Inhibiting tumor growth and metastasis in highly aggressive breast cancers with centrosome amplification

项目摘要 中心体扩增（CA）在癌症中非常普遍，并与肿瘤进展密切相关 在几种不同的癌症中，包括乳腺癌、前列腺癌、卵巢癌和肺癌，预后更差。中心体 扩增的细胞还表现出增加的运动性和侵袭性，导致转移。此外，CA是 与遗传畸变相关，如p53突变，通常在侵袭性形式中观察到 三阴性乳腺癌（TNBC）。我们的长期目标是实现增长， 具有CA的侵袭性乳腺肿瘤的转移性扩散，最终将导致患者 结果。该项目的总体目标是（i）在CA乳腺癌模型中抑制肿瘤生长 通过在靶向转化酸性蛋白质时诱导中心体去聚集和多极纺锤体的形成， 卷曲螺旋3（TACC 3），和（ii）阻断CA模型中的局部侵袭和转移性播散， 在抑制TACC 3后防止细胞极化、迁移和侵袭。核心假设是， 一方面，TACC 3抑制将阻止扩增的中心体主动聚集成两个纺锤体极 在有丝分裂细胞分裂期间，导致多极有丝分裂和p53改变的细胞中的凋亡性细胞死亡， 另一方面破坏了中心体和高尔基体的重新定向，微管成核和细胞极化， 间期细胞导致迁移减少。该项目的基本原理是TACC 3抑制 代表了一个独特的机会，消除最具侵略性的肿瘤， 中心体，而保留正常细胞。中心假设将通过追求两个具体的测试， 目标：1.）通过靶向TACC 3抑制具有CA的侵袭性乳腺癌模型中的肿瘤生长，和2.） 通过靶向TACC 3预防CA侵袭性乳腺癌模型的转移。State-of-the-art 将采用具有可平移方法的实验设置，包括肿瘤类器官，患者， 衍生的异种移植物和免疫活性转基因小鼠模型， 以CA为特征。本项目提出的研究是创新的，因为它旨在研究 TACC 3的有丝分裂/间期特异性相互作用组，其对于TACC 3介导的细胞分裂是必需的， 有丝分裂细胞和细胞极性的间期细胞，我们建议使用我们的新的和高效的阻断 TACC 3抑制剂以及使用CRISPR介导的敲除。该项目意义重大，因为 它有望提供关键的机制和表型临床前数据，以支持靶向 TACC 3同时抑制具有CA的乳腺癌模型中的肿瘤生长和转移，这将随后 显著降低了高度侵袭性癌症患者亚群的死亡率。

PROJECT SUMMARY Centrosome amplification (CA) is highly prevalent in cancer and strongly associated with tumor progression and worse prognosis in several different cancers, including breast, prostate, ovarian and lung. Centrosome amplified cells also demonstrate increased motility and invasiveness leading to metastasis. Moreover, CA is associated with genetic aberrations, such as p53 mutation that are commonly observed in aggressive forms of cancers, such as the triple negative breast cancer (TNBC). Our long-term goal is to target the growth and metastatic dissemination of aggressive breast tumors with CA that will ultimately lead to improved patient outcome. The overall objectives of this project are to (i) inhibit tumor growth in breast cancer models with CA via inducing centrosome de-clustering and formation of multipolar spindles upon targeting transforming acidic coiled-coil 3 (TACC3), and to (ii) block local invasion and metastatic dissemination in CA models by preventing cell polarization, migration and invasion upon inhibition of TACC3. The central hypothesis is that TACC3 inhibition will, on one hand, prevent active clustering of amplified centrosomes into two spindle poles during mitotic cell division leading to multipolar mitosis and apoptotic cell death in p53 altered cells, and on the other hand disrupts centrosome and Golgi re-orientation, microtubule nucleation and cell polarization in interphase cells leading to decreased migration. The rationale for this project is that TACC3 inhibition represents a unique opportunity to eliminate the most aggressive tumors that have supernumerary centrosomes, while sparing the normal cells. The central hypothesis will be tested by pursing two specific aims: 1.) To inhibit tumor growth in aggressive breast cancer models with CA by targeting TACC3, and 2.) To prevent metastasis in aggressive breast cancer models with CA by targeting TACC3. State-of-the-art experimental settings with translatable approaches will be employed, including tumor organoids, patient- derived xenografts and immunocompetent transgenic mouse model of advanced breast cancer that we characterized in terms of CA. The research proposed in this project is innovative as it aims to study mitosis/interphase-specific interactomes of TACC3 that are essential for TACC3-mediated cell division in mitotic cells and cell polarity in interphase cells that we propose to block using our novel and highly potent TACC3 inhibitor as well as using CRISPR-mediated knock-out. The proposed project is significant because it is expected to provide key mechanistic and phenotypic pre-clinical data to support the notion that targeting TACC3 concomitantly inhibits tumor growth and metastasis in breast cancer models with CA, which will then dramatically reduce mortality rates among patient subpopulation with highly aggressive cancers.