INJURY-RELATED MORPHOGENIC PATHWAY SIGNALING AND HEPATOCARCINOGENESIS

这个子项目是许多研究子项目中利用 资源由NIH/NCRR资助的中心拨款提供。子项目和 调查员(PI)可能从NIH的另一个来源获得了主要资金， 并因此可以在其他清晰的条目中表示。列出的机构是 该中心不一定是调查人员的机构。 目的#1：扩展对小动物模型的分析，以证明/反驳两个形态发生信号通路(Hedgehog(HH)和Wnt)的活性变化是否(以及为什么)导致肝癌的出现和/或转移。 A)根据正在进行的工作和其他实验室发表的文章，我们知道mdr2(mdr2-/-)基因纯合的小鼠在2周大的时候就有证据表明持续的肝损伤，这导致至少一个形态发生信号通路(HH)的持续激活，并最终导致肝硬化和大约1岁左右的肝细胞癌的发展。因此，我们将治疗4组小鼠，每组至少4只，最低年龄为1岁。第一组将接受HH途径的抑制剂；第二组将给予Wnt途径的抑制剂，我们也拥有这种抑制剂，并已成功地用于诱导结肠癌异种移植瘤的消退。最后两组为对照组。所有注射都将是腹膜内注射，连续3周，每天给药。治疗前，我们将对每组患者进行核磁共振检查，以确定肿瘤的大小、数量和位置(包括有无转移)。在治疗结束时，我们将对小鼠进行再次磁共振成像，以评估治疗效果。核磁共振成像是这项研究成功的核心和关键。正如目标2所示，总体目标是开发潜在的治疗方法，用于人类肝细胞癌患者或被认为是肝细胞癌高危患者。在如何对接受这些治疗的人类患者进行间歇性治疗效果评估方面，MRI是至关重要的翻译相关性。 B)(A)中建议的工作将在尚未发展为肝细胞癌的肝硬变小鼠身上重复。我们将用上述形态发生途径抑制剂或赋形剂对照治疗小鼠(在肝癌出现前一年)。小鼠将在开始治疗前接受MRI成像，以确保它们没有肿瘤，并在治疗结束时再次接受MRI成像，以评估肿瘤预防情况。在a)中提议的工作完成之前，不会开始实现这一目标的工作，因此，这项协作建议主要针对a)中所列的工作。本研究的目的是确定两种形态通路抑制剂作为肝癌的化学预防药物和/或治疗药物的安全性和有效性。 目的#2：应用在目的#1中获得的知识来确定形态发生途径活性依赖的致癌机制是否跨物种保守，从而作为开发新的肝细胞癌生物标志物或治疗的潜在靶点。这一目标的工作将在来自不同阶段的纤维化和/或肝细胞癌患者的人类活组织标本上进行。目标1的成功对于我们完成这一目标的能力是必要的，因为人类样本的可用性非常有限，而且在这一目标下建议进行的工作(微阵列分析、QRT-PCR和免疫组织化学)非常耗时和昂贵。作为一个整体，这项研究的目的是提供一个令人信服的理由，以加快对已经有肿瘤的患者或那些认为有高风险发展为肝细胞癌的患者进行新的肝细胞癌治疗的临床试验。

This subproject is one of many research subprojects utilizing the resources provided by a Center grant funded by NIH/NCRR. The subproject and investigator (PI) may have received primary funding from another NIH source, and thus could be represented in other CRISP entries. The institution listed is for the Center, which is not necessarily the institution for the investigator. Aim #1: Extend analysis of a small animal model to prove/disprove if (and why) changes in the activity of two morphogenic signaling pathways (Hedgehog (Hh) and Wnt) cause HCC to emerge and/or metastasize. a) Based on ongoing work and publications from other labs, we know that mice homozygous for a targeted deletion in mdr2 (mdr2-/-) have evidence of ongoing liver injury at as young as 2-weeks of age, which leads to sustained activation of at least one morphogenic signaling pathway (Hh) and culminates in liver cirrhosis and the outgrowth of HCC at around 1-year of age. Therefore, we will treat 4 groups of at least 4 mice each with a minimum age of 1 year. The 1st group will receive an inhibitor to the Hh pathway; the 2nd will be given an inhibitor to the Wnt pathway, which we also have in our possession ,and have successfully used to induce regression of colon cancer xenografts. The final 2 groups will be controls. All injections will be intraperitoneal, given daily for 3 weeks. Prior to treatment, we will MRI each group to characterize size, number, and location of tumors (including presence or absence of metastases). We will re-MRI the mice at treatment conclusion to evaluate for treatment effect prior to sacrifice. MRI imaging is central and critical to the study's success. As indicated in Aim 2, the overall goal is the development of potential treatments to use in human patients with HCC or felt to be at high risk for HCC. MRI is crucial in its translational relevance to how human patients receiving these treatments would be intervally evaluated for treatment effect. b) The work proposed in (a) will be repeated in cirrhotic mice who have yet to develop HCC. We will treat mice <1 year (prior to emergence of HCC) with the above-mentioned morphogenic pathway inhibitors or vehicle control. Mice will undergo MRI imaging prior to initiation of treatment to ensure they do not have tumor and again at conclusion of treatment to assess tumor prevention. Work on this aim will not begin until work proposed in a) is completed and therefore this proposal for collaboration is primarily directed to the work listed in a). The goal of this aim is to determine the safety and efficacy of two morphogenic pathway inhibitors as chemopreventive agents and/or treatments for HCC. Aim #2: Apply knowledge gained in Aim #1 to determine if morphogenic pathway activity-dependent carcinogenic mechanisms are conserved across species and therefore serve as potential targets for development of novel HCC biomarkers or treatments. Work in this aim will take place on human biopsy specimens from patients with various stages of fibrosis and/or HCC. The success of Aim #1 is necessary for our ability to complete this Aim as the availability of human samples is very limited and the work proposed to take place under this Aim (microarray analysis, QRT-PCR, and immunohistochemistry) is very time consuming and costly. The goal of this study as a whole is to provide a compelling rationale for expedited clinical trials for new treatments of HCC in patients who already have tumor or in those felt to be high risk for the development of HCC.