Molecular Pathology Research for Cancer Diagnostics and Biomarkers

开展研究是为了确定和开发新的人类疾病动物模型，并开发更好地确定模型相关性的手段，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志物。在开发癌症诊断方法和开发和表征人类癌症新模型的研究资源方面取得了进展。该研究项目包括开发癌症模型的分子诊断能力，开发用于定量病理的癌症标本的自动形态学图像分析方法，以及粘膜黑色素瘤靶向治疗的临床前开发。开发了独特的空间光谱图像分析算法，用于应用自动模式识别形态学图像分析来量化生物标本组织切片中的组织学肿瘤和非肿瘤区域。在肺癌、甲状腺癌、前列腺癌和转移癌的算法开发和验证方面取得了新的进展。利用定量光学成像技术开发和验证了对理解耐药机制重要的新模型。进行的调查从涉及几种癌症类型的新模型中获得了新的见解。这些研究包括前列腺癌中的碳水化合物代谢、血胎盘屏障的耐药机制、皮肤鳞状癌中的MET信号传导、EGFR驱动的肺癌以及顺铂介导的卵巢癌细胞毒性模型。黑色素瘤治疗的新模式正在开发中。黑色素瘤是人类和狗的一种重要的恶性肿瘤。与基因工程模型不同，自然发生的散发性犬黑色素细胞肿瘤与人类疾病有几个共同特征，这可能使犬类研究成为更相关的临床前模型。犬黑色素瘤很少出现在阳光照射的地方，大多数自发发生在口腔。自然发生的犬类黑色素细胞瘤，和人类一样，包括一些类似痣的良性病变，也包括侵袭性原发性黑色素瘤和广泛转移。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也可能存在于宠物狗中。与人类皮肤黑色素瘤相比，犬和人粘膜黑色素瘤（MM）似乎不常见BRAF、NRAS和c-kit突变，尽管这两个物种都具有AKT和MAPK信号激活。我们得出结论，犬类自然发生的MM和人类自然发生的MM的临床和组织病理学特征有显著的重叠。犬类自然发生的口腔黑色素瘤作为人类黑色素瘤的临床前模型正在进一步探索。大多数人类和犬MM表现出RAS/ERK和/或PI3K/mTOR信号通路激活。犬MM细胞系具有不同的ERK和AKT/mTOR激活水平，反映了犬自然存在的差异，对MEK抑制和PI3K/mTOR双抑制敏感。与caspase 3/7激活、细胞周期调节蛋白和Bcl-2家族蛋白表达的改变有关，两药联合可协同降低细胞存活率。联合用药后，两种药物靶向各自的信号通路，在体外增强通路介质p-ERK、p-AKT、p-S6和4E-BP1的减少，并显著抑制小鼠MM异种移植物实体瘤的生长。这些发现为多种介质同时靶向Ras/ERK和PI3K/mTOR通路激活的黑色素瘤提供了协同治疗效果的证据。CCR比较肿瘤学项目和校外研究人员组织了一个类似的跨学科方法来比较自然发生的犬脑肿瘤对人类脑癌研究的适用性。与美国食品和药物管理局合作开展的一项多中心临床诊断研究正在为临床癌症诊断的全切片数字病理组织图像平台的验证提供有用的标准。

Research is conducted to characterize and develop new animal models of human disease and to develop the means to better characterize a model's relevance, addressing critical barriers to research progress. Additional aims include the development of new research technologies for the evaluation and application of disease biomarkers. Progress was made in developing cancer diagnostics and in research resources useful in developing and characterizing new models of human cancer. 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, and preclinical development of targeted therapy for mucosal melanoma. Unique spatial-spectral image analysis algorithms were developed for applying automated pattern recognition morphometric image analysis to quantify histologic tumor and non-tumor areas in biospecimen tissue sections. Additional progress was made in developing and validating algorithms for cancers of lung, thyroid, prostate gland and cancer metastasis. Contributions to new models important for understanding mechanisms of drug resistance were developed and validated using quantitative optical imaging. Investigations were conducted that led to new insight from novel models involving several cancer types. These included carbohydrate metabolism in prostate cancer, drug resistance mechanisms at the blood placental barrier, MET signaling in cutaneous squamous cancer, EGFR driven lung cancer, and an ovarian cancer cell model of cisplatin-mediated cytotoxicity. New model development is taking place for melanoma treatment. Melanoma represents a significant malignancy in humans and dogs. 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, may also exist in pet dogs. Both canine and human mucosal melanomas (MM) appear to harbor BRAF, NRAS and c-kit mutations uncommonly, compared to human cutaneous melanomas, although both species share AKT and MAPK signaling activation. We conclude that there is significant overlap in the clinical and histopathological features of naturally occurring canine and human MM. Naturally occurring canine oral cavity melanoma is being explored further as a pre-clinical model for human melanoma. The majority of human and canine MM evaluated exhibited RAS/ERK and/or PI3K/mTOR signaling pathway activation. Canine MM cell lines, with varying ERK and AKT/mTOR activation levels reflective of naturally occurring differences in dogs, were sensitive to MEK inhibition and dual PI3K/mTOR inhibition. Two-drug combination synergistically decreased cell survival in association with caspase 3/7 activation, as well as altered expression of cell cycle regulatory proteins and Bcl-2 family proteins. In combination, the two drugs targeted their respective signaling pathways, potentiating reduction of pathway mediators p-ERK, p-AKT, p-S6, and 4E-BP1 in vitro, and in association with significantly inhibited solid tumor growth in MM xenografts in mice. These findings provide evidence of synergistic therapeutic efficacy when multiple mediators are simultaneously targeted in melanoma with Ras/ERK and PI3K/mTOR pathway activation. An analogous interdisciplinary approach to comparing the suitability of naturally occurring canine brain tumors to inform human brain cancer research was organized with the CCR Comparative Oncology Program and extramural investigators. Conduct of a multi-center clinical diagnostic study in collaboration with the US Food and Drug Administration is leading to useful criteria for validation of whole slide digital pathology tissue image platforms for clinical cancer diagnosis.