Molecular Imaging to Evaluate EGFR and SRC Targeted Therapies in Colorectal....

概述--成像数据以及它与细胞凋亡改变的关系本可以更强烈 声称，调查人员证明了他们有能力进行拟议的广泛成像实验， 但它们并没有真正解释为什么它们是必要的，也没有解释它们将如何帮助计划中的人体试验。我们 同意在先前的申请中，我们没有为所提议的成像提供充分的生物学基础 方式(包括细胞凋亡、增殖、EGFR表达和VEGFR表达)。在这里，我们有 修改了背景和意义部分，以明确说明每一种 关于所使用的治疗的建议的成像指标。这一理论基础得到了进一步强调 纳入了自上一年以来产生的额外的体外和体内初步数据 呈件。虽然超出了拟议的临床前调查的范围，但我们高度设想 有效的无创影像读数(S)将为治疗反应提供有价值的信息 优化患者个体化治疗。更直接的是，可以使用非侵入性成像度量 在临床上作为治疗反应的标志，取代侵入性活组织检查程序。因此，在修订后的 应用，我们已经解决了这里开发的成像度量的临床效用和预期用途，以及 确定其临床实施的优先顺序。 连续的CT研究和辐射剂量的潜在影响没有讨论，应该要么 放弃或进一步考虑。序列CT的潜在影响，以及有效辐射剂量， 在修订后的申请(背景和意义以及研究设计和方法)中进行了讨论，以及 我们现在强调，在研究期间，每只动物只会收集两次CT扫描。这些扫描将是 用于与PET图像([18F]-FLT或[18F]-FDG)进行解剖配准，就像在PET/CT中常规进行的那样。 ‘确定成像变量与反应关系的标准没有得到很好的描述，而且 似乎对统计输入的使用是有限的。在修改后的实验计划中，我们仔细描述了 将用于验证所建议的成像指标的程序，特别是在组织水平上使用 几种公认的免疫组织化学(IHC)方法。凯伊·华盛顿博士，GL病理学专家， 已被增加为关键人员，将负责组织的IHC分析。此外，我们的 统计分析计划已作了相当大的修订，并将因新的统计核心而得到加强 将由余雪博士执导。 “人们对研究项目1缺乏重点的热情降低了。”我们做了一件大事 共同努力专注于项目1的目标。我们假设最相关的生物学读数是 研究项目1包括细胞凋亡、增殖、EGFR表达和VEGFR表达。因此，我们 通过减少研究的成像指标总数，大大集中了该项目，消除了 切线相关指标，例如使用NIR-conPK11195和DCE MRI的PBR成像。此外，我们 集中了我们的治疗策略。以前，AIM 1中描述的治疗方案依赖于 假设顺序给予两种机械上不同的EGFR轴抑制剂将导致 提高了治疗效果。此外，目标2专注于EGFR和HDAC的新组合 抑制力。虽然我们对这些创新战略持乐观态度，但我们认识到，重点是 这种应用的基础是新的成像方法的发展，而不是创新的建立 治疗方案。因此，我们的实验计划现在使用了一种简单得多的治疗策略 建立了临床反应的读数。

Overview- 'The imaging data and how it will be related to apoptosis alteration could have been more strongly stated, and 'Investigators demonstrate their ability to carry out the extensive imaging experiments proposed, but they do not really explain why they are necessary or how they will help in the planned human trials.' We agree that in the previous application we did not provide adequate biological rationale for the proposed imaging modalities (including apoptosis, proliferation, EGFR expression, and VEGFR expression). Here, we have revised the Background and Significance section to explicitly state the biological rationale for each of the proposed imaging metrics with respect to the utilized treatments. This rationale is further emphasized by the inclusion of additional preliminary in vitro and in vivo data that has been generated since the previous submission. Although beyond the scope of the proposed pre-clinical investigations, we envision that highly effective non-invasive imaging readout(s) of treatment response will provide valuable information towards optimizing individualized patient therapy. More directly, non-invasive imaging metrics could be employed clinically as markers of treatment response in place of invasive biopsy procedures. Thus, in the revised application, we have addressed the clinical utility and intended use of the imaging metrics developed here and prioritize their clinical implementation. 'The potential impact of sequential CT studies and radiation dose was not discussed and should either be dropped or further considered.' The potential impact of sequential CT, as well as the effective radiation dose, is discussed in the revised application (Background and Significance and Research Design and Methods), and we now emphasize that only two CT scans will be collected per animal during the study. These scans will be used for anatomical co-registration with PET images ([18F]-FLT or [18F]-FDG) as are routinely done in PET/CT. 'Criteria for determining the relationships of imaging variables to response are not well described, and there appears to be limited use of statistical input.' In the revised experimental plan, we have carefully described procedures that will be used to validate the proposed imaging metrics, particularly at the tissue level using several well-accepted immunohistochemical (IHC) methods. Dr. Kay Washington, an expert in Gl pathology, has been added as key personnel and will be responsible for the IHC analysis of tissues. Furthermore, our statistical analysis plan has been revised considerably and will be strengthened by the new Statistics Core that will be directed by Dr. Yu Shyr. 'Reduced enthusiasm [was expressed] for the lack of focus in Research Project 1.' We have made a major concerted effort to focus the aims of Project 1. We hypothesize that the most relevant biological readouts to be studied in Project 1 include apoptosis, proliferation, EGFR expression and VEGFR expression. Therefore, we have considerably focused the project by reducing the total number of imaging metrics studied, eliminating tangentially relevant metrics such as PBR imaging with NIR-conPK11195 and DCE MRI. Furthermore, we have focused our therapeutic strategies. Previously, the therapeutic regimen described in Aim 1 relied on the hypothesis that sequential administration of two mechanistically distinct EGFR axis inhibitors would result in improved therapeutic efficacy. Additionally, Aim 2 focused on a novel combination of EGFR and HDAC inhibition. While we remain optimistic with respect to these innovative strategies, we recognize that the focus of this application is upon the development of novel imaging methods and not the establishment of innovative therapeutic regimens. Therefore, our experimental plan now utilizes a much simpler therapeutic strategy with established readouts of clinical response.