Ovarian Cancer Chemoprevention

 描述(申请人提供)：卵巢癌是妇科癌症中最致命的一种，因为它通常在晚期被发现，而且对其病因知之甚少。目前的一种理论认为，最常见的卵巢癌--高级别浆液性卵巢癌起源于输卵管。我们建议研究我们的无毒口服化学预防药剂NSC 721689(SHetA2)在输卵管组织和细胞中的药物和化学预防活性。我们的药物干扰了一种名为mortalin的分子伴侣的功能，导致线粒体肿胀和有丝分裂，在癌细胞中过渡到凋亡，而正常细胞对这些作用具有抵抗力。理解这种差异效应的原因的一个线索是一种改变的死亡蛋白形式，它出现在经NSC-721689处理的正常上皮细胞培养物中，并随着培养物经历停滞、永生化和转化而逐渐消失。我们观察到Mortalin在卵巢癌肿瘤微环境中过表达。NSC 721689还通过磷酸化、泛素化和细胞周期蛋白D1的降解导致G1细胞周期停滞；在体外癌症和非癌症细胞中发生的事件，以及在体内的小鼠化学预防模型中发生的事件。NCI对我们的药物进行了广泛的临床前研究，结果表明，我们的药物在体外和体内具有化学预防活性，缺乏诱变代谢物、致突变性或致畸性，没有毒性，并且具有适合口服化学预防药物的药理特征。我们的目标是研究NSC 721689在卵巢癌化学预防中的作用，产生所需的数据：1)将这种有前途的药物用于卵巢癌化学预防的临床试验；2)开发可能适用于所有癌症的其他化学预防策略。目的1将进行0期临床试验，以确定在健康志愿者的血液和计划切除子宫的患者的输卵管组织中达到微摩尔药物水平所需的口服神经干细胞721689胶囊的数量。目的建立Dicerflx/Flox Ptenflx/Flox Amhr2cre/+双基因敲除小鼠，研究口服神经干细胞721689对输卵管肿瘤组织学和多发性的影响。目的3将从临床试验和动物模型中测量NSC 721689对细胞周期调节蛋白和死亡蛋白的影响。重要的结果将在从临床标本建立的人输卵管分泌上皮细胞(HFTSECs)的培养中得到验证。目的3还将确定神经干细胞721689诱导的死亡蛋白改变的身份，并将研究如何获得对神经干细胞721689诱导的自噬和凋亡的敏感性，而神经干细胞721689诱导的死亡蛋白改变随着hFTSECs的传代、永生化和转化而丧失。组织芯片(TMA)和图像分析分别用于评估癌细胞和间质细胞、逆相蛋白阵列(RPPA)和hFTSEC/成纤维细胞共培养，将用于研究mortalin和其他NSC 721689改变的蛋白质在药物敏感性中的作用。

 DESCRIPTION (provided by applicant): Ovarian cancer is the most deadly of gynecologic cancers because it is most often detected at late stage and little is known of its etiology. A current theory is that high grade serous, the most common ovarian cancer, originates in the fallopian tube. We propose to study pharmaceutical and chemoprevention activity of our non-toxic oral chemoprevention agent, NSC 721689 (SHetA2) in fallopian tube tissue and cells. Our drug interferes with the function of a molecular chaperone called mortalin leading to mitochondrial swelling and mitophagy that transitions to apoptosis in cancer cells, while normal cells are resistant to these effects. A clue to understanding the reason for this differential effet is an altered form of mortalin that appears in NSC-721689-treated normal epithelial cell cultures and is gradually lost as the cultures undergo stasis, immortalization and transformation. We observed mortalin overexpression in the ovarian cancer tumor microenvironment. NSC 721689 also causes G1 cell cycle arrest through phosphorylation, ubiquitination and degradation of cyclin D1; events that occur in both cancer and non- cancer cells in vitro, and in a murine chemoprevention model in vivo. Extensive preclinical studies of our drug conducted by the NCI demonstrated chemoprevention activity in vitro and in vivo, lack of mutagenic metabolites, mutagenicity or teratogenicity, no toxicity and a pharmacologic profile suitable for an oral chemoprevention agent. Our objectives are to study NSC 721689 in ovarian cancer chemoprevention producing data needed to: 1) bring this promising drug to clinical trials for ovarian cancer chemoprevention and 2) develop additional chemoprevention strategies that may be applicable to all cancers. Aim 1 will conduct a Phase 0 clinical trial to determine the number of oral NSC 721689 capsules needed to achieve micromolar drug levels in the blood of healthy volunteers and in fallopian tube tissues of patients scheduled for hysterectomy. Aim 2 will generate Dicer-Pten Double Knock Out [Dicerflox/flox Ptenflox/flox Amhr2cre/+] mice, which have been shown to develop fallopian tube tumors that spread to the ovary, and study the effects of oral NSC 721689 on fallopian tube tumor histology and multiplicity. Aim 3 will measure NSC 721689 effects on cell cycle regulatory proteins and mortalin in specimens from the clinical trial and animal model. Significant results will be validated in cultures of human fallopian tube secretory epithelial cells (hFTSECs) established from the clinical specimens. Aim 3 will also determine the identity of the NSC 721689-induced alteration in mortalin and will study how sensitivity to NSC 721689-induced autophagy and apoptosis is gained, while NSC 721689-induced mortalin alteration is lost, as hFTSECs are passaged, immortalized and transformed. Tissue microarrays (TMAs) and image analysis to separately evaluate cancer and stromal cells, reverse phase protein arrays (RPPAs) and hFTSEC/fibroblast co-cultures will be used to study the roles of mortalin and other NSC 721689 altered proteins in drug sensitivity.