Development of a Multi-Panel Multiplex Immunofluorescence Breast Cancer Immunophenotyping Assay

 说明书(由申请人提供)拟议的研究和商业化计划的总体目标是开发一种技术，为乳腺癌的临床诊断和研究提供一种定量的、多元化的IHC免疫表型分析方法。这些检测将使肿瘤学家获得最大量的信息，以最少的组织作为诊断和治疗计划的基础，允许在临床部位之间进行更大的重复性测试，并在组织形态的背景下提供可量化的信息。这一平台技术与乳腺癌检测的验证将使我们能够将该技术的使用扩展到其他多路标记物面板，从而使一系列测试能够提供分支诊断，从而做出更好的治疗决策。然后，这一平台技术可以扩展到其他癌症和疾病，既包括临床诊断，也包括研究试剂。在我们最初的“原则证明”研究的基础上，我们将使用我们现有的检测抗体/技术，在浸润性乳腺癌活检组织中演示使用我们的UltraPlex-BC1小组(ER-雌激素受体、PR-孕激素受体、HER2和Ki-67)的多重荧光染色，同时开发第二代检测试剂，以扩展我们未来的复合能力。UltraPlex-Bc1小组将乳腺活检组织分成4种主要亚型，其中一种特别具有侵袭性，缺乏主要的ER/PR/HER2标记物，称为三阴性乳腺癌。我们的第二个小组，Ultraplex-BC2旨在将三重阴性肿瘤细分为更具侵袭性的基底肿瘤和侵袭性较弱的非基底肿瘤。使用我们的第二代检测试剂，我们将演示浸润性乳腺肿瘤的灵敏多重IHC染色，并将其与目前使用单一染色/酶标记检测试剂的技术进行比较。最后，我们将使用一系列分型的乳腺肿瘤组织来验证这两种分析方法。

 DESCRIPTION (provided by applicant) The overall aim of the proposed research and commercialization plan is to develop a technology that will yield a quantitative, multiplex IHC immunophenotyping assays for breast cancer clinical diagnosis and research. These assays will allow the oncologist to accrue the maximum amount of information on which to base diagnosis and treatment plans using a minimum amount of tissue, allow greater reproducibility of testing between clinical sites and provide quantifiable information within the context of tissue morphology. Validation of this platform technology with the breast cancer assays will allow us to expand use of the technology to other multiplex marker panels, allowing the possibility of a series of tests providing branched diagnosis resulting in better treatment decisions. This platform technology can then be expanded to other cancers and diseases encompassing both clinical diagnostics as well as research reagents. Expanding on our initial "proof-of-principle" studies, we will demonstrate multiplex fluorescent staining using our UltraPlex-BC1 panel (ER - estrogen receptor, PR - progesterone receptor, HER2 and Ki-67) on invasive breast cancer biopsies using our existing detection antibodies/technology whilst at the simultaneously develop second-generation detection reagents to expand our future plexing capacity. The UltraPlex-BC1 panel provides subtyping of breast biopsies into 4 major subtypes, one of which is a particularly aggressive form lacking the main ER/PR/HER2 markers, called triple negative breast cancer. Our second panel, Ultraplex-BC2 aims to subdivide the triple negative tumors into the more aggressive basal and less aggressive non-basal forms. Using our second-generation detection reagents we will demonstrate sensitive multiplex IHC staining of invasive breast tumors and compare this to current technology that employs single staining/enzyme labeled detection reagents. Finally we will validate both assays using a series of typed breast tumor tissues.