INJURY-RELATED MORPHOGENIC PATHWAY SIGNALING AND HEPATOCARCINOGENESIS

该子项目是利用 由NIH/NCRR资助的中心赠款提供的资源。子项目和 研究者（PI）可能从另一个NIH来源获得主要资金， 因此可以在其他CRISP条目中表示。列出的机构是 中心，不一定是研究者的机构。 目标一：扩展对小动物模型的分析，以证明/反驳两种形态发生信号通路（Hedgehog（Hh）和Wnt）的活性变化是否（以及为什么）导致HCC出现和/或转移。 a）基于其他实验室正在进行的工作和出版物，我们知道mdr 2靶向缺失纯合子（mdr 2-/-）小鼠在2周龄时就有持续肝损伤的证据，这导致至少一种形态发生信号通路（Hh）的持续激活，并在1岁左右达到肝硬化和HCC的发展。因此，我们将处理4组小鼠，每组至少4只，最小年龄为1岁。第一组将接受Hh通路的抑制剂;第二组将接受Wnt通路的抑制剂，我们也拥有这种抑制剂，并已成功用于诱导结肠癌异种移植物的消退。最后2组为对照组。所有注射均为腹膜内注射，每日一次，持续3周。治疗前，我们将对每组进行MRI，以表征肿瘤的大小、数量和位置（包括是否存在转移）。 我们将在处理结束时对小鼠进行重新MRI，以在处死前评价处理效果。MRI成像是研究成功的核心和关键。如目标2所示，总体目标是开发用于患有HCC或被认为具有HCC高风险的人类患者的潜在治疗方法。 MRI在其翻译相关性方面至关重要，因为接受这些治疗的人类患者将如何间隔评估治疗效果。 B）（a）中提出的工作将在尚未发展为HCC的阿尔茨海默病小鼠中重复。我们将用上述形态发生途径抑制剂或载体对照治疗<1岁（在HCC出现之前）的小鼠。小鼠将在开始治疗前进行MRI成像以确保它们没有肿瘤，并在治疗结束时再次进行MRI成像以评估肿瘤预防。在a）中提议的工作完成之前，不会开始这方面的工作，因此，本合作建议主要针对a）中列出的工作。本研究的目的是确定两种形态发生途径抑制剂作为HCC的化学预防剂和/或治疗的安全性和有效性。 目标二：应用目标#1中获得的知识，确定形态发生途径活性依赖性致癌机制是否在物种间保守，并因此作为开发新型HCC生物标志物或治疗的潜在靶点。这方面的工作将在来自不同阶段纤维化和/或HCC患者的人类活检标本上进行。 目标#1的成功对于我们完成该目标的能力是必要的，因为人类样本的可用性非常有限，并且在该目标下进行的工作（微阵列分析，QRT-PCR和免疫组织化学）非常耗时且昂贵。本研究的总体目标是为在已经患有肿瘤或被认为具有HCC发展高风险的患者中加速HCC新治疗的临床试验提供令人信服的理由。

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.