Tumor-promoting functions of TMPRSS13 in breast cancer progression

摘要：TMPRSS13是一种细胞表面锚定的丝氨酸蛋白酶，在人类乳腺中表达上调。 癌细胞与正常乳腺上皮细胞的比较。使用TMPRSS13缺陷小鼠 模型中，我们发现TMPRSS13在乳腺癌的发生中起着因果作用，在这一过程中它起到了 对原发肿瘤的起始、生长和肺部转移有显著影响。在培养的人乳腺癌中 细胞中，TMPRSS13的沉默导致细胞增殖减少和细胞凋亡增加。首要目标是 目的：验证TMPRSS13作为乳腺癌治疗新靶点的可能性。我们将执行这两项功能 和机制实验以确定TMPRSS13在乳腺癌中的作用，使用平行和 以假设为导向的实验和不偏不倚的方法。 有待检验的中心假设是TMPRSS13通过激活PRO-13促进肿瘤进展。 生存和前侵袭信号，代表了乳腺癌治疗的新靶点。我们制定了 三个独立的具体目标是检验这一假说。在目标1中，我们将确定TMPRSS13如何促进 支持生存和入侵。我们发现上皮向间充质转化的两个中枢调节因子 (EMT)，以及Snail锌指转录因子家族成员Snail和Slug受 TMPRSS13.这一发现的意义以及TMPRSS13和TMPRSS13的功能关系 蜗牛/鼻涕虫将在各种细胞分析中进行测试。同时，最先进的RNA-Seq分析和 将进行定量质谱分析，以发现差异表达的转录本和蛋白质， 以确定TMPRSS13介导的功能的关键通路。在目标2中，我们将发展和 在乳腺癌细胞模型中测试TMPRSS13的新抑制剂。药物化学将被用于识别 TMPRSS13抑制剂在我们收集的丝氨酸蛋白酶模拟肽化合物中抑制TMPRSS13 活动。化合物将在每个位置进行修饰，以增加效力、选择性和稳定性。 TMPRSS13抑制剂将在乳腺癌的2D和3D细胞培养模型中得到验证 增殖/存活/凋亡和侵袭试验评估抗癌效果。在目标3中，我们将评估 新型TMPRSS13抑制剂在患者来源异种移植(PDX)模型中的体内疗效。传统型 化疗药物通常会导致严重的副作用，并具有产生耐药性的重大风险。 初步数据表明，TMPRSS13靶向导致乳腺癌细胞对化疗的敏感性增加。 因此，TMPRSS13抑制作为单用药以及与TMPRSS13联合治疗的疗效。 体内化疗药物的护理标准将进行评估。 将最先进的小鼠模型与基于2D和3D细胞培养的检测相结合 转录组和蛋白质组分析和药物发现包括一种创新和有效的战略 用于研究人类癌症疾病。

Abstract: TMPRSS13 is a cell-surface anchored serine protease that is up-regulated in human breast carcinoma cancer cells compared to normal breast epithelial cells. Using a TMPRSS13 deficient mouse model, we found that TMPRSS13 plays a causal role in mammary carcinogenesis, where it contributes significantly to primary tumor initiation, growth, and metastasis to the lungs. In cultured human breast cancer cells, silencing of TMPRSS13 causes decreased proliferation and increased apoptosis. The overarching goal is to validate TMPRSS13 as a potential new therapeutic target in breast cancer. We will perform both functional and mechanistic experiments to pinpoint the role of TMPRSS13 in breast cancer using parallel and complimentary hypothesis-driven experiments and unbiased approaches. The central hypothesis to be tested is that TMPRSS13 promotes tumor progression by activating pro- survival and pro-invasive signaling and represents a novel target for breast cancer treatment. We formulated three independent specific aims to test this hypothesis. In Aim 1, we will determine how TMPRSS13 promotes pro-survival and invasion. We discovered that two central regulators of epithelial-to-mesenchymal transition (EMT), and members of the Snail zinc-finger transcription factor family, Snail and Slug, are regulated by TMPRSS13. The significance of this finding and the functional relationship between TMPRSS13 and Snail/Slug will be tested in a variety of cellular assays. In parallel, state-of-the-art RNA-Seq analysis and quantitative mass spectrometry will be performed to uncover differentially expressed transcripts and proteins, respectively, to identify pathways critical for TMPRSS13-mediated functions. In Aim 2, we will develop and test new inhibitors of TMPRSS13 in breast cancer cellular models. Medicinal chemistry will be used to identify TMPRSS13 inhibitors in our collection of serine protease peptidomimetic compounds that inhibit TMPRSS13 activity. Compounds will be modified at each position to increase potency, selectivity, and stability. TMPRSS13 inhibitors will be validated in 2D and 3D cell culture models of breast cancer using quantitative proliferation/survival/apoptosis, and invasion assays to assess anticancer effects. In Aim 3, we will evaluate the efficacy of novel TMPRSS13 inhibitors in vivo using patient derived xenograft (PDX) models. Conventional chemotherapeutics often cause severe side effects and carry a significant risk of developing resistance. Preliminary data indicate that TMPRSS13 targeting leads to increased chemosensitivity in breast cancer cells. Therefore, the therapeutic effect of TMPRSS13 inhibition as mono-therapy as well as combination-therapy with standard of care chemotherapy drugs in vivo will be evaluated. The combination of state-of-the art mouse models and 2D and 3D cell culture based assays with transcriptome and proteome profiling and drug discovery encompasses an innovative and impactful strategy for studying human cancerous disease.