Transglutaminase Modulated Tumor-Stroma Interaction in Pancreatic Cancer

转谷氨酰胺酶调节胰腺癌中的肿瘤-基质相互作用

• 批准号: 8114594
• 负责人: Daniela E Matei
• 金额: $ 16.08万
• 依托单位: INDIANA UNIV-PURDUE UNIV AT INDIANAPOLIS
• 依托单位国家: 美国
• 财政年份: 2011
• 资助国家: 美国
• 起止时间: 2011-09-06 至 2013-08-31
• 项目状态: 已结题
• 来源: https://reporter.nih.gov/project-details/8114594
• 关键词: Address; Adenocarcinoma Cell; Antineoplastic Agents; Apoptosis; Apoptotic; Biology; Cancer Etiology; Cancer Patient; Cell Survival; Clinical; Coculture Techniques; Collagen; Conditioned Culture Media; Cues; Data; Desmoplastic; Detection; Development; Disease Resistance; Drug Delivery Systems; Endothelial Cells; Environment; Enzyme-Linked Immunosorbent Assay; Enzymes; Epithelial; Epithelium; Etiology; Exploratory/Developmental Grant; Extracellular Matrix; Fibroblast Growth Factor; Fibroblasts; Fibrosis; Foundations; Future; Glutamine; Growth Factor; High Pressure Liquid Chromatography; Image; Lesion; Lysine; Malignant - descriptor; Malignant Neoplasms; Malignant neoplasm of pancreas; Mass Spectrum Analysis; Measurement; Measures; Modeling; Monitor; Myofibroblast; Neoplasm Metastasis; Outcome; P-Glycoprotein; Pancreas; Pancreatic Ductal Adenocarcinoma; Patients; Penetrance; Permeability; Pilot Projects; Platelet-Derived Growth Factor; Play; Positioning Attribute; Process; Proteins; RNA; Radiation; Relative (related person); Reporting; Research; Resistance; Role; Stress; Stromal Cells; Testing; Therapeutic; Tissues; Transfection; Transglutaminases; Trichrome stain; Tumor Tissue; Work; Xenograft procedure; angiogenesis; base; cancer cell; cell stroma; cellular engineering; chemotherapy; clinically relevant; crosslink; cytokine; density; gemcitabine; inhibitor/antagonist; innovation; knock-down; neoplastic cell; novel; overexpression; pancreas xenograft; pancreatic cancer cells; pancreatic neoplasm; research study; resistance mechanism; response; stellate cell; tumor; tumor growth; tumor progression; uptake

DESCRIPTION (provided by applicant): The interaction between cancer cells and the pancreatic stroma has been implicated in pancreatic cancer progression and resistance to chemotherapy and radiation. Based on our preliminary results demonstrating prominent tissue transglutaminase (TG2) expression and activity in the pancreatic tumor millieu, we hypothesized that TG2 secreted by pancreatic cancer cells modulates stroma, promotes cancer cell survival, and impairs chemotherapy delivery. We propose to test the hypothesis by three specific aims: Aim 1: Measure proliferation and activation of pancreatic stromal cells in response to TG2 secreted by pancreatic cancer cells. Aim 2: Demonstrate that TG2 knockdown or its enzymatic inhibition regulate stroma composition in pancreatic tumors. Aim 3: Demonstrate that TG2 knockdown or its enzymatic inhibition increases gemcitabine delivery into the pancreatic milieu. Aim 1 will use co-culture experiments to measure the effects of secreted TG2 on proliferation, survival and activation of fibroblasts and endothelial cells. Cytokines and growth factors secreted by fibroblasts will be measured in media conditioned by stromal and cancer cells by using Multiplex ELISA. Aims 2 and 3 will use orthotopic pancreatic xenografts generated from pancreatic cancer cells in which TG2 was knocked down by using stable transfection of sh-RNA or antisense construct targeting TG2, or TG2 will be inhibited by using KCC009, a specific enzymatic inhibitor. Endpoints of the experiments are measurement and comparison of: fibrosis (immunostaining for collagen and trichrome stain), number and proliferation of stromal cells (myofibroblasts and pancreatic stellate cells), angiogenesis (microvessel density), survival and proliferation of pancreatic cancer cells, and penetrance of chemotherapy into pancreatic tissue. For the latter endpoint, we will measure the concentration of gemcitabine within pancreatic tissue derived from cancer cells expressing normal or low TG2 levels by using HPLC mass spectrometry. In summary, successful demonstration of the role of tissue transglutaminase in modulation of the pancreatic stroma will provide a strong rationale for investigating further TG2 inhibition as an anti-cancer strategy or as means to sensitize pancreatic tumors to chemotherapy. These studies have direct clinical relevance to pancreatic cancer patients. PUBLIC HEALTH RELEVANCE: The study of TG2 as a modulator of the tumor cell-stroma interaction in pancreatic cancer is novel and has direct therapeutic implications. We hypothesized that modulation of the pancreatic stroma by TG2 plays a critical role in tumor progression and resistance to chemotherapy. Successful demonstration of this process will provide the rationale for pursuing TG2 inhibition to block pancreatic tumor progression, enhance uptake of chemotherapeutics within tumors, and increase response to anti-cancer agents in pancreatic cancer. Ultimately, such TG2-directed strategies will increase the sensitivity of pancreatic tumors to chemotherapy and impact directly the clinical outcome of this deadly cancer.

描述（由申请人提供）：癌细胞和胰腺间质之间的相互作用与胰腺癌的进展和对化疗和放疗的耐药性有关。基于我们的初步结果显示，组织转谷氨酰胺酶（TG2）在胰腺肿瘤中显著表达和活性，我们假设胰腺癌细胞分泌的TG2调节基质，促进癌细胞存活，并损害化疗递送。我们提出通过三个特定目的来验证这一假设：目的1：测量胰腺癌细胞分泌TG2时胰腺基质细胞的增殖和激活。目的2：证明TG2敲低或其酶抑制调节胰腺肿瘤的基质组成。目的3：证明TG2敲低或其酶促抑制增加吉西他滨进入胰腺环境的递送。目的1将使用共培养实验来测量分泌TG2对成纤维细胞和内皮细胞的增殖、存活和活化的影响。在基质细胞和癌细胞调节的培养基中，采用多重ELISA法测定成纤维细胞分泌的细胞因子和生长因子。Aims 2和3将使用由胰腺癌细胞生成的原位胰腺异种移植物，其中TG2通过稳定转染sh-RNA或靶向TG2的反义构建物而被敲除，或者TG2将被KCC009（一种特异性酶抑制剂）抑制。实验的终点是测量和比较：纤维化（胶原和三色染色的免疫染色），基质细胞（肌成纤维细胞和胰腺星状细胞）的数量和增殖，血管生成（微血管密度），胰腺癌细胞的存活和增殖，以及化疗进入胰腺组织的外显率。对于后一个终点，我们将使用HPLC质谱法测量来自表达正常或低TG2水平的癌细胞的胰腺组织中吉西他滨的浓度。总之，成功证明组织转谷氨酰胺酶在胰腺间质调节中的作用将为进一步研究TG2抑制作为抗癌策略或作为胰腺肿瘤对化疗敏感的手段提供强有力的理论依据。这些研究对胰腺癌患者有直接的临床意义。