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)是最常见的非霍奇金淋巴瘤。它是一种侵袭性和破坏性的癌症。尽管联合化疗有可能治愈，但超过三分之一的患者会死于他们的疾病。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。某些亚型的DLBCL已经被发现对所有现有的治疗方法反应特别差。因此，迫切需要开发更有效和更好耐受性的“靶向”治疗方法。在过去的十年里，我们对这种疾病的潜在生物学的理解已经有了相当大的进步。这在很大程度上是因为研究了不同亚型淋巴瘤之间基因队列的活性有何不同。这些“基因表达”研究主要检查了每个细胞中的总信使核糖核酸。信使分子是一种信使分子，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，更活跃的基因会产生更多的信使核糖核酸，这就是为什么信使核糖核酸通常被用作基因活性的代用品。然而，最近已经清楚的是，并不是所有的信使核糖核酸分子都能以同样的效率“翻译”成蛋白质。此外，癌症中的一个常见发现是，携带有利于癌症的指令的某些信使核糖核酸分子可能优先被肿瘤翻译。现在，最新的技术使人们有机会测量每个单独的信使核糖核酸的翻译速率。我将使用这一技术来识别那些优先由不同亚型的DLBCL翻译的mRNAs。那些翻译率变化最大的基因/mRNA将在细胞培养系统中进行测试，以确定对淋巴瘤的发展和生长贡献最大的基因/mRNAs。最终，旨在抑制这些基因的药物可能会被证明对淋巴瘤的治疗有用。我预计，这种方法将为抗淋巴瘤药物治疗的发展确定新的靶点。此外，我还将研究淋巴瘤细胞优先翻译某些信使核糖核酸的机制。我将使用一种计算方法来筛选优先翻译的mRNA，以确定淋巴瘤细胞可能用来控制翻译的共同序列。然后，我将用这些序列作为诱饵，找出导致淋巴瘤翻译改变的“翻译因素”。我将研究这些翻译因子的活性是如何通过淋巴瘤特定的信号通路或突变来控制的。事实上，已经在已知的调节淋巴瘤细胞翻译的因素中发现了突变，但这些突变的功能意义尚未被调查。总的来说，这个项目将提供一个详细的理解，淋巴瘤细胞是如何破坏通常受到严格调控的基因活动的。通过这样做，该项目可能会为淋巴瘤的新形式的靶向治疗提供机会。此外，我们了解到的许多关于淋巴瘤翻译调控机制的知识可能被证明与其他疾病，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.