MALDI Imaging of Cancer Signaling Signatures

癌症信号特征的 MALDI 成像

• 批准号: 7659682
• 负责人: Stephen J. Kron
• 金额: $ 20.79万
• 依托单位: UNIVERSITY OF CHICAGO
• 依托单位国家: 美国
• 财政年份: 2008
• 资助国家: 美国
• 起止时间: 2008-07-16 至 2011-06-30
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/7659682
• 关键词: Acrylamides; Antibodies; Architecture; Biological Assay; Biopsy; Biosensor; Cell Extracts; Cell Separation; Cells; Characteristics; Cleaved cell; Cytolysis; Detection; Diagnosis; Diagnostic Neoplasm Staging; Drug Delivery Systems; Emerging Technologies; Engineering; Epidermal Growth Factor Receptor; Epidermal Growth Factor Receptor Tyrosine Kinase Inhibitor; Erlotinib; Excision; Family; Frozen Sections; Gefitinib; Gene Expression; Head and Neck Neoplasms; Head and Neck Squamous Cell Carcinoma; Heterogeneity; Histology; Human; Hydrogels; Image; Ions; Island; Label; Lasers; Link; Malignant Neoplasms; Measurement; Measures; Methods; Microdissection; Modeling; Modification; Monitor; Neoplasm Metastasis; Nude Mice; Oncogenic; Operative Surgical Procedures; Outcome; Patients; Pattern; Peptide Mapping; Peptides; Phase; Phosphorylation; Phosphotransferases; Population; Printing; Protein Tyrosine Kinase; Proteins; Proteomics; Radio; Radioisotopes; Reaction; Reading; Relative (related person); Resistance; Resolution; Route; Sampling; Scanning; Sepharose; Signal Transduction; Signal Transduction Pathway; Signaling Protein; Solid; Spatial Distribution; Spectrometry, Mass, Matrix-Assisted Laser Desorption-Ionization; Squamous cell carcinoma; Stromal Cells; Surface; Technology; Therapeutic; Thick; Tissue Sample; Tissues; Translating; Tumor Tissue; Work; base; biochip; cancer cell; cancer diagnosis; cancer imaging; detector; immunocytochemistry; inhibitor/antagonist; innovation; leukemia; molecular marker; neoplastic cell; outcome forecast; prognostic; response; sensor; surface coating; tissue/cell culture; tool; tumor; tumor xenograft; two-dimensional

DESCRIPTION (provided by applicant): Detection of the activation of cellular kinases associated with oncogenic signaling can serve as a valuable molecular marker for cancer diagnosis and as a predictive tool for selection of therapy. In human squamous cell carcinoma of the head and neck (SCCHN), EGFR activation is associated with therapeutic resistance, increased metastasis and poor outcomes. Histology and tumor architecture provide complementary and critical information about cancer stage and grade. By adapting and extending emerging technologies for kinase sensor biochips, we propose to develop the capability to image the distribution of cancer signaling in tumor tissue obtained by biopsy or surgical excision. We and others have developed robust, sensitive and specific biochip-based assays for kinase activity in cellular lysates, using immunodetection, radionuclide incorporation or MALDI-TOF MS analysis as a read-out. We now intend to adapt these methods to create multiplexed assays whereby multiple peptides, serving as specific substrates for kinase involved in oncogenic signaling, will be linked reversibly to the surface of a biochip. This multiplexed biosensor will be exposed to thick sections of tumor biopsies by "tissue print" to allow phosphorylation of the peptides. The cellular material will be washed away and the biochip will be interrogated by MALDI-TOF MS imaging to detect relative peptide phosphorylation, creating an image of the multiple kinase activities across the tumor section. These kinase activity images can be correlated with conventional histology and immunocytochemistry to enhance diagnosis and prognosis. To establish proof-of-principle in the R21 phase, methods will be developed using human SCCHN tissue culture cells and xenograft tumors of grown in athymic nude mice. We will take advantage of the activated EGFR kinase characteristic of this tumor and the availability of the specific EGFR inhibitors gefitinib and erlotinib to develop and validate our biochip sensor and imaging capabilities. We anticipate achieving sufficient sensitivity and resolution to detect SCCHN tumor islands embedded in extensive stroma.

描述（由申请人提供）：检测与致癌信号传导相关的细胞激酶的活化可以作为癌症诊断的有价值的分子标志物和治疗选择的预测工具。在人头颈部鳞状细胞癌（SCCHN）中，EGFR活化与治疗耐药性、转移增加和预后不良相关。组织学和肿瘤结构提供了有关癌症分期和分级的补充和关键信息。通过适应和扩展激酶传感器生物芯片的新兴技术，我们建议开发成像活检或手术切除获得的肿瘤组织中的癌症信号传导的分布的能力。我们和其他人已经开发了强大的，敏感的和特定的基于生物芯片的细胞裂解物中的激酶活性测定，使用免疫检测，放射性核素掺入或MALDI-TOF MS分析作为读出。我们现在打算调整这些方法来创建多路复用的测定，从而多个肽，作为特定的底物激酶参与致癌信号，将可逆地连接到生物芯片的表面。这种多重生物传感器将通过“组织印迹”暴露于肿瘤活检的厚切片，以允许肽的磷酸化。细胞物质将被冲洗掉，生物芯片将通过MALDI-TOF MS成像进行询问，以检测相对的肽磷酸化，从而创建跨肿瘤切片的多种激酶活性的图像。这些激酶活性图像可以与传统的组织学和免疫细胞化学，以提高诊断和预后。为了在R21阶段建立原理验证，将使用人SCCHN组织培养细胞和在无胸腺裸鼠中生长的异种移植肿瘤开发方法。我们将利用这种肿瘤的活化EGFR激酶特性以及特异性EGFR抑制剂吉非替尼和厄洛替尼的可用性来开发和验证我们的生物芯片传感器和成像能力。我们期望达到足够的灵敏度和分辨率来检测嵌入广泛基质中的SCCHN肿瘤岛。

项目成果

Photocleavable peptide-oligonucleotide conjugates for protein kinase assays by MALDI-TOF MS.

• DOI: 10.1039/c2mb25163a
• 发表时间: 2012-09
• 期刊: Molecular bioSystems
• 作者: Zhou G;Khan F;Dai Q;Sylvester JE;Kron SJ
• 通讯作者: Kron SJ