Prediction of Therapy Response in Colorectal Cancer

结直肠癌治疗反应的预测

• 批准号: 8552855
• 负责人: Thomas Ried
• 金额: $ 46.14万
• 依托单位: DIVISION OF BASIC SCIENCES - NCI
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/8552855
• 关键词: Academic Medical Centers; Address; Adverse effects; Aftercare; Anti-CEA Antibody; Award; BCL2 gene; BIRC5 gene; Behavior; Biological; Biological Assay; Biological Markers; Biopsy; CYP19A1 gene; Caliber; Cancer Biology; Cancer Patient; Cancer cell line; Cell Cycle Progression; Cell Line; Climate; Clinical; Clinical Management; Clinical Research; Clinical Trials; Clinical assessments; Collaborations; Colorectal Cancer; Combined Modality Therapy; Complement; DNA Double Strand Break; DNA Sequence Analysis; Data Set; Detection; Development; Disease; Dose; Double Strand Break Repair; Enrollment; Fluorouracil; Funding; Gene Expression; Gene Expression Profile; Gene Expression Profiling; Gene Expression Regulation; Gene Silencing; Gene Targeting; Genes; Genetic Polymorphism; Genetic Variation; Genomics; German population; Germany; Goals; Image; Immunohistochemistry; In complete remission; Individual; Interdisciplinary Study; Investigation; Laboratories; Large Intestine Carcinoma; Left; Link; M Phase Arrest; MKI67 gene; Magnetic Resonance Imaging; Measurement; Measures; Mediating; MicroRNAs; Micrometastasis; Mismatch Repair; Modality; Modification; Molecular Profiling; Molecular Target; Monitor; Mucous Membrane; Neoadjuvant Therapy; Operative Surgical Procedures; PET/CT scan; Participant; Pathway interactions; Patients; Pattern; Pharmaceutical Preparations; Phase III Clinical Trials; Positioning Attribute; Postdoctoral Fellow; Predictive Value; Primary Neoplasm; Process; Proteins; Publishing; RNA Interference; Radiation; Radiation therapy; Radiosensitization; Randomized; Recruitment Activity; Rectal Adenocarcinoma; Rectal Cancer; Recurrence; Reporter; Reporting; Research; Research Design; Research Infrastructure; Resistance; Roentgen Rays; Sampling; Signal Transduction; Specificity; Staging; Staining method; Stains; Stratification; System; TCF7L2 gene; TP53 gene; Testing; Therapeutic; Thymidylate Synthase; Tissue-Specific Gene Expression; Toxic effect; Treatment Failure; Ultrasonography; Validation; Visceral; X-Ray Computed Tomography; arm; base; beta catenin; cancer cell; clinical practice; clinically relevant; cohort; cytotoxic; design; established cell line; follow-up; holistic approach; improved; in vivo; insight; interest; neoplastic cell; novel; outcome forecast; oxaliplatin; prospective; protein function; rectal; research study; response; success; survivin; tool; transcription factor; treatment response; tumor; two-arm study

The clinical study CAO-/ARO-/AIO-94 of the German Rectal Cancer Study Group led clinicians around the world to embrace neoadjuvant chemoradiotherapy as the treatment of choice for patients with locally advanced rectal cancer because it reduces the rate of local recurrence. A follow-up prospective randomized phase III clinical trial is now in place (CAO-/ARO-/AIO-04). The goal of this trial is to address whether tumor response and prognosis can be further improved by the addition of oxaliplatin to the existing combined modality therapy (5-FU and radiation). The gene expression analyses, aCGH experiments, and measurement of miRNA profiles that we propose in Project 3 will be done on patients enrolled in this trial. Our collaborations with Gttingen are integrated with a Clinical Research Unit with the title The biological basis of individual tumor response in patients with rectal cancer (http://www.kfo179.de/). The establishment of such Clinical Research Units in Germany is highly competitive and is aimed at establishing a climate and an infrastructure conducive to interdisciplinary research that is focused on clinically relevant questions. The Clinical Research Units consists of eight subprojects. We are actively involved in Subproject 1, Gene expression signature and genetic polymorphisms for response prediction of rectal carcinomas to preoperative chemoradiotherapy and Subproject 2, Functional validation of genes involved in resistance of rectal carcinomas to preoperative chemoradiation. The other subprojects analyze other pertinent aspects of rectal cancer biology and relevant clinical parameters, including the detection of micrometastases, the development of an anti-CEA-based immuno-PET/CT in vivo imaging system for therapy monitoring, the analysis of mismatch-repair status as it relates to therapy response, and the identification of biomarkers (including specific SNPs in genes in drug metabolizing pathways) to assess treatment-related toxicity. Together, we are able to pursue a holistic approach to the problem of treatment failure and disease prognostication in patients with rectal cancer.There is a wide spectrum of tumor responsiveness of rectal adenocarcinomas to preoperative chemoradiotherapy ranging from complete response to complete resistance. We were therefore interested to investigate whether parallel gene expression profiling of the primary tumor can contribute to stratification of patients into groups of responders or non-responders. We have collected pre-therapeutic biopsies from 30 locally advanced rectal carcinomas (determined by rectal ultrasound as uT3 and uT4) and analyzed these samples for gene expression signatures using microarrays. All patients were participants of a phase III clinical trial (CAO/ARO/AIO-94, German Rectal Cancer Trial) and were randomized to receive a preoperative combined modality therapy including 5-fluorouracil and radiation. In an initial set of 23 patients responders and non-responders (measured by T-level down-sizing) showed significantly different expression levels for 54 genes (p<0.001). The ability to predict response to therapy using gene expression profiles was rigorously evaluated using leave-one-out cross-validation (LOOCV). Tumor behavior was correctly predicted in 83% of patients (p=0.02). Sensitivity (correct prediction of response) was 78% and specificity (correct prediction of non-response) was 86% with a positive predictive value of 78% and negative predictive value of 86%. These results suggest that pre-therapeutic gene expression profiling may assist in response prediction of rectal adenocarcinomas to preoperative chemoradiotherapy. The implementation of gene expression profiles for treatment stratification and clinical management of cancer patients requires validation in large, independent studies, which are now under way. In parallel to the validation of the 54 gene set in Gttingen we plan to profile all 300 rectal cancer samples (and matched normal mucosa) using the Agilent 44K expression platform at the NCI, a number required for a test robust enough to enter clinical practice. To date, 107 tumor samples and matched mucosa have already been successfully processed. Obviously, these analyses not only provide insight into mechanisms of response to chemoradiotherapy but also into basic pathways disturbed in rectal carcinomas and are thus thematically linked to Project 1 and 2. We consider the availability of matched normal mucosa as a definitive enrichment of the dataset. Depending on the genes that we identify as differentially expressed, we will consider whether DNA sequence analysis might be appropriate to identify potential polymorphisms associated with protein function. We hypothesize that modification of expression levels of genes involved in resistance to chemoradiotherapy could be explored for the purpose of sensitizing a priori resistant tumors to treatment. For instance, silencing of genes that are upregulated in resistant tumors in cell lines established from these tumor samples could potentially result in increased sensitivity. This line of research will be pursued in Gttingen as subproject 2 of the Clinical Research Unit. Dr. Marian Grade, who was a postdoctoral fellow in my laboratory, was awarded a Junior Group Leader position to focus on the functional validation of response genes. Due to our expertise we will establish cell lines from resistant and responsive tumors in my laboratory for functional analysis and will continue to collaborate with Dr. Grade. These analyses will be complemented by establishing microRNA expression profiles from responsive and non-responsive patients. Since one function of miRNA is the regulation of gene expression, we anticipate that differences in the expression profiles could indeed reveal biomarkers of response. In addition, we have begun to identify cell lines derived from primary colorectal carcinomas that show differential sensitivity to 5-FU and radiation. We anticipate that differential gene expression profiles after treatment can help to identify genes involved in resistance which can then be validated in primary tumors.A considerable percentage of rectal cancers are resistant to standard preoperative chemoradiotherapy. Because patients with a priori resistant tumors do not benefit from multimodal treatment, understanding and overcoming this resistance remains of utmost clinical importance. We recently reported over-expression of the Wnt transcription factor TCF4, also known as TCF7L2, in rectal cancers that were resistant to 5-fluorouracil-based chemoradiotherapy. Because Wnt signaling has not been associated with treatment response, we aimed to investigate whether TCF4 mediates chemoradioresistance. RNA-interference mediated silencing of TCF4 was employed in three colorectal cancer cell lines, and sensitivity to (chemo-) radiotherapy was assessed using a standard colony formation assay. Silencing of TCF4 caused a significant sensitization of colorectal cancer cells to clinically relevant doses of X-rays. This effect was restricted to tumor cells with high TCF reporter activity, but, presumably, in a beta-catenin independent manner. Radiosensitization was the consequence of (i) a transcriptional deregulation of Wnt/TCF4 target genes, (ii) a silencing-induced G2/M phase arrest, (iii) an impaired ability to adequately halt cell cycle progression after radiation, and (iv) a compromised DNA double strand break repair as assessed by gamma-H2AX staining. Taken together, our results indicate a novel mechanism through which the Wnt transcription factor TCF4 mediates chemoradioresistance. Moreover, they suggest that TCF4 is a promising molecular target to sensitize resistant tumor cells to (chemo-) radiotherapy.

德国直肠癌研究小组的临床研究CAO-/ARO-/AIO-94导致世界各地的临床医生接受新辅助化学疗法，作为对局部晚期直肠癌患者的选择的治疗方法，因为它降低了局部复发的速度。 现在已经进行了前瞻性随机临床试验（CAO-/ARO-/AIO-04）。 该试验的目的是解决通过在现有的合并情态疗法（5-FU和辐射）中添加奥沙利铂（5-FU和辐射），是否可以进一步改善肿瘤反应和预后。 基因表达分析，ACGH实验和我们在项目3中提出的miRNA谱的测量将对参加该试验的患者进行。 我们与Gttingen的合作与临床研究单元的标题集成在一起，标题是直肠癌患者（http://www.kfo179.de/）的个体肿瘤反应的生物学基础。 在德国建立此类临床研究单位具有很高的竞争力，旨在建立一个有利于跨学科研究的气候和基础设施，该研究旨在介绍临床相关问题。 临床研究单元由八个副标题组成。 我们积极参与了次要注射1，基因表达特征和遗传多态性，用于对直肠癌对术前化学放疗的响应预测和副标理2，对直肠癌抗性的基因的功能验证。 The other subprojects analyze other pertinent aspects of rectal cancer biology and relevant clinical parameters, including the detection of micrometastases, the development of an anti-CEA-based immuno-PET/CT in vivo imaging system for therapy monitoring, the analysis of mismatch-repair status as it relates to therapy response, and the identification of biomarkers (including specific SNPs in genes in drug metabolizing pathways) to评估与治疗有关的毒性。 总之，我们能够在直肠癌患者的治疗衰竭和疾病预后的问题上采取整体方法。在直肠腺癌的肿瘤反应性方面广泛，到术前术治疗疗法，范围从完全反应到完全抗性。 因此，我们有兴趣研究原发性肿瘤的平行基因表达分析是否可以将患者分为反应者或非反应者组。 我们已经从30个局部直肠癌（由直肠超声为UT3和UT4确定）中收集了治疗前活检，并使用微阵列分析了这些样品的基因表达特征。 所有患者均为III期临床试验的参与者（CAO/ARO/AIO-94，德国直肠癌试验），并随机分配以接受术前合并的模态疗法，包括5-氟尿嘧啶和辐射。在最初的23例患者反应者和非反应者（通过T级尺寸测量）中，54个基因的表达水平显着不同（P <0.001）。 使用剩余的交叉验证（LOOCV）对使用基因表达谱预测治疗反应的能力进行了严格评估。 在83％的患者中正确预测了肿瘤行为（P = 0.02）。 灵敏度（正确预测反应）为78％，特异性（正确预测无响应）为86％，正预测值为78％，负预测值为86％。 这些结果表明，治疗前基因表达分析可能有助于响应直肠腺癌对术前化学放疗。 用于癌症患者治疗分层和临床管理的基因表达谱的实施需要在正在进行的大型独立研究中进行验证。与Gttingen中的54个基因验证的验证，我们计划使用NCI的Agilent 44K表达平台介绍所有300个直肠癌样品（并匹配正常粘膜），这是一个足够可靠的测试所需的数字，足以进入临床实践。 迄今为止，已经成功处理了107种肿瘤样品和匹配的粘膜。 显然，这些分析不仅可以深入了解对化学疗法的反应机制，而且还介绍了直肠癌中受干扰的基本途径，因此与项目1和第2项目相关。我们认为匹配的正常粘膜的可用性是数据集的明确富集。 根据我们鉴定为差异表达的基因，我们将考虑是否可能适合鉴定与蛋白质功能相关的潜在多态性。我们假设可以探索参与化学放疗的抗性基因表达水平的修饰，目的是为了使先验耐药性肿瘤对治疗敏感。 例如，在这些肿瘤样品中建立的细胞系中耐药性肿瘤中上调的基因沉默可能会导致敏感性提高。 这项研究将在Gttingen进行，作为临床研究部门的亚第2条。 玛丽安·坡（Marian Grade）博士是我实验室的博士后研究员，被授予初级小组领导者职位，专注于响应基因的功能验证。 由于我们的专业知识，我们将在我的实验室中建立耐药和反应性肿瘤的细胞系，以进行功能分析，并将继续与Dr. Grade合作。 这些分析将通过建立反应性和无反应性患者的microRNA表达谱而得到补充。 由于miRNA的一个功能是基因表达的调节，因此我们预计表达谱的差异确实可以揭示出反应的生物标志物。 此外，我们已经开始鉴定出源自原代结直肠癌的细胞系，这些细胞系对5-FU和辐射显示出差异的敏感性。 我们预计治疗后的差异基因表达谱可以帮助鉴定抗抗性的基因，然后在原发性肿瘤中进行验证。大量的直肠癌对标准术前术治疗疗法具有抵抗力。 由于具有先验抗性肿瘤的患者并不能受益于多模式治疗，因此理解并克服这种抗性仍然是最重要的临床重要性。 我们最近报道了Wnt转录因子TCF4的过表达，也称为TCF7L2，在对基于5-氟尿嘧啶的化学放疗的直肠癌中。 由于Wnt信号传导与治疗反应尚未相关，因此我们旨在研究TCF4是否介导化学降低剂。在三种结直肠癌细胞系中使用了RNA干扰介导的TCF4沉默，并使用标准菌落形成测定法评估了对（化学 - ）放疗的敏感性。 TCF4的沉默导致大肠癌细胞对临床相关剂量的X射线的显着敏化。 这种作用仅限于具有高TCF报告基因活性的肿瘤细胞，但大概是以β-catenin独立的方式进行的。 放射敏化是（i）Wnt/TCF4靶基因的转录放松管制，（ii）沉默诱导的G2/M相倒置，（iii）辐射后充分停止细胞周期进展的能力受损，（iv）（iv）（iv）（iv）DNA双链损坏的dna双链损坏，以通过GAMMA-HAMMA-HAMMA-HAMMA-HAMMA-HAMMA-HAMBA-HAMMA-HAMBA-HAMBA-HAMBA-HAMBA-HAMBA-HAMMA-HAMMA-HAMMA-HAMMA-HAMMA-HAMMA-HAMMA-HAMMA-HAMMA-HAMMA 2AXS STENANGINASENATINAD。综上所述，我们的结果表明了一种新的机制，通过该机制Wnt转录因子TCF4介导了化学放电量。 此外，他们认为TCF4是将抗性肿瘤细胞敏感到（化学）放疗的有前途的分子靶标。