Computational Molecular Pathology Research for Cancer Diagnostics and Biomarkers

进行研究以表征和开发新的人类疾病动物模型 并开发更好地表征模型中产生的疾病相关性的方法， 解决研究进展的关键障碍。其他目标包括： 新的研究技术和手段，询问癌症组织进行评估， 疾病生物标志物的应用。该研究还旨在将方法转化为 用于临床应用的癌症治疗。在发展研究方面取得了进展 用于开发和表征人类癌症和癌症新模型的资源 诊断该研究产生了方法和计算能力， 检测并定量核膜中促进转移的蛋白， 对细胞感应和转移的机械洞察。乳腺癌建模工作的重点是 CLIC 4在人乳腺癌中的表达升高及其与早期侵袭的关系， 在女性中效果不佳。宿主微环境中的CLIC 4是肺转移所需的， 而宿主CLIC 4的缺失导致原发性肿瘤中的微环境， 转移前肺，这是不利的肿瘤活力和肺定植在 模型特别是CLIC 4缺陷小鼠体内的原发性肿瘤微环境 出现坏死和血管异常，通过计算分子组织检测 分析。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 浸润肺部的免疫细胞的种类或它们的激活状态。这一帧 参考其潜在的临床相关性。我们侧写了几个肺 腺癌的免疫细胞浸润，主要评估巨噬细胞和T 细胞亚群，以及这些肿瘤中的反应性肿瘤相关纤维增生。中 一个小的，高度策划的样本集，所有标本都有一定程度的免疫反应 癌细胞之间的浸润。我们没有观察到肿瘤之间的明显差异， 有限的参数集（CD8、CD68、CD4、FOXP3、PDL 1），其中突变型EGFR是驱动因素 癌基因与突变型KRAS。然而，纤维化的程度似乎与 淋巴细胞浸润这组分析有力地证明了免疫系统的免疫性。 在肿瘤异质性的背景下细胞浸润的复杂性。最后，调查 提供了调节性T淋巴细胞的存在、高CCL5表达和 人肺腺癌中的CD8+效应子功能。关于毒品的工作也在继续 用于粘膜黑色素瘤的联合靶向治疗。临床和病理 人类和犬粘膜黑色素瘤之间的相关性是实质性的， 犬中自发性自然发生的粘膜黑色素瘤的发生率较高， 预测建模的好机会。犬和人的粘膜黑色素瘤 与人类皮肤黑色素瘤相比， 尽管两种物种共享AKT和MAPK信号传导激活。人类的基因组景观 和犬粘膜黑色素瘤表现出高度多样性，通常缺乏复发热点 与皮肤黑色素瘤相关的突变。尽管仍有许多问题有待确定， 有证据表明Ras/MAPK和/或PI3K/AKT/mTOR信号通路的激活是常见的， 在这两个物种，并可能代表治疗干预的目标。通过本文的研究 sapanisertib是一种mTORC1/2抑制剂，选自PI3K/mTOR抑制剂文库， 与MEK抑制协同作用;后者的临床前疗效已在之前得到证实 用于治疗犬粘膜黑色素瘤使用曲美替尼和曲美替尼联合抑制MEK和mTORC 1/2 sapanisertib，产生细胞凋亡和细胞周期改变，协同减少细胞 不同基础信号激活水平的犬粘膜黑色素瘤细胞系的存活率。 与单个抑制剂相比，交错的sapanisertib剂量，加上每日 曲美替尼对于限制小鼠中原发性粘膜黑素瘤异种移植物生长是最佳的， 转移模型中的肿瘤扩散，同时最小化血液和肾脏方面 方面的影响.抑制剂下调各自的信号传导靶标， 另外还抑制了通路相互串扰。药代动力学的进一步研究 两种药物的联合治疗正在进行中。

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.