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

理由：在人类中，DLBCL已被证明具有遗传异质性和miRNA谱预测疾病预后。在狗身上，淋巴瘤的研究有限，遗传特征也相对较少，这意味着很难发现它与人类疾病的相关性。然而，在人类和狗中，有很大一部分淋巴瘤是难以治疗的。总体目的：我们建议通过整合a)高级图像分析b)基因组测序和c)总RNA测序（包括微RNA测序），结合更准确的淋巴瘤表型，更好地定义犬淋巴瘤的遗传亚型和表征对不同亚型治疗的反应。假设：犬淋巴瘤可以通过分子、遗传学和基于图像的协同分析更准确地诊断、治疗和预测。目的1：通过机器学习和人工智能方法对数字全幻灯片图像（WSIs）进行图像分析，确定犬淋巴瘤的形态特征；目的2：通过全基因组关联研究和CNV分析，确定DLBCL易感性的遗传标记；通过mRNAseq和小RNA-seq确定犬DLBCL中mRNA和miRNA的表达谱，并研究是否可以在外周血血清中检测到肿瘤相关的miRNA。目的4：比较1-3例化疗反应良好的病例和治疗难治的病例的遗传和形态学特征。

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.