Computational Molecular Pathology Research for Cancer Diagnostics and Biomarkers

进行研究是为了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。其他目标包括开发新的研究技术，以评估和应用疾病生物标记物，并将癌症治疗方法转化为临床应用。在开发可用于开发和表征人类癌症和癌症诊断新模型的研究资源方面取得了进展。这项研究项目包括发展癌症模型的分子诊断能力，通过企业规模的图像分析平台和数据库的合作，开发用于定量病理的癌症标本的自动形态图像分析方法。在表征癌症疾病的新动物模型方面，针对粘膜黑色素瘤的靶向治疗的临床前开发取得了进展。与基因工程模型不同，零星的自然发生的犬类黑素细胞肿瘤与人类疾病有几个共同的特征，这可能使对狗的研究成为更相关的临床前模型。犬类黑色素瘤很少发生在阳光暴晒的部位，大多数自发发生在口腔。自然发生的犬类黑素细胞瘤的谱系，在人类中是真实的，包括一些类似痣的良性病变，以及侵袭性的原发黑色素瘤，以及广泛的转移。与人类一样，不同的黑色素瘤亚型在体细胞和易感生殖系遗传变化、细胞起源、流行病学、与紫外线辐射的关系以及从良性到恶性肿瘤的进展方面也存在自然存在的宠物狗。自发性黏膜黑色素瘤是一种罕见的侵袭性亚型。需要新的治疗方法，但积累足够的证据来改善患者结果是困难的。人和犬粘膜黑色素瘤之间的临床和病理相关性很大，而犬自发性自然发生的粘膜黑色素瘤的相对较高的发病率代表了使用比较肿瘤学方法进行预测建模的一个有希望的机会。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。人类和犬类黏膜黑色素瘤的基因组图谱呈现出高度的多样性。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MEK抑制；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转导靶点，联合使用还抑制了通路的相互串扰。联合用药不会显著改变血浆沙尼司替布的药代动力学；然而，单次给药后曲美替尼的曲线下面积在沙尼司替布的存在下增加。靶向RAS/MAPK和PI3K/AKT/mTOR信号转导通路有望成为治疗犬和人粘膜黑色素瘤的理想靶点。

Research is conducted to characterize and develop new animal models of human disease and to develop the means to better characterize a model's disease relevance, addressing critical barriers to research progress. Additional aims include the development of new research technologies for the evaluation and application of disease biomarkers and 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 cancer diagnostics. This research project included developing capabilities in molecular diagnostics for cancer models, developing methods for automated morphometric image analysis of cancer specimens for quantitative pathology through collaboration for enterprise scale of image analysis platforms and database. In characterizing new animal models of cancer disease, advancement was made in preclinical development of targeted therapy for mucosal melanoma. Distinct from genetically engineered models, sporadic naturally occurring canine melanocytic neoplasms share several characteristics with human disease that could make investigation in dogs a more relevant pre-clinical model. Canine melanomas rarely arise in sun-exposed site and most occur spontaneously in the oral cavity. The spectrum of naturally occurring canine melanocytic neoplasia, as is true in people, includes benign lesions with some analogy to nevi, as well as invasive primary melanoma, and widespread metastasis. As in humans, distinct melanoma subtypes differing in somatic and predisposing germ-line genetic alterations, cell of origin, epidemiology, relationship to ultraviolet radiation and progression from benign to malignant tumors, also exist naturally pet dogs. Melanomas arising spontaneously in the mucous membranes are a rare and aggressive subtype. New treatment approaches are needed, yet accumulating sufficient evidence to improve patient outcomes is difficult. 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 using a comparative oncology approach. Both canine and human mucosal melanomas appear to harbor BRAF, NRAS and c-kit mutations uncommonly, compared to human cutaneous melanomas. The genomic landscapes of human and canine mucosal melanomas appear highly diverse. 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, the mTORC1/2 inhibitor sapanisertib 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 down-modulated respective signaling targets and the combination additionally suppressed pathway reciprocal crosstalk. The combination did not significantly change plasma sapanisertib pharmacokinetics; however, trametinib area under the curve following a single dose was increased in the presence of sapanisertib. Targeting Ras/MAPK and PI3K/ AKT/mTOR signal transduction pathways appear rational therapies for canine and human mucosal melanoma.