Implementation of phosphoprotein preservation technology for cancer biospecimens

癌症生物样本磷蛋白保存技术的实现

• 批准号: 8311650
• 负责人: Lance Allen Liotta
• 金额: $ 30.95万
• 依托单位: GEORGE MASON UNIVERSITY
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-08-15 至 2014-07-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8311650
• 关键词: Address; Adoption; Advanced Development; Antigens; Architecture; Archives; Biological Markers; Biological Preservation; Biopsy Specimen; Blinded; Breast; Buffers; Cancer Histology; Cancer Patient; Cell Count; Cell membrane; Cells; Chemistry; Clinical; Clinical Research; Collagen Type IV; Collection; Colon; Community Hospitals; Cytokeratin; DNA; Diagnosis; Diagnostic; ERBB2 gene; Endometrium; Epidermal Growth Factor Receptor; Epitopes; Equipment; Estrogen Receptors; Female; Fixatives; Formalin; Freezing; Funding; Generations; Goals; Guidelines; Health Care Costs; Hematoxylin and Eosin Staining Method; Histocompatibility Testing; Histology; Human; Immunohistochemistry; Informed Consent; International; Ischemia; Knowledge; Liver parenchyma; Malignant Neoplasms; Measurement; Measures; Molecular; Molecular Profiling; Molecular Target; Morphology; Mucous Membrane; Mus; Nuclear; One-Step dentin bonding system; Operative Surgical Procedures; Organ; Paraffin; Paraffin Embedding; Pathologist; Pathway interactions; Phase; Phosphoproteins; Phosphotransferases; Process; Progesterone Receptors; Prostate; Protein Kinase; Protein Microchips; Proteins; RNA; Research; Shipping; Ships; Side; Signal Pathway; Specimen; Staining method; Stains; Structure of parenchyma of lung; Technology; Testing; Therapeutic; Time; Tissue Preservation; Tissue Procurements; Tissues; United States National Institutes of Health; Validation; cost; diagnostic accuracy; inhibitor/antagonist; innovation; kinase inhibitor; laser capture microdissection; lymph nodes; male; novel; sample fixation; tissue fixing; tissue processing; tumor

DESCRIPTION (provided by applicant): An urgent clinical goal is to identify molecular networks associated with subpopulations of cancer patients who may respond individually to molecular targeted inhibitors. Current molecular targeted therapeutics is directed at protein kinases and/or their phosphorylated substrates. Therefore, measurement of this new class of phosphoprotein signal pathway epitopes in tumor biopsy samples is crucial for individualizing molecular targeted therapies. Phosphoprotein antigen epitopes are not adequately preserved by formalin fixation and paraffin embedding, and freezing of tissue is very expensive and compromises diagnostic accuracy. We propose the advanced development and clinical validation of an innovative and transformative technology for preserving tissue phosphoproteins and diagnostic histomorphology for clinical cancer molecular profiling. Applying knowledge gained under an NIH R21 funded study, we created a novel tissue preservation chemistry that stabilizes all classes of phosphoproteins, is compatible with paraffin embedding, while maintaining complete diagnostic histomorphology, and fully preserving critical diagnostic immunohistology (IHC) antigens including Estrogen Receptor, Progesterone Receptor, HER2, and Ki-67. These IHC antigens are not preserved by special research fixatives used for tissue RNA preservation. Our new non-formalin tissue preservative, termed Biomarker and Histology Preservative (BHP) can be seamlessly introduced into the current community hospital clinical diagnostic workflow with no additional steps or equipment. At the time of procurement, tissue can be immersed directly in the new fixative and processed into a paraffin block for routine diagnosis, obviating the need for costly freezing during shipping or storage. BHP offers the potential for substantial improvements over conventional formalin fixation. In the present application we propose the blinded clinical validation of our novel preservation chemistry in community hospital settings, utilizing a team of international pathologists for validation. The goal of the project is one-step paraffin block stabilization of all classes of cellular phosphoproteins, diagnostic histomorphology, and diagnostic immunohistochemistry antigens, while at the same time maintaining full diagnostic morphology equivalent or superior to standard formalin fixation. We will collect fresh surgical tissue, under informed consent, covering a broad variety of organs and cancer histology to develop an archive of 150 cases of matched paraffin and frozen specimens. We will measure 100 validated phosphoprotein epitopes spanning membrane, cytoplasmic and nuclear compartments from extracted paraffin sections using Reverse Phase Protein Microarray (RPMA) and Laser Capture Microdissection (LCM) technology. Following objective independent validation by diagnostic pathologists, this transformative technology will be ready for widespread clinical and research use. Adoption of the technology would mean that only one diagnostic paraffin block could be used for all classes of molecular profiling rather than the current requirement for multiple blocks. This would increase diagnostic accuracy while substantially reducing costs.

描述（由申请人提供）：紧急临床目标是确定与可能单独对分子靶向抑制剂反应的癌症患者亚群有关的分子网络。当前的分子靶向疗法针对蛋白激酶和/或其磷酸化底物。因此，肿瘤活检样品中这种新型的磷酸蛋白信号途径表位的测量对于个体化分子靶向疗法至关重要。通过福尔马林固定和石蜡嵌入，磷酸蛋白抗原表位不能充分保存，并且组织的冷冻非常昂贵，并且会损害诊断准确性。我们提出了一种创新和变革性技术的高级开发和临床验证，用于保存组织磷蛋白以及用于临床癌分子分析的诊断组织形态学。在NIH R21资助的研究下应用知识，我们创建了一种新型的组织保存化学，稳定所有类别的磷酸蛋白，与石蜡嵌入兼容，同时保持完整的诊断性组织形态学，并完全保留关键的诊断诊断免疫学（IHC）抗原抗体，包括雌激素受体，雌激素受体，孕激素，kergesteron，267。这些IHC抗原不能由用于组织RNA保存的特殊研究固定剂保存。我们的新的非正素组织防腐剂，称为生物标志物和组织学防腐剂（BHP）可以无缝地引入当前的社区医院临床诊断工作流程中，而没有其他步骤或设备。在采购时，可以将组织直接浸入新的固定剂中，并加工到石蜡块中以进行常规诊断，从而避免了在运输或存储期间昂贵的冷冻需求。必和必拓提供了对常规福尔马林固定的实质性改进的潜力。在本应用程序中，我们提出了在社区医院环境中对新型保存化学的盲目临床验证，利用一组国际病理学家进行验证。该项目的目的是对所有类别的细胞磷蛋白，诊断性组织形态学和诊断性免疫组织化学抗原抗原的一级石蜡稳定，同时保持完全诊断的形态等效或优于标准福尔马林固定。我们将在知情同意下收集新鲜的手术组织，涵盖各种各样的器官和癌症组织学，以开发150例匹配的石蜡和冷冻标本的档案。我们将使用反相蛋白微阵列（RPMA）和激光捕获微分辨率（LCM）技术测量跨膜，细胞质和核区室的100个经过验证的磷酸蛋白表位。经过诊断病理学家的客观独立验证，该变革性技术将为广泛的临床和研究用途做好准备。该技术的采用将意味着只有一个诊断石蜡可以用于所有类别的分子分析，而不是当前对多个块的需求。这将提高诊断准确性，同时大大降低成本。