Development of a 3D Imaging Diagnostic Tool for the Improved Characterization of Metastatic Melanoma

总结 该II期项目的目的是通过以下方式改善黑色素瘤患者的结局和生存率： 减少漏诊转移性疾病的假阴性前哨淋巴结活检的数量 通过常规的组织处理和组织学表征。据估计，4,200名患者 每年在美国以这种方式误诊，这导致更保守的治疗方案， 这些患者比需要有效地治疗他们的疾病。未接受治疗的患者的结局 适当的治疗方案是，他们经历了三年减少十年生存， 调整后的生命年换算成12亿美元的经济损失。为了解决这个问题，我们 开发了一种三维组织成像和数字分析方法， 前哨淋巴结活检组织的表征以及分离的肿瘤细胞和微 传统组织病理学通常会遗漏的转移。我们已经通过我们的 初步研究表明，使用这种方法，我们可以识别这些癌细胞在组织中， 特征为节点阴性，并且我们可以区分真阴性和假阴性样本。 该项目的重点是采用这种方法，并将其转化为CLIA诊断检测， 作为实验室开发的测试提供给前哨淋巴结活检阴性的患者， 第二阶段的工作。最初，该测试将在传统的组织评估后提供，但最终可能 可用作所有黑色素瘤前哨淋巴结活检组织的主要诊断方法。的 将通过与合作伙伴开展回顾性临床研究来完成该检测方法的开发 Cedars-Sinai与存档的阴性前哨淋巴结活检组织块， 用于表征这些样品的整体特征。通过这项研究，我们将证明的准确性， 检测的特异性和敏感性，并将能够量化它降低发生率的程度。 前哨淋巴结活检的假阴性。通过执行本临床 研究中，我们将建立一个统计模型，根据样本的基本三个阈值来确定样本的阈值- 三维特征（例如癌细胞总数、癌细胞聚集体体积、癌细胞密度） 并将其分类为“不存在转移”（即真阴性）或“存在转移”（即真阳性）。 此外，我们开发的软件将为阳性样本描述切片应取自何处 用于由Visikol病理学家使用传统组织病理学确认的样品， 该分析适合于传统的分类范例。最后，我们将把我们的数字病理学软件 分析方法，以符合21 CFR第11部分的应用程序，这将需要在结束时， 项目开始提供作为CLIA测试的检测。为了支持这一做法，并允许启动一个 实验室开发的测试，我们还将建立验证文件包和相关测试 证明试验的范围、专属性、重现性、准确度和精密度所需。因此，我们认为， 在本项目结束时，我们将建立并验证一种可以开始提供给 CLIA实验室扩建后的患者和配备临床人员。

Summary The objective of this Phase II project is to improve outcomes and survival for patients with melanoma by reducing the number of false negative sentinel lymph node biopsies wherein metastatic disease is missed through conventional tissue processing and histological characterization. It is estimated that 4,200 patients are misdiagnosed in this manner in the US per year which results in more conservative treatment regimens for these patients than is required to effectively treat their disease. The outcome of patients not receiving the proper treatment regimen is that they experience a three-year reduction in ten-year survival which on a quality adjusted life year basis translates into a $1.2 billion economic loss. To address this problem, we have developed a three-dimensional tissue imaging and digital analysis approach which allows for the complete characterization of sentinel lymph node biopsy tissues and the identification of isolated tumor cells and micro- metastases that are commonly missed with traditional histopathology. We have demonstrated through our preliminary studies that using this approach we can identify these cancer cells in tissues that were previously characterized as node negative and that we can differentiate true negative from false negative samples. The focus of this project is to take this approach and transform it into a CLIA diagnostic assay that can be offered as a laboratory developed test to patients with negative sentinel lymph node biopsies at the conclusion of this Phase II work. Initially, the test will be offered following traditional tissue evaluation but eventually could be used as a primary diagnostic approach for all melanoma sentinel lymph node biopsy tissues. The development of this assay will be completed through conducting a retrospective clinical study in partnership with Cedars-Sinai with archived negative sentinel lymph node biopsy tissue blocks where the approach will be used to characterize these samples in their entirety. Through this study, we will demonstrate the accuracy, specificity and sensitivity of the test and will be able to quantify the extent to which it reduces the incidence of false negatives in the characterization of sentinel lymph node biopsies. Through the execution of this clinical study, we will build a statistical model that will threshold samples based upon their underlying three- dimensional features (e.g. total number of cancer cells, cancer cell aggregate volume, density of cancer cells) and classify them as ‘no metastases present’ (i.e. true negative) or ‘metastases present’ (i.e. true positive). Furthermore, the software we develop will for positive samples describe where a section should be taken from the sample for confirmation by a Visikol pathologist using traditional histopathology such that the report from the assay fits into the traditional classification paradigm. Lastly, we will transform our digital pathology software analysis approach into a 21 CFR part 11 compliant application which will be required at the conclusion of the project to start offering the assay as a CLIA test. To support this approach and to allow for the launch of a laboratory developed test, we will also build the validation documentation package and associated tests required to demonstrate the range, specificity, reproducibility, accuracy and precision of the assay. Therefore, at the conclusion of this project we will have built and validated an assay which can start to be offered to patients following the build-out of a CLIA lab and staffing with clinical personnel.