CHARACTERIZATION OF BREAST CANCER DORMANCY IN BONE

这项建议的目的是确定在癌症中处于休眠状态的癌细胞之间的遗传差异。 骨头和那些已经从休眠状态“重新唤醒”的人，并发现负责的机制。 这些研究将为目前在美国的近300万乳腺癌患者确定潜在的治疗靶点。 缓解。有足够的证据表明，缺氧（缺氧）刺激甲状旁腺相关的 蛋白（PTHrP）并抑制乳腺癌细胞中的白血病抑制因子（LIF）信号传导，这可能控制 重新觉醒的过程。LIFR的缺失或PTHrP的过表达导致弱表达的转化。 转移性乳腺癌细胞的侵袭性和高转移性细胞，这导致了假设，缺氧 促进骨转移部分通过下调LIFR信号，这刺激癌细胞， 通过血管内皮生长因子（VEGF）的产生退出休眠。这将使用几个 目前的乳腺癌模型，其中LIF和PTHrP信号传导被破坏，以确定它们对mRNA水平的影响 乳腺癌细胞从休眠到增殖的变化。弱骨转移性人 MCF-7和小鼠D2.0R癌细胞系将用作骨中乳腺癌休眠的模型。这些 细胞将通过LIF信号传导抑制和PTHrP过表达而被驱离体内休眠。GFP标记 乳腺癌细胞将被RFP染色，RFP随着细胞分裂而丢失，这些标签将用于 用于RNA测序的骨髓中的间歇性休眠与休眠后（增殖）细胞。该提案 还将研究缺氧诱导因子（HIF）和LIF信号在骨原发肿瘤部位的作用 使用乳腺癌的MMTV-PyMT自发模型和缺乏HIF 1 α和HIF 2 α的小鼠， 乳腺 博士约翰逊的近期目标是为骨骼中的乳腺癌细胞建立休眠特征 在多个模型中保持一致。拟议的项目将确定基因畸变在休眠vs 为约翰逊博士在一个学术机构建立一个独立的实验室奠定了基础。 研究机构。她的研究培训计划包括参加相关的研讨会，讲习班， 由斯坦福大学主办的课程，以及共同导师和受人尊敬的职业顾问的监督 她成立了一个委员会来监督职业和科学发展。

This objective of this proposal is to identify genetic differences between cancer cells that lie dormant in the bone and those that have “re-awakened” from a dormant state, and to discover the mechanisms responsible. These studies will identify potential therapeutic targets for the nearly 3 million breast cancer patients currently in remission. There is sufficient evidence that hypoxia (a lack of oxygen) stimulates parathyroid hormone-related protein (PTHrP) and inhibits leukemia inhibitor factor (LIF) signaling in breast cancer cells, which may control the re-awakening process. Deletion of LIFR or over-expression of PTHrP results in the conversion of weakly metastatic breast cancer cells to invasive and highly metastatic cells, which leads to the hypothesis that hypoxia promotes bone metastasis in part through down-regulation of LIFR signaling, which stimulates cancer cells to exit dormancy through vascular endothelial growth factor (VEGF) production. This will be tested using several current breast cancer models in which LIF and PTHrP signaling is disrupted to determine their effect on mRNAlevel changes in breast cancer cells as they evolve from dormant to proliferative. Weakly bone metastatic human MCF-7 and mouse D2.0R cancer cell lines will be used as models of breast cancer dormancy in bone. These cells will be driven out of dormancy in vivo through LIF signalling inhibition and PTHrP over-expression. GFPlabelled breast cancer cells will be dyed with RFP that is lost as cells divide, and these tags will be used to fractionate dormant vs post-dormant (proliferative) cells in the bone marrow for RNA sequencing. The proposal will also examine the role of hypoxia inducible factor (HIF) and LIF signaling at the primary tumor site on bone metastasis using the MMTV-PyMT spontaneous model of breast cancer and mice lacking HIF1α and HIF2α in the mammary gland. Dr. Johnson’s immediate goal is to establish a dormancy signature for breast cancer cells in the bone that is consistent across multiple models. The proposed project will identify genetic aberrations in dormant vs proliferative cells and lay the foundation for Dr. Johnson to establish an independent laboratory at an academic research institution. Her research training program includes attendance at relevant seminars, workshops, and courses hosted by Stanford University, as well as oversight from co-mentors and a respected career advisory committee she has formed to monitor both career and scientific development.