Computational Molecular Pathology Research for Cancer Diagnostics and Biomarkers

开展研究以表征和开发人类疾病的新动物模型，并开发更好地表征模型疾病相关性的方法，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，用于评估和应用疾病生物标志物，并将癌症治疗方法转化为临床应用。在开发研究资源方面取得了进展，这些研究资源可用于开发和描述人类癌症和癌症诊断的新模型。该研究项目包括开发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合作开发用于定量病理学的癌症标本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，在粘膜黑色素瘤的靶向治疗的临床前开发方面取得了进展。与基因工程模型不同，散发性自然发生的犬黑色素细胞肿瘤与人类疾病有几个共同特征，这可能使犬研究成为更相关的临床前模型。犬黑色素瘤很少发生在阳光照射的部位，大多数自发发生在口腔。与人类一样，犬黑素细胞瘤的自然发生谱包括与痣类似的良性病变，以及侵袭性原发性黑色素瘤和广泛转移。在人类中，不同的黑色素瘤亚型在体细胞和易感性生殖系遗传改变、细胞起源、流行病学、与紫外线辐射的关系以及从良性肿瘤到恶性肿瘤的进展方面不同，也自然存在宠物狗。黑色素瘤自发性发生在粘膜是一种罕见的和积极的亚型。需要新的治疗方法，但积累足够的证据来改善患者的预后是困难的。人类和犬粘膜黑色素瘤之间的临床和病理学相关性是实质性的，并且犬中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有希望的机会。与人类皮肤黑色素瘤相比，犬和人类粘膜黑色素瘤似乎罕见地具有BRAF、NRAS和c-kit突变。人类和犬粘膜黑色素瘤的基因组景观呈现高度多样性。尽管仍有许多有待确定，但证据表明Ras/MAPK和/或PI 3 K/AKT/mTOR信号通路激活在两个物种中是常见的，并且可能代表治疗干预的靶点。通过本研究，从PI 3 K/mTOR抑制剂库中选择mTORC 1/2抑制剂sapanisertib与MEK抑制剂协同作用;后者的临床前疗效先前已在犬粘膜黑色素瘤中得到证实。使用曲美替尼和sapanisertib联合抑制MEK和mTORC 1/2，产生细胞凋亡和细胞周期改变，协同降低具有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。与单个抑制剂相比，交错剂量的sapanisertib联合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和转移模型中的肿瘤扩散是最佳的，同时最大限度地减少血液学和肾脏副作用。抑制剂下调各自的信号传导靶标，并且组合另外抑制通路相互串扰。该组合并未显着改变血浆sapanisertib的药代动力学;然而，在存在sapanisertib的情况下，单次给药后曲美替尼的曲线下面积增加。靶向Ras/MAPK和PI 3 K/ AKT/mTOR信号转导通路是治疗犬和人粘膜黑色素瘤的合理疗法。

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.