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: 8079819
• 负责人: Lance Allen Liotta
• 金额: $ 32.74万
• 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/8079819
• 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. PUBLIC HEALTH RELEVANCE: Our technology for preserving proteins, at the time of collection, will enable analysis of molecular targeted kinase inhibitors for individualized therapy, while considerably reducing healthcare costs, and increasing diagnostic accuracy. Our new non-formalin tissue preservative can be seamlessly introduced into the current community hospital clinical diagnostic workflow with no additional steps or equipment.

描述（由申请人提供）：一个紧迫的临床目标是鉴定与可能对分子靶向抑制剂有个体应答的癌症患者亚群相关的分子网络。目前的分子靶向治疗针对蛋白激酶和/或其磷酸化底物。因此，在肿瘤活检样本中测量这类新的磷蛋白信号通路表位对于个体化分子靶向治疗至关重要。磷蛋白抗原表位不能通过福尔马林固定和石蜡包埋充分保存，并且组织的冷冻非常昂贵并且损害诊断准确性。我们提出了一种创新和变革性技术的先进开发和临床验证，用于保存组织磷蛋白和临床癌症分子谱的诊断组织形态学。应用NIH R21资助研究获得的知识，我们创建了一种新型组织保存化学，可稳定所有类别的磷蛋白，与石蜡包埋相容，同时保持完整的诊断组织形态学，并完全保留关键的诊断免疫组织学（IHC）抗原，包括雌激素受体，孕激素受体，HER 2和Ki-67。这些IHC抗原不被用于组织RNA保存的特殊研究固定剂保存。我们新的非福尔马林组织防腐剂，称为生物标志物和组织学防腐剂（BHP），可以无缝地引入到当前的社区医院临床诊断工作流程中，无需额外的步骤或设备。在采购时，组织可以直接浸入新的固定剂中，并加工成石蜡块用于常规诊断，从而避免了在运输或储存期间进行昂贵的冷冻。BHP提供了比传统福尔马林固定有实质性改进的潜力。在本申请中，我们提出在社区医院环境中对我们的新型保存化学进行盲法临床验证，利用国际病理学家团队进行验证。该项目的目标是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的一步石蜡块稳定化，同时保持与标准福尔马林固定等同或上级的完全诊断形态学。我们将在知情同意的情况下收集新鲜手术组织，涵盖各种器官和癌症组织学，以建立150例匹配石蜡和冷冻标本的档案。我们将使用反相蛋白微阵列（RPMA）和激光捕获显微切割（LCM）技术从提取的石蜡切片中测量100个经过验证的跨膜、细胞质和核区室的磷蛋白表位。经过诊断病理学家的客观独立验证，这项变革性技术将准备用于广泛的临床和研究。采用该技术将意味着只有一个诊断石蜡块可以用于所有类别的分子谱分析，而不是目前需要的多个块。这将提高诊断的准确性，同时大大降低成本。 公共卫生关系：我们在收集蛋白质时保存蛋白质的技术将使分子靶向激酶抑制剂的分析能够用于个体化治疗，同时大大降低医疗成本，提高诊断准确性。我们的新型非福尔马林组织防腐剂可以无缝地引入到当前的社区医院临床诊断工作流程中，无需额外的步骤或设备。