor initiating cell populations. In vitro assays of self-renewal and invasion potential, and a novel in vivo method recently developed by our laboratory, the MIND (mouse intraductal) model, will be utilized in this proposal to test our hypothesis. The MIND technique is a xenograft model of human DCIS in which primary human DCIS cells are engrafted intraductally into mammary glands of immunodeficient mice. This model mimics DCIS in vivo in the mouse to the closest condition that it is found in the human since DCIS initiates inside the ducts. Therefore the mammary ducts provide a natural microenvironment for DCIS cell growth. Specific Aim 1 is to identify and characterize the stem cell population's role in growth and invasion potential among DCIS subtypes in vivo and in vitro. Primary human cells of each DCIS subtype (Luminal A & B, basal, and HER2 over expressing) will be used to examine self-renewal ability, invasion potential, and the epithelial mesenchymal transition (EMT) phenotype of potential stem cell populations using both in vivo and in vitro techniques. Specific Aim 2 is to determine the influence of ER and PR paracrine signaling on primary human DCIS cells stem growth, self- renewal, and invasion potential using mammosphere assays and the MIND model. We will assess the paracrine effects of steroid hormone treatment on self-renewal ability, invasive progression, and EMT by in vitro mammospheres assays and in vivo MIND xenotransplantation. The molecular characterization of tumor initiating cells and the signaling pathways controlling their fate is promising to lead to the development of individualized therapeutic strategies for prevention of DCIS and/or invasive progression. My ultimate goal is to use our novel model of human primary DCIS, which mimics the heterogeneity of human non-invasive breast lesions, to study the mechanisms underlying progression to invasive breast cancer. In the long term, these studies should facilitate more accurate risk assessment and the design of therapeutic strategies for prevention based on a better understanding of the distinct mechanisms of malignant progression. The data obtain will be the foundation of my future grant proposals and completion of these proposed experiments is essential to the development of my own independent research program.

职位描述(申请人提供)：我的目标是领导一个专注于乳腺癌预防的翻译研究小组。促进多样性的职业发展奖(K22)将使我能够在人类乳腺癌前病变领域获得额外的研究培训，并收集关键的初步数据，这些数据将成为我追求独立研究职位时未来提案的基础。在收到这个享有盛誉的奖项后，我预计在两年内获得一个职位，并寻求独立的资金，如RO1。在获奖期的第一年，我将专注于作为一名翻译研究人员的培训，参加临床和转化性癌症研究入门课程，并参加乳腺癌工作组，这是一个旨在将长凳科学家、临床研究人员、患者护理人员和公共卫生研究人员聚集在一起，确定合作研究方向并实现一个主要目标--根除乳腺癌--的会议。在我准备领导自己的独立研究小组时，为了发展我的领导力和管理技能，我将参加全年提供的专业发展系列课程。我会每周与我的导师见面，讨论我的研究进展、潜在的资助机会以及我寻找独立职位的问题。在我的第二年，我计划争取独立的资金，并申请教职。我预计已经从下面提出的实验中收集了足够的数据，为我自己的研究计划提供了坚实的基础。这一点，加上K22已经到位的资金，应该会让我在开始申请教职员工职位时处于有利地位。导管原位癌(DCIS)是一种非浸润性癌病变，大约一半的时间进展为浸润性癌。预测DCIS恶性进展的能力可以显著提高患者的治疗和存活率。然而，我们目前还不能区分那些具有侵袭潜力的DCIS病变。研究已经确定了乳腺癌病变中似乎驱动转移和上皮-间充质转化(EMT)的细胞亚群。这些启动肿瘤的干细胞或癌症干细胞具有许多正常干细胞的特性，例如长期自我更新的潜力。此外，几个研究小组最近证实，卵巢类固醇激素间接影响乳腺干细胞数量和癌症进展。这项应用的目的是描述参与异常干细胞自我更新和侵袭的类固醇激素旁分泌信号通路，这些信号通路有可能发展亚型特异性治疗。我们的假设是，DCIS恶性进展的风险是由DCIS肿瘤启动细胞群体中异常的自我更新途径决定的。体外检测自我更新和侵袭潜能，以及一种新的 我们实验室最近开发的活体方法，即Mind(小鼠导管内)模型，将在这一提议中用于验证我们的假设。Mind技术是一种人类DCIS的异种移植模型，在该模型中，原代人类DCIS细胞被植入免疫缺陷小鼠的乳腺内。该模型在小鼠体内模拟DCIS，与在人类中发现的情况最接近，因为DCIS起始于导管内。因此，乳腺导管为DCIS细胞的生长提供了自然的微环境。具体目标1是确定和表征干细胞群体在体内和体外DCIS亚型之间的生长和侵袭潜力中的作用。每种DCIS亚型的原代人类细胞(Lumina A&B、基础和HER2过表达)将用于检测潜在干细胞群体的自我更新能力、侵袭潜力和上皮间充质转化(EMT)表型。具体目标2是利用乳房分析和心智模型来确定ER和PR旁分泌信号对原代人类DCIS细胞干细胞生长、自我更新和侵袭潜力的影响。我们将通过体外乳房试验和体内异种精神移植来评估类固醇激素治疗对自我更新能力、侵袭进展和EMT的旁分泌作用。肿瘤起始细胞和控制其命运的信号通路的分子特征有望导致针对DCIS和/或侵袭性进展的个体化治疗策略的发展。我的最终目标是使用我们新的人类原发DCIS模型，它模拟了人类非侵袭性乳腺病变的异质性，来研究浸润性乳腺癌进展的机制。从长远来看，这些研究应该有助于更准确地进行风险评估，并在更好地了解恶性进展的不同机制的基础上设计预防的治疗策略。获得的数据将是我未来拨款申请的基础，完成这些拟议的实验对我自己的独立研究计划的发展至关重要。