Identification of oncogenic drivers in aggressive B cell Lymphoma by ribosome profiling and a novel primary human lymphocyte transformation assay

弥漫性大B细胞淋巴瘤（DLBCL）是非霍奇金淋巴瘤的最常见形式。它是一种侵略性和毁灭性的癌症。虽然联合化疗有可能治愈，但超过三分之一的患者会死于疾病。重要的是，DLBCL的发病率随着年龄的增长而增加，许多老年患者根本无法耐受所需的化疗。已鉴定出DLBCL的某些亚型对所有现有疗法的反应特别差。因此，迫切需要开发更有效和耐受性更好的“靶向”治疗。在过去的十年里，我们对这种疾病的潜在生物学的理解有了相当大的进展。这在很大程度上源于研究不同亚型淋巴瘤之间基因组的活性如何不同的研究。这些“基因表达“研究主要检查每个细胞中mRNA的总量。mRNA是一种信使分子，它携带来自基因DNA的指令，然后被翻译成最终的活性产物，称为蛋白质。一般来说，更活跃的基因产生更多的mRNA，这就是为什么mRNA通常被用作基因活性的代表。然而，最近已经清楚的是，并非所有的mRNA分子都以相同的效率“翻译”成蛋白质。此外，在癌症中的一个常见发现是，携带对癌症有利的指令的某些mRNA分子可能优先被肿瘤翻译。最近的技术现在允许测量每个单独mRNA的翻译速率的机会。我将使用这种技术来识别那些优先被DLBCL不同亚型翻译的mRNA。将在细胞培养系统中测试翻译速率变化最大的那些基因/mRNA，以鉴定对淋巴瘤的发展和生长贡献最大的那些基因/mRNA。最终，设计用于抑制这些基因的药物可能被证明在淋巴瘤的治疗中是有用的。我预计这种方法将为抗淋巴瘤药物治疗的发展确定新的目标。此外，我将调查的机制，其中一些mRNA是优先翻译的淋巴瘤细胞。我将使用计算方法筛选优先翻译的mRNA，以确定淋巴瘤细胞可能用于控制翻译的共同序列。然后，我将使用这些序列作为诱饵，以确定“翻译因子”的改变翻译淋巴瘤负责。我将研究这些翻译因子的活性是如何控制的，无论是淋巴瘤特异性信号通路或突变。事实上，突变已经被确定在已知的调节淋巴瘤细胞中的翻译的因素，但这些突变的功能意义还有待调查。总体而言，该项目将提供一个详细的了解淋巴瘤细胞如何破坏其基因的正常严格调控的活动。在这样做的过程中，该项目可能会揭示新形式的淋巴瘤靶向治疗的机会。此外，我们所了解的淋巴瘤中受损的翻译调节机制可能与其他疾病，特别是其他类型的癌症具有更广泛的相关性。

Diffuse Large B Cell Lymphoma (DLBCL) is the most common form of non-Hodgkin Lymphoma. It is an aggressive and devastating form of cancer. Although potentially curable with combination chemotherapy, more than a third of patients will succumb to their disease. Importantly, the incidence of DLBCL increases with age and many older patients are simply unable to tolerate the required chemotherapy. Certain subtypes of DLBCL have been identified that respond particularly poorly to all existing therapies. Thus there is a pressing need for the development of more effective and better tolerated, "targeted" treatments. Over the last decade there have been considerable advances in our understanding of underlying biology of this disease. Much of this has arisen from studies that examine how the activity of cohorts of genes differs between different subtypes of lymphoma. These "gene expression " studies have predominantly examined the total amount mRNA in each cell. mRNA is a messenger molecule that carries instructions from the gene's DNA before being translated in to the final active product, termed the protein. In general more active genes make more mRNA, which is why mRNA has generally been used as a proxy for gene activity. However, recently it has become clear that not all mRNA molecules are "translated" into protein with equal efficiency. Furthermore, a frequent finding in cancer is that certain mRNA molecules that carry instructions advantageous to the cancer may be preferentially translated by the tumor. Recent technology now allows the opportunity to measure the translational rate for each individual mRNA. I will use this technique to identify those mRNAs that are preferentially translated by the different subtypes of DLBCL. Those genes / mRNAs with the greatest changes in translational rate will be tested in cell culture systems to identify those that contribute most to the development and growth of the lymphoma. Ultimately drugs designed to inhibit these genes may prove useful in the treatment of lymphoma. I anticipate that this approach will identify new targets for the development of anti-lymphoma drug treatments. In addition I will investigate the mechanism by which some mRNA is preferentially translated by lymphoma cells. I will use a computational approach to screen preferentially translated mRNA to identify common sequences that the lymphoma cells may use to control the translation. I will then use these sequences as bait to identify the "translation factors' responsible for the altered translation in lymphoma. I will investigate how the activity of these translation factors is controlled, either by lymphoma specific signaling pathways or by mutation. Indeed, mutations have already been identified in factors known to regulate translation in lymphoma cells but the functional significance of these mutations is yet to be investigated.Overall this project will provide a detailed understanding of how lymphoma cells corrupt the normally tightly regulated activity of their genes. In doing so this project may reveal opportunities for new forms of targeted treatment in lymphoma. In addition much of what we learn about the mechanisms of corrupted translational regulation in lymphoma may prove to be of broader relevance to other diseases, in particular other types of cancer.