A Multi-disciplinary Approach to Investigating Canine Diffuse Large B cell Lymphoma

原理：在人类中，DLBCL已被证明具有遗传异质性，miRNA谱可预测疾病预后。在狗中，淋巴瘤的研究有限，遗传特征相对较少，这意味着很难找到与人类疾病的相关性。然而，在人类和狗中，有一个重要的淋巴瘤亚组是难治性的。总体目标：我们建议更好地定义犬淋巴瘤的遗传亚型，并通过整合a）高级图像分析B）基因组测序和c）总RNA测序，通过更准确的淋巴瘤表型组合来表征对不同亚型治疗的反应（包括micro-RNA测序）。犬淋巴瘤可以更准确地诊断，治疗和诊断，通过协同分子，遗传学和图像为基础的分析。通过机器学习和人工智能方法对数字全载玻片图像（WSI）进行图像分析，定义犬淋巴瘤的形态学特征目标2：通过全基因组关联研究和CNV分析识别DLBCL易感性的遗传标记目标3：通过mRNAseq和small RNA-seq确定犬DLBCL中的mRNA和miRNA表达谱，目的4：比较1-3中的基因和形态学特征，以区分对化疗有良好反应的病例和对治疗无反应的病例。

RATIONALE: In humans, DLBCL has been shown to demonstrate genetic heterogeneity and miRNA profiles predicting disease prognosis. In dogs there are limited studies and relatively little genetic characterisation of lymphoma, meaning that it is difficult to find correlation with the human disease. Nevertheless, in both humans and dogs, there is a significant subset of lymphomas that are refractory to treatment. OVERALL AIM: We propose to better define genetic subtypes of canine lymphoma and characterising response to treatment of the different subtypes through a combination of more accurate phenotyping of lymphomas by integrating a) advanced image analysis b) genome sequencing, and c) total RNA sequencing (including micro-RNA sequencing).HYPOTHESIS: Canine lymphoma can be more accurately diagnosed, treated and prognosticated through a synergistic molecular, genetics, and image based analysis.SPECIFIC PROJECT OBJECTIVESOBJECTIVE 1: Define morphometric characteristics of canine lymphoma through image analysis of digital whole slide images (WSIs) via machine learning and artificial intelligence methodsOBJECTIVE 2: Identify genetic markers for DLBCL susceptibility through genome wide association studies and CNV analysesOBJECTIVE 3: Define mRNA and miRNA expression profiles in canine DLBCL through mRNAseq and small RNA-seq, and investigate if tumour-associated miRNA can be detected in peripheral blood serumOBJECTIVE 4: Genetic and morphological signatures from 1-3 will be compared between cases that show good response to chemotherapy and those that are refractory to treatment.