Computational Molecular Pathology Research for Cancer Diagnostics and Biomarkers

癌症诊断和生物标志物的计算分子病理学研究

• 批准号: 10703030
• 负责人: Robert Simpson
• 金额: $ 143.47万
• 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/10703030
• 关键词: Address; Adenocarcinoma; Animal Model; Apoptosis; Applied Research; Automobile Driving; BRAF gene; Biological Markers; Biological Models; Breast Cancer Model; C-KIT Mutation; CD8B1 gene; CLIC4 gene; Cancer Diagnostics; Cancer Model; Canis familiaris; Cell Cycle; Cell Line; Cell Survival; Cells; Clinical; Comprehension; Coupled; Cutaneous Melanoma; Development; Diagnosis; Diagnostic; Diagnostic Neoplasm Staging; Disease; Disease model; Dose; Drug Combinations; Drug Kinetics; Epidermal Growth Factor Receptor; Evaluation; FOXP3 gene; Fibrosis; Genomics; Growth; Hematology; Human; Immune; Immune response; Incidence; Individual; Infiltration; Investigation; Investigational Therapies; KRAS2 gene; Kidney; Libraries; Lung; Lung Adenocarcinoma; Lymphocyte; MAP Kinase Gene; MEK inhibition; Malignant Neoplasms; Medical; Medicine; Metastatic Neoplasm to the Lung; Methods; Modeling; Molecular; Molecular Computations; Monitor; Mus; Mutation; Necrosis; Neoplasm Metastasis; Nuclear Envelope; Oncogenes; Outcome; PI3K/AKT; Pathogenesis; Pathologic; Pathology; Pathway interactions; Phenotype; Primary Neoplasm; Proteins; Proto-Oncogene Proteins c-akt; RANTES; Reagent; Research; Research Subjects; Resources; Sampling; Signal Transduction; Specimen; T-Lymphocyte Subsets; Techniques; Technology; Testing; Therapeutic Intervention; Tissues; Translating; Tumor-infiltrating immune cells; Woman; Work; Xenograft procedure; anticancer research; biomarker discovery; cancer biomarkers; cancer cell; cancer therapy; cancer type; carcinogenesis; clinical application; clinically relevant; design; drug development; human disease; human model; improved; inhibitor; insight; lung colonization; mTOR Inhibitor; mTOR Signaling Pathway; macrophage; malignant breast neoplasm; molecular pathology; mucosal melanoma; mutant; novel; preclinical efficacy; preclinical trial; predictive modeling; programmed cell death ligand 1; response; side effect; targeted treatment; technology development; therapeutic development; tool; tumor; tumor heterogeneity; tumor immunology; tumor microenvironment; vascular abnormality

Research is conducted to characterize and develop new animal models of human disease and to develop the means to better characterize disease relevance produced in the model, addressing critical barriers to research progress. Additional aims include the development of new research technologies and means to interrogate cancer tissues for the evaluation and application of disease biomarkers. The research is also aimed to translate approaches to cancer treatment intended for clinical application. Progress was made in developing research resources useful in developing and characterizing new models of human cancer and in cancer diagnostics. The research resulted in methods and computational capabilities utilized to detect and quantify a metastasis promoting protein in the nuclear membrane that contributed to mechanistic insight in cell sensing and metastasis. Work in modeling breast cancer focused on elevated CLIC4 expression in human breast cancers and its alignment with early invasion and poor outcome in women. CLIC4 in the host microenvironment is required for lung metastasis, while the absence of host CLIC4, results in a microenvironment in the primary tumor and the pre-metastatic lungs that is unfavorable for tumor viability and lung colonization in the model. In particular the primary tumor microenvironment within mice deficient in CLIC4 develops necrosis and vascular abnormalities, detected through computational molecular tissue analyses. Research also focused on a highly topical and outstanding issue in tumor immunology. In pulmonary adenocarcinoma, we collaboratively posed whether the driver oncogene influences the kinds of immune cells that infiltrate the lung or their activation status. this frame of reference was undertaken for its potential clinical relevance. We profiled several lung adenocarcinomas for their immune cell infiltrations, evaluating primarily macrophages and T cell subsets, in addition to reactive tumor-associated fibroplasia in these tumors. In a small, highly curated set of samples, all specimens had some degree of immune response infiltration amid cancer cells. We did not observe a clear difference among the tumors using a limited set of parameters (CD8, CD68, CD4, FOXP3, PDL1), where mutant EGFR was the driving oncogene, versus mutant KRAS. However, the degree of fibrosis did appear to correlate with lymphocyte infiltration. This set of analyses provides a strong demonstration of the immune cell infiltration complexity in the context of tumor heterogeneity. Finally, the investigation provided correlates between presence of T regulatory lymphyocytes, high CCL5 expression and CD8+ effector function in human pulmonary adenocarcinomas. Work also continued on drug development for combined targeted therapy for mucosal melanomas. Clinical and pathological correlates between human and canine mucosal melanomas are substantial, and the relatively greater incidence of spontaneous naturally occurring mucosal melanoma in dogs represents a promising opportunity for predictive modeling. Both canine and human mucosal melanomas appear to harbor BRAF, NRAS and c-kit mutations uncommonly, compared to human cutaneous melanomas, although both species share AKT and MAPK signaling activation. The genomic landscapes of human and canine mucosal melanoma appear highly diverse and generally lack recurring hotspot mutations associated with cutaneous melanomas. Although much remains to be determined, evidence indicates that Ras/MAPK and/or PI3K/AKT/mTOR signaling pathway activations are common in both species and may represent targets for therapeutic intervention. Through this research sapanisertib, an mTORC1/2 inhibitor, was selected from a PI3K/mTOR inhibitor library to collaborate with MEK inhibition; the latter preclinical efficacy was demonstrated previously for canine mucosal melanoma. Combined inhibition of MEK and mTORC1/2, using trametinib and sapanisertib, produced apoptosis and cell-cycle alteration, synergistically reducing cell survival in canine mucosal melanoma cell lines with varying basal signaling activation levels. Compared with individual inhibitors, a staggered sapanisertib dose, coupled with daily trametinib, was optimal for limiting primary mucosal melanoma xenograft growth in mice, and tumor dissemination in a metastasis model, while minimizing hematologic and renal side effects. Inhibitors downmodulated respective signaling targets and the combination additionally suppressed pathway reciprocal crosstalk. Further studies in the pharmacokinetics of the two-drug combination are ongoing.

开展研究是为了描述和开发新的人类疾病动物模型，并开发更好地描述模型中产生的疾病相关性的方法，以解决研究进展的关键障碍。其他目标包括开发新的研究技术和手段，以审问癌症组织，以评估和应用疾病生物标志物。这项研究还旨在将癌症治疗方法转化为临床应用。在开发有益于开发和表征新的人类癌症模型和癌症诊断方面的研究资源方面取得了进展。这项研究的结果是利用方法和计算能力来检测和量化核膜中的转移促进蛋白，这有助于从机制上洞察细胞感知和转移。乳腺癌模型的工作集中在人类乳腺癌中CLIC4的高表达，以及它与女性早期侵袭和不良预后的一致性。肺转移需要宿主微环境中的CLIC4，而宿主CLIC4的缺失会导致原发肿瘤和转移前肺的微环境不利于肿瘤的存活和肺的定植。特别是，通过计算分子组织分析检测到，缺乏CLIC4的小鼠体内的原始肿瘤微环境会发展为坏死和血管异常。研究还集中在肿瘤免疫学中一个高度热门和突出的问题上。在肺腺癌中，我们共同提出了驱动癌基因是否影响渗透到肺部的免疫细胞的种类或它们的激活状态。这个参考框架是为了它潜在的临床相关性而采用的。我们分析了几种肺腺癌的免疫细胞浸润情况，主要评估了巨噬细胞和T细胞亚群，以及这些肿瘤中的反应性肿瘤相关纤维增生。在一组小的、高度精选的样本中，所有样本都在癌细胞中有一定程度的免疫反应渗透。使用一组有限的参数(CD8、CD68、CD4、FOXP3、PDL1)，我们没有观察到肿瘤之间的明显差异，其中突变的EGFR是驱动癌基因，而突变的KRAS是驱动癌基因。然而，纤维化的程度似乎确实与淋巴细胞浸润有关。这一组分析有力地证明了在肿瘤异质性的背景下免疫细胞渗透的复杂性。最后，本研究提供了人类肺腺癌中T调节性淋巴细胞的存在、CCL5的高表达和CD8+效应分子功能之间的相关性。针对黏膜黑色素瘤联合靶向治疗的药物开发工作也在继续。人和犬粘膜黑色素瘤之间的临床和病理相关性很大，而犬自然发生的粘膜黑色素瘤的相对较高的发病率为预测性建模提供了一个有希望的机会。与人类皮肤黑色素瘤相比，犬和人粘膜黑色素瘤似乎都罕见地含有BRAF、NRAS和c-kit突变，尽管这两个物种都有AKT和MAPK信号激活。人类和犬类黏膜黑色素瘤的基因组图谱表现出高度的多样性，通常缺乏与皮肤黑色素瘤相关的复发热点突变。尽管仍有许多有待确定的地方，但有证据表明，RAS/MAPK和/或PI3K/AKT/mTOR信号通路的激活在这两个物种中都很常见，可能是治疗干预的靶点。通过这项研究，从PI3K/mTOR抑制剂文库中选择了mTORC1/2抑制剂Sapanisertib来协同抑制MEK；后者的临床前疗效先前已被证明对犬粘膜黑色素瘤有效。联合使用曲美替尼和沙巴尼司替布抑制MEK和mTORC1/2，可导致细胞凋亡和细胞周期改变，协同降低不同基础信号激活水平的犬粘膜黑色素瘤细胞系的细胞存活率。与单独的抑制剂相比，交错剂量的Sapanisertib与每日一次的曲美替尼相结合，是限制小鼠原发黏膜黑色素瘤异种移植瘤生长和转移模型中肿瘤扩散的最佳方案，同时将血液和肾脏副作用降至最低。抑制剂下调了各自的信号靶标，并且联合使用还抑制了通路的相互串扰。对这两种药物联合的药物动力学的进一步研究正在进行中。